荣筋拈痛方对白细胞介素-1β诱导的体外大鼠软骨细胞自噬相关基因表达的影响

来源:用户上传      作者:潘丹虹 李路 王文义 林晴 付长龙 吴广文 叶锦霞

　　
　　 【关键词】 膝骨关节炎;荣筋拈痛方;软骨细胞;自噬;白细胞介素-1β;大鼠
　　expression of autophagy related gene of interleukin-1β-induced rat chondrocytes in vitro and its mechanism in the prevention and treatment of knee osteoarthritis.Methods：The knee cartilages of 4-week-old SD rats were isolated and cultured in vitro.The second-generation chondrocytes were stained with toluidine blue for cell identification.Via IL-1β 10 ng・mL-1，The chondrocyte inflammation model in vitro was induced.After successful induction，Rongjin Niantong Fang（50，100，200 μg・mL-1）was used for incubation for 48 h.The cells were divided into a blank control group，a model group and the low，medium and high dose groups.The effect of Rongjin Niantong Fang on chondrocyte activity was detected by the CCK-8 method;The mRNA expressions of LC3Ⅱ and Beclin 1 in chondrocytes were detected by qPCR;The ratio of autophagy protein LC3Ⅱ/Ⅰ and the protein expression level of Beclin 1 were observed by Western Blot.Results：Compared with the model control group，the cell viability in the low，medium and high dose groups of Rongjin Niantong Fang significantly increased in a dose-dependent manner（P < 0.01）;The mRNA of LC3Ⅱ and Beclin 1 and the protein expression of LC3Ⅱ/Ⅰ and Beclin 1 in the model control group were higher than those in the blank control group（P < 0.05）;After the intervention of Rongjin Niantong Fang（100 μg・mL-1），the mRNA of LC3Ⅱ and Beclin 1 and the protein expression levels of LC3Ⅱ/Ⅰ and Beclin 1 decreased（P < 0.05）.Conclusion：Rongjin Niantong Fang may delay the degeneration of knee osteoarthritis by inhibiting the level of autophagy.
　　 【Keywords】 knee osteoarthritis;Rongjin Niantong Fang（s筋拈痛方）;chondrocytes;autophagy;interleukin-1β;rats
　　 膝骨关节炎（knee osteoarthritis，KOA）是一种以关节软骨退行性改变和关节炎症为特征的慢性骨关节疾病[1]，多见于中老年人，年龄≥60岁的人群中约有80%的人出现骨关节炎（osteoarthritis，OA）[2]。自噬是细胞生存、分化和发育必不可少的途径，在炎症反应中具有双向调节作用，在促进和抑制炎症反应方面同等重要。白细胞介素-1β（IL-1β）被普遍认为是引起关节软骨基质降解的主要促炎症细胞因子，常作为OA体外退变软骨细胞模型诱导剂。前期研究表明，荣筋拈痛方具有多靶点成分，临床防治KOA效果明显[3-5]。因此，本研究选用荣筋拈痛方作为干预媒介，以体外退变软骨细胞作为研究载体，观察荣筋拈痛方对退变软骨细胞自噬的影响，为临床应用提供实验依据。

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