Objective There is a lack of effective and long-term safe drugs for the treatment of osteoarthritis(OA).Tetrandrine(Tet)has been approved and used to treat rheumatoid arthritis for several decades,but its effect on OA...Objective There is a lack of effective and long-term safe drugs for the treatment of osteoarthritis(OA).Tetrandrine(Tet)has been approved and used to treat rheumatoid arthritis for several decades,but its effect on OA has not been investigated.Herein,we explored the effect of Tet on OA and its underlying mechanism.Methods OA was induced using destabilization of the medial meniscus(DMM)in C57BL/6J mice.The animals were randomly divided into sham,DMM,Tet,celecoxib(CXB),and indomethacin(INDO)groups.Each group was given solvent or corresponding drugs by gavage for 7 weeks after convalescence.Pathological staining,OARSI scores,micro-computed tomography and behavior tests were performed to evaluate the effects of Tet.Results Tet remarkably alleviated cartilage injury in the knee joint,limited bone remodeling in the subchondral bone,and delayed progression of OA.Tet also significantly relieved joint pain and maintained function.Further mechanistic studies revealed that Tet lowered inflammatory cytokine levels and selectively suppressed gene and protein expression of cyclooxygenase(COX)-2 but not COX-1(P<0.01).Tet also reduced the production of prostaglandin E2 without damaging the gastric mucosa.Conclusion We found that Tet could selectively inhibit COX-2 gene expression and decrease cytokine levels in mice,thus reducing inflammation and improving OA without obvious gastric adverse events.These results provide a scientific basis for the clinical application of Tet in the treatment of OA.展开更多
The effect of tetrandrine (Tet) on the infarction area and volume of rat brain induced by middle cerebral artery occlusion (MCAO) was investigated. The treatment with Tet 7.5, 12.0 or 15.0 mg·kg 1 , or with...The effect of tetrandrine (Tet) on the infarction area and volume of rat brain induced by middle cerebral artery occlusion (MCAO) was investigated. The treatment with Tet 7.5, 12.0 or 15.0 mg·kg 1 , or with fructose 1,6 diphosphate (FDP) 200 and 350 mg·kg 1 ip immediately after MCAO, respectively, significantly reduced the infarction area and volume in a dose dependent manner. MK801 and FDP also displayed a protective effect on brain ischemia. A combination of Tet and FDP administered immediately after MCAO, produced a more potent protective effect than those treated with Tet or FDP alone. When Tet or FDP was administered 1 h and 2 h after MCAO, respectively, they could still significantly reduce the infarction area and volume of brain tissue. But, there was no significant protective effect when these two compounds were given 3 h after MCAO.展开更多
\ The effects of tetrandrine (Tet) on cytosolic free calcium ([Ca2+]i) in subcultured bovine aortic smooth muscle cells (SMC) were studied by Fura2 and ARCMMIC cation measurement system. Tet (1～100 μmol·L-1) ...\ The effects of tetrandrine (Tet) on cytosolic free calcium ([Ca2+]i) in subcultured bovine aortic smooth muscle cells (SMC) were studied by Fura2 and ARCMMIC cation measurement system. Tet (1～100 μmol·L-1) had no effect on the resting [Ca2+]i, but had inhibitory effects on [Ca2+]i elevation induced by high K+, 5HT, ATP, Ang II and NE in the presence of extracellular Ca2+. High concentration of Tet also inhibited Pheinduced [Ca2+]i elevation in absence of extracellular Ca2+. Tet (1～100 μmol·L-1) inhibited KCl (60 mmol·L-1) induced [Ca2+]i elevation in dosedependent manner, the IC50 value was 9.2 (95% confidence limits: 5.7～14.9) mmol·L-1. The results suggested that Tet had blocking effects on both VOC and ROC in bovine aortic SMC. It appears that the mechanisms of blocking effect of Tet on ROC might be primarily due to its Ca2+ entry blocking effects.展开更多
It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and ...It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and different 5-HT receptors on pentobarbital-induced sleep by using the loss-of-righting reflex method. The results showed that augmentative effect of tetrandrine on pentobarbital hypnosis in mice were potentiated by the p-MPPI （5-HT1A receptor antagonist） （1 mg/kg, i.p.） and ketanserin （5-HT2A/2C receptor antagonist） （1.5 mg/kg, i.p.）, respectively. Pretreatment with either 8-OH-DPAT （5-HT1A receptor agonist） （0.1 mg/kg, s.c.） or DOI （5-HT2A/2C receptor agonist） （0.2 mg/kg, i.p.） significantly decreased pentobarbital-induced sleep time, and tetrandrine （60 mg/kg, i.g.） significantly reversed this effect. These results suggest that both the 5-HTLA and 5-HT2A/2C subfamily may be involved in the potentiating mechanism of tetrandrine＇s effects on pantobarbital hypnosis.展开更多
AIM： To investigate the therapeutic effect of tetrandrine on liver fibrosis induced by thioacetamide in rats in vivo and in vitro. METHODS： In vitro study： we investigated the effect of tetrandrine on the apoptosis...AIM： To investigate the therapeutic effect of tetrandrine on liver fibrosis induced by thioacetamide in rats in vivo and in vitro. METHODS： In vitro study： we investigated the effect of tetrandrine on the apoptosis of rat hepatic stellate cells transformed by simian virus 40 （T-HSC/CI-6）, which retains the features of activated cells. In vivo study： hepatic fibrosis was induced in rats by thioacetamide. Tetrandrine was given orally to rats at doses of 5, 10 or 20 mg/kg for 4 wk compared with intraperitoneal injection of interferon-r. RESULTS： In vitro study： 5, 10 or 25 μg/mL of tetrandrine-induced activation of caspase-3 in t-HSC/CI-6 cells occurred dose-dependently. In vivo study： tetrandrine treatment as well as interferon-r significantly ameliorated the development of fibrosis as determined by lowered serum levels of aspartate aminotransferase （AST）, alanine aminotransferase （ALT）, total bilirubin （T-Bil） and the levels of liver hydroxyproline （Hyp）, hyaluronic acid （HA）, laminin （LN） and also improved histological findings. The effects of tetrandrine at the concentration of 20 mg/kg were better than the other concentration groups. CONCLUSION： Tetrandrine promotes the apoptosis of activated HSCs in vitro. Tetrandrine administration can prevent liver fibrosis and liver damage induced by thioacetamide in rats in vivo, indicating that it might exert a direct effect on rat HSCs.展开更多
AIM: To investigate the effect of various concentrations of tetrandrine on activation of quiescent rat hepatic stellate cells (HSCs) and transforming growth factor-β (TGF-β) signaling in vitro.METHODS: HSCs were iso...AIM: To investigate the effect of various concentrations of tetrandrine on activation of quiescent rat hepatic stellate cells (HSCs) and transforming growth factor-β (TGF-β) signaling in vitro.METHODS: HSCs were isolated from rats by in situperfusion of liver and 18% Nycodenz gradient centrifugation, and primarily cultured on uncoated plastic plates for 24 hwith DMEM containing 20% fetal bovine serum (FBS/DMEM) before the culture medium was substituted with 2% FBS/DMEM for another 24 h. Then, the HSCs were cultured in 2% FBS/DMEM with tetrandrine (0.25, 0.5, 1,2 mg/L, respectively). Cell morphological features were observed under an inverted microscope, smooth muscleα-actin (α-SMA) was detected by immunocytochemistry and image analysis system, laminin (LN) and type Ⅲprocollagen (PCⅢ) in supernatants were determined byradioimmunoassay. TGF-β1 mRNA, Smad 7 mRNA and Smad 7 protein were analyzed with RT-PCR and Western blotting, respectively.RESULTS: Tetrandrine at the concentrations of 0.25-2 mg/L prevented morphological transformation of HSC from the quiescent state to the activated one, while α-SMA, LN and PCⅢ expressions were inhibited. As estimated by gray values, the expression of α-SMA in tetrandrine groups (0.25, 0.5, 1, 2 mg/L) was reduced from 21.3% to 42.2%(control: 0.67, tetrandrine groups: 0.82, 0.85, 0.96, or 0.96, respectively, which were statistically different from the control, P＜0.01), and the difference was more significant in tetrandrine at 1 and 2 mg/L. The content of LN in supernatants was significantly decreased in tetrandrine groups to 58.5%, 69.1%, 65.8% or 60.0% that of the control respectively, and that of PCⅢ to 84.6%, 81.5%,75.7% or 80.7% respectively (P＜0.05 vs control), with no significant difference among tetrandrine groups. RTPCR showed that TGF-β1 mRNA expression was reduced by tetrandrine treatments from 56.56% to 87.90% in comparison with the control, while Smad 7 mRNA was increased 1.4-4.8 times. The TGF-β1 mRNA and Smad 7 mRNA expression was in a significant negative correlation (r= -0.755, P＜0.01), and both were significantly correlated with α-SMA protein expression (r = -0.938, P＜0.01;r = 0.938, P＜0.01, respectively). The up-regulation of Smad 7 protein by tetrandrine (1 mg/L)was confirmed by Western blotting as well.CONCLUSION: Tetrandrine has a direct inhibiting effect on the activation of rat HSCs in culture. It up-regulates the expression of Smad 7 which in turn blocks TGF-β1 expression and signaling.展开更多
