补肾强筋胶囊对膝骨关节炎的治疗作用及分子机制研究  

Study on the Effect and Molecular Mechanism of Bushenqiangjin Capsule on Knee Osteoarthritis

作　　者：彭莎 姚楠[1,2,3] 卢岩岩 许学猛[1,2] 吴淮[1,2] 刘文刚 黄雪君[1,2,3] 陈国材 PENG Sha;YAO Nan;LU Yanyan;XU Xuemeng;WU Huai;LIU Wengang;HUANG Xuejun;CHEN Guocai(The Fifth Clinical College of Guangzhou University of Chinese Medicine,Guangzhou 510095,China;Guangdong Second Traditional Chinese Medicine Hospital(Guangdong Province Engineering Technology Research Institute of Traditional Chinese Medicine),Guangzhou 510095,China;Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine,Guangzhou 510095,China)

机构地区：[1]广州中医药大学第五临床医学院,广州510095 [2]广东省第二中医院(广东省中医药工程技术研究院) [3]广东省中医药研究开发重点实验室

出　　处：《中国中医骨伤科杂志》2021年第4期13-19,共7页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：广东省基础与应用基础研究基金项目(2018A030313584);广东省中医药局科研项目(20194002,20183001)。

摘　　要：目的:探讨补肾强筋胶囊对膝骨关节炎(KOA)的治疗作用及相关分子机制。方法:将40只SD大鼠随机分为4组,每组10只。采用关节腔注射木瓜蛋白酶和半胱氨酸建立KOA大鼠模型。正常组和模型组给予生理盐水灌胃,补肾高、低剂量组分别灌胃0.486 g/kg和0.243 g/kg补肾强筋胶囊。6周后处死大鼠并采集膝关节软骨和血清进行检测。苏木精-伊红(HE)染色和番红-固绿染色观察各组大鼠膝关节软骨病理变化并进行改良Mankin评分;酶联免疫吸附测定(ELISA)检测大鼠血清白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平;实时荧光定量PCR和蛋白免疫印迹(Western Blot)检测MMP13、Beclin1、LC3B、ULK1、p-AMPK、p-mTOR mRNA和蛋白的表达。结果:与模型组相比,补肾高、低剂量组大鼠膝关节软骨退变程度明显减轻且Mankin评分显著降低,差异有统计学意义(P<0.01)。补肾高、低剂量组大鼠血清IL-1β和TNF-α水平显著降低,差异有统计学意义(P<0.01)。补肾高、低剂量组大鼠软骨MMP13 mRNA和蛋白表达显著降低,Beclin1、LC3B-Ⅱ/Ⅰ、ULK1 mRNA和蛋白表达显著升高,p-mTOR蛋白表达显著降低,差异有统计学意义(P<0.01)。补肾高剂量组大鼠软骨p-AMPK蛋白表达显著升高,差异有统计学意义(P<0.01)。结论:补肾强筋胶囊可能通过AMPK/mTOR通路上调KOA模型大鼠膝关节软骨自噬水平从而延缓病程。Objective:To explore the effect and relative molecular mechanism of Bushenqiangjin capsule on knee osteoarthritis(KOA).Methods:40 SD rats were randomly divided into 4 groups, with 10 rats in each group.The KOA rat model was established by injection of papain and cysteine into joints.After 4 weeks, normal group and model group were given oral administration of normal saline, the high and low dose groups of bushenqiangjin capsule were given oral administration of 0.486 g/kg and 0.243 g/kg Bushenqiangjin capsule respectively.After 6 weeks, all the rats were killed, knee cartilage and serum were collected and tested.Hematoxylin-eosin(HE) staining and Safranin o-fast green staining were used to observe the pathological changes of knee cartilage in each group and modified Mankin score was calculated.The levels of serum interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) in rats were detected by enzyme linked immunosorbent assay(ELISA).The mRNA and protein expressions of MMP13,Beclin1,LC3 B,ULK1,p-AMPK and p-mTOR were detected by Real-time PCR and Western Blot.Results:Compared with the model group, the degeneration degree of knee cartilage and Mankin score of rats in the high and low dose groups of bushenqiangjin capsule were reduced significantly(P<0.01).Compared with the model group, the levels of serum IL-1β and TNF-α in the high and low dose groups of bushenqiangjin capsule were decreased significantly(P<0.01).Compared with the model group, the mRNA and protein expression of MMP13 were decreased significantly, the mRNA and protein expressions of Beclin1,LC3 B-Ⅱ/Ⅰ and ULK1 were increased significantly, and the protein expression of p-mTOR was decreased significantly in the high and low dose groups of bushenqiangjin capsule(P<0.01).In addition, the protein expression of p-AMPK was increased significantly in the high dose group of bushenqiangjin capsule(P<0.01).Conclusion:Bushenqiangjin capsule may up-regulate the autophagy level of knee cartilage in KOA model rats through AMPK/mTOR pathway, which delays

关 键 词：补肾强筋胶囊 膝骨关节炎 自噬 分子机制 

分 类 号：R684.3[医药卫生—骨科学]