摘要
目的探索程氏蠲痹汤(JBT)对胶原蛋白诱导关节炎(CIA)大鼠治疗的可能机制。方法将雌性SD大鼠分为正常组、CIA模型组、甲氨蝶呤(MTX)组、不同剂量JBT组、LY294002(PI3K阻断剂)组。评估JBT对实验大鼠治疗前后足趾肿胀度、关节炎指数测定的影响,HE染色检测滑膜组织病变情况,ELISA检测大鼠滑膜组织匀浆中白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)水平。实时定量PCR检测磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)、beclin-1、P62的mRNA表达水平,Western blot法检测AKT、磷酸化的AKT(p-AKT)、mTOR、磷酸化的mTOR(p-mTOR)、PI3K、磷酸化的PI3K(p-PI3K)、P62、beclin-1、微管相关蛋白1轻链3B(LC3B)蛋白表达。结果与正常组相比,给药前1 h,其余各组大鼠足趾明显肿胀。与给药前1 h相比,高剂量JBT组、MTX组、LY294002组给药30 d后,采血前2 h足趾肿胀明显缓解。JBT能显著减轻大鼠足趾肿胀度、关节炎指数评分、滑膜组织的破坏。高剂量JBT组大鼠滑膜样本IL-1β、TNF-α水平以及PI3K、AKT、mTOR、P62的mRNA表达水平降低,p-PI3K、p-AKT、p-mTOR、P62蛋白水平明显降低,beclin-1 mRNA和蛋白表达、LC3B蛋白水平明显升高。结论JBT可能通过抑制PI3K/AKT/mTOR信号通路的激活减轻关节炎症,高剂量JBT的治疗效果与MTX、LY294002的治疗效果相当。
Objective To investigate the potential mechanism of Cheng’s Juanbi Decoction(JBT)for treating collageninduced arthritis(CIA)in rats.Methods Female SD rats were divided into normal group,CIA model group,methotrexate(MTX)group,JBT group with different doses,and LY294002(PI3K blocker)group.The effects of JBT on toe swelling and arthritis index of rats before and after treatment were evaluated.HE staining was used to observe the pathological changes of synovial tissues.ELISA was used to determine the levels of interleukin-1β(IL-1β)and tumor necrosis factorα(TNF-α)in synovium of rats.Real-time quantitative PCR was used to detect mRNA expression levels of phosphatidylinositol 3 kinase(PI3K),protein kinase B(AKT),mammalian target of rapamycin(mTOR),beclin-1,and P62.The expressions of AKT,phosphorylated AKT(p-AKT),mTOR,phosphorylated mTOR(p-mTOR),PI3K,phosphorylated PI3K(p-PI3K),P62,beclin-1,and microtubule-associated protein 1 light chain 3B(LC3B)were detected by Western blot analysis.Results Compared with the normal group,the toe of other groups was significantly swollen 1 hour before administration.Compared with the conditions 1 hour before administration,toe swelling in the high-dose JBT group,MTX group,and LY294002 group was significantly relieved 2 hours before blood collection after 30 days of administration.JBT can significantly reduce the degree of toe swelling,arthritis index(AI)score,and the destruction of synovial tissue.The levels of IL-1β,TNF-α,mRNA expression of PI3K,AKT,mTOR and P62,and protein levels of p-PI3K,p-AKT,p-mTOR,and P62 in synovium samples of rats in the high-dose JBT group were significantly decreased.Beclin-1 mRNA and protein expression and LC3B protein level were significantly increased.Conclusion JBT may alleviate joint inflammation by inhibiting the activation of the PI3K/AKT/mTOR signaling pathway,and the therapeutic effect of high-dose JBT is comparable to that of MTX and LY294002.
作者
孙广瀚
朱俊
许霞
万磊
南淑玲
王玉凤
赵黎
程卉
王坤
刘莹
欧阳增
SUN Guanghan;ZHU Jun;XU Xia;WAN Lei;NAN Shuling;WANG Yufeng;ZHAO Li;CHENG Hui;WANG Kun;LIU Ying;Ouyang Zeng(School of Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230012;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031;Scientific Research and Technology Center of Anhui University of Chinese Medicine,Hefei 230021,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2023年第11期961-966,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
2020年度国家中医药管理局“中医药古籍文献和特色技术传承专项”(GZY-KJS-2020-044)
2020年安徽省高等学校自然科学研究项目(KJ2020A0378)
2019年度安徽省社科规划项目(AHSKY2019D070)
全国中医药创新骨干人才培训项目(国中医药人教函[2019]128号)
2019年安徽省高校优秀青年骨干人才项目(gxgwfx2019024)
安徽省省级质量工程教学研究项目(2018jyxm1068)。