In the screening tests of drugs for silicosis in our laboratory, we found that TT, a type of alkaloid isolated from Stephania tetrandra, could inhibit the development of experimental silicosis of rats and the synthesi...In the screening tests of drugs for silicosis in our laboratory, we found that TT, a type of alkaloid isolated from Stephania tetrandra, could inhibit the development of experimental silicosis of rats and the synthesis of collagen in rat lung. Chest X-rays of silicotic patients treaied with TT for 1-3 years showed obvious changes. The silicotic nodules became smallel and shadows became clearer. PVNO was proved to have anti-silicotic effect on animal and clinically. This presentation reports the effect of them on collagen mRNA.Dot blot results showed that 1 (Ⅰ) and 1 (Ⅲ) mRNA levels increased significantly at 60 and 120 days after the rats were exposed to silica dust. The mRNA levels went down at 1 and 3 months after treated by TT and PVNO. In situ hybridization observation revealed that the silver grains of Type Ⅰand Type Ⅲ collagen were scattered within the fibroblasts in cellular nodules and in thickened interstitium of silicosis tissue. The amounts of mRNA silver grains decreased in the lung tissue treated by TT and PVNO. It was suggested that TT and PVNO may inhibil the gene expression of collagen during silicosis展开更多
AIM: To investigate the effect of Tetrandrine (Tet) on LPS-induced NF-κB activation and cell injury in pancreatic acinar cells and to explore the mechanism of Tetrandrine preventing LPS-induced acinar cell injury. ME...AIM: To investigate the effect of Tetrandrine (Tet) on LPS-induced NF-κB activation and cell injury in pancreatic acinar cells and to explore the mechanism of Tetrandrine preventing LPS-induced acinar cell injury. METHODS: Male rat pancreatic acinar cells were isolated by collagenase digestion, then exposed to LPS (10 mg/L), Tet (50μmol/L, 100μmol/L) or normal media. At different time point (30 min, 1 h, 4 h, 10 h) after treatment with the agents, cell viability was determined by MTT, the product and nuclear translocation of subunit p65 of NF-κB was visualized by immunofluorescence staining and nuclear protein was extracted to perform EMSA which was used to assay the NF-κB binding activity. RESULTS: LPS induced cell damage directly in a time dependent manner and Tet attenuated LPS-induced cell damage (50μmol/L, P < 0.05; 100μmol/L, P < 0.01). NF-κB p65 immunofluorescence staining in cytoplasm increased and began showing its nuclear translocation within 30 min and the peak was shown at 1 h of LPS 10 mg/L treatment. NF-κB DNA binding activity showed the same alteration pattern as p65 immunofluorescence staining. In Tet group, the immunofluorescence staining in cytoplasm and nuclear translocation of NF-κB were inhibited significantly. CONCLUSION: NF-κB activation is an important early event that may contribute to inflammatory responses and cell injury in pancreatic acinar cells. Tet possesses the protective effect on LPS-induced acinar cell injury by inhibiting NF-κB activation.展开更多
Objective To assess the ability of tetrandrine （Tet） to enhance the sensitivity to irradiation and its mechanism in cell lines of human breast cancer p53-mutant MCF-7/ADR, p53-wild-type MCF-7 and human colon carcino...Objective To assess the ability of tetrandrine （Tet） to enhance the sensitivity to irradiation and its mechanism in cell lines of human breast cancer p53-mutant MCF-7/ADR, p53-wild-type MCF-7 and human colon carcinoma p53-mutant HT-29 as well as in C26 colorectal carcinoma-bearing BALB/c mice. Methods MCF-7/ADR, HT-29 and MCF-7 cells were exposed to irradiation in the absence or presence of tetrandrine. The effect of Tet on the cytotoxicity of X-irradiation in these three cells was determined and the effect of tetrandrine on cell cycle arrest induced by irradiation in its absence or presence was studied by flow cytometry. Moreover, mitotic index measurement determined mitosis of cells to enter mitosis. Western blotting was employed to detect cyclin B1 and Cdc2 proteins in extracts from irradiated or non-irradiated cells of MCF-7/ADR, HT-29 and MCF-7 treated with tetrandrine at various concentrations. Tumor growth delay assay was conducted to determine the radio-sensitization of tetrandrine in vivo. Results Clonogenic assay showed that tetrandrine markedly enhanced the lethal effect of X-rays on p53-mutant MCF-7/ADR and HT-29 cells and the sensitization enhancement ratio （SER） of tetrandrine was 1.51 and 1.63, but its SER was only 1.1 in p53-wt MCF-7 cells. Irradiated p53-mutant MCF-7/ADR and HT-29 cells were only arrested in G2/M phase while MCF-7 cells were arrested in G1 and G2/M phases. Radiation-induced G2 phase arrests were abrogated by tetrandrine in a concentration-dependent manner in MCF-7/ADR and HT-29 cells, whereas redistribution within MCF-7 cell cycle changed slightly. The proportion of cells in M phase increased from 1.3% to 14.7% in MCF-7/ADR cells, and from 1.5% to 13.2% in HT-29 cells, but 2.4% to 7.1% in MCF-7 cells. Furthermore, the levels of cyclin B 1 and Cdc2 expression decreased after X-irradiation in MCF-7/ADR and HT-29 cells, and the mitotic index was also lower. Tet could reverse the decrease and induce the irradiated cells to enter mitosis （M phase）. Endosomatic experiment showed that tetrandrine caused tumor growth delay in irradiated mice. Conclusion Tetrandrine boosts the cell killing activity of irradiation both in vitro and in vivo. Tetrandrine is a potent abrogator for G2 checkpoint control and can sensitize the cells to radiation.展开更多
Tetrandrine is one of the major active ingredients in Menispermaceae Stephania tetrandra S.Moore,and has specific therapeutic effects in ischemic cerebrovascular disease.Its use in vascular dementia has not been studi...Tetrandrine is one of the major active ingredients in Menispermaceae Stephania tetrandra S.Moore,and has specific therapeutic effects in ischemic cerebrovascular disease.Its use in vascular dementia has not been studied fully.Here,we investigated whether tetrandrine would improve behavioral and cellular impairments in a two-vessel occlusion rat model of chronic vascular dementia.Eight weeks after model establishment,rats were injected intraperitoneally with 10 or 30 mg/kg tetrandrine every other day for 4 weeks.Behavioral assessment in the Morris water maze showed that model rats had longer escape latencies in training trials,and spent less time swimming in the target quadrant in probe trials,than sham-operated rats.However,rats that had received tetrandrine showed shorter escape latencies and longer target quadrant swimming time than untreated model rats.Hematoxylin-eosin and Nissl staining revealed less neuronal necrosis and pathological damage,and more living cells,in the hippocampus of rats treated with tetrandrine than in untreated model rats.Western blot assay showed that interleukin-1β expression,and phosphorylation of the N-methyl-D-aspartate 2B receptor at tyrosine 1472,were lower in model rats that received tetrandrine than in those that did not.The present findings suggest that tetrandrine may be neuroprotective in chronic vascular dementia by reducing interleukin-1β expression,N-methyl-D-aspartate receptor 2B phosphorylation at tyrosine 1472,and neuronal necrosis.展开更多
