Background:Puerarin(Pue)has been reported to be a natural active ingredient with multiple antifibrotic properties.This work aimed at exploring the function of Pue in oral submucousfibrosis(OSF)treatment.Methods:Human or...Background:Puerarin(Pue)has been reported to be a natural active ingredient with multiple antifibrotic properties.This work aimed at exploring the function of Pue in oral submucousfibrosis(OSF)treatment.Methods:Human oral mucosafibroblasts(hOMF)were induced with transforming growth factor beta1(TGF-β1)and intervened with Pue.Expressions offibrosis-related markers were analyzed by Western blot and IF staining.Cell viability was characterized by the CCK-8 assay.Expressions of miR-30 family members were quantified by qRT-PCR.The correlation betweenfibroblast activation protein(FAP)and miR-30 family expression was evaluated by the Pearson correlation coefficient.Bioinformatics prediction and dual-luciferase reporter assay were employed to verify the regulation between FAP and miR-30b-5p.The specific mechanism of Pue on OSF was explored through the promotion or inhibition of miR-30b-5p.Results:After induction by TGF-β1,hOMF showed upregulated Collagen I,Collagen III,and FAP expressions,while miR-30 family expression was downregulated with miR-30b-5p being the most significant.Pue intervention inhibited the excessive proliferation of TGF-β1-induced hOMF,downregulated FAP,collagen type 3(COL3A1),collagen type 1(COL1A1),matrix metalloproteinase 1(MMP1),and matrix metalloproteinase 3(MMP3)expressions,and restored miR-30 family expression.Bioinformatics prediction and dual-luciferase reporter assay revealed that miR-30b-5p selectively inhibited FAP expression.Mechanistically,miR-30b-5p mimic suppressed the excessive proliferation of TGF-β1-induced hOMF and declinedfibrosis levels.Pue intervention significantly reversed the promotion of TGF-β1-induced OSF by miR-30b-5p inhibition.Conclusion:Pue mediated miR-30b-5p targeting FAP against OSF,which provided a theoretical basis for the pathogenesis research and Pue application in OSF.展开更多
Objective:To investigate the effect of puerarin on inflammation and alveolar bone resorption in rats with experimental periodontitis.Methods:Thirty-six 7-week-old Sprague-Dawley rats were randomly divided into three g...Objective:To investigate the effect of puerarin on inflammation and alveolar bone resorption in rats with experimental periodontitis.Methods:Thirty-six 7-week-old Sprague-Dawley rats were randomly divided into three groups(n=12).Twelve rats as the control group and the remaining rats were to establish a chronic periodontitis model ligated with orthodontic ligature wire on the cervical part of the left maxillary first molar under general anesthesia.All three groups were administered by gavage for 14 days:equal amounts of saline were given to the control and periodontitis groups,and 400 mg/kg concentration of puerarin was given to the puerarin group.All rats were anesthetized after 24 h of the last administration,blood was collected from the abdominal aorta,and the serum was centrifuged for the detection of IL-4,IL-10,IL-6,and IFN-γin peripheral blood,then all rats were executed,some rats separated from the left maxilla,fresh gingival tissue removed from the buccal-palatal side of the left maxillary first molar,and the remaining maxillary bone tissue used for the detection of Micro-CT;some rats subjected to the left maxillary bone specimens taken with the gingival tissues,used for HE staining detection.Results:HE staining showed that inflammatory cell infiltration was significantly reduced in the puerarin group compared to the periodontitis group.ELISA analysis showed that puerarin promoted IL-4 and IL-10 expression and decreased IL-6 and IFN-γexpression levels in serum(P<0.05).Micro-CT showed that puerarin significantly reduced alveolar bone resorption compared to the periodontitis group(P<0.05).Conclusion:Puerarin may inhibit inflammation and alveolar bone resorption in experimental periodontitis rats.展开更多
Objective:To investigate the effects of puerarin on the activity of superoxide dismutase(SOD), and expressions of advanced glycation end-product(AGE) receptor(RAGE) and vascular endothelial growth factor(VEGF) in reti...Objective:To investigate the effects of puerarin on the activity of superoxide dismutase(SOD), and expressions of advanced glycation end-product(AGE) receptor(RAGE) and vascular endothelial growth factor(VEGF) in retinas of streptozotocin(STZ)-induced early diabetic rats. Methods:Diabetic rat models were established by inducing diabetes via intra-peritoneal injection of STZ.Rats were randomly divided into normal(control),diabetic(DM),and DM+ puerarin groups.After intra-gastric administration of puerarin(500 mg/kg/day for 4 weeks),levels of SOD and malondialdehyde(MDA) were determined in serum and retina.mRNA and protein expression levels of RAGE and VEGF in retinas were determined by real-lime polymerase chain reaction(RT-PCR)(mRNA) and Western blot analysis(protein levels).Results:There was significantly lower SOD activity and significantly higher MDA in serum and retinas of the DM group compared with the two other groups(P【0.05).After treatment with puerarin,SOD activity increased and MDA content decreased in this group(P【0.05).mRNA and protein expression levels of RACE and VECF in the DM group were significantly higher than those of the other groups (P【0.05),and decreased after puerarin treatment(P【0.05).Conclusions:Puerarin is able to enhance SOD activity,and inhibit RAGE and VEGF expressions in retinas of STZ-induced early diabetic ruts.展开更多
AIM To reveal the protective mechanism of the combined use of vitamin D and puerarin in the progression of hepatic fibrosis induced by carbon tetrachloride(CCl4).METHODS Eight-week-old male Wistar rats were randomly d...AIM To reveal the protective mechanism of the combined use of vitamin D and puerarin in the progression of hepatic fibrosis induced by carbon tetrachloride(CCl4).METHODS Eight-week-old male Wistar rats were randomly divided into a normal control group(C group), a CCl4 group(CCl4 group), a vitamin D group(V group), a puerarin group(P group), and a combined group of vitamin D and puerarin(V + P group), each of which contained ten rats. In this way, we built a rat model of CCl4-induced hepatic fibrosis with intervention by vitamin D, puerarin, or a combination of the two. After eight weeks, the mice were sacrificed to collect serum and liver specimens. Blood was collected to detect the hyaluronic acid(HA). We also measured hydroxyproline(Hyp) and prepared paraffin sections of liver. After Sirius red staining, the liver specimens were observed under a microscope. RT-PCR and western blot analysis were adopted to detect the mRNA and the proteinlevels of Collagen I, Collagen III, Wnt1, and β-catenin in the liver tissues, respectively.RESULTS Hepatic fibrosis was observed in the CCl4 group. In comparison, hepatic fibrosis was attenuated in the V, P, and V + P groups: the HA level in blood and the Hyp level in liver were reduced, and the mRNA levels of Collagen I, Collagen III, Wnt, and β-catenin in liver were also decreased, as well as the protein levels of Wnt1 and β-catenin. Among these groups, the V + P group demonstrated the greatest amelioration of hepatic fibrosis.CONCLUSION The combined application of vitamin D and puerarin is capable of alleviating CCl4-induced hepatic fibrosis of rats. As to the mechanism, it is probably because the combined use is able to silence the Wnt1/β-catenin pathway, suppress the activation of hepatic stellate cells, and reduce the secretion of collagen fibers, therefore improving the anti-hepatic fibrosis effect.展开更多
