The therapeutic actions of Qing Luo Yin (QLY清络饮) with heat property and Wen Luo Yin (WLY温络饮) with cold property on pain, swelling of the ankle, arthritis index and ultrastructures of synoviocytes were compared i...The therapeutic actions of Qing Luo Yin (QLY清络饮) with heat property and Wen Luo Yin (WLY温络饮) with cold property on pain, swelling of the ankle, arthritis index and ultrastructures of synoviocytes were compared in rats of type II collagen-induced arthritis (CIA), with tripterygium glycosidorum (TG) used as control. The results indicated that both QLY and WLY could reduce pain, swelling of the ankle and the arthritis index of CIA, and QLY had better effects in reducing the swelling of the ankle and controlling the secondary pathological lesions as compared with WLY. Investigation on the ultrastructures of synoviocytes indicated that both QLY and WLY could reduce the number of Golgi apparatus, rough surface endoplasmic reticulum, dense bodies, matrix filaments and vacuoles so as to suppress the excessive secretion of synoviocytes in rats of CIA.展开更多
Triptolide(TP),a major active component of Tripterygium wilfordii Hook.F.(TWHF),is used to treat rheumatoid arthritis(RA).However,it has a narrow therapeutic window due to its serious toxicities.To increase the therap...Triptolide(TP),a major active component of Tripterygium wilfordii Hook.F.(TWHF),is used to treat rheumatoid arthritis(RA).However,it has a narrow therapeutic window due to its serious toxicities.To increase the therapeutic index,a new triptolide-loaded transdermal delivery system,named triptolide-loaded liposome hydrogel patch(TP-LHP),has been developed.In this paper,we used a micro-needle array to deliver TP-LHP to promote transdermal absorption and evaluated this treatment on the pharmacokinetics and pharmacodynamics of TP-LHP in a rat model of collagen-induced arthritis(CIA).The pharmacokinetic results showed that transdermal delivery of microneedle TP-LHP yielded plasma drug levels which fit a onecompartment open model.The relationship equation between plasma concentration and time was C=303.59(e 0.064 t e 0.287t).The results of pharmacodynamic study demonstrated that TP-LHP treatment mitigated the degree of joint swelling and suppressed the expressions of fetal liver kinase-1,fetal liver tyrosine kinase-4 and hypoxia-inducible factor-1α in synovium.Other indicators were also reduced by TP-LHP,including hyperfunction of immune,interleukin-1β and interleukin-6 levels in serum.The therapeutic mechanism of TP-LHP might be regulation of the balance between Th1 and Th2,as well as inhibition of the expression and biological effects of vascular endothelial growth factor.展开更多
Objective:SC-E3 is a polyherbal formula that contains five medicinal herbs used frequently in traditional herbal medicine.In our previous study,we demonstrated the antioxidant and anti-inflammatory effects of SC-E3.Th...Objective:SC-E3 is a polyherbal formula that contains five medicinal herbs used frequently in traditional herbal medicine.In our previous study,we demonstrated the antioxidant and anti-inflammatory effects of SC-E3.The present study examined the effects of SC-E3 in a mouse model of type-II collagen-induced arthritis(CIA).Methods:In vivo,male DBA/1 J mice were immunized by intradermal injection of bovine type-II collagen and complete or incomplete Freund’s adjuvant,to induce arthritis.SC-E3 was orally administered daily for 23 days.In vitro,bone marrow-derived macrophages(BMMs)were treated with macrophage colony-stimulating factor(M-CSF)and receptor activator of nuclear factor-j B ligand(RANKL)in the absence or presence of SC-E3.Results:Administrations of SC-E3 were found to have anti-arthritic effects in the joints of CIA mice,as evidenced by reduced paw swelling,bone erosion and deformation,inflammatory cell infiltration,and inflammation in synovial membrane.SC-E3 also reduced serum levels of tumor necrosis factor-a,interleukin-1 b,aspartate aminotransferase and alanine aminotransferase.Furthermore,tartrateresistant acid phosphatase-positive osteoclast numbers in the joints were significantly lower in SC-E3-treated CIA mice than in CIA mice.In addition,the differentiations of BMMs to multinucleated osteoclasts induced by M-CSF and RANKL stimulation were dose-dependently reduced by SC-E3.Conclusion:These results suggest that SC-E3 possesses substantial anti-arthritic activity because it inhibits pro-inflammatory cytokines and osteoclastogenesis,and that SC-E3 has potential therapeutic use for the treatment of rheumatoid arthritis.展开更多