AIM To study the effects of tetrandrine on portal hypertensive gastric mucosal lesions. METHODS Portal hypertensive models were induced in Wistar rats by 60% CCl 4 3ml/kg body weight subcutaneous injection, once e...AIM To study the effects of tetrandrine on portal hypertensive gastric mucosal lesions. METHODS Portal hypertensive models were induced in Wistar rats by 60% CCl 4 3ml/kg body weight subcutaneous injection, once every 4 days for 56 days. The animals were randomly divided into portal hypertension, tetrandrine and propranolol groups and subsequently, treated by physiological saline, tetrandrine and propranolol respectively for 15 days. Some healthy rats were used as normal group. Portal venous pressure (PVP), gastric mucosal prostaglandin E 2 (PGE 2) content, gastric mucosal blood flow (GMBF), gastric adherent mucus (GAM), ALT, ALP and serum total bilirubin (STB), were measured and liver tissues were observed histologically. RESULTS In tetrandrine group and propranolol group, PVP was significantly lower (1 43±0 13, 1 45±0 12 vs 1 89±0 18kPa; P <0 01) and gastric mucosal PGE 2 content (138 59±12 68, 129 98±14 31 vs 104 65±12 97pg/mg; P <0 01), GMBF (11 80±3 47, 10 54±3 63 vs 6 61±2 82ml·h·kg; P <0 05) and GAM (3 01±0 15, 2 98±0 21 vs 2 24mg±0 26mg; P <0 01) was significantly higher than that in portal hypertension control group. In tetrandrine group intrahepatic proliferative fibrous tissues were reduced and serum ALT (47 67±25 90 vs 189 33±41 21 King U; P <0 01), ALP (0 22±0 04 vs 0 31±0 06μmol·s -1 /L; P <0 01) and STB (4 75±0 76 vs 11 12±2 93μmol/L; P <0 01) were lowered as compared with these in portal hypertension control group. ALT (209 34±36 91 vs 189 33±41 21 King U; P >0 05) and STB (11 63±3 01 vs 11 12±2 93μmol/L; P >0 05) in propranolol group were not different from that in portal hypertension controls group, but it showed more marked hepatocellular degeneration and necrosis and elevation of ALP (0 46±0 05 vs 0 31±0 06μmol·s -1 /L; P <0 01). CONCLUSION Tetrandrine can improve the functions of gastric mucosa and liver, and facilitate the absorption of intrahepatic proliferative fibrous tissues. Propranolol can aggravate hepatosis though it may improve portal hypertensive gastric mucosal lesions.展开更多
The effect of surface charges on the cellular uptake rate and drug release profile of tetrandrine-loaded poly（lactic-co-glycolic acid）（PLGA） nanoparticles（TPNs） was studied. Stabilizer-free nanoprecipitation met...The effect of surface charges on the cellular uptake rate and drug release profile of tetrandrine-loaded poly（lactic-co-glycolic acid）（PLGA） nanoparticles（TPNs） was studied. Stabilizer-free nanoprecipitation method was used in this study for the synthesis of TPNs. A typical layer-by-layer approach was applied for multi-coating particles＇ surface with use of poly（styrene sulfonate） sodium salt（PSS） as anionic layer and poly（allylamine hydrochloride）（PAH） as cationic layer. The modified TPNs were characterized by different physicochemical techniques such as Zeta sizer, scanning electron microscopy and transmission electron microscopy. The drug loading efficiency, release profile and cellular uptake rate were evaluated by high performance liquid chromatography and confocal laser scanning microscopy, respectively. The resultant PSS/PAH/PSS/PAH/TPNs（4 layers） exhibited spherical-shaped morphology with the average size of 160.3±5.165 nm and zeta potential of –57.8 m V. The encapsulation efficiency and drug loading efficiency were 57.88% and 1.73%, respectively. Multi-layer coating of polymeric materials with different charges on particles＇ surface could dramatically influence the drug release profile of TPNs（4 layers vs. 3 layers）. In addition, variable layers of surface coating could also greatly affect the cellular uptake rate of TPNs in A549 cells within 8 h. Overall, by coating particles＇ surface with those different charged polymers, precise control of drug release as well as cellular uptake rate can be achieved simultaneously. Thus, this approach provides a new strategy for controllable drug delivery.展开更多
Objective: This study aims to clarify the effect of the active components puerarin and tetrandrine on the chondrogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods: Using network pharmacology, ...Objective: This study aims to clarify the effect of the active components puerarin and tetrandrine on the chondrogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods: Using network pharmacology, protein targets of puerarin and tetrandrine were predicted, and a database of cartilage formation targets was established. The protein target information related to disease was then collected, and the drug-targeting network was constructed by analyzing the protein–protein interactions. Genes related to chondrogenesis induced by puerarin and tetrandrine and chondroblast differentiation signaling pathways were searched. Finally, potential drug-and disease-related genes,as well as proteins, were screened and verified using real-time RT-PCR and western blotting.Results: Network pharmacological studies have shown that puerarin and tetrandrine are involved in BMSCs cartilage differentiation. The experimental results showed that puerarin and tetrandrine could regulate the expression of cartilage differentiation-related genes and proteins. Puerarin increased the protein expression of COL2 A1, COL10 A1, MMP13, and SOX-9,as well as the gene expression of Col2 a1, Mmp13, Tgfb1, and Sox-9. Tetrandrine increased the protein expression of COL2 A1,COL10 A1, MMP13, and SOX-9, as well as the gene expression of Col10 a1, Tgfb1, Sox-9, and Acan. The combination of puerarin and tetrandrine increased the protein expression of COL2 A1, COL10 A1, MMP13, and SOX-9 and the gene expression of Col2 a1,Col10 a1, Sox-9, and Acan.Conclusions: Puerarin, tetrandrine, and their combination can promote the proliferation of BMSCs and induce their differentiation into chondrocytes, and they are thus expected to be inducers of chondrogenic differentiation. These results suggest that puerarin and tetrandrine have potential therapeutic effects on osteoarthritis.展开更多
The changes of the cytoplasmic free calcium level in the neutrophils after smoke in-halation injury were observed in rabbits and then the effects of tetrandrine,a calcium antago-nist,on the changes of free calcium lev...The changes of the cytoplasmic free calcium level in the neutrophils after smoke in-halation injury were observed in rabbits and then the effects of tetrandrine,a calcium antago-nist,on the changes of free calcium level were studied.It was found that the number of neu-trophils increased significantly preceded by a transient decrease in the blood and also increasedin the bronehoalveolar lavage fluid after smoke inhalation.and the level of cytoplasmic free calci-um in the blood neutrophil increased likewise.Administration of tetrandrine resulted in a reduc-tion of the neutrophils number in the lungs and the free calcium level in the blood neutrophils toalleviate the pulmonary injury due to smoke inhalation.It is believed that there is a close rela-tionship between the activation of neutrophils and the pathophysiological changes of the lungs,and tetrandrine can exert its therapeutic effects on the injury by decreasing the free calcium levelin the neutrophils to modulate their functions.展开更多
