SIRT1/FOXO3a/p27信号通路在白藜芦醇抑制肺动脉平滑肌细胞增殖中的作用  

Resveratrol suppresses proliferation of pulmonary arterial smooth muscle cells via SIRT1/FOXO3a/p27 signaling pathway

作　　者：柯蕊[1] 和平[1] 吴媛媛[1] 张永红[1] 张伟[1] 刘原[1] KE Rui;HE Ping;WU Yuanyuan;ZHANG Yonghong;ZHANG Wei;LIU Yuan(The Second Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710004,China)

机构地区：[1]西安交通大学第二附属医院,西安710004

出　　处：《山东医药》2021年第5期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金项目(81800030)。

摘　　要：目的探讨白藜芦醇(RES)是否通过影响沉默信息调节因子2相关酶1(SIRT1)/叉头蛋白O3a(FOXO3a)/p27信号通路抑制肺动脉平滑肌细胞(PASMC)增殖,为肺动脉高压新药研发提供一定的理论和实验依据。方法体外培养大鼠PASMC并随机分为三组,对照组不处理,5-HT刺激组加1μmol/L 5-HT,不同浓度RES干预组在加入5-HT前用10、30、100、300μmol/L RES预处理1 h,作用24 h后采用BrdU掺入法检测各组细胞增殖情况并筛选RES作用浓度;为研究机制进一步将PASMCs分为六组,对照组、5-HT刺激组、RES干预组(筛选出的RES作用浓度)处理同前,RES+SIRT1抑制剂干预组、RES+FOXO3a抑制剂干预组在加入1μmol/L 5-HT前用RES分别联合1μmol/L EX527、AS1842856预处理1 h,RES+p27 siRNA转染组在加入1μmol/L 5-HT前预先转染p27 siRNA 24 h后联合RES;作用24 h后采用BrdU掺入法检测各组细胞增殖情况,采用Western blotting法检测对照组、5-HT刺激组、RES干预组、RES+SIRT1抑制剂干预组的FOXO3a、p27蛋白及RES+FOXO3a抑制剂干预组的p27蛋白。结果PASMC增殖率5-HT刺激组高于对照组,100、300μmol/L RES干预组均低于5-HT刺激组(P均<0.05),故以100μmol/L作为后续实验中RES干预的作用浓度。PASMC增殖率5-HT刺激组高于对照组,RES干预组低于5-HT刺激组;RES+SIRT1抑制剂干预组、RES+FOXO3a抑制剂干预组、RES+p27 siRNA转染组均高于RES干预组(P均<0.05),三组间差异无统计学意义。PASMC中FOXO3a、p27蛋白表达量5-HT刺激组均低于对照组,RES干预组均高于5-HT刺激组,RES+SIRT1抑制剂干预组均低于RES干预组(P均<0.05);RES+FOXO3a抑制剂干预组p27蛋白表达量低于RES干预组(P<0.05)。结论RES可通过激活SIRT1上调FOXO3a,进而上调细胞周期抑制蛋白p27,最终抑制PASMC增殖。Objective To investigate whether resveratrol inhibits pulmonary arterial smooth muscle cells(PASMCs)proliferation by affecting silent mating type information regulation 2 homolog-1(SIRT1)/forkhead box protein O3a(FOXO3a)/p27 signaling pathway,and to provide a certain theoretical and experimental basis for the development of new drugs for pulmonary hypertension(PH).Methods Primary PASMCs were randomly divided;cells in the control group were not treated,cells in the 5-HT group were stimulated with 1μmol/L 5-HT for 24 h,and cells in the resveratrol groups were pre-treated with different concentrations(10,30,100,and 300μM)of resveratrol for 1 h before incubation with 1μmol/L 5-HT for another 24 h;the cell proliferation was measured by BrdU incorporation assay.To further verify the regu⁃latory mechanisms,we randomly divided PASMCs into six groups;cells in the control group,5-HT group,and resveratrol group were treated as the above;cells in the resveratrol+SIRT1 inhibitor group and resveratrol+FOXO3a inhibitor group were pre-treated with resveratrol accompanied with 1μmol/L EX527 or AS1842856,and cells in the resveratrol+p27 siR⁃NA group were transfected with p27 siRNA for 24 h before resveratrol treatment,followed by incubation with 1μmol/L 5-HT for 24 h;the cell proliferation was assayed by BrdU incorporation.The expression of p27 protein was measured in the resveratrol+FOXO3a inhibitor group and the protein levels of FOXO3a and p27 were determined in other groups by West⁃ern blotting.Results The proliferation rates of PASMCs in the 100 mol/L and 300 mol/L resveratrol groups were lower than those in the 5-HT group(all P<0.05).Resveratrol(100 mol/L)with effective inhibition rate was used for the further study.PASMC proliferation rates were higher in the 5-HT group than in the control group,which were lower in the resvera⁃trol group than in the 5-HT group,and they were higher in the resveratrol+SIRT1 inhibitor group,resveratrol+FOXO3a inhibitor group and resveratrol+p27 siRNA group than in the resveratrol

关 键 词：肺动脉平滑肌细胞 细胞增殖 白藜芦醇 SIRT1蛋白 FOXO3a蛋白 P27蛋白 

分 类 号：R543.2[医药卫生—心血管疾病]