Objective:To investigate the effects of the active ingredients combined therapy on inflammatory factors interleukin 1 beta(IL-1β)and neuropeptide Y(NPY)based on pharmacodynamics in rats.Methods:The animal model was b...Objective:To investigate the effects of the active ingredients combined therapy on inflammatory factors interleukin 1 beta(IL-1β)and neuropeptide Y(NPY)based on pharmacodynamics in rats.Methods:The animal model was built by transient middle cerebral artery occlusion(MCAO).The method for evaluating the concentrations of the FA-Pr-AI components in rat plasma was established by using HPLC and the expression levels of IL-1βand NPY were determined by ELISA.A new mathematics method of the trend of percentage rate of change(PRC)was used to assess the correlation between pharmacokinetics(PK)and pharmacodynamics(PD).Results:FA-Pr-Al in combination reduced neurological deficits,decreased infarct volume and inhibited the expression levels of IL-1βand NPY(all P<0.05)compared with the model group.FA,Pr and Al all displayed two compartment open models in rats.Clockwise hysteresis loops were obtained by time-concentration-effect curves.IL-1βand NPY level changes in the plasma followed an opposite trend to the plasma concentration tendency after C_(max)was reached.Astragaloside's PRC value was significantly higher than those of FA and puerarin between 120 to 180 min.Conclusions:The pharmacokinetics of FA-PrAl in combination were closely related its pharmacodynamics in treating ischemia/reperfusion injury,and the components of FA-Pr-Al may have a synergistic pharmacological effect.Astragaloside may play a more pronounced role in regulating IL-1βand NPY levels compared with puerarin or FA.展开更多
Summary: To evaluate the protective effect of puerarin on ischemic myocardium in dogs with acute myocardial infarction (AMI) and to reveal its possible mechanism, 10 dogs were randomly divided into puerarin group (gro...Summary: To evaluate the protective effect of puerarin on ischemic myocardium in dogs with acute myocardial infarction (AMI) and to reveal its possible mechanism, 10 dogs were randomly divided into puerarin group (group G) and control group (group C). AMI model was established in all dogs. Puerarin or saline was administered over a period of 21 days. Coronary angiography was performed before and after ligation of coronary artery. Eight hemorheological parameters were examined before and 22 days after the operation. The infarct area and vessel density of myocardium were assessed. The infarct area in group G was smaller than that in group C. Angiography 2 h and 22 d after ligation of coronary artery revealed significant augmentation of collateral vessels in group G as compared with control group. The platelet aggregation and the blood viscosity were in- creased during AMI when compared with control phase, and the increased indexes during AMI would be inhibited when puerarin were given. Capillaries and distribution vessel density in is- chemic zone on day 22 showed statistically significant augmentation in group G as compared with control group. Puerarin might improve the opening and formation of coronary collateral circula- tion, and might inhibit the increase of platelet aggregation and the blood viscosity during AMI, and thereby improve microcirculation and restrict myocardial infarct area.展开更多
Objective To investigate the effects of puerarin (Pur) on myocardial perfusion and ventricular wall motion in patients with acute coronary syndrome (ACS).Methods Thirty-seven patients with ACS were randomly divided in...Objective To investigate the effects of puerarin (Pur) on myocardial perfusion and ventricular wall motion in patients with acute coronary syndrome (ACS).Methods Thirty-seven patients with ACS were randomly divided into two groups:conventional treatment group (n= 17,11 males,range of age:32-80 years,average age:60.9±4.9 years) and Pur treatment group (n=20,12 males,range of age:40-76 years,average age:62.7±3.5 years).Patients in the conventional treatment group received standard treatment according to the current guidelines,while patients in the Pur treatment group received intravenous administration of Pur (500 mg/day) for 10 days plus conventional treatment.Real-time myocardial contrast echocardiography (RT-MCE) was performed to evaluate the change in myocardial perfusion index (MPI) and ventricular wall motion index (VWMI) at admission and 10 days after treatment.Results At 10 days after treatment,MPI was significantly higher (P【0.01) and VWMI significantly lower (P【0.01) in the Pur group comparing with those in the conventional group.Conclusions Puerarin might improve myocardial microcirculation perfusion and ventricular wall motion in patients with ACS.展开更多
AIM: To evaluate the toxicity of endogeneous peroxynitrite on transplanted retinal pigment epithelial (RPE) sheets and the effect of puerarin on their survival in the C57BL/6 mice after RPE sheets have been transplant...AIM: To evaluate the toxicity of endogeneous peroxynitrite on transplanted retinal pigment epithelial (RPE) sheets and the effect of puerarin on their survival in the C57BL/6 mice after RPE sheets have been transplanted into SD rats' subretinal space .METHODS: C57BL/6 mice eyes were used to culture RPE cells.Ninety-six SD rats were involved in the experiment.They were divided into control(block control) ,streptozotocin(STZ,negative control),untransplanted RPE (positive control) and transplanted RPE groups respectively.Diabetes was induced in SD rats by intra-peritoneal STZ injection in the latter three groups.Saline was injected into the subretinal space of 24 SD rats in the untransplanted RPE group and primary RPE sheets were injected into the subretinal space of 24 SD rats in the transplanted RPE group.Puerarin (45 mg/kg) was administrated into both untransplanted RPE and transplanted RPE groups of diabetic rats through intraperitoneal injection route after RPE sheets transplantation.At 20,40,60 days after surgery,Western blotting analysis,DNA ladder and RT-PCR were used for determining the differences in expression of nitrotyrosine (NT,the foot print of peroxynitrite ),apoptosis and iNOS mRNA in the control,STZ,untransplanted RPE and transplanted RPE groups respectively.HE staining was used for determining the RPE survival in the subretinal space of the transplanted RPE group.RESULTS: Apoptosis and expression of NT and iNOS mRNA were observed in STZ,untransplanted RPE and transplanted RPE groups,but were delayed in untransplanted RPE and transplanted RPE groups in a time-dependent manner compared with control and STZ groups (P<0.01).There were no differences between the two groups (P >0.01).NT,DNA ladder,iNOS mRNA were down-regulated,which were associated with the decrease of expression of peroxynitrite.Numerous pigmented cells emerged and increased in number in the subretinal space during the 60-day observation period after transplantation.On day 20,heavily pigmented cells were visible at the transplant site;On day 40,monolayer and multilayered transplant was visible in the subretinal space;On day 60,heavily pigmented monolayer and multilayered transplants with round apical profile were present along Bruch's membrane.CONCLUSION: Puerarin increased the 60-day survival of C57BL/6 mice RPE xenografts in the SD rats' subretinal space,which may be related to its direct inhibition of apoptosis of RPE cells and antagnism of damage of peroxynitrite to RPE cells.展开更多
A type of triangular pyramid-shaped microparticles of puerarin was synthesized by using oil-in-oil microemulsion approach which is simple and economical under the action of copper substrate.The pyramid-shaped micropar...A type of triangular pyramid-shaped microparticles of puerarin was synthesized by using oil-in-oil microemulsion approach which is simple and economical under the action of copper substrate.The pyramid-shaped microparticles would be made up of deposit of nanospheres or nanorods and have two significant characters.One is its complex surface morphology like coral reef.The other is a lot of nanopores in existence in the microparticle body.Two possible formation routes were speculated.展开更多