To examine the role and effect of nitric oxide synthase type Ⅱ(NOSⅡ) in cirrhotic rats, expression of NOSⅡ mRNA was detected by real time RT-PCR. The enzymatic activity of nitric oxide synthase and the circulating ...To examine the role and effect of nitric oxide synthase type Ⅱ(NOSⅡ) in cirrhotic rats, expression of NOSⅡ mRNA was detected by real time RT-PCR. The enzymatic activity of nitric oxide synthase and the circulating levels of NO, systemic and portal hemodynamics and quantification of cirrhosis were measured. Chinese traditional medicine was used to treat cirrhotic rats and the effect of NO was evaluated. Double-blind method was used in experiment. Our results showed the concentration of NO and the enzymatic activity of NOS increased markedly at all stages of cirrhosis and iNOSmRNA was strongly expressed. Meanwhile, the portal-venous-pressure (PVP) and portal-venous-flow (PVF) were significantly increased. NO, NOS and iNOSmRNA were positively correlated to the degree of hepatic fibrosis. Tetrandrine significantly inhibited NO production and the expression of iNOSmRNA. Our results suggested that increased hepatic expression of NOSⅡ is one of the important factors causing cirrhosis and portal hypertension. Tetrandrine can significantly ameliorate cirrhosis and portal hypertension.展开更多
A rat model of pleurisy was established with an intrapleural injection carrageenan(Car) in the dose of 10 mg/kg to study the anti-inflammatory action of tetrandrine(Tet) and its mechanism. At the 8th hour after inject...A rat model of pleurisy was established with an intrapleural injection carrageenan(Car) in the dose of 10 mg/kg to study the anti-inflammatory action of tetrandrine(Tet) and its mechanism. At the 8th hour after injection, there was an increase of cytosolic free calcium level in the neutrophils (Neu[Ca￣(2+]_i) and intensification of calmodulin activity of the neutrophils(Neu-CaMA) in the pleural exudate. When Tet was given intragastrically in the doses of 10, 20 , 40 and 80 mg/kg 30 min before and 4 h after the injection of Car, the amount, protein content and neutrophil count of the pleural exudate were significantly reduced, Neu[Ca2+]_i decreased and Neu-CaMA inhibited. In addition, calcium(32 μmol/L) was able to antagonize the inhibition of Tet(20～160 μmol/L) on Neu-CaMA in vitro. On the basis of these findings, it is believed that the excellent anti-inflammatory effect of Tet might be related to its antagonism to calcium-calmodulin system of the body and the inhibition of Tet on Neu-CaMA is mediated through the reduction of Neu-[Ca2+]_i.展开更多
Objective:Exploring the key pathways affecting the development of early silicosis based on bioinformatics and in vitro experiments.Method:Collecting differentially expressed genes in silicosis patients through literat...Objective:Exploring the key pathways affecting the development of early silicosis based on bioinformatics and in vitro experiments.Method:Collecting differentially expressed genes in silicosis patients through literature mining;Collecting differentially expressed genes in silicon dioxide infusion mice by using a high-throughput gene expression database(GEO);Obtaining disease targets related to silicosis by means of online human Mendelian genetic database(OMIM),GeneCards and comparative toxicgenomics database(CTD);differentially expressed genes and disease targets were subjected to gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genome(KEGG)enrichment analysis via R-package and Metascape platforms,respectively.The Schrödinger and Pymol software were used for molecular docking and modification.Silicon dioxide-stimulated macrophages and epithelial cells were modeled and analyzed by PCR and western blot(WB).Result:2065 differentially expressed genes in silicosis patients,2291 differentially expressed genes in rat infused with silicon dioxide,and 803 targets for silicosis-related diseases were screened out.GO enrichment analysis mainly involves G protein-coupled receptor binding,the regulation of inflammatory response,and participation in immune response.The enrichment analysis of KEGG pathway mainly included ECM-receptor interaction,TNF signaling pathway,and IL-17 signaling pathway.IL-17 signaling pathway was screened out from different genes and disease targets,indicating that IL-17 signaling pathway might be the key pathway for the development of silicosis.Molecular docking results showed that the silicosis drug tetrandrine had good binding effect with the RAF/MEK/ERK pathway in the IL-17 signaling pathway.Cellular experiments showed that tetrandrine reduced the expression of inflammatory factors such as IL-6 and TGF-βin macrophages by regulating the RAF/MEK/ERKpathway,and inhibited the epithelialmesenchymal transition and expression of inflammatory factors in epithelial cells.Conclusion:Tetrandrine regulates the inflammatory response and epithelial-mesenchymal transition(EMT)through the RAF/MEK/ERK pathway and thus affects the early progression of silicosis.展开更多
Objective： To study the effects of tetrandrine （Tet） on phenotypic modulation of vascular smooth muscle cells （VSMCs） and expression of p38 mitogen-activated protein kinase （p38MAPK） as well as mitogen-activate...Objective： To study the effects of tetrandrine （Tet） on phenotypic modulation of vascular smooth muscle cells （VSMCs） and expression of p38 mitogen-activated protein kinase （p38MAPK） as well as mitogen-activated protein kinase phosphatase-1 （MKP-1） after vascular intimal injury. Methods： HE staining was used to analyze vascular morphology of sham-injured group, injured group and Tet-treated group at day 28. lmmunohistochemistry, Western blot and RT-PCR were respectively used to detect the expression change of smooth muscle a-actin （SMa-actin）, proliferation cell nuclear antigen （PCNA）, p38MAPK and MKP-1 of injured group and Tet group at days 7, 14 and 28 after balloon injury. Results： ① All layers of vascular wall in sham-injured group were intact at day 28. The neointimal area was significantly increased and the lumen area notably decreased in injured group at day 28. The neointimal proliferation in Tet treated group was less than that in injured group, and the lumen area of Tet group was significantly increased than that of injured group at day 28. ②Compared with the injured group, the expression of SMa-actin, PCNA, p38MAPK and MKP-1 of vascular wall in Tet group was no difference, and the neointimal proliferation condition was also basically as same as injured group at day 7 after injury. The expression of PCNA and p38MAKP in Tet group was obviously lower than that in injured group, and the expression of MKP-1 in Tet group was obviously higher than that in injured group at days 14 and 28 after injury. The expression of SMa-actin in Tet group was slightly higher than that in injured group at days 14 and 28 after injury. Conclusions： Tet could reduce neointimal proliferation by inhibiting VSMCs phenotypic modulation and p38MAPK signaling transduction pathway as well as its down regulation.展开更多
ObjectiveTo elucidate the effect of tetrandrine on acute ischemic renal injury and its relation with apoptosis. MethodsA model for bilateral post ischemic renal injury in rats was developed by clamping renal pedicles...ObjectiveTo elucidate the effect of tetrandrine on acute ischemic renal injury and its relation with apoptosis. MethodsA model for bilateral post ischemic renal injury in rats was developed by clamping renal pedicles for 45 min. Renal tissular DNA fragmentation analysis and renal tissular HE staining were used. Also quantitative analysis of apoptosis in injured renal tubular epithelium was carried out by using TdT mediated dUTP nick and labeling (TUNEL). ResultsApoptosis of renal tubular epithelium increased in acute ischemic renal injury. Tetrandrine could remarkably decrease the level of apoptosis in injured renal tubule while protecting renal tissue against the ischemic injuries. ConclusionTetrandrine could adjust the level of apoptosis in renal tubular epithelium and alleviate renal tissular injury.展开更多
基金This study was supported by the Natural Science Foundation of Hubei Province(No.2020CFB868).