Rheumatoid arthritis(RA)is a chronic autoimmune disorder marked by persistent synovial inflammation and joint degradation,posing challenges in the development of effective treatments.Nuciferine,an alkaloid found in lo...Rheumatoid arthritis(RA)is a chronic autoimmune disorder marked by persistent synovial inflammation and joint degradation,posing challenges in the development of effective treatments.Nuciferine,an alkaloid found in lotus leaf,has shown promising anti-inflammatory and anti-tumor effects,yet its efficacy in RA treatment remains unexplored.This study investigated the antiproliferative effects of nuciferine on the MH7A cell line,a human RA-derived fibroblast-like synoviocyte,revealing its ability to inhibit cell proliferation,promote apoptosis,induce apoptosis,and cause G1/S phase arrest.Additionally,nuciferine significantly reduced the migration and invasion capabilities of MH7A cells.The therapeutic potential of nuciferine was further evaluated in a collagen-induced arthritis(CIA)rat model,where it markedly alleviated joint swelling,synovial hyperplasia,cartilage injury,and inflammatory infiltration.Nuciferine also improved collagen-induced bone erosion,decreased pro-inflammatory cytokines and serum immunoglobulins(IgG,IgG1,IgG2a),and restored the balance between T helper(Th)17 and regulatory T cells in the spleen of CIA rats.These results indicate that nuciferine may offer therapeutic advantages for RA by decreasing the proliferation and invasiveness of FLS cells and correcting the Th17/Treg cell imbalance in CIA rats.展开更多
To develop a simple and highly sensitive high performance liquid chromatography with electrospray ionization mass spectrometric(LC-ESI-MS) method for the simultaneous determination of madecassoside and its major metab...To develop a simple and highly sensitive high performance liquid chromatography with electrospray ionization mass spectrometric(LC-ESI-MS) method for the simultaneous determination of madecassoside and its major metabolite madecassic acid in rat plasma, and compare the pharmacokinetics of the two compounds in normal and collagen-induced arthritis(CIA) rats. Glycyrrhetinic acid was used as the internal standard(IS). Chromatographic separation was accomplished on an Inertsil ODS-3 column, using a gradient elution with the mobile phase composed of acetonitrile and water acidified with 0.1%(V/V) formic acid. Detection was achieved by ESI-MS under the negative selected ion monitoring(SIM) mode. In normal and CIA rats, madecassoside(30 mg·kg-1) was orally administered for 21 consecutive days from the day of arthritis onset. For madecassoside, the linear range was 10–1 000 ng·mL-1 with the square regression coefficient(r) of 0.998 9, while for madecassic acid, the linear range was 10–500 ng·mL-1 with the square regression coefficient(r) of 0.996 1. The lower limit of quantification was 10 ng·mL-1 for both analytes. The intra- and inter-day precision ranged from 1.78% to 13.42% for madecassoside and 2.30% to 14.90% for madecassic acid, and the accuracy was between –0.95% and 6.30% for madecassoside and between –1.48% and 5.34% for madecassic acid. The average recoveries of madecassoside, madecassic acid and IS from spiked plasma samples were > 81%. The developed method was successfully applied to the pharmacokinetic study of madecassoside and madecassic acid in rats after an oral administration of madecassoside. During initial 7 days of dosing, the cmax and AUC of madecassoside were greatly decreased and Vd/F was markedly increased in CIA rats, and no significant difference was observed on the first day of dosing. In contrast, the T1/2, cmax and AUC of madecassic acid were significantly increased, and Ke of madecassic acid was greatly decreased in CIA rats compared with normal rats. Along with repeated administration of madecassoside, the differences of pharmacokinetic parameters of both madecassoside and madecassic acid between CIA and normal rats gradually subsided. The pharmacokinetic characteristics of both madecassoside and madecassic acid in rats were significantly altered by arthritis status, and the differences of pharmacokinetic parameters between arthritis and normal rats coincide with the severity of arthritis.展开更多
Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis(RA),but the relationship between the two phenomena remains unclear.We explored whether and how cholinergic dysfunction ...Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis(RA),but the relationship between the two phenomena remains unclear.We explored whether and how cholinergic dysfunction accelerates protein citrullination and consequently drives the development of RA.Cholinergic function and protein citrullination levels in patients with RA and collageninduced arthritis(CIA)mice were collected.In both neuron-macrophage coculture system and CIA mice,the effect of cholinergic dysfunction on protein citrullination and expression of peptidylarginine deiminases(PADs)was assessed by immunofluorescence.The key transcription factors for PAD4 expression were predicted and validated.Cholinergic dysfunction in the patients with RA and CIA mice negatively correlated with the degree of protein citrullination in synovial tissues.The cholinergic or alpha7 nicotinic acetylcholine receptor(a7nAChR)deactivation and activation resulted in the promotion and reduction of protein citrullination in vitro and in vivo,respectively.Especially,the activation deficiency of a7nAChR induced the earlier onset and aggravation of CIA.Furthermore,deactivation of a7nAChR increased the expression of PAD4 and specificity protein-3(SP3)in vitro and in vivo.Our results suggest that cholinergic dysfunction-induced deficient a7nAChR activation,which induces the expression of SP3 and its downstream molecule PAD4,accelerating protein citrullination and the development of RA.展开更多