This work was done to investigate succinylated commercial whey protein isolate(S-WPI)as an oral sustained-release delivery carrier for puerarin 5(PR-5). The succinylation condi-tions were established for S-WPIs by opt...This work was done to investigate succinylated commercial whey protein isolate(S-WPI)as an oral sustained-release delivery carrier for puerarin 5(PR-5). The succinylation condi-tions were established for S-WPIs by optimization of single factor study and Box–Beehnkendesign. The effect of succinylation degree on S-WPIs solubility was evaluated. Physicochem-ical properties of S-WPIs dried by different three methods on their flow ability, particle size,morphology and in vitro release behavior were studied. After preparing PR-5 sustained re-lease protein tablets with S-WPIs as the carrier by direct powder compression method, thedrug release were studied in vitro and the oral pharmacokinetics and bioavailability wasevaluated using in vivo dog model. It was observed that concentration of substrate has asignificant effect on succinylation. Release behavior in vitro showed spry dried S-WPIs with100% succinylation rate and 30% drug loading would be applied to the preparation of PR-5 sustained-release protein tablets based on the swelling mechanism(protein loss). Comparedwith PR-5 conventional tablet with oral administration, T max value of PR-5 sustained-releaseprotein tablets was approximately 1.58 fold greater than those of the conventional tablets asfurther evidenced by the significantly prolonged MRT and T 1/2. The findings demonstratedthat spray-dried S-WPIs has potential as a promising functional excipient for the design of PR-5 oral sustained-release tablets which can fully improve sustained-release effect and oral bioavailability.展开更多
Objective:In this study,the influence of puerarin,paeoniflorin,and menthol on the structure and barrier function of tight junctions(TJs)in MadineDarby canine kidney epithelial(MDCK)and MDCK-multi-drug resistance 1(MDR...Objective:In this study,the influence of puerarin,paeoniflorin,and menthol on the structure and barrier function of tight junctions(TJs)in MadineDarby canine kidney epithelial(MDCK)and MDCK-multi-drug resistance 1(MDR1)cells was evaluated to determine the mechanisms by which the drugs cross the bloodebrain barrier(BBB).Method:Cells were treated with puerarin,paeoniflorin,and menthol followed by immunohistochemical staining with occludin,claudin-1,and F-actin.The cells were then observed using laser-scanning confocal microscopy.Average optical density(AOD)of the immunofluorescence images of the proteins were analyzed using ImageJ software while Transepithelial electrical resistance(TEER)was measured using an epithelial voltohmmeter.Results:Confocal microscopy revealed that puerarin-and paeoniflorin-treated tight junction proteins were conspicuous while menthol suppressed their expression.Correspondingly,AOD values of cells treated with puerarin or paeoniflorin,or both showed no difference compared to the control group(P>.05)while the menthol group value was downregulated.In 3 h,TEER of cells not treated with menthol were similar to the control group,while treatment with menthol significantly decreased TEER value(P<.05).In addition,application of menthol decreased TEER in MDCK cells earlier than in MDCK-MDR1 cells.Conclusion:Menthol but not puerarin and paeoniflorin may enhance paracellular transport and improve drug penetration of the BBB by disrupting the structure and,thereby,weakening the barrier function of TJs.展开更多
The purpose of this study was to investigate the influence of the supercritical CO2 processing on the particle size and morphology of puerarin crystals. The process parameters included solvents, temperature, pressures...The purpose of this study was to investigate the influence of the supercritical CO2 processing on the particle size and morphology of puerarin crystals. The process parameters included solvents, temperature, pressures, antisolvent times, addition volumes, antisolvent addition rates and solute concentrations. After being processed, the dramatic reduction of the dimensions and the change of the crystal shape were noticed. Decreasing the antisolvent addition rate, increasing the temperature and the addition volume below 50 ml led to a decrease in size. The new crystal of puerarin generated at the optimal conditions was 30.34 μm.The solvent of methanol and the concentration of 60 mg/ml were found to determine the type and degree of crystallinity of particles. These results showed that this process has the potential to produce a drug recrystallization product with newly generated crystal forms and the size of drug particles could be controlled through the tuning of various experimental conditions.展开更多
OBJECTIVE The aim of this study was to investigate the protective effect of puerarin on alcoholtoxicity in rat pheochromocytoma cell line(PC12). METHODS The PC12 cells were incubated with different concentrations of p...OBJECTIVE The aim of this study was to investigate the protective effect of puerarin on alcoholtoxicity in rat pheochromocytoma cell line(PC12). METHODS The PC12 cells were incubated with different concentrations of puerarin in advance. The protective effects of the puerarin on alcohol induced PC12 cel impairment were evaluated according to the fol owing approach: the viability of PC12 cel was determined by MTT assay and the impairment level was evaluated by analysis the leakage content of the lactate dehydrogenase(LDH). The cel apoptosis degree and the pro-apoptotic p53 protein expression were measured by flow cytometry. RESULTS Alcohol significantly impaired PC12 cel viability(P<0.05),and increased LDH leakage(P<0.05),induced cell apoptosis and upregulated expression of p53(P<0.05).While Puerarin significantly reversed these changes(P<0.05). CONCLUSION Puerarin might exert protection effect against ethanol-induced neurotoxicity via inhibition the expression of p53 protein.展开更多
NLRP3 inflammasome-mediated cell pyroptosis aggravates the development of cerebral ischemia/reperfusion(I/R)injury,and the aim of this study is to investigate the potential utilization of the Chinese medicine,Puerarin...NLRP3 inflammasome-mediated cell pyroptosis aggravates the development of cerebral ischemia/reperfusion(I/R)injury,and the aim of this study is to investigate the potential utilization of the Chinese medicine,Puerarin,in treating this disease.Through conducting in vitro and in vivo experiments,the present study illustrated that Puerarin regulated LncRNA double homeobox A pseudogene 8(DUXAP8)/miR-223-3p axis to inactivate NLRP3-mediated pyroptotic cell death,resulting in the attenuation of I/R injury.Specifically,the cerebral I/R injury in rat models and hypoxia/reoxygenation(H/R)in primary hippocampus neuron(PHN)cells were inducted,which were subsequently exposed to Puerarin treatment.As expected,we validated that Puerarin suppressed cell pyroptosis and rescued cell viability in I/R rat hippocampus tissues and H/R PHN cells.Next,through bioinformatics analysis,we noticed that miR-223-3p targeted both LncRNA DUXAP8 and NLRP3 mRNA,and both LncRNA DUXAP8 ablation and miR-223-3p overexpression inactivate NLRP3-mediated cell pyroptosis to rescue cell viability in H/R PHN cells.Interestingly,we evidenced that Puerarin restrained LncRNA DUXAP8 expressions,but upregulated miR-223-3p in I/R rat tissues and H/R PHN cells,and the protective effects of Puerarin on H/R PHN cells were abrogated by overexpressing LncRNA DUXAP8 and silencing miR-223-3p.Collectively,we concluded that Puerarin regulated LncRNA DUXAP8/miR-223-3p/NLRP3 signaling cascade to attenuate I/R injury.展开更多
