血府逐瘀合剂对PDGF-BB诱导的血管平滑肌细胞增殖、迁移及PI3K/AKT/GSK-3β/β-catenin信号通路的影响  

Effects of Xuefu Zhuyu Mixture on PDGF-BB-induced Proliferation and Migration of Vascular Smooth Muscle Cells and PI3K/AKT/GSK-3β/β-Catenin Signal Pathway

作　　者：肖遥[1] 方丽 胡有志[1] Xiao Yao;Fang Li;Hu Youzhi(Department of Cardiology,Hubei Hospital of Traditional Chinese Medicine,Wuhan 430061,China;Second Staff Hospital,Wuhan Iron and Steel(Group)Company)

机构地区：[1]湖北省中医院心内科,武汉430061 [2]武汉钢铁(集团)公司第二职工医院

出　　处：《中国药师》2022年第8期1310-1316,共7页China Pharmacist

基　　金：湖北省卫生健康委员会中医药科研项目(编号:ZY2021M047)。

摘　　要：目的:探究血府逐瘀合剂对血小板衍生生长因子(PDGF)-BB诱导的血管平滑肌细胞增殖、迁移及磷脂酰肌醇3-激酶/丝氨酸/苏氨酸蛋白激酶/糖元合成酶激酶3β/β-连环蛋白(PI3K/AKT/GSK-3β/β-catenin)信号通路的影响。方法:利用重组鼠PDGF-BB体外刺激大鼠胸大动脉平滑肌细胞A7R5诱导增殖、迁移模型,分为6组:对照组、PDGF-BB组、低剂量血府逐瘀合剂+PDGF-BB组、高剂量血府逐瘀合剂+PDGF-BB组、PI3K激活剂+PDGF-BB组、PI3K激活剂+高剂量血府逐瘀合剂+PDGF-BB组,CCK-8法检测A7R5细胞增殖活力;流式细胞术检测A7R5细胞周期;细胞划痕实验检测A7R5细胞迁移情况;免疫印迹检测A7R5细胞增殖、迁移相关蛋白及PI3K/AKT/GSK-3β/β-catenin信号通路相关蛋白。结果:与对照组比较,PDGF-BB组A7R5细胞增殖率、迁移面积、S期比例,PCNA、cyclin D1、MMP9蛋白及p-PI3K/PI3K、p-AKT/AKT、p-GSK-3β/GSK-3β、胞核β-catenin/总β-catenin蛋白比值显著增加,G0/G1期比例、E-cadherin蛋白显著减少(P<0.05);与PDGF-BB组比较,低、高剂量血府逐瘀合剂+PDGF-BB组A7R5细胞增殖率、迁移面积、S期比例,PCNA、cyclin D1、MMP9蛋白及p-PI3K/PI3K、p-AKT/AKT、p-GSK-3β/GSK-3β、胞核β-catenin/总β-catenin蛋白比值显著减少,G0/G1期比例、E-cadherin蛋白显著增加,且高剂量血府逐瘀合剂+PDGF-BB组优于低剂量血府逐瘀合剂+PDGF-BB组(P<0.05);与高剂量血府逐瘀合剂+PDGF-BB组比较,PI3K激活剂+高剂量血府逐瘀合剂+PDGF-BB组A7R5细胞增殖率、迁移面积、S期比例,PCNA、cyclin D1、MMP9蛋白,p-PI3K/PI3K、p-AKT/AKT、p-GSK-3β/GSK-3β、胞核β-catenin/总β-catenin蛋白比值显著增加,G0/G1期比例、E-cadherin蛋白显著减少(P<0.05)。结论:血府逐瘀合剂可抑制PDGF-BB诱导的血管平滑肌细胞增殖、迁移,可能通过抑制PI3K/AKT/GSK-3β/β-catenin信号通路而实现。Objective: To explore the effects of Xuefu Zhuyu mixture on platelet-derived growth factor(PDGF)-BB-induced proliferation and migration of vascular smooth muscle cells and phosphatidylinositol 3-kinase/protein kinase B/glycogen synthase kinase-3β/β-catenin(PI3 K/AKT/GSK-3β/β-catenin) signaling pathway. Methods: Recombinant rat PDGF-BB was used to stimulate rat thoracic artery smooth muscle cells A7 R5 to induce proliferation and migration in vitro. The experiment included six groups: control group, PDGF-BB group, low-dose Xuefu Zhuyu mixture+PDGF-BB group, high-dose Xuefu Zhuyu mixture+PDGF-BB group, PI3 K activator+PDGF-BB group, and PI3 K activator+high-dose Xuefu Zhuyu mixture+PDGF-BB group, CCK-8 method was performed to measure the A7 R5 cells proliferation activity;flow cytometry was performed to measure the A7 R5 cells cycle;cell scratch test was performed to measure the migration of A7 R5 cells;Western blot was performed to detect the A7 R5 cells proliferation and migration related proteins and PI3 K/AKT/GSK-3β/β-catenin signaling pathway related proteins. Results: Compared with the control group, the PDGF-BB group had significantly higher A7 R5 cells proliferation rate, migration area, S phase ratio, PCNA, cyclin D1, MMP9 proteins, p-PI3 K/PI3 K, p-AKT/AKT, p-GSK-3β/GSK-3β, and nuclear β-catenin/total β-catenin protein ratio, and significantly lower G0/G1 phase ratio and E-cadherin protein(P<0.05);compared with the PDGF-BB group, the low-and high-dose Xuefu Zhuyu mixture+PDGF-BB groups had significantly lower A7 R5 cells proliferation rate, migration area, S phase ratio, PCNA, cyclin D1, MMP9 proteins, p-PI3 K/PI3 K, p-AKT/AKT, p-GSK-3β/GSK-3β, and nuclear β-catenin/total β-catenin protein ratio, and significantly higher G0/G1 phase ratio and E-cadherin protein, and the high-dose Xuefu Zhuyu mixture+PDGF-BB group was better than the low-dose Xuefu Zhuyu mixture+PDGF-BB group(P<0.05);compared with the high-dose Xuefu Zhuyu mixture+PDGF-BB group, the PI3 K activator+high-dose Xuefu Zhuyu mixtu

关 键 词：血府逐瘀合剂 大鼠胸大动脉平滑肌细胞A7R5 血小板衍生生长因子-BB 增殖 迁移 细胞周期 磷脂酰肌醇3-激酶/丝氨酸/苏氨酸蛋白激酶/糖元合成酶激酶3β/β-连环蛋白信号通路 

分 类 号：R285.5[医药卫生—中药学]