Rheumatoid arthritis(RA)is a chronic inflammatory autoimmune disease.It is known that aucubin(AU)exerts anti-inflammatory activity,but its effects and mechanisms in RA are unclear.This study investigated the anti-infl...Rheumatoid arthritis(RA)is a chronic inflammatory autoimmune disease.It is known that aucubin(AU)exerts anti-inflammatory activity,but its effects and mechanisms in RA are unclear.This study investigated the anti-inflammatory effects and mechanisms of AU in vivo and in vitro.Human fibroblast-like synoviocyte cells from patients with RA(HFLS-RA),RAW264.7 cells,and MC3T3-E1 cells were used to evaluate the effects of AU on migration,invasion,apoptosis,osteoclast differentiation and production.Immunofluorescence was used to observe nuclear translocation of nuclear factor(NF)-/cB,the double luciferase reporter gene method was used to observe NF-/cB-p65 activity in AU-treated MC3T3-E1 cells.RT-qPCR was used to measure expression of bone metabolism and inflammation-related genes,and western blot was used to measure bone metabolism and NF-a:B protein expression levels.Collagen-induced arthritis(CIA)rat model was used for pharmacodynamics study.Arthritis indexes were measured in the ankle and knee,histological staining and Micro-computed tomography were performed on the ankle joints.Also,inflammatory factor gene expression and the levels of NF-/cB-related proteins were detected as in vitro.AU effectively inhibited HFLS-RA cell migration and invasion,promoted apoptosis,and inhibited RAW264.7 cell differentiation into osteoclasts,as well as inhibited NF-/cB-p65 activity in MC3T3-E1 cells.Notably,AU significantly reduced the gene expression levels of three cell-related inflammatory factors and bone metabolism factors,effectively inhibited the expression of p-lKKafi,p-lA:Ba,and p-p65 proteins.In vivo,AU relieved joint inflammation,reduced related inflammatory factors,and inhibited NF-/cB signaling.It could be used to treat RA-related synovial inflammation and bone destruction through the NF-/cB pathway.展开更多
Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) has been implicated in the regulation ofinflammation in rheumatoid arthritis (RA), primarily due to its ability to promote apoptosis in synoviocyte...Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) has been implicated in the regulation ofinflammation in rheumatoid arthritis (RA), primarily due to its ability to promote apoptosis in synoviocytes andinfiltrating lymphocytes. The aim of this study was to investigate the immunomodulatory mechanism and role ofTRAIL in inflammatory arthritis. We created an animal model of inflammatory arthritis and demonstrated that TRAILsignificantly inhibited joint inflammation and reduced the severity of arthritis. The suppression of joint inflammationwas not due to the TRAIL-mediated induction of apoptosis in T cells, macrophages or synovial fibroblasts. Incontrast, TRAIL directly inhibited T-cell proliferation and suppressed the production of cytokines, which indicatedthat TRAIL exerted its anti-inflammatory effects by direct inhibition of T-cell activation. Moreover, TRAIL receptor(TRAIL-R)-knockout mice developed more severe disease, and the protective effects of TRAIL were abolished in theexperimental arthritis model in TRAIL-R knockout mice. From these results, we conclude that TRAIL suppressesjoint inflammation via an apoptosis-independent pathway and directly inhibits T-cell activation. Our results providea novel apoptosis-independent, immune regulatory role for TRAIL in suppressing inflammatory arthritis and shedlight on the development of effective new therapies for autoimmune inflammatory diseases.展开更多
Background:Gingiva-derived mesenchymal stem cells(GMSCs)suppress immune and inflammatory responses in experimental arthritis models.Here,we determined whether GMSCs suppress the proliferation and invasion of rheumatoi...Background:Gingiva-derived mesenchymal stem cells(GMSCs)suppress immune and inflammatory responses in experimental arthritis models.Here,we determined whether GMSCs suppress the proliferation and invasion of rheumatoid arthritis fibroblast-like synoviocytes(RA FLSs).Methods:Surface markers of GMSCs were analyzed by flow cytometry.mRNA expression of matrix metalloproteinases and pro-inflammatory cytokine in RA FLSs was measured by quantitative polymerase chain reaction(PCR).RA FLS proliferation was analyzed by a 5-ethynyl-2′-deoxyuridine assay.To explore the molecular mechanisms,we assessed the effect of GMSCs on RA FLS proliferation by adding indoleamine-2,3-dioxygenase(IDO),CD39,or CD73 inhibitors.The invasion was analyzed by a transwell assay.The anti-inflammatory effects of GMSCs were assessed in a typeⅡcollagen-induced arthritis(CIA)mouse model.Results:Compared with human dermal fibroblast,GMSCs displayed a higher expression of CD39.Interleukin(IL)-8,IL-6,MMP-1,MMP-3,MMP-13,and monocyte chemoattractant protein-1 mRNA were all decreased in RA FLSs after incubation with GMSCs.GMSCs significantly reduced RA FLS proliferation in a dose-dependent manner in vitro,which was partly dependent on CD39/CD73 signaling.GMSCs significantly impeded the invasive capacity of RA FLSs in a dose-dependent manner in vitro.GMSCs infusion delayed arthritis onset and reduced arthritis scores in the CIA model.Conclusion:GMSCs are effective to inhibit the aggressive behavior of RA FLSs and treat experimental arthritis,implying their therapeutic potential in RA patients.展开更多
