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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 observe the effect of matrine on the collagen secretion and phenotype of cardiac fibroblasts (CFs) induced by platelet-derived growth factor (PDGF), and to explore the role of PI3K / Akt pathway in it Role. Methods: CFs were isolated, purified and cultured by trypsin digestion and differential adherent method. The CFs were randomly divided into 5 groups: blank control group (without drugs); PDGF group (10μg / L PDGF); Matrine low dose group (10μg / L PDGF + 0.25mmol / L matrine); matrine medium dose group (10μg / LPDGF + 0.5mmol / L matrine); matrine high dose group (10μg / L PDGF + 1.0 mmol / L matrine). The contents of collagen in supernatants were detected by ELISA. The expressions of α-SMA, PI3K and Akt mRNA were determined by RT-PCR. The expressions of p-Akt and α-SMA were detected by Western Blot. Results: Compared with the blank control group, the content of collagen in the supernatant of PDGF group increased significantly (P <0.05), and the expression of α-SMA, PI3K, Akt mRNA and α-SMA and p- ). Compared with PDGF group, the contents of collagen in supernatant of CFs of matrine small, medium and high dose groups were significantly decreased (P <0.05), and the expressions of α-SMA, PI3K, Akt mRNA and α-SMA and p- (P <0.05). Conclusion: Matrine may play an anti-myocardial fibrosis effect by decreasing PI3K / Akt pathway to reduce collagen secretion and inhibit phenotype conversion.