Pueraria mirifica(PM)has traditionally been used to relieve postmenopausal symptoms.Recently,its extract has been developed into various cosmetic products to promote skin rejuvenation and youthfulness.This study inves...Pueraria mirifica(PM)has traditionally been used to relieve postmenopausal symptoms.Recently,its extract has been developed into various cosmetic products to promote skin rejuvenation and youthfulness.This study investigated the phytochemicals of PM tuber and compared between the tuber flesh and its outer peel.Puerarin which is one of the major isoflavones and being considered as the marker compound was used to determine the presence of PM extract in local cosmetic products.Puerarin could be ionized by a mass spectrometer at both negative and positive modes.The peak ionized at the negative mode showed to have a narrower peak width(0.2 min)and higher signal-to-noise ratio(30)for pueararin(1 mg/L).The results also found PM extract contained many C-and O-glycosylated isoflavones,especially from its peel extract.This explains the peel extract showed to have four times higher antiradical activity than those of flesh extract.Puerarin from the cosmetic products was recovered via successive methanolic sonication and followed by liquid-liquid extraction using ethyl acetate.Puerarin was successfully partitioned from the highly complex chemical mixture of cosmetic products with the recovery ranged from 89.1%to 115%.Hence,isoflavones was found to be higher at the outer peels than its tuber flesh.A simple and reliable method has been developed to analyse the presence of PM extract in cosmetic products based on the detection of puerarin after successive extraction via methanolic sonication and ethyl acetate partition.展开更多
Objective To investigate whether the protective effects of puerarine(Pur)against cerebral ischemia is associated with depressing the extracellular levels of amino acid transmitters in brain of rats.Methods Male Spragu...Objective To investigate whether the protective effects of puerarine(Pur)against cerebral ischemia is associated with depressing the extracellular levels of amino acid transmitters in brain of rats.Methods Male Sprague-Dawley rats were subjected to transient middle cerebral artery occlusion(MCAO)for 60 min followed by 24 h reperfusion.Pur(50,100 mg/kg,i.p.)was administered at the onset of MCAO.The infarct rate and edema rate were detected on TTC(2,3,5-triphenyltetrazolium chloride)-stained coronal sections.The extracellular levels of amino acid transmitters were monitored in striatum of rats with ischemic/reperfusion injury using in vivo microdialysis technique.Furthermore,the protective effects of Pur against glutamate-induced neurotoxicity were detected.Glutamate-induced apoptotic and necrotic cells in hippocampus were estimated by flow cytometric analysis of Annexin-V and PI labeling cells.Results Pur(100 mg/kg)significantly decreased infarct size by 31.6%(P<0.05),reduced edema volume(P<0.05),and improved neurological functions(P<0.05)following MCAO.In these rats,the ischemia-induced extracellular levels of aspartate(Asp),glutamate(Glu),γ-aminobutyric acid(GABA),and taurine(Tau)were significantly reduced in striatum of vehicle-treated animals by 54.7%,56.7%,75.8%,and 68.1%(P<0.01 and P<0.05).Pur reduced the peak values of Glu and Asp more obviously than those of GABA and Tau,and the rate of Glu/GABA during MCAO markedly decreased in Pur-treated MCAO rats,compared with the vehicle-treated MCAO rats.Meanwhile,apoptosis and necrosis induced by Glu in cultured hippocampal neurons were significantly reduced after Pur treatment.Conclusion Acute treatment with Pur at the onset of occlusion significantly depresses ischemia-induced efflux of amino acids,especially,excitotoxicity in the striatum,a mechanism underlying the neuroprotective effect on cellular survival.展开更多
A novel compound, diethyl puerarin-7-yl phosphate, was synthesized through a simplified Atheron-Todd reaction for the first time. The structure of this compound was elucidated by IR, ESI-MS and NMR. Two conformations ...A novel compound, diethyl puerarin-7-yl phosphate, was synthesized through a simplified Atheron-Todd reaction for the first time. The structure of this compound was elucidated by IR, ESI-MS and NMR. Two conformations of the compound were testified by 2D NMR (HSQC and HMBC) and dynamic NMR. Furthermore, we carried out the conformational analysis using chemical calculation by the Gaussian 03. Finally, we obtained two preferred conformations and energy values.展开更多
Increasing evidence indicates that disruption of normal iron homeostasis may contribute to pathological development of Alzheimer's disease.Icariin,astragalus,and puerarin have been shown to suppress iron overload ...Increasing evidence indicates that disruption of normal iron homeostasis may contribute to pathological development of Alzheimer's disease.Icariin,astragalus,and puerarin have been shown to suppress iron overload in the cerebral cortex and improve spatial learning and memory disorders in Alzheimer's disease mice,although the underlying mechanism remains unclear.In the present study,APPswe/PS1ΔE9 transgenic mice were administered icariin,astragalus,and puerarin(120,80,and 80 mg/kg,respectively,once a day,for 3 months).Iron levels were detected by flame atomic absorption spectroscopy.Interleukin-1β,interleukin-6,and tumor necrosis factor-α levels were measured in the cerebral cortex by enzyme linked immunosorbent assay.Glutathione peroxidase and superoxide dismutase activity and malondialdehyde content were determined by colorimetry.Our results demonstrate that after treatment,iron levels and malondialdehyde content are decreased,while glutathione peroxidase and superoxide dismutase activities are increased.Further,interleukin-1β,interleukin-6,and tumor necrosis factor-α levels were reduced.These results confirm that compounds of icariin,astragalus,and puerarin may alleviate iron overload by reducing oxidative stress and the inflammatory response.展开更多
Summary: The protective effect of puerarin on the Parkinson disease (PD) mice with decreased es-trogen level was investigated in order to develop a new potential medicine as a substitute for estrogenfor preventing and...Summary: The protective effect of puerarin on the Parkinson disease (PD) mice with decreased es-trogen level was investigated in order to develop a new potential medicine as a substitute for estrogenfor preventing and treating PD. By using immunohistochemical method of avidinbiotin peroxidasecomplex (ABC), the distribution of the cells positive for tyrosine hydroxylase (TH) and fibres in thesubstantia nigra of the mouse were observed. These mice were divided into three groups randomly:group A, normal-female-mouse models; group B containing three subgroups, B1 (normal saline), B2(estrogen), B3 (puerarin); group C containing three sub groups, C1 (normal saline), C2(estrogen), C3 (puerarin). By using TUNEL the numbers of apoptosis cells in every visual field wascounted and the difference between the experimental group and control group was compared. The re-sults showed the numbers of the cells positive for TH were more and the numbers of apoptosis cellswere less in the normal-female-mouse models group than in the group of model made after ovar-iosteresis and the group of model made before ovariosteresis (P＜0.05), respectively. However,there was no significant difference, between the group given estrogen/puerarin and the controls, andbetween the group given estrogen and given puerarin. (P＞0. 05). It was suggested that puerarinmay have protective effect on the nigral neurons to PD. Moreover, the protective effect might serveas a surrogate of estrogen and be associated with the apoptosis.展开更多
基金This work was supported by the National Natural Science Foundation of China(Nos.81874496,82374530)the Clinical Medical Technology Innovation Guide Project of Hunan Province(No.2020SK53206)+3 种基金the Natural Science Foundation of Hunan Province(No.2021JJ70062)the Changsha Natural Science Foundation Project(No.kq2014019)the Health Special Fund Research Project of Hunan Province(No.B2020-07)the Clinical Pharmaceutical Research Fund of Hunan Medical Association(No.B202012).