Objective:To demonstrate the rheumatoid arthritis(RA)mechanisms by determining the biochemical changes.To investigate the therapeutic mechanism of moxibustion in RA-model rats using a gas chromatography-mass spectrome...Objective:To demonstrate the rheumatoid arthritis(RA)mechanisms by determining the biochemical changes.To investigate the therapeutic mechanism of moxibustion in RA-model rats using a gas chromatography-mass spectrometry(GC-MS)metabolomics approach.Methods:A total of 24 rats were divided into three groups as follows:normal control group,model group and moxibustion group.Rats in model group and moxibustion group were set up collagen-induced arthritis(CIA)model.Rats in moxibustion group were treated by moxibustion.After 3 weeks of intervention,right ankle joint,serum and articular synovium samples were collected.Right ankle joint samples were used for histopathological evaluation between 3 groups to get the pathological changes of tissues and cells.Serum and articular synovium samples were used to analyze the changed metabolites of moxibustion on RA rats by the GC-MS based metabolomics.Results:Treatment of moxibustion not only significantly increased the weight of CIA rats,reduced the swelling of hind paw,arthritic scores,IL-1β,TNF-αbut also improved histopathological evaluation in right ankle joint samples.Sixteen significantly altered metabolites were found in RA rats as potential biomarkers of arthritis.Thirteen metabolites,significantly adjusted by moxibustion to help relieve arthritis,were selected out as biomarkers of antiarthritic mechanism of moxibustion,which were mainly involved in phenylalanine,tyrosine and tryptophan biosynthesis,glycine,serine and threonine metabolism,phenylalanine metabolism,alanine,aspartate and glutamate metabolism,glyoxylate and dicarboxylate metabolism and aminoacyl-tRNA biosynthesis.Conclusions:We have indicated moxibustion treatment is able to resist inflammation in CIA rats effectively.Using GC-MS metabolomics technique,we detect novel metabolites in the moxibustion antiarthritic process,which may aid in advanced understanding of arthritis and therapeutic mechanism of moxibustion.展开更多
There has been an increase in interest in the use of altered peptides as antigen-specific therapeutic agents in autoimmune diseases.Here we investigated the inhibitory effect and possible mechanism of an altered influ...There has been an increase in interest in the use of altered peptides as antigen-specific therapeutic agents in autoimmune diseases.Here we investigated the inhibitory effect and possible mechanism of an altered influenza virus haemagglutinin(HA)-derived peptide in collagen-induced arthritis(CIA).CIA was induced in DBA/1 mice by immunisation with type II collagen(CII).Altered HA308-317,wild-type HA308-317 or irrelevant peptide was administered intranasally beginning from arthritis onset.Clinical and histological scores were assessed,and cytokine levels in the serum or supernatants from splenocytes were determined.The percentages of Th1 and Th2 cells in response to different peptides were analysed by FACS both in vivo and in vitro.Our results showed that intranasal administration of altered HA308-317 peptide significantly ameliorated CIA.The therapeutic effect of altered HA308-317 peptide was associated with a substantial decrease in production of interferon(IFN)-c,interleukin(IL)-6,monocyte chemoattractant protein(MCP)-1,anti-CII IgG,IgG1 and IgG2a antibodies,and an markedly increase in production of IL-10 and IL-4 in serum or supernatants from splenocytes treated with altered HA308-317 peptide.The percentage of Th2(CD41IL-41)cells was upregulated significantly by altered HA308-317 peptide with a decreased percentage of Th1(T helper 1;CD41INF-c1)cells both in vivo and in vitro.These findings suggest that altered HA308-317 peptide might be a promising candidate for rheumatoid arthritis(RA)treatment.展开更多
文摘The therapeutic actions of Qing Luo Yin (QLY清络饮) with heat property and Wen Luo Yin (WLY温络饮) with cold property on pain, swelling of the ankle, arthritis index and ultrastructures of synoviocytes were compared in rats of type II collagen-induced arthritis (CIA), with tripterygium glycosidorum (TG) used as control. The results indicated that both QLY and WLY could reduce pain, swelling of the ankle and the arthritis index of CIA, and QLY had better effects in reducing the swelling of the ankle and controlling the secondary pathological lesions as compared with WLY. Investigation on the ultrastructures of synoviocytes indicated that both QLY and WLY could reduce the number of Golgi apparatus, rough surface endoplasmic reticulum, dense bodies, matrix filaments and vacuoles so as to suppress the excessive secretion of synoviocytes in rats of CIA.
