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目的观察磷脂酰肌醇3激酶特异性抑制剂(LY294002),对血小板源生长因子(PDGF-BB)促大鼠肝星状细胞(HSC)增殖与Ⅰ型胶原表达的影响,并探讨其作用机制。方法用不同浓度的PDGF-BB和LY294002对大鼠肝星状细胞株(HSC-T6)进行处理,MTT法检测细胞的增殖,逆转录聚合酶链反应方法(RT-PCR)检测HSC-T6内I型胶原mRNA的表达。结果PDGF-BB在浓度1~10 ng/ml时呈剂量依赖性地促进HSC细胞增殖(P<0.05),当浓度大于10 ng/ml时处理组间无显著差异(P>0.05),其促增殖作用达到饱和。5μmol/L、10μmol/L、25μmol/L的LY294002均能显著且剂量依赖性地抑制PDGF-BB刺激的HSC-T6增殖(P<0.001)。PDGF-BB能促进HSCⅠ型胶原mRNA表达,24 h、48 h时相对于对照组的表达量为112.2%,182.5%。LY294002能显著抑制PDGF-BB促Ⅰ型胶原表达,24 h4、8 h时相对表达量为76.4%,52.6%。结论磷脂酰肌醇3激酶信号途径是调节血小板源生长因子促肝星状细胞增殖与分泌胶原的重要通道,抑制该通路可以抑制肝星状细胞增殖和胶原表达,具有抗肝纤维化的作用,对防治肝纤维化有一定的意义。
Objective To investigate the effects of phosphatidylinositol 3-kinase-specific inhibitor (LY294002) on proliferation and type Ⅰ collagen expression of rat hepatic stellate cells (HSC) stimulated by platelet derived growth factor (PDGF-BB) and its mechanism of action . Methods Rat hepatic stellate cell line (HSC-T6) was treated with different concentrations of PDGF-BB and LY294002. The proliferation of HSC-T6 cells was detected by MTT assay and the expression of HSC-T6 was detected by reverse transcription polymerase chain reaction Type I collagen mRNA expression. Results PDGF-BB promoted the proliferation of HSC cells in a dose-dependent manner (P <0.05) at the concentration of 1 ~ 10 ng / ml, while there was no significant difference (P> 0.05) when the concentration of PDGF-BB was higher than 10 ng / Proliferation reached saturation. LY294002 (5μmol / L, 10μmol / L, 25μmol / L) significantly and dose-dependently inhibited PDGF-BB-stimulated HSC-T6 proliferation (P <0.001). PDGF-BB could promote the expression of HSC typeⅠcollagen mRNA, which was 112.2% and 182.5% at 24 h and 48 h, respectively. LY294002 could significantly inhibit the expression of type I collagen induced by PDGF-BB. The relative expression of LY294002 at 24 h and 4 h was 76.4% and 52.6%, respectively. Conclusions Phosphatidylinositol 3 - kinase signaling pathway is an important channel that regulates proliferation and secretion of collagen by platelet - derived growth factor (IGF - 1) and inhibits hepatic stellate cell proliferation and collagen expression by inhibiting this pathway. It has anti - hepatic fibrosis effect, The prevention and treatment of liver fibrosis have some significance.