通痹健骨方水提物对脂多糖诱导人软骨细胞自噬的影响  

Effects of aqueous extract of Tongbi Jiangu Formula on lipopolysaccharide induced autophagy of human chondrocytes

作　　者：徐斌 李盛华[3] XU Bin;LI Sheng-hua(Xi’an Hospital of Traditional Chinese Medicine,Xi’an 710021,China;Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu Provincial Hospital of TCM,Lanzhou 730050,China)

机构地区：[1]西安市中医医院,西安710021 [2]甘肃中医药大学,兰州730000 [3]甘肃省中医院,兰州730050

出　　处：《中华中医药杂志》2023年第3期1222-1226,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：甘肃省科技计划项目(No.18JR2FA009)。

摘　　要：目的:探讨通痹健骨方水提物对脂多糖(LPS)诱导人软骨细胞的影响及作用机制。方法:提取人软骨细胞及传代培养,采用甲苯胺蓝染色法和Ⅱ型胶原蛋白免疫荧光染色法对细胞进行鉴定,LPS诱导软骨细胞模拟膝骨关节炎(KOA)软骨细胞模型,设对照组,模型组,通痹健骨方低、中、高剂量组,ELISA法检测肿瘤坏死因子(TNF)-α、白细胞介素(IL)-10、IL-1β的水平;Western Blot、qRT-PCR法检测肌球蛋白样BCL2结合蛋白(Beclin-1)、微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)、泛素结合蛋白P62(P62)蛋白及mRNA表达情况。结果:提取的细胞符合软骨细胞特征。与模型组比较,通痹健骨方中剂量组TNF-α、IL-1β、P62蛋白及mRNA显著降低(P<0.05,P<0.01),IL-10含量和LC3Ⅱ、Beclin-1蛋白及mRNA的表达均显著升高(P<0.01)。结论:通痹健骨方水提物可通过调节软骨细胞自噬,提高软骨细胞的抗应激能力和生存能力,抑制炎症反应,可能是通痹健骨方治疗KOA的机制之一。Objective:To explore the effects of water extract of Tongbi Jiangu Formula on human chondrocytes induced by lipopolysaccharide(LPS)and its mechanism.Methods:Human chondrocytes were extracted and subcultured.The cells were identified by toluidine blue staining and typeⅡcollagen immunofluorescence staining.Chondrocytes were induced by LPS to simulate knee osteoarthritis(KOA)chondrocytes model,and divided into control group,model group,Tongbi Jiangu Formula low-dose,medium-dose and high-dose groups.The levels of TNF-α,IL-10 and IL-1βwere detected by ELISA.Protein and mRNA expressions of Beclin-1,LC3Ⅱand P62 were detected by Western Blot and qRT-PCR.Results:The extracted cells were consistent with the characteristics of chondrocytes.Compared with model group,the contents of TNF-α,IL-1βand protein and mRNA expressions of P62 in Tongbi Jiangu Formula medium-dose group were significantly decreased(P<0.05,P<0.01),while the IL-10 content and LC3Ⅱ,Beclin-1 protein and mRNA expressions were significantly increased(P<0.01).Conclusion:The water extract of Tongbi Jiangu Formula can improve the anti-stress ability and survival ability of chondrocytes by regulating autophagy,and inhibit inflammatory response,which is one of the mechanisms of Tongbi Jiangu Formula in treating KOA.

关 键 词：通痹健骨方 人软骨细胞 自噬 膝骨关节炎 机制 

分 类 号：R285[医药卫生—中药学]