基金supported by National Science and Technology Major Project on Significant Creation of New Drugs of China(2009ZX09502-019)
文摘Triptolide(TP),a major active component of Tripterygium wilfordii Hook.F.(TWHF),is used to treat rheumatoid arthritis(RA).However,it has a narrow therapeutic window due to its serious toxicities.To increase the therapeutic index,a new triptolide-loaded transdermal delivery system,named triptolide-loaded liposome hydrogel patch(TP-LHP),has been developed.In this paper,we used a micro-needle array to deliver TP-LHP to promote transdermal absorption and evaluated this treatment on the pharmacokinetics and pharmacodynamics of TP-LHP in a rat model of collagen-induced arthritis(CIA).The pharmacokinetic results showed that transdermal delivery of microneedle TP-LHP yielded plasma drug levels which fit a onecompartment open model.The relationship equation between plasma concentration and time was C=303.59(e 0.064 t e 0.287t).The results of pharmacodynamic study demonstrated that TP-LHP treatment mitigated the degree of joint swelling and suppressed the expressions of fetal liver kinase-1,fetal liver tyrosine kinase-4 and hypoxia-inducible factor-1α in synovium.Other indicators were also reduced by TP-LHP,including hyperfunction of immune,interleukin-1β and interleukin-6 levels in serum.The therapeutic mechanism of TP-LHP might be regulation of the balance between Th1 and Th2,as well as inhibition of the expression and biological effects of vascular endothelial growth factor.
基金supported by the National Research Foundation of Korea grant funded by the Korea government(No.2019R1F1A1062998)。
文摘Objective:SC-E3 is a polyherbal formula that contains five medicinal herbs used frequently in traditional herbal medicine.In our previous study,we demonstrated the antioxidant and anti-inflammatory effects of SC-E3.The present study examined the effects of SC-E3 in a mouse model of type-II collagen-induced arthritis(CIA).Methods:In vivo,male DBA/1 J mice were immunized by intradermal injection of bovine type-II collagen and complete or incomplete Freund’s adjuvant,to induce arthritis.SC-E3 was orally administered daily for 23 days.In vitro,bone marrow-derived macrophages(BMMs)were treated with macrophage colony-stimulating factor(M-CSF)and receptor activator of nuclear factor-j B ligand(RANKL)in the absence or presence of SC-E3.Results:Administrations of SC-E3 were found to have anti-arthritic effects in the joints of CIA mice,as evidenced by reduced paw swelling,bone erosion and deformation,inflammatory cell infiltration,and inflammation in synovial membrane.SC-E3 also reduced serum levels of tumor necrosis factor-a,interleukin-1 b,aspartate aminotransferase and alanine aminotransferase.Furthermore,tartrateresistant acid phosphatase-positive osteoclast numbers in the joints were significantly lower in SC-E3-treated CIA mice than in CIA mice.In addition,the differentiations of BMMs to multinucleated osteoclasts induced by M-CSF and RANKL stimulation were dose-dependently reduced by SC-E3.Conclusion:These results suggest that SC-E3 possesses substantial anti-arthritic activity because it inhibits pro-inflammatory cytokines and osteoclastogenesis,and that SC-E3 has potential therapeutic use for the treatment of rheumatoid arthritis.