文摘Background:Puerarin(Pue)has been reported to be a natural active ingredient with multiple antifibrotic properties.This work aimed at exploring the function of Pue in oral submucousfibrosis(OSF)treatment.Methods:Human oral mucosafibroblasts(hOMF)were induced with transforming growth factor beta1(TGF-β1)and intervened with Pue.Expressions offibrosis-related markers were analyzed by Western blot and IF staining.Cell viability was characterized by the CCK-8 assay.Expressions of miR-30 family members were quantified by qRT-PCR.The correlation betweenfibroblast activation protein(FAP)and miR-30 family expression was evaluated by the Pearson correlation coefficient.Bioinformatics prediction and dual-luciferase reporter assay were employed to verify the regulation between FAP and miR-30b-5p.The specific mechanism of Pue on OSF was explored through the promotion or inhibition of miR-30b-5p.Results:After induction by TGF-β1,hOMF showed upregulated Collagen I,Collagen III,and FAP expressions,while miR-30 family expression was downregulated with miR-30b-5p being the most significant.Pue intervention inhibited the excessive proliferation of TGF-β1-induced hOMF,downregulated FAP,collagen type 3(COL3A1),collagen type 1(COL1A1),matrix metalloproteinase 1(MMP1),and matrix metalloproteinase 3(MMP3)expressions,and restored miR-30 family expression.Bioinformatics prediction and dual-luciferase reporter assay revealed that miR-30b-5p selectively inhibited FAP expression.Mechanistically,miR-30b-5p mimic suppressed the excessive proliferation of TGF-β1-induced hOMF and declinedfibrosis levels.Pue intervention significantly reversed the promotion of TGF-β1-induced OSF by miR-30b-5p inhibition.Conclusion:Pue mediated miR-30b-5p targeting FAP against OSF,which provided a theoretical basis for the pathogenesis research and Pue application in OSF.
基金Hainan Provincial Financial Science and Technology Program Funded Project-Hainan Provincial Key Research and Development Social Development Project 2020(No.ZDYF2020166)Hainan Provincial Financial Science and Technology Program funded project-Hainan Provincial Key Research and Development Social Development Project 2023(No.ZDYF2023SHFZ095)。
文摘Objective:To investigate the effect of puerarin on inflammation and alveolar bone resorption in rats with experimental periodontitis.Methods:Thirty-six 7-week-old Sprague-Dawley rats were randomly divided into three groups(n=12).Twelve rats as the control group and the remaining rats were to establish a chronic periodontitis model ligated with orthodontic ligature wire on the cervical part of the left maxillary first molar under general anesthesia.All three groups were administered by gavage for 14 days:equal amounts of saline were given to the control and periodontitis groups,and 400 mg/kg concentration of puerarin was given to the puerarin group.All rats were anesthetized after 24 h of the last administration,blood was collected from the abdominal aorta,and the serum was centrifuged for the detection of IL-4,IL-10,IL-6,and IFN-γin peripheral blood,then all rats were executed,some rats separated from the left maxilla,fresh gingival tissue removed from the buccal-palatal side of the left maxillary first molar,and the remaining maxillary bone tissue used for the detection of Micro-CT;some rats subjected to the left maxillary bone specimens taken with the gingival tissues,used for HE staining detection.Results:HE staining showed that inflammatory cell infiltration was significantly reduced in the puerarin group compared to the periodontitis group.ELISA analysis showed that puerarin promoted IL-4 and IL-10 expression and decreased IL-6 and IFN-γexpression levels in serum(P<0.05).Micro-CT showed that puerarin significantly reduced alveolar bone resorption compared to the periodontitis group(P<0.05).Conclusion:Puerarin may inhibit inflammation and alveolar bone resorption in experimental periodontitis rats.
文摘Objective:To investigate the effects of puerarin on the activity of superoxide dismutase(SOD), and expressions of advanced glycation end-product(AGE) receptor(RAGE) and vascular endothelial growth factor(VEGF) in retinas of streptozotocin(STZ)-induced early diabetic rats. Methods:Diabetic rat models were established by inducing diabetes via intra-peritoneal injection of STZ.Rats were randomly divided into normal(control),diabetic(DM),and DM+ puerarin groups.After intra-gastric administration of puerarin(500 mg/kg/day for 4 weeks),levels of SOD and malondialdehyde(MDA) were determined in serum and retina.mRNA and protein expression levels of RAGE and VEGF in retinas were determined by real-lime polymerase chain reaction(RT-PCR)(mRNA) and Western blot analysis(protein levels).Results:There was significantly lower SOD activity and significantly higher MDA in serum and retinas of the DM group compared with the two other groups(P【0.05).After treatment with puerarin,SOD activity increased and MDA content decreased in this group(P【0.05).mRNA and protein expression levels of RACE and VECF in the DM group were significantly higher than those of the other groups (P【0.05),and decreased after puerarin treatment(P【0.05).Conclusions:Puerarin is able to enhance SOD activity,and inhibit RAGE and VEGF expressions in retinas of STZ-induced early diabetic ruts.
