来源：风湿病与关节炎,2022,11（2）:1-5,15.

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

（基础研究）

潘丹虹¹，李　路¹，王文义¹，林　晴¹，付长龙²，吴广文¹，叶锦霞¹

　　【摘　要】目的：观察荣筋拈痛方对白细胞介素-1β（IL-1β）诱导的体外大鼠软骨细胞自噬相关基因表达的影响，探讨其防治膝骨关节炎的作用机制。方法：取4周龄SD大鼠膝关节软骨，进行软骨细胞的体外分离培养，选取第2代软骨细胞甲苯胺蓝染色进行细胞鉴定。经IL-1β 10 ng·mL^-1诱导建立体外软骨细胞炎症模型，诱导成功后，给予荣筋拈痛方（50，100，200 μg·mL^-1）孵育48 h。将细胞分为空白对照组，模型对照组，荣筋拈痛方低、中、高剂量组。分别采用CCK-8法检测荣筋拈痛方对软骨细胞活性的影响，qPCR法检测软骨细胞中LC3Ⅱ、Beclin 1的mRNA表达，Western Blot法观察自噬蛋白LC3Ⅱ/Ⅰ比值、Beclin 1的蛋白表达水平。结果：与模型对照组比较，荣筋拈痛方低、中、高剂量组均显著提高细胞活力，呈剂量依赖性增加（P < 0.01）；模型对照组较空白对照组LC3Ⅱ、Beclin 1的mRNA及LC3Ⅱ/Ⅰ、Beclin 1的蛋白表达水平升高（P < 0.05）；经100 μg·mL^-1荣筋拈痛方干预后LC3Ⅱ、Beclin 1的mRNA及LC3Ⅱ/Ⅰ、Beclin 1的蛋白表达水平降低（P < 0.05）。结论：荣筋拈痛方可能通过抑制自噬水平，从而具有延缓膝骨关节炎退变的作用。

　　【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

Effect of Rongjin Niantong Fang（荣筋拈痛方）on the Expression of Autophagy Related Gene of Interleukin-1β-induced Rat Chondrocytes in Vitro

PAN Dan-hong,LI Lu,WANG Wen-yi,LIN Qing,FU Chang-long,WU Guang-wen,YE Jin-xia

　　【ABSTRACT】Objective:To observe the effect of Rongjin Niantong Fang（荣筋拈痛方）on the

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（荣筋拈痛）;chondrocytes;autophagy;interleukin-1β;rats