基金supported by the National Natural Science Foundation of China(No.82274329,82304991)the China Postdoctoral Science Foundation(No,2023M732336)Shanghai Science and Technology Committee Sailing Program Foundation(No.23YF1442500)。
文摘Rheumatoid arthritis(RA)is a chronic autoimmune disorder marked by persistent synovial inflammation and joint degradation,posing challenges in the development of effective treatments.Nuciferine,an alkaloid found in lotus leaf,has shown promising anti-inflammatory and anti-tumor effects,yet its efficacy in RA treatment remains unexplored.This study investigated the antiproliferative effects of nuciferine on the MH7A cell line,a human RA-derived fibroblast-like synoviocyte,revealing its ability to inhibit cell proliferation,promote apoptosis,induce apoptosis,and cause G1/S phase arrest.Additionally,nuciferine significantly reduced the migration and invasion capabilities of MH7A cells.The therapeutic potential of nuciferine was further evaluated in a collagen-induced arthritis(CIA)rat model,where it markedly alleviated joint swelling,synovial hyperplasia,cartilage injury,and inflammatory infiltration.Nuciferine also improved collagen-induced bone erosion,decreased pro-inflammatory cytokines and serum immunoglobulins(IgG,IgG1,IgG2a),and restored the balance between T helper(Th)17 and regulatory T cells in the spleen of CIA rats.These results indicate that nuciferine may offer therapeutic advantages for RA by decreasing the proliferation and invasiveness of FLS cells and correcting the Th17/Treg cell imbalance in CIA rats.
基金financially supported by the National Natural Science Foundation of China(No.81374038,81173631)the Jiangsu Overseas Research&Training Program for University Prominent Young&Middle-aged Teachers and Presidents+1 种基金sponsored by Qing Lan Project,College Students Innovation Project for the R&D of Novel Drugs(No.J1030830)partially funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘To develop a simple and highly sensitive high performance liquid chromatography with electrospray ionization mass spectrometric(LC-ESI-MS) method for the simultaneous determination of madecassoside and its major metabolite madecassic acid in rat plasma, and compare the pharmacokinetics of the two compounds in normal and collagen-induced arthritis(CIA) rats. Glycyrrhetinic acid was used as the internal standard(IS). Chromatographic separation was accomplished on an Inertsil ODS-3 column, using a gradient elution with the mobile phase composed of acetonitrile and water acidified with 0.1%(V/V) formic acid. Detection was achieved by ESI-MS under the negative selected ion monitoring(SIM) mode. In normal and CIA rats, madecassoside(30 mg·kg-1) was orally administered for 21 consecutive days from the day of arthritis onset. For madecassoside, the linear range was 10–1 000 ng·mL-1 with the square regression coefficient(r) of 0.998 9, while for madecassic acid, the linear range was 10–500 ng·mL-1 with the square regression coefficient(r) of 0.996 1. The lower limit of quantification was 10 ng·mL-1 for both analytes. The intra- and inter-day precision ranged from 1.78% to 13.42% for madecassoside and 2.30% to 14.90% for madecassic acid, and the accuracy was between –0.95% and 6.30% for madecassoside and between –1.48% and 5.34% for madecassic acid. The average recoveries of madecassoside, madecassic acid and IS from spiked plasma samples were > 81%. The developed method was successfully applied to the pharmacokinetic study of madecassoside and madecassic acid in rats after an oral administration of madecassoside. During initial 7 days of dosing, the cmax and AUC of madecassoside were greatly decreased and Vd/F was markedly increased in CIA rats, and no significant difference was observed on the first day of dosing. In contrast, the T1/2, cmax and AUC of madecassic acid were significantly increased, and Ke of madecassic acid was greatly decreased in CIA rats compared with normal rats. Along with repeated administration of madecassoside, the differences of pharmacokinetic parameters of both madecassoside and madecassic acid between CIA and normal rats gradually subsided. The pharmacokinetic characteristics of both madecassoside and madecassic acid in rats were significantly altered by arthritis status, and the differences of pharmacokinetic parameters between arthritis and normal rats coincide with the severity of arthritis.
基金supported by the“Double First-Class”University Project(CPU2022QZ31,China)。
文摘Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis(RA),but the relationship between the two phenomena remains unclear.We explored whether and how cholinergic dysfunction accelerates protein citrullination and consequently drives the development of RA.Cholinergic function and protein citrullination levels in patients with RA and collageninduced arthritis(CIA)mice were collected.In both neuron-macrophage coculture system and CIA mice,the effect of cholinergic dysfunction on protein citrullination and expression of peptidylarginine deiminases(PADs)was assessed by immunofluorescence.The key transcription factors for PAD4 expression were predicted and validated.Cholinergic dysfunction in the patients with RA and CIA mice negatively correlated with the degree of protein citrullination in synovial tissues.The cholinergic or alpha7 nicotinic acetylcholine receptor(a7nAChR)deactivation and activation resulted in the promotion and reduction of protein citrullination in vitro and in vivo,respectively.Especially,the activation deficiency of a7nAChR induced the earlier onset and aggravation of CIA.Furthermore,deactivation of a7nAChR increased the expression of PAD4 and specificity protein-3(SP3)in vitro and in vivo.Our results suggest that cholinergic dysfunction-induced deficient a7nAChR activation,which induces the expression of SP3 and its downstream molecule PAD4,accelerating protein citrullination and the development of RA.
