基于MAPK信号通路探讨复方芪芎颗粒对IL-1β诱导的兔骨关节炎软骨细胞的作用及其机制  

The Mechanism of Compound Qixiong Granule(复方芪芎颗粒)on IL-1βInduced Osteoarthritis Chondrocytes Based on MAPK Signaling Pathway

作　　者：杨丽莎[1] 陈利锋 张传成 刘雪晴 谭张奎 YANG Lisha;CHEN Lifeng;ZHANG Chuancheng;LIU Xueqin;TAN Zhangkui(School of Medicine,Wuhan University of Science and Technology,Wuhan Hubei 430065,China;Central Theater Command General Hospital,Wuhan Hubei 430070,Chin;Huizhou Hospital Affiliated to Guangzhou Medical University,Guangzhou Guangdong 516001,China)

机构地区：[1]武汉科技大学医学部医学院,湖北武汉430065 [2]中部战区总医院,湖北武汉430070 [3]广州医科大学附属惠州医院,广东广州516001

出　　处：《中医药导报》2024年第8期1-7,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖北省医学青年拔尖人才;武汉市知识创新专项(2022020801010517)。

摘　　要：目的:基于MAPK信号通路探讨复方芪芎颗粒对IL-1β诱导的兔骨关节炎软骨细胞的作用及其机制。方法:选取5只清洁级新西兰大白兔,提取兔软骨细胞培养至第2代,通过IL-1β诱导软骨细胞,建立骨关节炎软骨细胞体外模型,将其分为空白对照组、IL-1β组、双醋瑞因组、复方芪芎颗粒低浓度组、复方芪芎颗粒中浓度组、复方芪芎颗粒高浓度组。空白对照组不进行任何干预,IL-1β组采用质量浓度为10μg/L的IL-1β处理,双醋瑞因组采用10-5mol/L双醋瑞因处理,复方芪芎颗粒低、中、高浓度组分别采用10 mg/mL、20 mg/mL、30 mg/mL复方芪芎颗粒处理。干预24、48、72 h后,采用实时荧光定量逆转录聚合酶链式反应(RT-qPCR)检测各组P38激酶、c-Jun氨基端激酶(JNK)、细胞外信号调节蛋白激酶(ERK)、P65激酶、基质金属蛋白酶13(MMP-13)基因表达水平;干预72h后,采用Western blotting法检测各组P38、JNK、ERK、P65、MMP-13蛋白表达水平。结果:干预24、48、72 h后,IL-1β组P38 mRNA、JNK mRNA、ERK mRNA、P65 m RNA、MMP-13 mRNA相对表达量高于空白对照组(P<0.05);双醋瑞因组和复方芪芎颗粒低、中、高浓度组P38 mRNA、JNK mRNA、ERK mRNA、P65 mRNA、MMP-13 mRNA相对表达量均低于IL-1β组(P<0.05);复方芪芎颗粒高浓度组P38 mRNA、JNK mRNA、ERK m RNA、P65 mRNA、MMP-13 mRNA相对表达量低于复方芪芎颗粒低浓度组(P<0.05)。随着药物干预时长的延长,双醋瑞因组和复方芪芎颗粒低、中、高浓度组P38 mRNA、JNK mRNA、ERK mRNA、P65 m RNA、MMP-13 mRNA相对表达量均呈降低趋势(P<0.05)。干预72 h后,IL-1β组P38、JNK、ERK、P65、MMP-13蛋白相对表达量高于空白对照组(P<0.01);双醋瑞因组和复方芪芎颗粒低、中、高浓度组P38、JNK、ERK、P65、MMP-13蛋白相对表达量均低于IL-1β组(P<0.01);复方芪芎颗粒高浓度组P38、JNK、ERK、P65、MMP-13蛋白相对表达量低于复方芪芎颗粒低浓度组(P<0.01)。结论:Objective:To investigate the mechanism of compound Qixiong granule on IL-1βinduced rabbit chondrocytes in osteoarthritis based on the MAPK signaling pathway.Methods:Totally five clean New Zealand white rabbits were selected.Chondrocytes were extracted from rabbit cartilage and cultured to the second generation.The chondrocytes were induced with IL-1βto establish an in vitro model of osteoarthritis chondrocytes.The chondrocytes were divided into blank control group,IL-1βgroup,Diacerein group,low-dose compound Qixong granules group(low-dose group),medium-dose compound Qixong granules group(medium-dose group)and high-dose compound Qixong granules group(high-dose group).The blank control group received no intervention.The IL-1βgroup was treated with IL-1βat a concentration of 10μg/L.The Diacerein group was treated with Diacerein at a concentration of 10-5 mol/L.The low,medium,and high-dose groups were treated with 10 mg/mL,20 mg/mL,and 30 mg/mL of compound Qixiong granules,respectively.After intervention for 24,48 and 72 hours,RT-qPCR was used to detect the gene expression levels of P38 kinase,c-Jun N-terminal kinase(JNK),extracellular signal-regulated kinase(ERK),P65 kinase and matrix metalloproteinase 13(MMP-13)in each group.After 72 hours of intervention,the protein expression levels of P38,JNK,ERK,P65 and MMP-13 in each group were detected by Western blotting.Results:After 24,48 and 72 hours of intervention,the IL-1βgroup showed higher relative expression levels of P38 mRNA,JNK mRNA,ERK mRNA,P65 mRNA and MMP-13 mRNA than blank control group(P<0.05);The Diacerein group,low-dose group,medium-dose group and high-dose group showed lower relative expression levels of P38 mRNA,JNK mRNA,ERK mRNA,P65 mRNA and MMP-13 mRNA than IL-1βgroup(P<0.05);The high-dose group showed lower relative expression levels of P38 mRNA,JNK mRNA,ERK mRNA,P65 mRNA and MMP-13 mRNA than low-dose group(P<0.05).With the prolongation of drug intervention time,the relative expression levels of P38 mRNA,JNK mRNA,ERK mRNA,P65 mRNA and MMP-13 m

关 键 词：骨关节炎 复方芪芎颗粒 白介素-1Β MAPK信号通路 软骨细胞 新西兰大白兔 

分 类 号：R285.5[医药卫生—中药学]