北五味子乙素对大鼠心脏成纤维细胞增殖的抑制作用及其PI3K/Akt/P27kip1信号通路机制  

Inhibitory effects of Schisandrin B on cardiac fibroblast proliferation and its PI3K/Akt/P27kip1 signaling pathway mechanism

作　　者：刘春旭 孙红霞 胡波[2] 姜恩平 LIU Chunxu;SUN Hongxia;HU Bo;JIANG Enping(Department of Pharmacology,School of Pharmacy,Beihua University,Jilin 132013,China;Department of Respiratory Medicine,Affiliated Hospital,Beihua University,Jilin 132002,China;Department of Pathology,Guangdong Medical University,Dongguan 523808,China)

机构地区：[1]北华大学药学院药理教研室,吉林吉林132013 [2]北华大学附属医院呼吸科,吉林吉林132011 [3]广东医科大学基础医学院病理教研室,广东东莞523808

出　　处：《吉林大学学报（医学版）》2022年第1期52-58,共7页Journal of Jilin University：Medicine Edition

基　　金：吉林省教育厅“十三五”科学技术项目(JJKH20200077KJ)。

摘　　要：目的:观察北五味子木脂素的单体成分之一五味子乙素(SchB)对血管紧张素Ⅱ(AngⅡ)诱导的新生SD大鼠心脏成纤维细胞(CFb)增殖、细胞周期及Ⅰ型胶原和Ⅲ型胶原蛋白表达水平的影响,探讨磷脂酰肌醇3-激酶/蛋白激酶B/细胞周期抑制蛋白P27(PI3K/Akt/P27kip1)信号通路的作用机制。方法:取出生1~3 d的SD大鼠乳鼠的心室组织,采用胰酶消化和差速贴壁法培养经鉴定为所需要的CFb。实验分为对照组、模型组、LY294002(PI3K抑制剂)组、SchB组和LY294002(提前30 min加入)+SchB组,采用0.1μmol·L^(-1)AngⅡ刺激CFb,作用24 h后用四氮唑盐(MTT)比色法检测各组CFb增殖抑制率,免疫细胞化学染色法检测各组CFb中Ⅰ型胶原和Ⅲ型胶原蛋白表达水平,流式细胞术检测各组不同细胞周期CFb百分率并计算增殖指数(PI),Western blotting法检测各组CFb中PI3K、Akt、磷酸化Akt(p-Akt)和P27kip1蛋白表达水平。结果:与对照组比较,模型组CFb增殖抑制率明显升高(P<0.01),CFb中Ⅰ型胶原和Ⅲ型胶原蛋白表达水平明显升高(P<0.01),G_(0)/G_(1)期CFb百分率明显降低(P<0.01),S期CFb百分率和PI明显升高(P<0.01),CFb中PI3K和p-Akt蛋白表达水平明显升高(P<0.01),P27kip1蛋白表达水平明显降低(P<0.01);与模型组比较,LY294002组、SchB组和LY294002+SchB组CFb增殖抑制率明显降低(P<0.05或P<0.01),CFb中Ⅰ型胶原和Ⅲ型胶原蛋白表达水平明显降低(P<0.05或P<0.01),G_(0)/G_(1)期CFb百分率明显升高(P<0.05或P<0.01),S期CFb百分率和PI明显降低(P<0.05或P<0.01),PI3K和p-Akt蛋白表达水平明显降低(P<0.05),P27kip1蛋白表达水平明显升高(P<0.05或P<0.01)。结论:SchB能抑制AngⅡ诱导的CFb增殖和胶原蛋白沉积,其机制可能与调控PI3K/Akt/P27kip1信号转导通路有关。Objective:To observe the effects of Schisandrin B(SchB),one of the monomers of lignans from Schisandra chinensis,on the proliferation,cell cycle,typeⅠcollagen and typeⅢcollagen expression levels in the neonatal SD rat cardiac fibroblasts(CFb)induced by angiotensinⅡ(AngⅡ),and to explore the mechanisms of phosphatidylinositol 3-kinase/protein kinase B/cell cycle inhibiting protein(P27kip1)(PI3K/Akt/P27kip1)signal pathways.Methods:The myocadium tissue was obtained from the neonatal 1-3 d of SD rats.The CFb were cultured by trypsin digestion and differential adhesion method and identified as the required CFb.The experiment was divided into control group,model group,LY294002(PI3K inhibitor)group,SchB group,LY294002+SchB group.The CFb were stimulated with 0.1μmol·L^(-1) AngⅡ,and the inhibitory rates of proliferation of CFb were detected by MTT colorimetry after treated for 24 h.The expression levels of typeⅠand typeⅢcollagen proteins were detected by immunohistochemistry method,the percentages of CFb in different cell cycle in various groups were detected by flow cytometry.The expression levels of PI3K,Akt,phosphorylated Akt(p-Akt)and P27kip1 proteins in the CFb in various groups were detected by Western blotting method.Results:Compared with control group,the inhibitory rate of proliferation of the CFb in model group was increased(P<0.01),the exspression levels of typeⅠcollagen and typeⅢcellagen proteins were increased(P<0.01),the percentage of CFb in G_(0)/G_(1) phase was decreased(P<0.01),the percentage of CFb in S phase and the proliferation index(PI)were increased(P<0.01),the expression levels of PI3K and Akt proteins in the CFb were increased(P<0.01),and the expression level of P27kip1 protein was decreased(P<0.01);compared with model group,the inhibitory rates of prolifenition of the CFb in LY294002,SchB,and LY294002+SchB groups were significantly decreased(P<0.05 or P<0.01),the expression levels of typeⅠand typeⅢcollagen proteins were significantly decreased(P<0.05 or P<0.01),the percenta

关 键 词：五味子乙素 磷脂酰肌醇3-激酶 蛋白激酶B 心脏成纤维细胞 信号通路 

分 类 号：R285.5[医药卫生—中药学] R542.2[医药卫生—中医学]