青藤碱通过PI3K/AKt-mTOR信号通路调节膝OA兔软骨自噬水平机制研究  

Sinomenine Regulates the Autophagy Level of Cartilage in Knee OA Rabbits through PI3K/AKt-mTOR Signaling Pathway

作　　者：郑洁[1] 袁普卫[1] 赵莉平[1] 边敏佳[1] 康武林[1] ZHENG Jie;YUAN Puwei;ZHAO Liping;BIAN Minjia;KANG Wulin(Shaanxi Universtiy of Chinese Medicine,Xianyang 712046,Shaanxi,China)

机构地区：[1]陕西中医药大学

出　　处：《辽宁中医药大学学报》2019年第8期30-33,共4页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：陕西省中医药管理局中医药科研计划项目(JCPT035);陕西省教育厅科研项目(17JK0195);陕西省科技厅自然科学基础项目(2016JQ8004);陕西省三秦学者创新团队项目

摘　　要：目的:观察不同剂量青藤碱(SIN)对膝OA兔关节软骨自噬水平以及自噬抑制信号通路PI3K/AKtmTOR关键分子表达的影响。方法:50只新西兰大白兔随机分为空白组、模型组、SIN低剂量组、SIN中剂量组和SIN高剂量组。空白组不予任何处理,模型组、SIN低剂量组、SIN中剂量组和SIN高剂量组通过Hulth法建立膝骨关节炎模型后,分别用生理盐水、SIN进行膝关节腔注射,共干预10次,SIN用量由低到高分别为0.2mL(5mg)、0.35mL(8.75mg)和0.5mL(12.5mg)。实时荧光定量PCR法检测关节软骨Atg-5和Atg-12的mRNA表达,Westernblot法检测关节软骨Beclin-1、LC3-Ⅱ、PI3K、AKt和mTOR的蛋白表达。结果:与空白组比较,模型组和SIN低剂量组Atg-5、Atg-12的mRNA表达以及Beclin-1、LC3-Ⅱ的蛋白表达显著下调(P<0.05),SIN中、高剂量组显著高于模型组,SIN中、高剂量两组之间比较无统计学差异(P>0.05);与空白组比较,模型组和SIN低剂量组PI3K、AKt和mTOR的蛋白表达明显上调(P<0.05),SIN中、高剂量组明显低于模型组(P<0.05),SIN中剂量组与高剂量组比较无统计学差异(P>0.05)。结论:中、高剂量的青藤碱可通过抑制PI3K/AKt-mTOR信号通路活性上调膝OA兔关节软骨自噬水平。Objective:To observe the effect of different doses of sinomenine(SIN) on autophagy level of knee OA rabbit cartilage and expression of key molecules of PI3 K/AKt-mTOR signal pathway. Methods:A total of 50 New Zealand white rabbits were randomly divided into 5 groups:control group,model group,SIN low dose group,SIN moderate dose group and SIN high dose group. Model group,SIN low dose group,SIN moderate dose group and SIN high dose group were established knee osteoarthritis model by Hulth method,SIN groups received intra-articular injection of SIN(0.2 mL,0.35 mL and 0.5 mL respectively),model group received intra-articular injection of 0.2 mL physiological saline. After 10 times of treament,qPCR was used to detect the mRNA level of Atg-5 and Atg-12,and Western blot was used to detect the protein level of Beclin-1,LC3-Ⅱ,PI3 K,AKt and mTOR. Results:Compared with control group,mRNA expression of Atg-5 and Atg-12 as well as protein expression of Beclin-1 and LC3-Ⅱ in model group and SIN low dose groups were all raised significantly(P<0.05),and that in SIN moderate group and high dose group were increased than model group,and there was no significant difference between SIN moderate dose group and SIN high dose group(P>0.05);compared with control group,protein expression of PI3 K,AKt and mTOR in model group and SIN low dose group were all increased obviously(P<0.05),and that in SIN moderate and high dose groups were decreased than model group(P<0.05),and there was no significant difference between SIN moderate dose group and SIN high dose group(P>0.05). Conclusion:Moderate and high dose of sinomenine can upregulate the level of autophagy of articular cartilage in knee OA rabbit by inhibiting the activity of PI3 K/akt-mtor signaling pathway.

关 键 词：青藤碱 骨性关节炎 自噬 PI3K/AKt-mTOR通路 

分 类 号：R285.5[医药卫生—中药学]