基于免疫印迹实验验证芍药甘草汤治疗骨关节炎关键靶点和通路  

Validation of Key Targets and Pathways of Shaoyao Gancao Decoction(芍药甘草汤)in the Treatment of Osteoarthritis Based on Western Blot Experiments

作　　者：宋庆慧[1] 李秋月 杨丽平[1] 朱嘉[1] 侯成志 张万强[1] SONG Qinghui;LI Qiuyue;YANG Liping;ZHU Jia;HOU Chengzhi;ZHANG Wanqiang(Wangjing Hospital,China Academy of Chinese Medical Sciences,Beijing 100102,China)

机构地区：[1]中国中医科学院望京医院,北京100102

出　　处：《辽宁中医药大学学报》2022年第9期10-13,共4页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：中国中医科学院优秀青年科技人才培养专项(ZZ13-YQ-036)。

摘　　要：目的基于免疫印迹实验深入探究芍药甘草汤对骨关节炎(osteoarthritis,OA)的药效机制。方法将40只大鼠随机分为4组,分别为假手术组(sham组)、模型组、芍药甘草汤(SGD)中剂量组(4.32 g/kg)、SGD高剂量组(8.64 g/kg)。除sham组以外的所有组大鼠使用内侧半月板失稳术(destabilization of the medial meniscus,DMM)构建骨关节炎模型,所有大鼠在手术造模后第1天开始灌胃给药,每日1次,持续8周,SGD组给予一定剂量的SGD,sham组和模型组给予等量的蒸馏水。给药8周后,对所有大鼠进行麻醉处死并取材。苏木素-伊红(HE)染色观察软骨组织形态学改变,免疫印迹法(Western blot)检测软骨组织中p38丝裂原活化蛋白激酶(MAPK)及C-myc蛋白的表达水平。结果与sham组比较,模型组、SGD中剂量组及高剂量组大鼠关节软骨的C-myc和p38分子蛋白表达水平明显升高(P<0.01),用芍药甘草汤干预后,其蛋白表达出现下降趋势,且下降幅度与芍药甘草汤浓度呈正相关,当芍药甘草汤浓度达到8.64 g/kg时,与模型组相比差异具有统计学意义(P<0.001)。结论芍药甘草汤能抑制OA大鼠软骨中p38和C-myc蛋白的过表达。Objective To explore the pharmacodynamic mechanism of Shaoyao Gancao Decoction(芍药甘草汤)on osteoarthritis based on western blot experiments.Methods Forty rats were randomly divided into 4 groups:sham group,model group,Shaoyao Gancao Decoction(SGD)medium-dose group(4.32 g/kg),and Shaoyao Gancao Decoction(SGD)high-dose group(8.64 g/kg).All rats except the sham group were used to establish osteoarthritis model by destabilization of the medial meniscus(DMM),for 8 weeks,the SGD group was given a certain dose of SGD,and the sham group and the model group were given the same amount of distilled water.After 8 weeks of administration,all rats were anesthetized and sacrificed and the materials were collected.The morphological changes of cartilage tissue were observed by hematoxylin-eosin(HE)staining,and the expression levels of p38 MAPK and C-myc protein in cartilage tissue were detected by Western blot.Results Compared with the sham group,the expression levels of C-myc and p38 molecular proteins in the articular cartilage of the rats in the model group,Shaoyao Gancao Decoction(SGD)medium-dose group,and Shaoyao Gancao Decoction(SGD)high-dose group were significantly increased(P<0.01),after intervention with Shaoyao Gancao Decoction,its protein expression showed a downward trend,and the decline was positively correlated with the concentration of Shaoyao Gancao Decoction.When the concentration of Shaoyao Gancao Decoction reached 8.64 g/kg,there was a difference compared with the model group.Statistically significant(P<0.001).Conclusion Shaoyao Gancao Decoction can inhibit the overexpression of p38 and C-myc proteins in cartilage of OA rats.

关 键 词：芍药甘草汤 骨关节炎 原癌基因C-MYC P38丝裂原活化蛋白激酶 

分 类 号：R684.3[医药卫生—骨科学]