基金Supported by Key Laboratory Cultivation Base of Universities in Guangxi(Guangxi Education Research)No.[2014] 6
文摘AIM To reveal the protective mechanism of the combined use of vitamin D and puerarin in the progression of hepatic fibrosis induced by carbon tetrachloride(CCl4).METHODS Eight-week-old male Wistar rats were randomly divided into a normal control group(C group), a CCl4 group(CCl4 group), a vitamin D group(V group), a puerarin group(P group), and a combined group of vitamin D and puerarin(V + P group), each of which contained ten rats. In this way, we built a rat model of CCl4-induced hepatic fibrosis with intervention by vitamin D, puerarin, or a combination of the two. After eight weeks, the mice were sacrificed to collect serum and liver specimens. Blood was collected to detect the hyaluronic acid(HA). We also measured hydroxyproline(Hyp) and prepared paraffin sections of liver. After Sirius red staining, the liver specimens were observed under a microscope. RT-PCR and western blot analysis were adopted to detect the mRNA and the proteinlevels of Collagen I, Collagen III, Wnt1, and β-catenin in the liver tissues, respectively.RESULTS Hepatic fibrosis was observed in the CCl4 group. In comparison, hepatic fibrosis was attenuated in the V, P, and V + P groups: the HA level in blood and the Hyp level in liver were reduced, and the mRNA levels of Collagen I, Collagen III, Wnt, and β-catenin in liver were also decreased, as well as the protein levels of Wnt1 and β-catenin. Among these groups, the V + P group demonstrated the greatest amelioration of hepatic fibrosis.CONCLUSION The combined application of vitamin D and puerarin is capable of alleviating CCl4-induced hepatic fibrosis of rats. As to the mechanism, it is probably because the combined use is able to silence the Wnt1/β-catenin pathway, suppress the activation of hepatic stellate cells, and reduce the secretion of collagen fibers, therefore improving the anti-hepatic fibrosis effect.
基金supported by the National Science Foundation of China(81274176)the Clinical Medicine Special Foundation of China(12012064)+1 种基金the National Science Foundation of Province(LY13H280008)the Science and Technology Department of public welfare project(2014C33212)
文摘Objective:To investigate the effects of the active ingredients combined therapy on inflammatory factors interleukin 1 beta(IL-1β)and neuropeptide Y(NPY)based on pharmacodynamics in rats.Methods:The animal model was built by transient middle cerebral artery occlusion(MCAO).The method for evaluating the concentrations of the FA-Pr-AI components in rat plasma was established by using HPLC and the expression levels of IL-1βand NPY were determined by ELISA.A new mathematics method of the trend of percentage rate of change(PRC)was used to assess the correlation between pharmacokinetics(PK)and pharmacodynamics(PD).Results:FA-Pr-Al in combination reduced neurological deficits,decreased infarct volume and inhibited the expression levels of IL-1βand NPY(all P<0.05)compared with the model group.FA,Pr and Al all displayed two compartment open models in rats.Clockwise hysteresis loops were obtained by time-concentration-effect curves.IL-1βand NPY level changes in the plasma followed an opposite trend to the plasma concentration tendency after C_(max)was reached.Astragaloside's PRC value was significantly higher than those of FA and puerarin between 120 to 180 min.Conclusions:The pharmacokinetics of FA-PrAl in combination were closely related its pharmacodynamics in treating ischemia/reperfusion injury,and the components of FA-Pr-Al may have a synergistic pharmacological effect.Astragaloside may play a more pronounced role in regulating IL-1βand NPY levels compared with puerarin or FA.
基金This project was supported by a grant from the Scientific Committee of Hubei Province!(No. 951p1502).
文摘Summary: To evaluate the protective effect of puerarin on ischemic myocardium in dogs with acute myocardial infarction (AMI) and to reveal its possible mechanism, 10 dogs were randomly divided into puerarin group (group G) and control group (group C). AMI model was established in all dogs. Puerarin or saline was administered over a period of 21 days. Coronary angiography was performed before and after ligation of coronary artery. Eight hemorheological parameters were examined before and 22 days after the operation. The infarct area and vessel density of myocardium were assessed. The infarct area in group G was smaller than that in group C. Angiography 2 h and 22 d after ligation of coronary artery revealed significant augmentation of collateral vessels in group G as compared with control group. The platelet aggregation and the blood viscosity were in- creased during AMI when compared with control phase, and the increased indexes during AMI would be inhibited when puerarin were given. Capillaries and distribution vessel density in is- chemic zone on day 22 showed statistically significant augmentation in group G as compared with control group. Puerarin might improve the opening and formation of coronary collateral circula- tion, and might inhibit the increase of platelet aggregation and the blood viscosity during AMI, and thereby improve microcirculation and restrict myocardial infarct area.
基金Jiangsu Bureau of Traditimal Chinese Medicine (No.H05105)
文摘Objective To investigate the effects of puerarin (Pur) on myocardial perfusion and ventricular wall motion in patients with acute coronary syndrome (ACS).Methods Thirty-seven patients with ACS were randomly divided into two groups:conventional treatment group (n= 17,11 males,range of age:32-80 years,average age:60.9±4.9 years) and Pur treatment group (n=20,12 males,range of age:40-76 years,average age:62.7±3.5 years).Patients in the conventional treatment group received standard treatment according to the current guidelines,while patients in the Pur treatment group received intravenous administration of Pur (500 mg/day) for 10 days plus conventional treatment.Real-time myocardial contrast echocardiography (RT-MCE) was performed to evaluate the change in myocardial perfusion index (MPI) and ventricular wall motion index (VWMI) at admission and 10 days after treatment.Results At 10 days after treatment,MPI was significantly higher (P【0.01) and VWMI significantly lower (P【0.01) in the Pur group comparing with those in the conventional group.Conclusions Puerarin might improve myocardial microcirculation perfusion and ventricular wall motion in patients with ACS.
基金Supported by Hebei Province Science Foundation, China (No.07276101D-3)Clinical Science Project Fund of the Ministry of Health in Hebei Province, China (No. 03078)Foreign Studying Project Fund in Hebei Province, China (No. 07-03)
文摘AIM: To evaluate the toxicity of endogeneous peroxynitrite on transplanted retinal pigment epithelial (RPE) sheets and the effect of puerarin on their survival in the C57BL/6 mice after RPE sheets have been transplanted into SD rats' subretinal space .METHODS: C57BL/6 mice eyes were used to culture RPE cells.Ninety-six SD rats were involved in the experiment.They were divided into control(block control) ,streptozotocin(STZ,negative control),untransplanted RPE (positive control) and transplanted RPE groups respectively.Diabetes was induced in SD rats by intra-peritoneal STZ injection in the latter three groups.Saline was injected into the subretinal space of 24 SD rats in the untransplanted RPE group and primary RPE sheets were injected into the subretinal space of 24 SD rats in the transplanted RPE group.Puerarin (45 mg/kg) was administrated into both untransplanted RPE and transplanted RPE groups of diabetic rats through intraperitoneal injection route after RPE sheets transplantation.At 20,40,60 days after surgery,Western blotting analysis,DNA ladder and RT-PCR were used for determining the differences in expression of nitrotyrosine (NT,the foot print of peroxynitrite ),apoptosis and iNOS mRNA in the control,STZ,untransplanted RPE and transplanted RPE groups respectively.HE staining was used for determining the RPE survival in the subretinal space of the transplanted RPE group.RESULTS: Apoptosis and expression of NT and iNOS mRNA were observed in STZ,untransplanted RPE and transplanted RPE groups,but were delayed in untransplanted RPE and transplanted RPE groups in a time-dependent manner compared with control and STZ groups (P<0.01).There were no differences between the two groups (P >0.01).NT,DNA ladder,iNOS mRNA were down-regulated,which were associated with the decrease of expression of peroxynitrite.Numerous pigmented cells emerged and increased in number in the subretinal space during the 60-day observation period after transplantation.On day 20,heavily pigmented cells were visible at the transplant site;On day 40,monolayer and multilayered transplant was visible in the subretinal space;On day 60,heavily pigmented monolayer and multilayered transplants with round apical profile were present along Bruch's membrane.CONCLUSION: Puerarin increased the 60-day survival of C57BL/6 mice RPE xenografts in the SD rats' subretinal space,which may be related to its direct inhibition of apoptosis of RPE cells and antagnism of damage of peroxynitrite to RPE cells.
