摘要
目的观察消肿止痛合剂对大鼠膝骨性关节炎软骨中LC3、Beclin1、caspase-9、单磷酸腺苷活化蛋白激酶(p-AMPK)、哺乳动物雷帕霉素靶蛋白(mTOR)表达的影响,探讨消肿止痛合剂干预膝骨性关节炎(knee osteoarthritis,KOA)形成的作用机制。方法将80只Wistar大鼠随机分为4组:观察组(消肿止痛合剂组)、对照组(硫酸氨基葡萄糖片组)、模型组、假手术组,每组20只,除假手术外,其余3组通过改良Hulth法行膝骨关节炎造模。药物连续干预8周后,取各组大鼠膝关节软骨,采用ELISA方法检测血清中白细胞介素-6 (IL-6)、基质金属蛋白酶-3(MMP-3)、环氧化酶-2(COX-2)水平,实时荧光定量聚合酶链式反应检测各组软骨中LC3、Beclin1、caspase-9 mRNA的表达情况,蛋白质印迹法(Western blot)检测各组软骨细胞中pAMPK、mTOR蛋白的表达。结果与假手术组比较,模型组IL-6、MMP-3、COX-2水平明显升高,下调LC3、Beclin1、p-AMPK的表达量,上调caspase-9、mTOR的表达量,差异有统计学意义(P<0. 05);与模型组比较,观察组、对照组降低IL-6、MMP-3、COX-2水平,显著上调LC3、Beclin1、p-AMPK的表达量,下调caspase-9、mTOR的表达量,差异有统计学意义(P<0. 05);其中观察组优于对照组,差异有统计学意义(P<0. 05)。结论消肿止痛合剂对KOA关节软骨损伤的保护可能与AMPK/mTOR信号通路相关。
Objective To observe the effect of the anti-swelling and analgesic mixture on the expressions of LC3,Beclin1,caspase-9,p-AMPK,and mammalian rapamycin target protein(mT OR) in the cartilage of the knee osteoarthritis(KOA) rats,and to explore the intervention mechanism of the formation of KOA with the anti-swelling and analgesic mixture. Methods Eighty Wistar rats were randomly divided into 4 groups: the observation group(anti-swelling and analgesic mixture group),the control group(glucosamine sulfate tablet group),the model group,and the sham operation group(20 rats in each group). Except for the false operation,KOA model was established with modified Hulth method in the other 3 groups. After 8 weeks of continuous drug intervention,the cartilage of the knee joint was collected. Serum levels of IL-6,MMP-3,and COX-2 were detected using ELISA method. The mR NA expressions of LC3,Beclin1,and caspase-9 in the cartilage of each group were detected with real-time quantitative polymerase chain reaction(Realtime PCR). Protein expressions of p-AMPK and mT OR in chondrocytes were detected with Western blotting. Results Compared with those in the sham operation group,the levels of IL-6,MMP-3,and COX-2 in model group increased significantly,the expressions of LC3,Beclin1,and p-AMPK were down-regulated,and the expressions of caspase-9 and mT OR were up-regulated(P < 0. 05).Compared with those in the model group,the levels of IL-6,MMP-3,and COX-2 were down-regulated in the observation group and control group,and LC3,Beclin-2 were up-regulated significantly(P< 0. 05). Conclusion The protective effect of anti-swelling and analgesic mixture on KOA articular cartilage injury may be related to AMPK/mT OR signaling pathway.
作者
李金鹏
刘涛
何志军
李岩
陈文
宋渊
LI Jinpeng;LIU Tao;HE Zhijun;LI Yan;CHEN Wen;SONG Yuan(Gansu Provincial Hospital of Traditional Chinese Medicine,Lanzhou 730050,China)
出处
《中国骨质疏松杂志》
CAS
CSCD
北大核心
2020年第2期231-235,共5页
Chinese Journal of Osteoporosis
基金
甘肃省自然科学基金(17JR5RA052)
甘肃省中医药管理局科研项(GZK-2017-33)
关键词
消肿止痛合剂
膝骨性关节炎
信号通路
自噬
大鼠
动物实验
anti-swelling and analgesic mixture
knee osteoarthritis
signaling pathway
autophagy
rats
animal experimentation
