降低脂筏内integrin β1减轻HPS大鼠血清诱导PMVECs增殖和迁移  

Decreased integrinβ1 in lipid raft inhibits proliferation and migration of HPS rat serum-induced PMVECs

作　　者：高静 周家奇 GAO Jing;ZHOU Jia-qi(Department of Anesthesiology, the Children Hospital, Zhejiang University School of Medicine, Hangzhou 310000;Department of ICU, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310000, China)

机构地区：[1]浙江大学医学院附属儿童医院麻醉科,浙江杭州310000 [2]浙江大学医学院第一附属医院重症医学科,浙江杭州310000

出　　处：《基础医学与临床》2021年第10期1417-1422,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81901937)。

摘　　要：目的探讨脂筏内整合素β1(integrinβ1)在肝肺综合征(HPS)大鼠血清诱导肺微血管内皮细胞(PMVECs)增殖和迁移中的作用机制。方法将PMVECs分为4组:对照组:健康大鼠血清刺激细胞;HPS组:HPS大鼠血清刺激细胞;MβCD组:事先用0.01 mol/L MβCD预处理细胞1 h,再用HPS大鼠血清刺激细胞;si-integrinβ1组:利用siRNA技术构建integrinβ1低表达细胞,再用HPS大鼠血清刺激细胞。采用Western blot分别检测integrinβ1、FAK和pY397FAK在PMVECs脂筏内外的表达情况;采用CCK-8法检测细胞增殖改变;细胞划痕实验检测细胞迁移改变;采用ELISA检测血管内皮生长因子(VEGF)的分泌情况。结果与对照组比较,HPS组脂筏内integrinβ1表达明显增加(P<0.05),黏着斑激酶(FAK)活化水平显著增加,细胞增殖、迁移和VEGF分泌显著增加(P<0.05);与HPS组比较,HPS+MβCD组在给予MβCD扰乱脂筏完整结构后,脂筏内integrinβ1表达受到明显抑制(P<0.05);与HPS组比较,在给予MβCD或小干扰RNA抑制integrinβ1表达后,FAK活化水平受到显著抑制,细胞增殖、迁移和VEGF分泌显著降低(P<0.05)。结论HPS大鼠血清通过促进PMVECs脂筏内integrinβ1表达上调从而激活integrinβ1/FAK通路进而促进细胞增殖、迁移和VEGF分泌,介导了HPS肺血管重塑效应。Objective To investigate the effects of integrinβ1 within lipid raft on PMVECs.Methods PMVECs were divided into four groups:control group;HPS group:HPS;group and MβCD group:the cells were pretreated with 0.01 mol/L MβCD for 1 hour,and then cultured with HPS rat serum;si-integrinβ1 group:construction of integrinβ1 low expression cells using siRNA technology,and then cultured with HPS rat serum.The expression of integrinβ1,FAK and pY397FAK in lipid rafts of PMVECs in each group was detected by Western blot;cell pro-liferation,migration were observed by CCK-8 and wound healing assay;the secretion of angiogenic factors(VEGF)was detected by ELISA.Results Compared with the control group,the expression of integrinβ1 and the level of activated FAK in the lipid rafts of HPS group were higher than that of the control group(P<0.05),PMVECs proliferation,migration and the level of secreted VEGF were significantly increased(P<0.05).Compared with the HPS group,the expression of integrinβ1 and the level of activated FAK in the lipid rafts were significantly reduced in the MβCD group or in siRNA integrinβ1 group(P<0.05),PMVECs proliferation,migration and secretion of VEGF were also significantly reduced(P<0.05).Conclusions The up-regulation of integrinβ1 within lipid raft activates integrinβ1/FAK pathway thereby promotes cell proliferation,migration and secretion of VEGF,which may mediate pulmonary vascular remodeling of HPS.

关 键 词：肝肺综合征(HPS) 整合素Β1 肺微血管内皮细胞(PMVECs) 血管内皮生长因子(VEGF) 

分 类 号：R73.3[医药卫生—肿瘤]