[Objectives]To study the effect and mechanism of baicalin on the activation of NLRP3 inflammasome in human fibroblast like synoviocytes of rheumatoid arthritis(HFLS-RA).[Methods]To confirm that baicalin alleviated the...[Objectives]To study the effect and mechanism of baicalin on the activation of NLRP3 inflammasome in human fibroblast like synoviocytes of rheumatoid arthritis(HFLS-RA).[Methods]To confirm that baicalin alleviated the activation of NLRP3 inflammasome in HFLS-RA,the expression of NLRP3 before and after baicalin treatment was observed by immunofluorescence.Western blot was used to detect the protein expression of p-PI3K,p-Akt,NF-κB p65,NLRP3,ASC and caspase-1 after baicalin treatment for 48 h,and the contents of IL-1 and IL-18 in the supernatents were detected by ELISA.In order to explore the mechanism of baicalin alleviating the activation of NLRP3 inflammasome,the corresponding relationship between let-7i-3p and PIK3CA was verified by double luciferin and Westen blot analysis.The expression of let-7i-3p and PI3K before and after baicalin intervention was detected by RT-qPCR.let-7i-3p interference was used to verify whether baicalin mitigated the activation of enhanced NLRP3 inflammasome.[Results]Baicalin(50 and 100 mg/L)significantly reduced the activation of NLRP3 inflammasome,inhibited the protein expressions of p-PI3K,p-Akt,NF-κB p65,NLRP3,ASC and caspase-1,and the secretion of IL-1 and IL-18.let-7i-3p and PIK3CA had a targeted correspondence,and baicalin up-regulated the expression of let-7i-3p and down-regulated the expression of PIK3CA.Baicalin attenuated the activation of NLRP3 inflammasome enhanced by let-7i-3p interference.[Conclusions]Baicalin can up-regulate let-7i-3p expression,inhibit PI3K/Akt/NF-κB signal transduction,and thus reduce the activation of NLRP3 inflammasome in HFLS-RA.展开更多
OBJECTIVE Aryl hydrocarbon receptor(Ahr)is thought to be a crucial factor that regulates immune responses,which may be involved in the pathogenesis of autoimmune inflammation including rheumatoid arthritis(RA).The res...OBJECTIVE Aryl hydrocarbon receptor(Ahr)is thought to be a crucial factor that regulates immune responses,which may be involved in the pathogenesis of autoimmune inflammation including rheumatoid arthritis(RA).The results of our group in recent years have shown that CP-25,a novel ester derivative of paeoniflorin,has a good effect on improving RA animal models.However,whether the anti-arthritis effect of CP-25 is related to Ahr remains unclear.METHODS CP-25 treatment ameliorated adjuvant-induced arthritis(AA),a mouse model of RA,by inhibiting Ahr-related activities in fibroblasts like synoviocytes(FLS).AA rats were treated with CP-25 or paroxetine from day 17 to 33 after immunization.RESULTS CP-25 alleviated arthritis symptoms and the pathological changes,decreased the expression of Ahr in the synovium and FLS of AA rats.Besides,treatment with CP-25 reduced the proliferation and migration of MH7A caused by Ahr activation.In addition,we also demonstrated that CP-25 down-regulated the co-expression and co-localization of Ahr and G protein-coupled receptor kinase 2(GRK2)in MH7A.CONCLUSION The data presented here demonstrated that CP-25 suppressed FLS dysfunction in rats with AA,which were associated with reduced Ahr activation and the interaction between Ahr and GRK2.展开更多
目的研究黄芩苷减轻类风湿关节炎成纤维样滑膜细胞(human fibroblast like synoviocytes of rheumatoid arthritis,HFLS-RA)NLRP3炎性小体活化的作用及机制。方法为证实黄芩苷减轻HFLS-RA中NLRP3炎性小体活化,采用免疫荧光观察黄芩苷处...目的研究黄芩苷减轻类风湿关节炎成纤维样滑膜细胞(human fibroblast like synoviocytes of rheumatoid arthritis,HFLS-RA)NLRP3炎性小体活化的作用及机制。方法为证实黄芩苷减轻HFLS-RA中NLRP3炎性小体活化,采用免疫荧光观察黄芩苷处理前后NLRP3的表达;Western blot检测黄芩苷处理48 h后,p-PI3K、p-Akt、NF-κB p65、NLRP3、ASC及caspase-1蛋白的表达;ELISA检测细胞上清中IL-1及IL-18的含量。为探究黄芩苷减轻NLRP3炎性小体活化的机制,采用双荧光素酶及Western blot验证let-7i-3p与PIK3CA的对应关系;RT-qPCR检测黄芩苷干预前后let-7i-3p及PI3K的表达;采用let-7i-3p干扰,验证黄芩苷是否减轻了增强的NLRP3炎性小体的活化。结果50、100 mg·L^(-1)黄芩苷明显减轻了NLRP3炎性小体的活化,抑制p-PI3K、p-Akt、NF-κB p65、NLRP3、ASC、caspase-1蛋白的表达,抑制IL-1、IL-18的分泌。let-7i-3p与PIK3CA存在靶向对应关系,黄芩苷上调了let-7i-3p的表达、下调了PIK3CA的表达;黄芩苷减轻了因let-7i-3p干扰而增强的NLRP3炎性小体的活化。结论黄芩苷经上调let-7i-3p表达、抑制PI3K/Akt/NF-κB信号转导,从而减轻NLRP3炎性小体的活化。展开更多
基金Supported by the National Natural Science Foundation of China(82360802):the Natural Science Foundation of Ningxia Province,China(2022AAC 03152).
