B族Ⅰ型清道夫受体过表达对血小板源性生长因子诱导的冠状动脉平滑肌细胞增殖和迁移的影响

Effect of scavenger receptor class B type Ⅰ overexpression on proliferation and migration of coronary artery smooth muscle cells induced by platelet-derived growth factor

目的探讨B族Ⅰ型清道夫受体(SR-BⅠ)在人冠状动脉平滑肌细胞(hCASMC)增殖和迁移过程中的作用及机制。方法常规培养hCASMC,根据处理方法将细胞分为空白对照组、5μg·L^(-1)血小板源性生长因子BB(PDGF-BB)组、10μg·L^(-1)PDGF-BB组、20μg·L^(-1)PDGF-BB组、20μg·L^(-1)PDGF-BB+腺病毒绿色荧光蛋白(AdGFP)组(PDGF-BB+Ad-GFP组)、20μg·L^(-1)PDGF-BB+Ad-GFP-SR-BⅠ组(PDGF-BB+Ad-GFP-SR-BⅠ组)。采用噻唑兰法检测各组细胞增殖情况,Western blot法检测各组细胞中SR-BⅠ表达,Transwell法检测各组细胞迁移情况。结果空白对照组及5、10、20μg·L^(-1)PDGF-BB组细胞中SR-BⅠ相对表达量分别为1.00±0.05、0.76±0.08、0.52±0.06、0.30±0.05,5、10、20μg·L^(-1)PDGF-BB组细胞SR-BⅠ相对表达量均显著低于空白对照组(P<0.05),10、20μg·L^(-1)PDGF-BB组细胞SR-BⅠ相对表达量均显著低于5μg·L^(-1)PDGF-BB组(P<0.05),20μg·L^(-1)PDGF-BB组细胞SR-BⅠ相对表达量显著低于10μg·L^(-1)PDGF-BB组(P<0.05)。培养24、48、72 h时,5、10、20μg·L^(-1)PDGF-BB组及PDGF-BB+Ad-GFP组、PDGF-BB+Ad-GFP-SR-BⅠ组细胞增殖能力均显著高于空白对照组(P<0.05),10、20μg·L^(-1)PDGF-BB组及PDGF-BB+Ad-GFP组、PDGF-BB+Ad-GFP-SR-BⅠ组细胞增殖能力显著高于5μg·L^(-1)PDGF-BB组(P<0.05),20μg·L^(-1)PDGF-BB组、PDGF-BB+Ad-GFP组、PDGF-BB+Ad-GFP-SR-BⅠ组细胞增殖能力显著高于10μg·L^(-1)PDGF-BB组(P<0.05); PDGF-BB+Ad-GFP-SR-BⅠ组细胞增殖能力低于20μg·L^(-1)PDGF-BB组和PDGF-BB+Ad-GFP组(P<0.05); 20μg·L^(-1)PDGF-BB组与PDGF-BB+Ad-GFP组细胞增殖能力比较差异无统计学意义(P>0.05)。空白对照组、20μg·L^(-1)PDGF-BB组、PDGF-BB+Ad-GFP组及PDGFBB+Ad-GFP-SR-BⅠ组细胞迁移数分别为24.2±3.6、76.6±4.2、75.2±4.8和60.6±3.6,各组细胞迁移数比较差异有统计学意义(F=40.695,P<0.01); 20μg·L^(-1)PDGF-BB组、PDGF-BB+Ad-GFP组及PDGF-BB+Ad-GFP-SR-BⅠ组细胞迁移数显著高于空白对照组,PDGF-BB+Ad-GFP-SR-BⅠ组细胞迁移数显著低于20μg·L^(-1)PDGF-BB组、PDGF-BB+Ad-GFP组(P<0.05); 20μg·L^(-1)PDGF-BB组与PDGF-BB+Ad-GFP组细胞迁移数比较差异无统计学意义(P>0.05)。结论 PDGF-BB能够促进h CASMC增殖和迁移,并减少SR-BⅠ表达,过表达SR-BⅠ能够抑制PDGFBB的作用,抑制h CASMC增殖和迁移。

