三七总皂苷通过激活PI3K/AKT/mTOR信号通路调控低氧高二氧化碳环境下大鼠PASMCs的自噬和增殖  

Panax notoginseng saponins regulate autophagy and proliferation of rat PASMCs under hypoxia and hypercapnia environment by activating PI3K/AKT/mTOR signaling pathways

作　　者：陈玲珑 武垣伶 张赛[2] 黄丹娜 王万铁[2] CHEN Ling-long;WU Yuan-ling;ZHANG Sai;HUANG Dan-na;WANG Wan-tie(Department of emergency medicine,wenzhou people's hospital,wenzhou 325000,China;Department of physiology and pathophysiology,wenzhou medical university,wenzhou 325035,China;Clinical lab,Shanxi Province Bethune Hospital,taiyuan 030000,China)

机构地区：[1]温州市人民医院急诊医学科,浙江温州325000 [2]温州医科大学生理学与病理生理学系,浙江温州325035 [3]山西白求恩医院检验科,山西太原030000

出　　处：《中国病理生理杂志》2021年第7期1246-1251,共6页Chinese Journal of Pathophysiology

基　　金：温州市基础性科研项目(No.Y20190390);浙江省介入肺脏病重点实验室建设项目(No.2019E10014)。

摘　　要：目的:探讨三七总皂苷(PNS)在低氧高二氧化碳(HH)环境下对大鼠肺动脉平滑肌细胞(PASMCs)自噬和增殖的作用及其机制。方法:将大鼠PASMCs随机分为5组:对照(CON)组、HH组、HH+PNS组、PI3K抑制剂LY294002组(HH+LY组)及HH+PNS+LY组。CON组置于常氧细胞培养箱(21%O_(2)、5%CO_(2))培养24 h,其余4组置于造模培养箱(5%O_(2)、6%CO_(2))并加入各组对应的药物培养24 h。CCK-8法检测细胞活力;RT-qPCR和Western blot法检测自噬相关指标AKT、mTOR、LC3和P62的mRNA和蛋白水平;Western blot检测增殖细胞核抗原(PCNA)的表达水平;通过透射电镜观察细胞内部自噬小体形成情况。结果:与CON组比,HH组PASMCs的活力减弱,AKT、mTOR和P62的mRNA及p-AKT、p-mTOR和P62的蛋白水平降低,而LC3的mRNA和蛋白水平升高,PCNA表达上调,自噬小体数量增多(P<0.05);与HH组比,HH+PNS组PASMCs的活力减弱,AKT、mTOR和P62的mRNA及p-AKT、p-mTOR和P62的蛋白水平升高,LC3的mRNA和蛋白水平降低,自噬小体数量减少,PCNA表达下调(P<0.05);与HH组相比,HH+LY组PASMCs的活力减弱,AKT、mTOR和P62的mRNA及p-AKT、p-mTOR和P62的蛋白水平降低,而LC3的mRNA和蛋白水平升高,自噬小体数量增多,PCNA表达上调(P<0.05);与HH+PNS组相比,HH+PNS+LY组AKT、mTOR和P62的mRNA及p-AKT、p-mTOR和P62的蛋白水平降低,LC3的mRNA和蛋白水平升高,自噬小体数量增加,PCNA表达上调(P<0.05)。结论:在HH环境下,PNS可能通过激活PI3K/AKT/mTOR信号通路抑制大鼠PASMCs的自噬和增殖。AIM:to investigate the effect of Panax notoginseng saponins(PNS)on autophagy and viability of rat pulmonary artery smooth muscle cells(PASMCs)under hypoxia and hypercapnia(HH)environment,and to explore the mechanism.METHODS:The rat PASMCs were randomly divided into 5 groups:control(CON)group,HH group,HH+PNS group,HH+LY294002(LY)group and HH+PNS+LY group.The cells in CON group were cultured in an nor-moxic cell incubator(21%O_(2),5%CO_(2))for 24 h,and the cells in other 4 groups were terated with corresponding drugs and cultured in a modeling incubator(5%O_(2),6%CO_(2))for 24 h.Cell viability was measured by CCK-8 assay.RT-qPCR was used to detect the mRNA levels of AKT,mTOR,LC3 and P62.The protein levels of p-AKT,p-mTOR,LC3B,P62 and proliferating cell nuclear antigen(PCNA)were detected by Western blot.Transmission electron microscopy was used to observed the changes in the internal ultrastructure of cells,such as autophagosomes.RESULTS:Compared with CON group,the viability of PASMCs in HH group was decreased,AKT,mTOR and P62 mRNA levels and p-AKT,p-mTOR and P62 protein levels were down-regulated,LC3 mRNA level and LC3B protein level were up-regulated,the number of autophagosomes was increased,and PCNA expression was up-regulated(P<0.05).Compared with HH group,the viabili-ty of PASMCs in HH+PNS group was decreased,AKT,mTOR and P62 mRNA levels and p-AKT,p-mTOR and P62 pro-tein levels were up-regulated,LC3 mRNA level and LC3B protein level were down-regulated(P<0.05),the number of au-tophagosomes was decreased,and PCNA expression was down-regulated(P<0.05).Compared with HH group,the viabili-ty of PASMCs in HH+LY group was decreased,AKT,mTOR and P62 mRNA levels and p-AKT,p-mTOR and P62 protein levels were down-regulated,LC3 mRNA level and LC3B protein level were up-regulated,the number of autophagosomes was increased,and PCNA expression was up-regulated(P<0.05).Compared with HH+PNS group,the viability of PASMCs in HH+PNS+LY group was decreased,AKT,mTOR and P62 mRNA levels and p-AKT,p-mTOR and P62 pro-tein levels were

关 键 词：低氧高二氧化碳性肺动脉高压 肺动脉平滑肌细胞 三七总皂苷 自噬 细胞增殖 PI3K/AKT/mTOR信号通路 

分 类 号：R363[医药卫生—病理学] R563[医药卫生—基础医学]