论文部分内容阅读
目的观察丹参素对PDGF-BB刺激下大鼠肝星状细胞活化增殖,Ⅰ、Ⅲ型胶原合成与分泌,ERK1/2和PI3K通道蛋白表达的影响,探讨丹参素抑制肝纤维化的分子机制。方法用不同浓度丹参素(0.250、0.125、0.062 mmol/L)、ERK通道特异性抑制剂UO126(20 nmol/L)和PI3K通道特异性抑制剂LY294002(20 nmol/L)分别作用于经血小板衍生生长因子PDGF-BB(10 ng/ml)处理的大鼠肝星状细胞(HSC)48 h,CCK8法检测HSC增殖活性,免疫化学染色法测定Ⅰ、Ⅲ型胶原合成,酶联免疫吸附法观察Ⅰ、Ⅲ型胶原分泌,Western blot测定ERK1/2、p-ERK1/2、AKT和p-AKT蛋白表达。结果 PDGF-BB 10 ng/ml处理后,HSC增殖明显增加(0.139±0.009),丹参素能抑制PDGF-BB刺激大鼠HSC增殖的作用,并随丹参素浓度增加(0.062、0.125、0.250 mmol/L)细胞增殖受到的抑制作用越强(0.102±0.008、0.083±0.007、0.066±0.014,P<0.05),Ⅰ、Ⅲ型胶原合成及分泌均显著减少(P<0.05),p-ERK1/2和p-AKT蛋白表达均显著降低(P<0.05)。结论丹参素可通过抑制PDGF-BB诱导的HSC增殖和活化,同时抑制与肝纤维化密切相关的MAPK、PI3K途径中ERK、AKT蛋白的磷酸化,负性调控肝纤维化过程,其机制可能与抑制ERK1/2和PI3K信号转导通路有关。
Objective To observe the effects of Danshensu on PDGF-BB-induced activation and proliferation of rat hepatic stellate cells, the synthesis and secretion of type Ⅰ and type Ⅲ collagen, the expression of ERK1 / 2 and PI3K channels, and explore the molecular mechanism of Danshensu on hepatic fibrosis. Methods Tanshinone (0.250,0.125 and 0.062 mmol / L), ERK channel specific inhibitor UO126 (20 nmol / L) and PI3K channel specific inhibitor LY294002 (20 nmol / L) The HSC proliferation of rat hepatic stellate cells (HSCs) treated with PDGF-BB (10 ng / ml) for 48 h was measured by CCK8 assay. The expressions of type I and type III collagen were measured by immunochemical staining and enzyme-linked immunosorbent assay The expressions of ERK1 / 2, p-ERK1 / 2, AKT and p-AKT were detected by Western blot. Results Danshensu could significantly inhibit the proliferation of HSC induced by PDGF-BB in the presence of 10 ng / ml PDGF-BB (0.139 ± 0.009). With increasing concentrations of Danshensu (0.062,0.125 and 0.250 mmol / (P <0.05). The expression of p-ERK1 / 2 was significantly decreased (P <0.05), and the inhibition of p-ERK1 / 2 And p-AKT protein expression were significantly lower (P <0.05). Conclusion Danshensu can inhibit the proliferation and activation of HSC induced by PDGF-BB and inhibit the phosphorylation of MAPK and ERK, AKT protein in PI3K pathway, which may be closely related to hepatic fibrosis, and may be related to the negative regulation of hepatic fibrosis Inhibition of ERK1 / 2 and PI3K signal transduction pathway.