Objective To investigate the role and mechanism of scavenger receptor class B typeⅠ(SR-BⅠ) in the proliferation and migration of human coronary artery smooth muscle cells (hCASMC ). Methods hCASMC were routinely cultured and divided into blank control group,5 μg·L^-1 platelet-derived growth factor BB (PDGF-BB) group,10 μg·L^-1 PDGF-BB group,20 μg·L^-1 PDGF-BB group,20 μg·L^-1 PDGF-BB+adenovirus green fluorescent protein (Ad-GFP) group (PDGF-BB+Ad-GFP group) and20 μg·L^-1 PDGF-BB +Ad-GFP-SR-BⅠ group according to the treatment method.The cell proliferation was detected by thiazolam method,SR-BⅠ expression was detected by Western blot and the cell migration was detected by Transwell method. Results The relative expression of SR-BⅠ in the blank control group and5,10,20 μg·L^-1 PDGF-BB group was1.00±0.05,0.76±0.08,0.52±0.06 and 0.30±0.05,respectively.There was significant difference in the relative expression of SR-BⅠ in the each group.The relative expression of SR-BⅠ in the5,10,20 μg·L^-1 PDGF-BB group was significantly lower than that in the blank control group ( P <0.05).The relative expression of SR-BⅠ in the10,20 μg·L^-1 PDGF-BB group was significantly lower than that in the5 μg·L^-1 PDGF-BB group ( P <0.05),and the relative expression of SR-BⅠ in the20 μg·L^-1 PDGF-BB group was significantly lower than that in the10 μg·L^-1 PDGF-BB group ( P <0.05).After culturing for24,48,72 h,the proliferative ability in the5,10,20 μg·L^-1 PDGF-BB group,PDGF-BB+Ad-GFP group and PDGF-BB+Ad-GFP-SR-BⅠ group was significantly higher than that in the blank control group ( P <0.05);the proliferative ability in the10,20 μg·L^-1 PDGF-BB group was significantly higher than that in the5 μg·L^-1 PDGF-BB group ( P <0.05);the proliferative ability in the20 μg·L^-1 PDGF-BB group was significantly higher than that in the10 μg·L^-1 PDGF-BB group ( P <0.05);the proliferative ability in the PDGF-BB+Ad-GFP-SR-BⅠ group was lower than that in the20 μg·L^-1 PDGF-BB group and PDGF-BB+Ad-GFP group ( P <0.01);but there was no significant difference between the20 μg·L^-1 PDGF-BB group and PDGF-BB+Ad-GFP group ( P >0.05).The number of migrating cells in the blank control group,20 μg·L^-1 PDGF-BB group,PDGF-BB+Ad-GFP group and PDGF-BB +Ad-GFP-SR-BⅠ group was24.2±3.6,76.6±4.2,75.2±4.8 and 60.6±3.6;there was significant difference in the number of migrating cells among the groups( F =40.695, P <0.01);the number of migrating cells in the20 μg·L^-1 PDGF-BB group,PDGF-BB+Ad-GFP group and PDGF-BB+Ad-GFP-SR-BⅠ group was significantly higher than that in the blank control group;and the number of migrating cells in the PDGF-BB+Ad-GFP-SR-BⅠ group was lower than that in the20 μg·L -1 PDGF-BB group and PDGF-BB+Ad-GFP group ( P <0.05);but there was no significant difference between the20 μg·L^-1 PDGF-BB group and PDGF-BB+Ad-GFP group ( P >0.05). Conclusion PDGF-BB can promote the proliferation and migration of hCASMC,and reduce the expression of SR-BⅠ.SR-BⅠ overexpression can inhibit the effect of PDGF-BB and the proliferation and migration of PDGF-BB and hCASMC cells.