文摘Objective There is a lack of effective and long-term safe drugs for the treatment of osteoarthritis(OA).Tetrandrine(Tet)has been approved and used to treat rheumatoid arthritis for several decades,but its effect on OA has not been investigated.Herein,we explored the effect of Tet on OA and its underlying mechanism.Methods OA was induced using destabilization of the medial meniscus(DMM)in C57BL/6J mice.The animals were randomly divided into sham,DMM,Tet,celecoxib(CXB),and indomethacin(INDO)groups.Each group was given solvent or corresponding drugs by gavage for 7 weeks after convalescence.Pathological staining,OARSI scores,micro-computed tomography and behavior tests were performed to evaluate the effects of Tet.Results Tet remarkably alleviated cartilage injury in the knee joint,limited bone remodeling in the subchondral bone,and delayed progression of OA.Tet also significantly relieved joint pain and maintained function.Further mechanistic studies revealed that Tet lowered inflammatory cytokine levels and selectively suppressed gene and protein expression of cyclooxygenase(COX)-2 but not COX-1(P<0.01).Tet also reduced the production of prostaglandin E2 without damaging the gastric mucosa.Conclusion We found that Tet could selectively inhibit COX-2 gene expression and decrease cytokine levels in mice,thus reducing inflammation and improving OA without obvious gastric adverse events.These results provide a scientific basis for the clinical application of Tet in the treatment of OA.
文摘The effect of tetrandrine (Tet) on the infarction area and volume of rat brain induced by middle cerebral artery occlusion (MCAO) was investigated. The treatment with Tet 7.5, 12.0 or 15.0 mg·kg 1 , or with fructose 1,6 diphosphate (FDP) 200 and 350 mg·kg 1 ip immediately after MCAO, respectively, significantly reduced the infarction area and volume in a dose dependent manner. MK801 and FDP also displayed a protective effect on brain ischemia. A combination of Tet and FDP administered immediately after MCAO, produced a more potent protective effect than those treated with Tet or FDP alone. When Tet or FDP was administered 1 h and 2 h after MCAO, respectively, they could still significantly reduce the infarction area and volume of brain tissue. But, there was no significant protective effect when these two compounds were given 3 h after MCAO.
文摘\ The effects of tetrandrine (Tet) on cytosolic free calcium ([Ca2+]i) in subcultured bovine aortic smooth muscle cells (SMC) were studied by Fura2 and ARCMMIC cation measurement system. Tet (1～100 μmol·L-1) had no effect on the resting [Ca2+]i, but had inhibitory effects on [Ca2+]i elevation induced by high K+, 5HT, ATP, Ang II and NE in the presence of extracellular Ca2+. High concentration of Tet also inhibited Pheinduced [Ca2+]i elevation in absence of extracellular Ca2+. Tet (1～100 μmol·L-1) inhibited KCl (60 mmol·L-1) induced [Ca2+]i elevation in dosedependent manner, the IC50 value was 9.2 (95% confidence limits: 5.7～14.9) mmol·L-1. The results suggested that Tet had blocking effects on both VOC and ROC in bovine aortic SMC. It appears that the mechanisms of blocking effect of Tet on ROC might be primarily due to its Ca2+ entry blocking effects.
基金National Natural Science Foundation of China(Grant No.30772556 and 30640070)Research Fund of Janssen Research Council and the‘985'Project in Peking University.
文摘It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and different 5-HT receptors on pentobarbital-induced sleep by using the loss-of-righting reflex method. The results showed that augmentative effect of tetrandrine on pentobarbital hypnosis in mice were potentiated by the p-MPPI （5-HT1A receptor antagonist） （1 mg/kg, i.p.） and ketanserin （5-HT2A/2C receptor antagonist） （1.5 mg/kg, i.p.）, respectively. Pretreatment with either 8-OH-DPAT （5-HT1A receptor agonist） （0.1 mg/kg, s.c.） or DOI （5-HT2A/2C receptor agonist） （0.2 mg/kg, i.p.） significantly decreased pentobarbital-induced sleep time, and tetrandrine （60 mg/kg, i.g.） significantly reversed this effect. These results suggest that both the 5-HTLA and 5-HT2A/2C subfamily may be involved in the potentiating mechanism of tetrandrine＇s effects on pantobarbital hypnosis.
基金the National Natural Science Foundation of China, No. 32060127 and No. 30660225
文摘AIM： To investigate the therapeutic effect of tetrandrine on liver fibrosis induced by thioacetamide in rats in vivo and in vitro. METHODS： In vitro study： we investigated the effect of tetrandrine on the apoptosis of rat hepatic stellate cells transformed by simian virus 40 （T-HSC/CI-6）, which retains the features of activated cells. In vivo study： hepatic fibrosis was induced in rats by thioacetamide. Tetrandrine was given orally to rats at doses of 5, 10 or 20 mg/kg for 4 wk compared with intraperitoneal injection of interferon-r. RESULTS： In vitro study： 5, 10 or 25 μg/mL of tetrandrine-induced activation of caspase-3 in t-HSC/CI-6 cells occurred dose-dependently. In vivo study： tetrandrine treatment as well as interferon-r significantly ameliorated the development of fibrosis as determined by lowered serum levels of aspartate aminotransferase （AST）, alanine aminotransferase （ALT）, total bilirubin （T-Bil） and the levels of liver hydroxyproline （Hyp）, hyaluronic acid （HA）, laminin （LN） and also improved histological findings. The effects of tetrandrine at the concentration of 20 mg/kg were better than the other concentration groups. CONCLUSION： Tetrandrine promotes the apoptosis of activated HSCs in vitro. Tetrandrine administration can prevent liver fibrosis and liver damage induced by thioacetamide in rats in vivo, indicating that it might exert a direct effect on rat HSCs.