文摘[Objectives]To study the effect and mechanism of baicalin on the activation of NLRP3 inflammasome in human fibroblast like synoviocytes of rheumatoid arthritis(HFLS-RA).[Methods]To confirm that baicalin alleviated the activation of NLRP3 inflammasome in HFLS-RA,the expression of NLRP3 before and after baicalin treatment was observed by immunofluorescence.Western blot was used to detect the protein expression of p-PI3K,p-Akt,NF-κB p65,NLRP3,ASC and caspase-1 after baicalin treatment for 48 h,and the contents of IL-1 and IL-18 in the supernatents were detected by ELISA.In order to explore the mechanism of baicalin alleviating the activation of NLRP3 inflammasome,the corresponding relationship between let-7i-3p and PIK3CA was verified by double luciferin and Westen blot analysis.The expression of let-7i-3p and PI3K before and after baicalin intervention was detected by RT-qPCR.let-7i-3p interference was used to verify whether baicalin mitigated the activation of enhanced NLRP3 inflammasome.[Results]Baicalin(50 and 100 mg/L)significantly reduced the activation of NLRP3 inflammasome,inhibited the protein expressions of p-PI3K,p-Akt,NF-κB p65,NLRP3,ASC and caspase-1,and the secretion of IL-1 and IL-18.let-7i-3p and PIK3CA had a targeted correspondence,and baicalin up-regulated the expression of let-7i-3p and down-regulated the expression of PIK3CA.Baicalin attenuated the activation of NLRP3 inflammasome enhanced by let-7i-3p interference.[Conclusions]Baicalin can up-regulate let-7i-3p expression,inhibit PI3K/Akt/NF-κB signal transduction,and thus reduce the activation of NLRP3 inflammasome in HFLS-RA.
基金National Nature Science Foundation of China(81573443,82173824,81973332)Anhui Province Natural Science Fund(170808J10)+1 种基金Anhui Provincial Natural Science Foundation(2108085MH320)and Collaborative Innovation Project of Key Scientific Research Platform in Anhui Universities(GXXT-2020-065)。
文摘OBJECTIVE Aryl hydrocarbon receptor(Ahr)is thought to be a crucial factor that regulates immune responses,which may be involved in the pathogenesis of autoimmune inflammation including rheumatoid arthritis(RA).The results of our group in recent years have shown that CP-25,a novel ester derivative of paeoniflorin,has a good effect on improving RA animal models.However,whether the anti-arthritis effect of CP-25 is related to Ahr remains unclear.METHODS CP-25 treatment ameliorated adjuvant-induced arthritis(AA),a mouse model of RA,by inhibiting Ahr-related activities in fibroblasts like synoviocytes(FLS).AA rats were treated with CP-25 or paroxetine from day 17 to 33 after immunization.RESULTS CP-25 alleviated arthritis symptoms and the pathological changes,decreased the expression of Ahr in the synovium and FLS of AA rats.Besides,treatment with CP-25 reduced the proliferation and migration of MH7A caused by Ahr activation.In addition,we also demonstrated that CP-25 down-regulated the co-expression and co-localization of Ahr and G protein-coupled receptor kinase 2(GRK2)in MH7A.CONCLUSION The data presented here demonstrated that CP-25 suppressed FLS dysfunction in rats with AA,which were associated with reduced Ahr activation and the interaction between Ahr and GRK2.
