基于PI3K/AKT信号传导通路探究骨痹合剂对膝关节骨关节炎小鼠细胞凋亡和相关蛋白水平表达的影响  

Exploring the effect of Gu Bi mixture on cell apoptosis and related protein expression in KOA mice based on PI3K/AKT signaling pathway

作　　者：廖建青 吕静 张英杰 谢邦 马富文 王琦 LIAO Jianqing;LV Jing;ZHANG Yingjie;XIE Bang;MA Fuwen;WANG Qi(Yunnan University of Traditional Chinese Medicine,Kunming 650500,China;Yunnan Hospital of Traditional Chinese Medicine,Kunming 650021,China;不详)

机构地区：[1]云南中医药大学,昆明650500 [2]云南省中医医院,昆明650021

出　　处：《实用医学杂志》2021年第15期1916-1920,共5页The Journal of Practical Medicine

基　　金：国家自然科学基金项目(编号:81560781);云南省内设研究研究基金(编号:2018NS0038);云南省中医药大学2020年度研究生科学研究基金(编号:2020Y09)。

摘　　要：目的观察具有活血通络作用的骨痹合剂对膝关节骨关节炎(knee osteoarthritis,KOA)小鼠关节软骨细胞凋亡和PI3K/AKT信号传导通路相关蛋白表达水平的影响。方法采用前交叉韧带切断法建立KOA模型小鼠,造模8周后分别予骨痹合剂、氨糖美辛肠溶片溶于同体积蒸馏水、等体积的生理盐水灌胃,取双膝关节软骨进行检测。通过甲苯胺蓝染色对KOA软骨进行形态学观察,TUNEL法检测各组软骨细胞凋亡情况、蛋白质免疫印迹法(Western blotting)检测各组软骨细胞中PI3K/AKT/mTOR蛋白的表达水平。结果与生理盐水组比较,骨痹合剂组与氨糖美辛组甲苯胺蓝染色较深,软骨组织有明显改善,细胞凋亡明显减少(P<0.01),其中骨痹合剂组小鼠关节软骨中PI3K、AKT和mTOR蛋白表达均下降(P<0.05);氨糖美辛组小鼠关节软骨中PI3K、AKT和mTOR蛋白表达均下降(P<0.01)。结论骨痹合剂和氨糖美辛对KOA小鼠软骨细胞都具有明显的改善作用,骨痹合剂能通过抑制PI3K/AKT信号通路,减少软骨细胞凋亡,缓解膝关节骨关节炎进一步发展。Objective To observe the effect of Gu Bi mixture,which has the function of promoting blood circulation on the apoptosis of chondrocytes and the expression level of PI3K/AKT signaling pathway-related proteins in knee osteoarthritis(KOA)mice.Methods To cut the anterior cruciate ligament was used to establish in KOA model mice.Eight weeks after the modeling,the Gu Bi mixture group used Gu Bi mixture to fill the stomach,Glucosamine Indomethacin Entric-coated Tablets group in mice with Glucosamine Indomethacin Entric-coated Tablets dissolved in same volume of distilled water to fill the stomach,and normal saline(NS)group used equal volume of NS for gastric lavage,and the cartilage of both knee joints were tested.The morphology of KOA cartilage model was observed by toluidine blue staining,and apoptosis of chondrocytes in each group was detected by TUNEL method and expression levels of PI3K/AKT and mTOR proteins in chondrocytes in each group were detected by Western blotting.Results Compared with the NS group,the toluidine blue staining was deeper and the cartilage tissue was significantly improved in the Gu Bi mixture group and the Glucosamine Indomethacin Entric-coated Tablets group.Apoptosis of chondrocyte was reduced and the expressions of PI3K,AKT and mTOR proteins in the articular cartilage of the mice in the Gu Bi mixture group were decreased(P<0.05);PI3K/AKT and mTOR protein expression in articular cartilage of mice in the Glucosamine Indomethacin Entric-coated Tablets group decreased(P<0.01).Conclusion Both Gu Bi mixture and Glucosamine Indomethacin Entric-coated Tablets had significant improvement effects on KOA mice chondrocytes,Gu Bi mixture could inhibit the PI3K/AKT signaling pathways,reduce apoptosis of chondrocytes,and to further remit knee osteoarthritis.

关 键 词：膝关节骨关节炎 软骨细胞凋亡 活血通络法 骨痹合剂 PI3K/AKT信号传导通路 

分 类 号：R684.3[医药卫生—骨科学]