基金Supported by the College Science and Technology Developing Foundation of Shanghai, No. 02BK14
文摘AIM: To investigate the effect of various concentrations of tetrandrine on activation of quiescent rat hepatic stellate cells (HSCs) and transforming growth factor-β (TGF-β) signaling in vitro.METHODS: HSCs were isolated from rats by in situperfusion of liver and 18% Nycodenz gradient centrifugation, and primarily cultured on uncoated plastic plates for 24 hwith DMEM containing 20% fetal bovine serum (FBS/DMEM) before the culture medium was substituted with 2% FBS/DMEM for another 24 h. Then, the HSCs were cultured in 2% FBS/DMEM with tetrandrine (0.25, 0.5, 1,2 mg/L, respectively). Cell morphological features were observed under an inverted microscope, smooth muscleα-actin (α-SMA) was detected by immunocytochemistry and image analysis system, laminin (LN) and type Ⅲprocollagen (PCⅢ) in supernatants were determined byradioimmunoassay. TGF-β1 mRNA, Smad 7 mRNA and Smad 7 protein were analyzed with RT-PCR and Western blotting, respectively.RESULTS: Tetrandrine at the concentrations of 0.25-2 mg/L prevented morphological transformation of HSC from the quiescent state to the activated one, while α-SMA, LN and PCⅢ expressions were inhibited. As estimated by gray values, the expression of α-SMA in tetrandrine groups (0.25, 0.5, 1, 2 mg/L) was reduced from 21.3% to 42.2%(control: 0.67, tetrandrine groups: 0.82, 0.85, 0.96, or 0.96, respectively, which were statistically different from the control, P＜0.01), and the difference was more significant in tetrandrine at 1 and 2 mg/L. The content of LN in supernatants was significantly decreased in tetrandrine groups to 58.5%, 69.1%, 65.8% or 60.0% that of the control respectively, and that of PCⅢ to 84.6%, 81.5%,75.7% or 80.7% respectively (P＜0.05 vs control), with no significant difference among tetrandrine groups. RTPCR showed that TGF-β1 mRNA expression was reduced by tetrandrine treatments from 56.56% to 87.90% in comparison with the control, while Smad 7 mRNA was increased 1.4-4.8 times. The TGF-β1 mRNA and Smad 7 mRNA expression was in a significant negative correlation (r= -0.755, P＜0.01), and both were significantly correlated with α-SMA protein expression (r = -0.938, P＜0.01;r = 0.938, P＜0.01, respectively). The up-regulation of Smad 7 protein by tetrandrine (1 mg/L)was confirmed by Western blotting as well.CONCLUSION: Tetrandrine has a direct inhibiting effect on the activation of rat HSCs in culture. It up-regulates the expression of Smad 7 which in turn blocks TGF-β1 expression and signaling.
文摘In the screening tests of drugs for silicosis in our laboratory, we found that TT, a type of alkaloid isolated from Stephania tetrandra, could inhibit the development of experimental silicosis of rats and the synthesis of collagen in rat lung. Chest X-rays of silicotic patients treaied with TT for 1-3 years showed obvious changes. The silicotic nodules became smallel and shadows became clearer. PVNO was proved to have anti-silicotic effect on animal and clinically. This presentation reports the effect of them on collagen mRNA.Dot blot results showed that 1 (Ⅰ) and 1 (Ⅲ) mRNA levels increased significantly at 60 and 120 days after the rats were exposed to silica dust. The mRNA levels went down at 1 and 3 months after treated by TT and PVNO. In situ hybridization observation revealed that the silver grains of Type Ⅰand Type Ⅲ collagen were scattered within the fibroblasts in cellular nodules and in thickened interstitium of silicosis tissue. The amounts of mRNA silver grains decreased in the lung tissue treated by TT and PVNO. It was suggested that TT and PVNO may inhibil the gene expression of collagen during silicosis
基金Supported by the National Natural Science Foundation of China, No. 30370643
文摘AIM: To investigate the effect of Tetrandrine (Tet) on LPS-induced NF-κB activation and cell injury in pancreatic acinar cells and to explore the mechanism of Tetrandrine preventing LPS-induced acinar cell injury. METHODS: Male rat pancreatic acinar cells were isolated by collagenase digestion, then exposed to LPS (10 mg/L), Tet (50μmol/L, 100μmol/L) or normal media. At different time point (30 min, 1 h, 4 h, 10 h) after treatment with the agents, cell viability was determined by MTT, the product and nuclear translocation of subunit p65 of NF-κB was visualized by immunofluorescence staining and nuclear protein was extracted to perform EMSA which was used to assay the NF-κB binding activity. RESULTS: LPS induced cell damage directly in a time dependent manner and Tet attenuated LPS-induced cell damage (50μmol/L, P < 0.05; 100μmol/L, P < 0.01). NF-κB p65 immunofluorescence staining in cytoplasm increased and began showing its nuclear translocation within 30 min and the peak was shown at 1 h of LPS 10 mg/L treatment. NF-κB DNA binding activity showed the same alteration pattern as p65 immunofluorescence staining. In Tet group, the immunofluorescence staining in cytoplasm and nuclear translocation of NF-κB were inhibited significantly. CONCLUSION: NF-κB activation is an important early event that may contribute to inflammatory responses and cell injury in pancreatic acinar cells. Tet possesses the protective effect on LPS-induced acinar cell injury by inhibiting NF-κB activation.
基金supported by a grant from the Jiangsu Natural Science Foundation (No. BK2005203).
文摘Objective To assess the ability of tetrandrine （Tet） to enhance the sensitivity to irradiation and its mechanism in cell lines of human breast cancer p53-mutant MCF-7/ADR, p53-wild-type MCF-7 and human colon carcinoma p53-mutant HT-29 as well as in C26 colorectal carcinoma-bearing BALB/c mice. Methods MCF-7/ADR, HT-29 and MCF-7 cells were exposed to irradiation in the absence or presence of tetrandrine. The effect of Tet on the cytotoxicity of X-irradiation in these three cells was determined and the effect of tetrandrine on cell cycle arrest induced by irradiation in its absence or presence was studied by flow cytometry. Moreover, mitotic index measurement determined mitosis of cells to enter mitosis. Western blotting was employed to detect cyclin B1 and Cdc2 proteins in extracts from irradiated or non-irradiated cells of MCF-7/ADR, HT-29 and MCF-7 treated with tetrandrine at various concentrations. Tumor growth delay assay was conducted to determine the radio-sensitization of tetrandrine in vivo. Results Clonogenic assay showed that tetrandrine markedly enhanced the lethal effect of X-rays on p53-mutant MCF-7/ADR and HT-29 cells and the sensitization enhancement ratio （SER） of tetrandrine was 1.51 and 1.63, but its SER was only 1.1 in p53-wt MCF-7 cells. Irradiated p53-mutant MCF-7/ADR and HT-29 cells were only arrested in G2/M phase while MCF-7 cells were arrested in G1 and G2/M phases. Radiation-induced G2 phase arrests were abrogated by tetrandrine in a concentration-dependent manner in MCF-7/ADR and HT-29 cells, whereas redistribution within MCF-7 cell cycle changed slightly. The proportion of cells in M phase increased from 1.3% to 14.7% in MCF-7/ADR cells, and from 1.5% to 13.2% in HT-29 cells, but 2.4% to 7.1% in MCF-7 cells. Furthermore, the levels of cyclin B 1 and Cdc2 expression decreased after X-irradiation in MCF-7/ADR and HT-29 cells, and the mitotic index was also lower. Tet could reverse the decrease and induce the irradiated cells to enter mitosis （M phase）. Endosomatic experiment showed that tetrandrine caused tumor growth delay in irradiated mice. Conclusion Tetrandrine boosts the cell killing activity of irradiation both in vitro and in vivo. Tetrandrine is a potent abrogator for G2 checkpoint control and can sensitize the cells to radiation.