基金This work was supported by the Natural Science Foundation of China(No.81773922)Shanghai Natural Science Foundation(No.19ZR1452000).
文摘Rheumatoid arthritis(RA)is a chronic inflammatory autoimmune disease.It is known that aucubin(AU)exerts anti-inflammatory activity,but its effects and mechanisms in RA are unclear.This study investigated the anti-inflammatory effects and mechanisms of AU in vivo and in vitro.Human fibroblast-like synoviocyte cells from patients with RA(HFLS-RA),RAW264.7 cells,and MC3T3-E1 cells were used to evaluate the effects of AU on migration,invasion,apoptosis,osteoclast differentiation and production.Immunofluorescence was used to observe nuclear translocation of nuclear factor(NF)-/cB,the double luciferase reporter gene method was used to observe NF-/cB-p65 activity in AU-treated MC3T3-E1 cells.RT-qPCR was used to measure expression of bone metabolism and inflammation-related genes,and western blot was used to measure bone metabolism and NF-a:B protein expression levels.Collagen-induced arthritis(CIA)rat model was used for pharmacodynamics study.Arthritis indexes were measured in the ankle and knee,histological staining and Micro-computed tomography were performed on the ankle joints.Also,inflammatory factor gene expression and the levels of NF-/cB-related proteins were detected as in vitro.AU effectively inhibited HFLS-RA cell migration and invasion,promoted apoptosis,and inhibited RAW264.7 cell differentiation into osteoclasts,as well as inhibited NF-/cB-p65 activity in MC3T3-E1 cells.Notably,AU significantly reduced the gene expression levels of three cell-related inflammatory factors and bone metabolism factors,effectively inhibited the expression of p-lKKafi,p-lA:Ba,and p-p65 proteins.In vivo,AU relieved joint inflammation,reduced related inflammatory factors,and inhibited NF-/cB signaling.It could be used to treat RA-related synovial inflammation and bone destruction through the NF-/cB pathway.
基金This work was supported by grants from the National Science Council,Taiwan(NSC 101-2321-B-002-008 and 104-2314-B-281-002-,MOST 105-2320-B-002-034-and 105-2320-B-038-065-)。
文摘Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) has been implicated in the regulation ofinflammation in rheumatoid arthritis (RA), primarily due to its ability to promote apoptosis in synoviocytes andinfiltrating lymphocytes. The aim of this study was to investigate the immunomodulatory mechanism and role ofTRAIL in inflammatory arthritis. We created an animal model of inflammatory arthritis and demonstrated that TRAILsignificantly inhibited joint inflammation and reduced the severity of arthritis. The suppression of joint inflammationwas not due to the TRAIL-mediated induction of apoptosis in T cells, macrophages or synovial fibroblasts. Incontrast, TRAIL directly inhibited T-cell proliferation and suppressed the production of cytokines, which indicatedthat TRAIL exerted its anti-inflammatory effects by direct inhibition of T-cell activation. Moreover, TRAIL receptor(TRAIL-R)-knockout mice developed more severe disease, and the protective effects of TRAIL were abolished in theexperimental arthritis model in TRAIL-R knockout mice. From these results, we conclude that TRAIL suppressesjoint inflammation via an apoptosis-independent pathway and directly inhibits T-cell activation. Our results providea novel apoptosis-independent, immune regulatory role for TRAIL in suppressing inflammatory arthritis and shedlight on the development of effective new therapies for autoimmune inflammatory diseases.
