补肾调肝方含药血清对白细胞介素1β诱导软骨细胞凋亡的影响  

Effect of serum containing Bushen Tiaogan prescription on interleukin-1beta-induced chondrocyte apoptosis

作　　者：范帅 林杰彬 吴春飞 徐兆辉 Fan Shuai;Lin Jiebin;Wu Chunfei;Xu Zhaohui(The Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510000,Guangdong Province,China;Shenzhen Luohu District Traditional Chinese Medicine Hospital,Shenzhen 518000,Guangdong Province,China)

机构地区：[1]广州中医药大学第三附属医院,广东省广州市510000 [2]深圳市罗湖区中医院,广东省深圳市518000

出　　处：《中国组织工程研究》2021年第35期5594-5598,共5页Chinese Journal of Tissue Engineering Research

基　　金：广东省自然科学基金项目(2018A0303130103);广东省中医药局科研项目(20201171),项目负责人:范帅;广州中医药大学第三附属医院创新基金(sy201701),项目负责人:范帅。

摘　　要：背景:前期研究发现,补肾调肝方能明显改善骨关节炎症状,临床疗效肯定,但缺乏相关的实验依据。目的:探索补肾调肝方含药血清对白细胞介素1β诱导的软骨细胞凋亡的影响及其分子机制。方法:取传代培养的第3代大鼠软骨细胞,随机分为4组:PBS组(对照组)、10μg/L白细胞介素1β组、10μg/L白细胞介素1β+5%补肾调肝方含药血清(5%补肾调肝方含药血清组)及10μg/L白细胞介素1β+10%补肾调肝方含药血清(10%补肾调肝方含药血清组),分别处理软骨细胞24 h。采用流式细胞术检测细胞凋亡;蛋白印迹分析SOX9、NF-κB P65和p-NF-κB P65蛋白的表达水平。实验方案经广州中医药大学动物实验伦理委员会批准。结果与结论:①与对照组相比,白细胞介素1β可诱导软骨细胞的凋亡(P<0.05);②与白细胞介素1β组比,5%和10%补肾调肝方含药血清组软骨细胞的凋亡率均下降,且呈剂量依赖性(均P<0.05);③与对照组相比,白细胞介素1β组SOX9的表达明显降低(P<0.05),p-P65/P65明显升高(P<0.05);5%和10%补肾调肝方含药血清组SOX9的表达明显高于白细胞介素1β组(P<0.05),10%补肾调肝方含药血清组p-P65/P65显著低于白细胞介素1β组(P<0.05);④结果说明,补肾调肝方可能是通过调控SOX9/NF-κB抑制白细胞介素1β的诱导大鼠骨关节软骨细胞凋亡,起到缓解关节软骨退变的作用。BACKGROUND:Previous studies have found that Bushen Tiaogan prescription can significantly improve the symptoms of osteoarthritis,with a positive clinical effect,but there is still a lack of relevant experimental evidence.OBJECTIVE:To explore the effect of serum containing Bushen Tiaogan prescription on interleukin(IL)-1β-induced chondrocyte apoptosis and its molecular mechanism.METHODS:Passage 3 chondrocytes from rats were randomly divided into four groups:PBS group(control group),10μg/L IL-1βgroup,10μg/L IL-1β+5%Bushen Tiaogan prescription group,10μg/L IL-1β+10%Bushen Tiaogan prescription group,in which chondrocytes were treated for 24 hours.Flow cytometry was used to detect cell apoptosis,and western blot was used to analyze the expression levels of SOX9,NF-κB P65 and p-NF-κB P65 proteins.An approval was obtained from the Animal Experimental Ethics Committee of Guangzhou University of Chinese Medicine.RESULTS AND CONCLUSION:Compared with the control group,IL-1βinduced chondrocyte apoptosis(P<0.05).Compared with the IL-1βgroup,the apoptosis rate of chondrocytes in the 10μg/L IL-1β+5%Bushen Tiaogan prescription and 10μg/L IL-1β+10%Bushen Tiaogan prescription groups significantly decreased in a dose-dependent manner(P<0.05).Compared with the control group,the expression of SOX9 in the IL-1βgroup was significantly lower than that of the control group(P<0.05),and the expression of p-P65/P65 was significantly higher than that of the control group(P<0.05).The expression of SOX9 in the 10μg/L IL-1β+5%Bushen Tiaogan prescription and 10μg/L IL-1β+10%Bushen Tiaogan prescription groups was significantly higher than that in the IL-1βgroup(P<0.05),and the expression of p-P65/P65 in the 10μg/L IL-1β+10%Bushen Tiaogan prescription group was significantly lower than that in the IL-1βgroup(P<0.05).To conclude,Bushen Tiaogan prescription may inhibit IL-1β-induced apoptosis of rat osteoarticular chondrocytes by regulating SOX9/NF-κB,and play a role in alleviating articular cartilage degeneration.

关 键 词：骨关节炎 补肾调肝方 关节软骨 SOX9/NF-κB 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]