基金supported by the National Natural Science Foundation of China,No.81070886
文摘Tetrandrine is one of the major active ingredients in Menispermaceae Stephania tetrandra S.Moore,and has specific therapeutic effects in ischemic cerebrovascular disease.Its use in vascular dementia has not been studied fully.Here,we investigated whether tetrandrine would improve behavioral and cellular impairments in a two-vessel occlusion rat model of chronic vascular dementia.Eight weeks after model establishment,rats were injected intraperitoneally with 10 or 30 mg/kg tetrandrine every other day for 4 weeks.Behavioral assessment in the Morris water maze showed that model rats had longer escape latencies in training trials,and spent less time swimming in the target quadrant in probe trials,than sham-operated rats.However,rats that had received tetrandrine showed shorter escape latencies and longer target quadrant swimming time than untreated model rats.Hematoxylin-eosin and Nissl staining revealed less neuronal necrosis and pathological damage,and more living cells,in the hippocampus of rats treated with tetrandrine than in untreated model rats.Western blot assay showed that interleukin-1β expression,and phosphorylation of the N-methyl-D-aspartate 2B receptor at tyrosine 1472,were lower in model rats that received tetrandrine than in those that did not.The present findings suggest that tetrandrine may be neuroprotective in chronic vascular dementia by reducing interleukin-1β expression,N-methyl-D-aspartate receptor 2B phosphorylation at tyrosine 1472,and neuronal necrosis.
文摘AIM To study the effects of tetrandrine on portal hypertensive gastric mucosal lesions. METHODS Portal hypertensive models were induced in Wistar rats by 60% CCl 4 3ml/kg body weight subcutaneous injection, once every 4 days for 56 days. The animals were randomly divided into portal hypertension, tetrandrine and propranolol groups and subsequently, treated by physiological saline, tetrandrine and propranolol respectively for 15 days. Some healthy rats were used as normal group. Portal venous pressure (PVP), gastric mucosal prostaglandin E 2 (PGE 2) content, gastric mucosal blood flow (GMBF), gastric adherent mucus (GAM), ALT, ALP and serum total bilirubin (STB), were measured and liver tissues were observed histologically. RESULTS In tetrandrine group and propranolol group, PVP was significantly lower (1 43±0 13, 1 45±0 12 vs 1 89±0 18kPa; P <0 01) and gastric mucosal PGE 2 content (138 59±12 68, 129 98±14 31 vs 104 65±12 97pg/mg; P <0 01), GMBF (11 80±3 47, 10 54±3 63 vs 6 61±2 82ml·h·kg; P <0 05) and GAM (3 01±0 15, 2 98±0 21 vs 2 24mg±0 26mg; P <0 01) was significantly higher than that in portal hypertension control group. In tetrandrine group intrahepatic proliferative fibrous tissues were reduced and serum ALT (47 67±25 90 vs 189 33±41 21 King U; P <0 01), ALP (0 22±0 04 vs 0 31±0 06μmol·s -1 /L; P <0 01) and STB (4 75±0 76 vs 11 12±2 93μmol/L; P <0 01) were lowered as compared with these in portal hypertension control group. ALT (209 34±36 91 vs 189 33±41 21 King U; P >0 05) and STB (11 63±3 01 vs 11 12±2 93μmol/L; P >0 05) in propranolol group were not different from that in portal hypertension controls group, but it showed more marked hepatocellular degeneration and necrosis and elevation of ALP (0 46±0 05 vs 0 31±0 06μmol·s -1 /L; P <0 01). CONCLUSION Tetrandrine can improve the functions of gastric mucosa and liver, and facilitate the absorption of intrahepatic proliferative fibrous tissues. Propranolol can aggravate hepatosis though it may improve portal hypertensive gastric mucosal lesions.
基金supported by grants from the National Natural Science Foundation of China(No.81101690)Natural Science Foundation of Hubei Province(No.2014CFB403)Applied Basic Research Foundation of Wuhan Science and Technology Committee(No.2014060101010034)
文摘The effect of surface charges on the cellular uptake rate and drug release profile of tetrandrine-loaded poly（lactic-co-glycolic acid）（PLGA） nanoparticles（TPNs） was studied. Stabilizer-free nanoprecipitation method was used in this study for the synthesis of TPNs. A typical layer-by-layer approach was applied for multi-coating particles＇ surface with use of poly（styrene sulfonate） sodium salt（PSS） as anionic layer and poly（allylamine hydrochloride）（PAH） as cationic layer. The modified TPNs were characterized by different physicochemical techniques such as Zeta sizer, scanning electron microscopy and transmission electron microscopy. The drug loading efficiency, release profile and cellular uptake rate were evaluated by high performance liquid chromatography and confocal laser scanning microscopy, respectively. The resultant PSS/PAH/PSS/PAH/TPNs（4 layers） exhibited spherical-shaped morphology with the average size of 160.3±5.165 nm and zeta potential of –57.8 m V. The encapsulation efficiency and drug loading efficiency were 57.88% and 1.73%, respectively. Multi-layer coating of polymeric materials with different charges on particles＇ surface could dramatically influence the drug release profile of TPNs（4 layers vs. 3 layers）. In addition, variable layers of surface coating could also greatly affect the cellular uptake rate of TPNs in A549 cells within 8 h. Overall, by coating particles＇ surface with those different charged polymers, precise control of drug release as well as cellular uptake rate can be achieved simultaneously. Thus, this approach provides a new strategy for controllable drug delivery.
文摘Objective: This study aims to clarify the effect of the active components puerarin and tetrandrine on the chondrogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods: Using network pharmacology, protein targets of puerarin and tetrandrine were predicted, and a database of cartilage formation targets was established. The protein target information related to disease was then collected, and the drug-targeting network was constructed by analyzing the protein–protein interactions. Genes related to chondrogenesis induced by puerarin and tetrandrine and chondroblast differentiation signaling pathways were searched. Finally, potential drug-and disease-related genes,as well as proteins, were screened and verified using real-time RT-PCR and western blotting.Results: Network pharmacological studies have shown that puerarin and tetrandrine are involved in BMSCs cartilage differentiation. The experimental results showed that puerarin and tetrandrine could regulate the expression of cartilage differentiation-related genes and proteins. Puerarin increased the protein expression of COL2 A1, COL10 A1, MMP13, and SOX-9,as well as the gene expression of Col2 a1, Mmp13, Tgfb1, and Sox-9. Tetrandrine increased the protein expression of COL2 A1,COL10 A1, MMP13, and SOX-9, as well as the gene expression of Col10 a1, Tgfb1, Sox-9, and Acan. The combination of puerarin and tetrandrine increased the protein expression of COL2 A1, COL10 A1, MMP13, and SOX-9 and the gene expression of Col2 a1,Col10 a1, Sox-9, and Acan.Conclusions: Puerarin, tetrandrine, and their combination can promote the proliferation of BMSCs and induce their differentiation into chondrocytes, and they are thus expected to be inducers of chondrogenic differentiation. These results suggest that puerarin and tetrandrine have potential therapeutic effects on osteoarthritis.