基金National Natural Science Foundation of China,Grant/Award Number:82171768Zhejiang Provincial Natural Science Foundation of China,Grant/Award Number:LQ17H100001。
文摘Background:Gingiva-derived mesenchymal stem cells(GMSCs)suppress immune and inflammatory responses in experimental arthritis models.Here,we determined whether GMSCs suppress the proliferation and invasion of rheumatoid arthritis fibroblast-like synoviocytes(RA FLSs).Methods:Surface markers of GMSCs were analyzed by flow cytometry.mRNA expression of matrix metalloproteinases and pro-inflammatory cytokine in RA FLSs was measured by quantitative polymerase chain reaction(PCR).RA FLS proliferation was analyzed by a 5-ethynyl-2′-deoxyuridine assay.To explore the molecular mechanisms,we assessed the effect of GMSCs on RA FLS proliferation by adding indoleamine-2,3-dioxygenase(IDO),CD39,or CD73 inhibitors.The invasion was analyzed by a transwell assay.The anti-inflammatory effects of GMSCs were assessed in a typeⅡcollagen-induced arthritis(CIA)mouse model.Results:Compared with human dermal fibroblast,GMSCs displayed a higher expression of CD39.Interleukin(IL)-8,IL-6,MMP-1,MMP-3,MMP-13,and monocyte chemoattractant protein-1 mRNA were all decreased in RA FLSs after incubation with GMSCs.GMSCs significantly reduced RA FLS proliferation in a dose-dependent manner in vitro,which was partly dependent on CD39/CD73 signaling.GMSCs significantly impeded the invasive capacity of RA FLSs in a dose-dependent manner in vitro.GMSCs infusion delayed arthritis onset and reduced arthritis scores in the CIA model.Conclusion:GMSCs are effective to inhibit the aggressive behavior of RA FLSs and treat experimental arthritis,implying their therapeutic potential in RA patients.
基金National Natural Science Foundation of China:81774383,81904274supported by"Nursing Advantageous Discipline Construction Project in Jiangsu Universities"of Nanjing University of Chinese medicine:2019YSHL008,2019YSHL021。
文摘Objective:To demonstrate the rheumatoid arthritis(RA)mechanisms by determining the biochemical changes.To investigate the therapeutic mechanism of moxibustion in RA-model rats using a gas chromatography-mass spectrometry(GC-MS)metabolomics approach.Methods:A total of 24 rats were divided into three groups as follows:normal control group,model group and moxibustion group.Rats in model group and moxibustion group were set up collagen-induced arthritis(CIA)model.Rats in moxibustion group were treated by moxibustion.After 3 weeks of intervention,right ankle joint,serum and articular synovium samples were collected.Right ankle joint samples were used for histopathological evaluation between 3 groups to get the pathological changes of tissues and cells.Serum and articular synovium samples were used to analyze the changed metabolites of moxibustion on RA rats by the GC-MS based metabolomics.Results:Treatment of moxibustion not only significantly increased the weight of CIA rats,reduced the swelling of hind paw,arthritic scores,IL-1β,TNF-αbut also improved histopathological evaluation in right ankle joint samples.Sixteen significantly altered metabolites were found in RA rats as potential biomarkers of arthritis.Thirteen metabolites,significantly adjusted by moxibustion to help relieve arthritis,were selected out as biomarkers of antiarthritic mechanism of moxibustion,which were mainly involved in phenylalanine,tyrosine and tryptophan biosynthesis,glycine,serine and threonine metabolism,phenylalanine metabolism,alanine,aspartate and glutamate metabolism,glyoxylate and dicarboxylate metabolism and aminoacyl-tRNA biosynthesis.Conclusions:We have indicated moxibustion treatment is able to resist inflammation in CIA rats effectively.Using GC-MS metabolomics technique,we detect novel metabolites in the moxibustion antiarthritic process,which may aid in advanced understanding of arthritis and therapeutic mechanism of moxibustion.
基金grants from the National Major Scientific and Technological Specialized Project(2009ZX09103-746)National Program on Key Basic Research Project(2010CB529100)National Natural Science Foundation of China(30700751).
文摘There has been an increase in interest in the use of altered peptides as antigen-specific therapeutic agents in autoimmune diseases.Here we investigated the inhibitory effect and possible mechanism of an altered influenza virus haemagglutinin(HA)-derived peptide in collagen-induced arthritis(CIA).CIA was induced in DBA/1 mice by immunisation with type II collagen(CII).Altered HA308-317,wild-type HA308-317 or irrelevant peptide was administered intranasally beginning from arthritis onset.Clinical and histological scores were assessed,and cytokine levels in the serum or supernatants from splenocytes were determined.The percentages of Th1 and Th2 cells in response to different peptides were analysed by FACS both in vivo and in vitro.Our results showed that intranasal administration of altered HA308-317 peptide significantly ameliorated CIA.The therapeutic effect of altered HA308-317 peptide was associated with a substantial decrease in production of interferon(IFN)-c,interleukin(IL)-6,monocyte chemoattractant protein(MCP)-1,anti-CII IgG,IgG1 and IgG2a antibodies,and an markedly increase in production of IL-10 and IL-4 in serum or supernatants from splenocytes treated with altered HA308-317 peptide.The percentage of Th2(CD41IL-41)cells was upregulated significantly by altered HA308-317 peptide with a decreased percentage of Th1(T helper 1;CD41INF-c1)cells both in vivo and in vitro.These findings suggest that altered HA308-317 peptide might be a promising candidate for rheumatoid arthritis(RA)treatment.
