摘要
目的:观察苦参碱对血小板源性生长因子(platelet-derived growth factor,PDGF)诱导心肌成纤维细胞(cardiac fibroblasts,CFs)胶原分泌及表型转化的影响,同时探讨PI3K/Akt通路在其中发挥的作用。方法:通过胰酶消化法和差速贴壁法分离、纯化及培养大鼠CFs,将CFs随机分为5组:空白对照组(培养时不加药物);PDGF组(10μg/L PDGF);苦参碱小剂量组(10μg/L PDGF+0.25mmol/L苦参碱);苦参碱中剂量组(10μg/LPDGF+0.5mmol/L苦参碱);苦参碱大剂量组(10μg/L PDGF+1.0mmol/L苦参碱)。ELISA法检测细胞上清液中胶原蛋白含量,RT-PCR法测定平滑肌激动蛋白-α(α-SMA)、PI3K和Akt mRNA的表达;Western Blot法测p-Akt及α-SMA蛋白的表达。结果:干预48h后,与空白对照组比较,PDGF组上清液胶原含量明显增加(P<0.05);α-SMA、PI3K、Akt mRNA及α-SMA、p-Akt蛋白表达上调(P<0.05);与PDGF组比较,苦参碱小、中和大剂量组CFs上清液胶原含量均明显减少(P<0.05),α-SMA、PI3K、Akt mRNA及α-SMA、p-Akt蛋白表达均下调(P<0.05)。结论:苦参碱可能通过下调PI3K/Akt通路减少胶原分泌、抑制表型转化,从而发挥抗心肌纤维化作用。
Objective：To investigate the effects of Matrine on collagen secretion and phenotype transformation in cardiac fibroblasts induced by platelet-derived growth factor（PDGF）,and to explore the role of PI3K/Akt pathway.Method：The cardiac fibroblasts were isolated and purified from rat hearts by enzymatic digestion and preplate technique.The cells were randomly divided into 5groups：blank control group（culture without drug）;PDGF group（10μg/L PDGF）;low matrine dose group（PDGF10μg/L＋0.25mmol/L matrine）;middle matrine dose group（10μg/LPDGF＋0.5mmol/L matrine）;high matrine dose group（10μg/LPDGF＋1.0mmol/L matrine）.After 48 hour Cell culture,ELISA was used to detect the collagen secretion in cell supernatant,the mRNA expressions of PI3 K,Akt and smooth muscle-Alpha excited protein（α-SMA）were measured by RT-PCR,and protein levels ofα-SMA,p-Akt were detected by Western blot analyses.Result：Compared with blank control group,the synthesis of collagen in PDGF group was significantly increased（P〈0.05）,the mRNA expressions of PI3 K,Akt,α-SMA and the levels ofα-SMA and p-Akt protein were obviously up-regulated（P〈0.05）.Compared with PDGF group,significant down-regulations of PI3 K,Akt,α-SMA mRNA andα-SMA,p-Akt protein were observed in low,middle,and high matrine dose groups respectively,and the synthesis of collagen was also lower than PDGF group.Conclusion：Matrine may reduce the collagen secretion and inhibit the phenotype transformation by down regulating the PI3K/Akt pathway,and thus play a role in antimyocardial fibrosis.
出处
《临床心血管病杂志》
CAS
CSCD
北大核心
2017年第6期592-595,共4页
Journal of Clinical Cardiology
