益肺活血颗粒对血小板衍生生长因子诱导大鼠肺动脉平滑肌细胞增殖及转化生长因子活性的影响

Effect of Yifei Huoxue Granule on the Proliferation and the TGF-β Activity of Rat Pulmonary Artery Smooth Muscle Cells Induced by Platelet-derived Growth Factor BB

目的观察益肺活血颗粒(Yifei Huoxue Granule,YFHXG)对血小板衍生生长因子(platelet-de-rived growth factor BB,PDGF-BB)刺激大鼠肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)增殖的抑制作用及对转化生长因子(transforming growth factor-β,TGF-β)活性的影响。方法采用组织块贴壁法原代培养大鼠PASMCs,取对数生长期PASMCs随机分为对照组,PDGF-BB组,YFHXG高、中、低剂量组(PDGF-BB+YFHXG,终浓度为7·5、1·5、0·3mg/mL)。噻唑蓝(MTT)比色法测定各组PASMCs的增值率,流式细胞仪测定细胞周期构成比和增殖指数(proliferation index,PI),链霉素抗生物素蛋白-过氧化酶连接法(SP法)测定胞内TGF-β蛋白的表达量。结果与对照组比较,PDGF-BB组PASMCs增殖明显活跃(P<0·01);与PDGF-BB组比较,随着YFHXG各剂量组药物浓度的升高,大鼠PASMCs的生长明显受抑制、G0/G1期比例显著升高,S+G2/M期比例显著降低,PI显著降低,且TGF-β蛋白的表达量降低并呈浓度依赖关系(P<0·05,P<0·01)。结论 PDGF-BB可以直接刺激PASMCs的增殖,YFHXG可以显著抑制PDGF-BB对大鼠PASMCs的促增殖作用,并可调节TGF-β蛋白的表达。

Objective To explore the effects of Yifei Huoxue Granule（YFHXG） on the proliferation and the transforming growth factor-β（TGF-β） activity of rat pulmonary artery smooth muscle cells（PASMCs） induced by platelet-derived growth factor BB（PDGF-BB）.Methods Using tissue block adhering wall method,the primary rat PASMCs were cultured.PASMCs at the log phase growth were randomly divided into the control group,the PDGF-BB group,the PDGF-BB＋high YFHXG group（at the final concentration of 7.5 mg/mL）,the PDGF-BB＋middle YFHXG group（at the final concentration of 1.5 mg/mL）,and the PDGF-BB＋low YFHXG group（at the final concentration of 0.3 mg/mL）,respectively.MTT assay were employed to determine the cell proliferation rate of each group.Flow cytometric analyses were used to detect the cell cycle constituent ratio and the proliferation index（PI）.In addition,TGF-β protein＇s expression was determined by immunocytochemical assay（SP method）.Results Compared with the control group,the proliferation of PASMCs in the PDGF-BB group was obviously active（P0.01）.But when compared with the PDGF-BB group,along with the increased concentration of YFHXG,the growth of PASMCs was obviously inhibited,the cell ratio of G0/G1 phase obviously increased,the cell ratio of S＋G2/M phase significantly decreased,and PI significantly decreased.Besides,the expression of TGF-β protein decreased in a dose-dependent manner（P0.05,P0.01）.Conclusions PDGF-BB could directly stimulate the proliferation of PASMCs.YFHXG had a significant inhibition on the proliferation of rat PASMCs induced by PDGF-BB and could regulate the expression of TGF-β.