文摘The changes of the cytoplasmic free calcium level in the neutrophils after smoke in-halation injury were observed in rabbits and then the effects of tetrandrine,a calcium antago-nist,on the changes of free calcium level were studied.It was found that the number of neu-trophils increased significantly preceded by a transient decrease in the blood and also increasedin the bronehoalveolar lavage fluid after smoke inhalation.and the level of cytoplasmic free calci-um in the blood neutrophil increased likewise.Administration of tetrandrine resulted in a reduc-tion of the neutrophils number in the lungs and the free calcium level in the blood neutrophils toalleviate the pulmonary injury due to smoke inhalation.It is believed that there is a close rela-tionship between the activation of neutrophils and the pathophysiological changes of the lungs,and tetrandrine can exert its therapeutic effects on the injury by decreasing the free calcium levelin the neutrophils to modulate their functions.
文摘To examine the role and effect of nitric oxide synthase type Ⅱ(NOSⅡ) in cirrhotic rats, expression of NOSⅡ mRNA was detected by real time RT-PCR. The enzymatic activity of nitric oxide synthase and the circulating levels of NO, systemic and portal hemodynamics and quantification of cirrhosis were measured. Chinese traditional medicine was used to treat cirrhotic rats and the effect of NO was evaluated. Double-blind method was used in experiment. Our results showed the concentration of NO and the enzymatic activity of NOS increased markedly at all stages of cirrhosis and iNOSmRNA was strongly expressed. Meanwhile, the portal-venous-pressure (PVP) and portal-venous-flow (PVF) were significantly increased. NO, NOS and iNOSmRNA were positively correlated to the degree of hepatic fibrosis. Tetrandrine significantly inhibited NO production and the expression of iNOSmRNA. Our results suggested that increased hepatic expression of NOSⅡ is one of the important factors causing cirrhosis and portal hypertension. Tetrandrine can significantly ameliorate cirrhosis and portal hypertension.
文摘A rat model of pleurisy was established with an intrapleural injection carrageenan(Car) in the dose of 10 mg/kg to study the anti-inflammatory action of tetrandrine(Tet) and its mechanism. At the 8th hour after injection, there was an increase of cytosolic free calcium level in the neutrophils (Neu[Ca￣(2+]_i) and intensification of calmodulin activity of the neutrophils(Neu-CaMA) in the pleural exudate. When Tet was given intragastrically in the doses of 10, 20 , 40 and 80 mg/kg 30 min before and 4 h after the injection of Car, the amount, protein content and neutrophil count of the pleural exudate were significantly reduced, Neu[Ca2+]_i decreased and Neu-CaMA inhibited. In addition, calcium(32 μmol/L) was able to antagonize the inhibition of Tet(20～160 μmol/L) on Neu-CaMA in vitro. On the basis of these findings, it is believed that the excellent anti-inflammatory effect of Tet might be related to its antagonism to calcium-calmodulin system of the body and the inhibition of Tet on Neu-CaMA is mediated through the reduction of Neu-[Ca2+]_i.
基金National Natural Science Foundation of China(No.81971483)Anhui University Collaborative Innovation Project(No.GXXT‑2020‑058)Anhui University of Science and Technology Innovation and Entrepreneurship Project(No.2021CX2125,2021CX2126,2021CX2124)。
文摘Objective:Exploring the key pathways affecting the development of early silicosis based on bioinformatics and in vitro experiments.Method:Collecting differentially expressed genes in silicosis patients through literature mining;Collecting differentially expressed genes in silicon dioxide infusion mice by using a high-throughput gene expression database(GEO);Obtaining disease targets related to silicosis by means of online human Mendelian genetic database(OMIM),GeneCards and comparative toxicgenomics database(CTD);differentially expressed genes and disease targets were subjected to gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genome(KEGG)enrichment analysis via R-package and Metascape platforms,respectively.The Schrödinger and Pymol software were used for molecular docking and modification.Silicon dioxide-stimulated macrophages and epithelial cells were modeled and analyzed by PCR and western blot(WB).Result:2065 differentially expressed genes in silicosis patients,2291 differentially expressed genes in rat infused with silicon dioxide,and 803 targets for silicosis-related diseases were screened out.GO enrichment analysis mainly involves G protein-coupled receptor binding,the regulation of inflammatory response,and participation in immune response.The enrichment analysis of KEGG pathway mainly included ECM-receptor interaction,TNF signaling pathway,and IL-17 signaling pathway.IL-17 signaling pathway was screened out from different genes and disease targets,indicating that IL-17 signaling pathway might be the key pathway for the development of silicosis.Molecular docking results showed that the silicosis drug tetrandrine had good binding effect with the RAF/MEK/ERK pathway in the IL-17 signaling pathway.Cellular experiments showed that tetrandrine reduced the expression of inflammatory factors such as IL-6 and TGF-βin macrophages by regulating the RAF/MEK/ERKpathway,and inhibited the epithelialmesenchymal transition and expression of inflammatory factors in epithelial cells.Conclusion:Tetrandrine regulates the inflammatory response and epithelial-mesenchymal transition(EMT)through the RAF/MEK/ERK pathway and thus affects the early progression of silicosis.
文摘Objective： To study the effects of tetrandrine （Tet） on phenotypic modulation of vascular smooth muscle cells （VSMCs） and expression of p38 mitogen-activated protein kinase （p38MAPK） as well as mitogen-activated protein kinase phosphatase-1 （MKP-1） after vascular intimal injury. Methods： HE staining was used to analyze vascular morphology of sham-injured group, injured group and Tet-treated group at day 28. lmmunohistochemistry, Western blot and RT-PCR were respectively used to detect the expression change of smooth muscle a-actin （SMa-actin）, proliferation cell nuclear antigen （PCNA）, p38MAPK and MKP-1 of injured group and Tet group at days 7, 14 and 28 after balloon injury. Results： ① All layers of vascular wall in sham-injured group were intact at day 28. The neointimal area was significantly increased and the lumen area notably decreased in injured group at day 28. The neointimal proliferation in Tet treated group was less than that in injured group, and the lumen area of Tet group was significantly increased than that of injured group at day 28. ②Compared with the injured group, the expression of SMa-actin, PCNA, p38MAPK and MKP-1 of vascular wall in Tet group was no difference, and the neointimal proliferation condition was also basically as same as injured group at day 7 after injury. The expression of PCNA and p38MAKP in Tet group was obviously lower than that in injured group, and the expression of MKP-1 in Tet group was obviously higher than that in injured group at days 14 and 28 after injury. The expression of SMa-actin in Tet group was slightly higher than that in injured group at days 14 and 28 after injury. Conclusions： Tet could reduce neointimal proliferation by inhibiting VSMCs phenotypic modulation and p38MAPK signaling transduction pathway as well as its down regulation.
基金This paper was supported by the Natural Science Foundation of Nanjing Medical University (NY970 55)
文摘ObjectiveTo elucidate the effect of tetrandrine on acute ischemic renal injury and its relation with apoptosis. MethodsA model for bilateral post ischemic renal injury in rats was developed by clamping renal pedicles for 45 min. Renal tissular DNA fragmentation analysis and renal tissular HE staining were used. Also quantitative analysis of apoptosis in injured renal tubular epithelium was carried out by using TdT mediated dUTP nick and labeling (TUNEL). ResultsApoptosis of renal tubular epithelium increased in acute ischemic renal injury. Tetrandrine could remarkably decrease the level of apoptosis in injured renal tubule while protecting renal tissue against the ischemic injuries. ConclusionTetrandrine could adjust the level of apoptosis in renal tubular epithelium and alleviate renal tissular injury.