基金Anhui Provincial Natural Science Foundation(No.08020303080) for financial support of this work
文摘A type of triangular pyramid-shaped microparticles of puerarin was synthesized by using oil-in-oil microemulsion approach which is simple and economical under the action of copper substrate.The pyramid-shaped microparticles would be made up of deposit of nanospheres or nanorods and have two significant characters.One is its complex surface morphology like coral reef.The other is a lot of nanopores in existence in the microparticle body.Two possible formation routes were speculated.
文摘This work was done to investigate succinylated commercial whey protein isolate(S-WPI)as an oral sustained-release delivery carrier for puerarin 5(PR-5). The succinylation condi-tions were established for S-WPIs by optimization of single factor study and Box–Beehnkendesign. The effect of succinylation degree on S-WPIs solubility was evaluated. Physicochem-ical properties of S-WPIs dried by different three methods on their flow ability, particle size,morphology and in vitro release behavior were studied. After preparing PR-5 sustained re-lease protein tablets with S-WPIs as the carrier by direct powder compression method, thedrug release were studied in vitro and the oral pharmacokinetics and bioavailability wasevaluated using in vivo dog model. It was observed that concentration of substrate has asignificant effect on succinylation. Release behavior in vitro showed spry dried S-WPIs with100% succinylation rate and 30% drug loading would be applied to the preparation of PR-5 sustained-release protein tablets based on the swelling mechanism(protein loss). Comparedwith PR-5 conventional tablet with oral administration, T max value of PR-5 sustained-releaseprotein tablets was approximately 1.58 fold greater than those of the conventional tablets asfurther evidenced by the significantly prolonged MRT and T 1/2. The findings demonstratedthat spray-dried S-WPIs has potential as a promising functional excipient for the design of PR-5 oral sustained-release tablets which can fully improve sustained-release effect and oral bioavailability.
文摘Objective:In this study,the influence of puerarin,paeoniflorin,and menthol on the structure and barrier function of tight junctions(TJs)in MadineDarby canine kidney epithelial(MDCK)and MDCK-multi-drug resistance 1(MDR1)cells was evaluated to determine the mechanisms by which the drugs cross the bloodebrain barrier(BBB).Method:Cells were treated with puerarin,paeoniflorin,and menthol followed by immunohistochemical staining with occludin,claudin-1,and F-actin.The cells were then observed using laser-scanning confocal microscopy.Average optical density(AOD)of the immunofluorescence images of the proteins were analyzed using ImageJ software while Transepithelial electrical resistance(TEER)was measured using an epithelial voltohmmeter.Results:Confocal microscopy revealed that puerarin-and paeoniflorin-treated tight junction proteins were conspicuous while menthol suppressed their expression.Correspondingly,AOD values of cells treated with puerarin or paeoniflorin,or both showed no difference compared to the control group(P>.05)while the menthol group value was downregulated.In 3 h,TEER of cells not treated with menthol were similar to the control group,while treatment with menthol significantly decreased TEER value(P<.05).In addition,application of menthol decreased TEER in MDCK cells earlier than in MDCK-MDR1 cells.Conclusion:Menthol but not puerarin and paeoniflorin may enhance paracellular transport and improve drug penetration of the BBB by disrupting the structure and,thereby,weakening the barrier function of TJs.
基金the Basic Research Program from Science,Industry,Trade and Information Technology Commission of Shenzhen Municipality(Grant no.JCYJ20130402145002398)National Natural Science Foundation of China(Grant no.81102824).
文摘The purpose of this study was to investigate the influence of the supercritical CO2 processing on the particle size and morphology of puerarin crystals. The process parameters included solvents, temperature, pressures, antisolvent times, addition volumes, antisolvent addition rates and solute concentrations. After being processed, the dramatic reduction of the dimensions and the change of the crystal shape were noticed. Decreasing the antisolvent addition rate, increasing the temperature and the addition volume below 50 ml led to a decrease in size. The new crystal of puerarin generated at the optimal conditions was 30.34 μm.The solvent of methanol and the concentration of 60 mg/ml were found to determine the type and degree of crystallinity of particles. These results showed that this process has the potential to produce a drug recrystallization product with newly generated crystal forms and the size of drug particles could be controlled through the tuning of various experimental conditions.
基金The project supported by National Training Program of Innovation and Entrepreneurship for Undergraduates(201510439085)
文摘OBJECTIVE The aim of this study was to investigate the protective effect of puerarin on alcoholtoxicity in rat pheochromocytoma cell line(PC12). METHODS The PC12 cells were incubated with different concentrations of puerarin in advance. The protective effects of the puerarin on alcohol induced PC12 cel impairment were evaluated according to the fol owing approach: the viability of PC12 cel was determined by MTT assay and the impairment level was evaluated by analysis the leakage content of the lactate dehydrogenase(LDH). The cel apoptosis degree and the pro-apoptotic p53 protein expression were measured by flow cytometry. RESULTS Alcohol significantly impaired PC12 cel viability(P<0.05),and increased LDH leakage(P<0.05),induced cell apoptosis and upregulated expression of p53(P<0.05).While Puerarin significantly reversed these changes(P<0.05). CONCLUSION Puerarin might exert protection effect against ethanol-induced neurotoxicity via inhibition the expression of p53 protein.
基金supported by the Project of Scientific Research Fund of Traditional Chinese Medicine of Zhejiang Province(No.2020ZB230).
文摘NLRP3 inflammasome-mediated cell pyroptosis aggravates the development of cerebral ischemia/reperfusion(I/R)injury,and the aim of this study is to investigate the potential utilization of the Chinese medicine,Puerarin,in treating this disease.Through conducting in vitro and in vivo experiments,the present study illustrated that Puerarin regulated LncRNA double homeobox A pseudogene 8(DUXAP8)/miR-223-3p axis to inactivate NLRP3-mediated pyroptotic cell death,resulting in the attenuation of I/R injury.Specifically,the cerebral I/R injury in rat models and hypoxia/reoxygenation(H/R)in primary hippocampus neuron(PHN)cells were inducted,which were subsequently exposed to Puerarin treatment.As expected,we validated that Puerarin suppressed cell pyroptosis and rescued cell viability in I/R rat hippocampus tissues and H/R PHN cells.Next,through bioinformatics analysis,we noticed that miR-223-3p targeted both LncRNA DUXAP8 and NLRP3 mRNA,and both LncRNA DUXAP8 ablation and miR-223-3p overexpression inactivate NLRP3-mediated cell pyroptosis to rescue cell viability in H/R PHN cells.Interestingly,we evidenced that Puerarin restrained LncRNA DUXAP8 expressions,but upregulated miR-223-3p in I/R rat tissues and H/R PHN cells,and the protective effects of Puerarin on H/R PHN cells were abrogated by overexpressing LncRNA DUXAP8 and silencing miR-223-3p.Collectively,we concluded that Puerarin regulated LncRNA DUXAP8/miR-223-3p/NLRP3 signaling cascade to attenuate I/R injury.
