基于AMPKα/mTOR/p70S6K通路探讨补肾抗衰片类脂联素受体激动剂样作用干预人主动脉平滑肌细胞增殖的机制  

Mechanism of Bushen Kangshuai Tablet anti-HASMC proliferation via intervening the pathway of AMPKα/mTOR/p70S6K based on its effect similar as adiponectin receptor agonists

作　　者：庞树朝[1,2] 陈美玲 张军平[1,2] PANG Shuchao;CHEN Meiling;ZHANG Junping(First Teaching Hospital of Tianjin University of Traditional Chinese Midicine,Tianjin 300381,China;National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300381,China;Tianjin University of Traditional Chinese Midicine,Tianjin 300381,China)

机构地区：[1]天津中医药大学第一附属医院,天津300381 [2]国家中医针灸临床医学研究中心,天津300381 [3]天津中医药大学,天津300381

出　　处：《中华中医药杂志》2023年第10期4711-4716,共6页China Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:基于AMPKα/mTOR/p70S6K通路探讨补肾抗衰片(以下简称中药)类脂联素受体激动剂(AdipoRon)样作用影响人主动脉平滑肌细胞(HASMC)增殖的可能机制。方法:复制PDGF诱导HASMC增殖模型。随机分为对照组、模型组、中药高剂量组、中药低剂量组、阿托伐他汀(以下简称西药)高剂量组和西药低剂量组。用Western Blot法检测药物对PDGF诱导的HASMC上细胞周期蛋白D1(Cyclin D1)、脂联素受体(AdipoR)以及AMPKα/mTOR/p70S6K通路相关蛋白表达的影响。结果:与对照组比较,模型组HASMC经PDGF诱导后,其Cyclin D1蛋白表达显著升高(P<0.01);AdipoR1、AdipoR2蛋白表达显著降低(P<0.01);p-AMPKα/AMPKα显著降低(P<0.01),p-mTOR/mTOR和p-p70S6K/p70S6K显著升高(P<0.01)。与模型组比较,中药高、低剂量组能够抑制PDGF诱导的HASMC增殖,并显著降低Cyclin D1蛋白的表达水平(P<0.01,P<0.05)。中药高剂量组能够显著增强AdipoR1和AdipoR2蛋白表达水平(P<0.01),激活AMPKα蛋白并促进其磷酸化,上调p-AMPKα/AMPKα(P<0.01),进而抑制mTOR、p70S6K蛋白表达,下调pmTOR/mTOR和p-p70S6K/p70S6K(P<0.01)。中药低剂量组能够增强AdipoR1和AdipoR2蛋白表达水平(P<0.01,P<0.05),能够抑制mTOR、p70S6K蛋白表达,下调pmTOR/mTOR和p-p70S6K/p70S6K(P<0.05,P<0.01);其虽可激活AMPKα蛋白表达,但无法上调pAMPKα/AMPKα(P<0.01)。结论:高剂量的补肾抗衰片能够通过干预AMPKα/mTOR/p70S6K通路进而抑制HASMC增殖,其具体机制依赖于补肾抗衰片对AdipoR1和AdipoR2表达的上调作用,提示补肾抗衰片具有类AdipoRon样作用。Objective:To investigate the role of Bushen Kangshuai Tablet(BKT)anti-Human aortic smooth muscle cell(HASMC)proliferation via intervening the pathway of AMPKα/mTOR/p7OS6K based on its effect similar as adiponectin receptor agonists(AdipoRon).Methods:HASMC proliferation model induced by PDGF was replicated.The cells were randomly divided into control group,model group,BKT high-dose group,low-dose group,atorvastatin(WM)high-dose group and WM low-dose group.Western Blot was used to detect the effects of PDGF on the expression of Cyclin D1,adiponectin receptor(AdipoR)and AMPKα/mTOR/p7OS6K pathway related proteins in HASMC.Results:Compared with the control group,the expression of Cyclin D1 protein in HASMC of the model group was significantly increased after PDGF induction(P<0.01);The expressions of AdipoR1 and AdipoR2 were significantly decreased(P<0.01);p-AMPKα/AMPKαratio decreased significantly(P<0.01),the ratio of p-mTOR/mTOR and p-P70S6K/p70S6K increased significantly(P<0.01).Compared with the model group,BKT highdose and low-dose group could inhibit the proliferation of HASMC induced by PDGF and reduce the expression level of Cyclin D1 protein(P<0.01,P<0.05).BKT high-dose group could significantly increase the protein expression levels of AdipoR1 and AdipoR2(P<0.01),activate AMPKα protein expression and promote its phosphorylation,up-regulate pAMPKα/AMPKa(P<0.01),and then inhibit the protein expression of mTOR and p70S6K.Down-regulation of pmTOR/mTOR and p-p70S6K/p70S6K(P<0.01).BKT low-dose group could enhance the protein expression levels of AdipoR1 and AdipoR2(P<0.01,P<0.05),inhibit the protein expression of mTOR and p70S6K,and down-regulate pmTOR/mTOR and p-p70S6K/p70S6K(P<0.05,P<0.01).Although it could activate AMPKα protein expression,it could not up-regulate pAMPKα/AMPKaα(P<0.01).Conclusion:High dose of BKT can inhibit the proliferation of HASMC by interfering the AMPKα/mTOR/p7OS6K pathway,and the specific mechanism depends on the up-regulation of AdipoR1 and AdipoR2 expression by BKT,which sugge

关 键 词：补肾抗衰片 脂联素抵抗 脂联素受体 人主动脉平滑肌细胞 AMPKα/mTOR/p70S6K通路 

分 类 号：R285[医药卫生—中药学]