文摘Pueraria mirifica(PM)has traditionally been used to relieve postmenopausal symptoms.Recently,its extract has been developed into various cosmetic products to promote skin rejuvenation and youthfulness.This study investigated the phytochemicals of PM tuber and compared between the tuber flesh and its outer peel.Puerarin which is one of the major isoflavones and being considered as the marker compound was used to determine the presence of PM extract in local cosmetic products.Puerarin could be ionized by a mass spectrometer at both negative and positive modes.The peak ionized at the negative mode showed to have a narrower peak width(0.2 min)and higher signal-to-noise ratio(30)for pueararin(1 mg/L).The results also found PM extract contained many C-and O-glycosylated isoflavones,especially from its peel extract.This explains the peel extract showed to have four times higher antiradical activity than those of flesh extract.Puerarin from the cosmetic products was recovered via successive methanolic sonication and followed by liquid-liquid extraction using ethyl acetate.Puerarin was successfully partitioned from the highly complex chemical mixture of cosmetic products with the recovery ranged from 89.1%to 115%.Hence,isoflavones was found to be higher at the outer peels than its tuber flesh.A simple and reliable method has been developed to analyse the presence of PM extract in cosmetic products based on the detection of puerarin after successive extraction via methanolic sonication and ethyl acetate partition.
基金This study was supported by the Natural Science Foundation of Zhejiang Province (No. M303042)the Key Laboratory for Biomedical Engineering of Ministry of Education of China.
文摘Objective To investigate whether the protective effects of puerarine(Pur)against cerebral ischemia is associated with depressing the extracellular levels of amino acid transmitters in brain of rats.Methods Male Sprague-Dawley rats were subjected to transient middle cerebral artery occlusion(MCAO)for 60 min followed by 24 h reperfusion.Pur(50,100 mg/kg,i.p.)was administered at the onset of MCAO.The infarct rate and edema rate were detected on TTC(2,3,5-triphenyltetrazolium chloride)-stained coronal sections.The extracellular levels of amino acid transmitters were monitored in striatum of rats with ischemic/reperfusion injury using in vivo microdialysis technique.Furthermore,the protective effects of Pur against glutamate-induced neurotoxicity were detected.Glutamate-induced apoptotic and necrotic cells in hippocampus were estimated by flow cytometric analysis of Annexin-V and PI labeling cells.Results Pur(100 mg/kg)significantly decreased infarct size by 31.6%(P<0.05),reduced edema volume(P<0.05),and improved neurological functions(P<0.05)following MCAO.In these rats,the ischemia-induced extracellular levels of aspartate(Asp),glutamate(Glu),γ-aminobutyric acid(GABA),and taurine(Tau)were significantly reduced in striatum of vehicle-treated animals by 54.7%,56.7%,75.8%,and 68.1%(P<0.01 and P<0.05).Pur reduced the peak values of Glu and Asp more obviously than those of GABA and Tau,and the rate of Glu/GABA during MCAO markedly decreased in Pur-treated MCAO rats,compared with the vehicle-treated MCAO rats.Meanwhile,apoptosis and necrosis induced by Glu in cultured hippocampal neurons were significantly reduced after Pur treatment.Conclusion Acute treatment with Pur at the onset of occlusion significantly depresses ischemia-induced efflux of amino acids,especially,excitotoxicity in the striatum,a mechanism underlying the neuroprotective effect on cellular survival.
文摘A novel compound, diethyl puerarin-7-yl phosphate, was synthesized through a simplified Atheron-Todd reaction for the first time. The structure of this compound was elucidated by IR, ESI-MS and NMR. Two conformations of the compound were testified by 2D NMR (HSQC and HMBC) and dynamic NMR. Furthermore, we carried out the conformational analysis using chemical calculation by the Gaussian 03. Finally, we obtained two preferred conformations and energy values.
基金supported by the National Natural Science Foundation of China,No.81273983
文摘Increasing evidence indicates that disruption of normal iron homeostasis may contribute to pathological development of Alzheimer's disease.Icariin,astragalus,and puerarin have been shown to suppress iron overload in the cerebral cortex and improve spatial learning and memory disorders in Alzheimer's disease mice,although the underlying mechanism remains unclear.In the present study,APPswe/PS1ΔE9 transgenic mice were administered icariin,astragalus,and puerarin(120,80,and 80 mg/kg,respectively,once a day,for 3 months).Iron levels were detected by flame atomic absorption spectroscopy.Interleukin-1β,interleukin-6,and tumor necrosis factor-α levels were measured in the cerebral cortex by enzyme linked immunosorbent assay.Glutathione peroxidase and superoxide dismutase activity and malondialdehyde content were determined by colorimetry.Our results demonstrate that after treatment,iron levels and malondialdehyde content are decreased,while glutathione peroxidase and superoxide dismutase activities are increased.Further,interleukin-1β,interleukin-6,and tumor necrosis factor-α levels were reduced.These results confirm that compounds of icariin,astragalus,and puerarin may alleviate iron overload by reducing oxidative stress and the inflammatory response.
文摘Summary: The protective effect of puerarin on the Parkinson disease (PD) mice with decreased es-trogen level was investigated in order to develop a new potential medicine as a substitute for estrogenfor preventing and treating PD. By using immunohistochemical method of avidinbiotin peroxidasecomplex (ABC), the distribution of the cells positive for tyrosine hydroxylase (TH) and fibres in thesubstantia nigra of the mouse were observed. These mice were divided into three groups randomly:group A, normal-female-mouse models; group B containing three subgroups, B1 (normal saline), B2(estrogen), B3 (puerarin); group C containing three sub groups, C1 (normal saline), C2(estrogen), C3 (puerarin). By using TUNEL the numbers of apoptosis cells in every visual field wascounted and the difference between the experimental group and control group was compared. The re-sults showed the numbers of the cells positive for TH were more and the numbers of apoptosis cellswere less in the normal-female-mouse models group than in the group of model made after ovar-iosteresis and the group of model made before ovariosteresis (P＜0.05), respectively. However,there was no significant difference, between the group given estrogen/puerarin and the controls, andbetween the group given estrogen and given puerarin. (P＞0. 05). It was suggested that puerarinmay have protective effect on the nigral neurons to PD. Moreover, the protective effect might serveas a surrogate of estrogen and be associated with the apoptosis.
