褪黑素对PDGF-BB诱导的HSC-T6细胞增殖的影响及机制

Effects of melatonin on the proliferation of HSC-T6 cells induced by PDGF-BB and its mechanism

目的探究褪黑素(Mel)对血小板衍生生长因子-BB(PDGF-BB)诱导的肝星状细胞-T6 (HSC-T6)增殖的影响及机制。方法将HSC-T6细胞分为对照组、模型组、实验组(Mel 1 nmol/L、1μmol/L、0. 1 mmol/L),MTT实验检测Mel对PDGF-BB激活的HSC-T6细胞增殖的影响;实时荧光定量PCR(q PCR)实验检测PDGF-BB、PDGFR-βmRNA的表达;免疫组化及Western blot检测α-平滑肌肌动蛋白(α-SMA)、细胞外调节蛋白激酶1/2(ERK1/2)、磷酸化细胞外调节蛋白激酶1/2(p-ERK1/2)蛋白的表达。结果与对照组比较,PDGF-BB能显著诱导HSC-T6细胞增殖,明显上调HSC-T6细胞PDGF-BB、PDGFR-βmRNA及α-SMA、ERK1/2、pERK1/2蛋白的表达;与模型组比较,Mel能够抑制PDGF-BB激活的HSC-T6细胞增殖,同时抑制PDGF-BB、PDGFR-βmRNA表达,并下调α-SMA、ERK1/2、p-ERK1/2蛋白的表达。结论 Mel可显著抑制PDGF-BB诱导的HSC-T6细胞的增殖与活化,其作用机制可能与抑制PDGF-BB、PDGFR-β表达,进而抑制ERK1/2信号通路相关。

Objective To investigate the molecular mechanism of anti-fibrosis of melatonin in HSC-T6 cells stimulated by PDGF-BB.Methods The HSC-T6 cells were divided into five groups:control group,model group,melatonin(1 nmol/L,1μmol/L,0.1 mmol/L)group.Cell proliferation was analyzed by MTT assay.The mRNA expression levels of PDGF-BB and PDGFR-βwere detected by qPCR.The protein expression levels ofα-SMA,ERK1/2 and p-ERK1/2 were detected by immunohistochemical and Western blot.Results The effects of different doses of melatonin on HSC-T6 cells demonstrated that melatonin could significantly reduce the proliferation of PDGF-BB-induced HSC-T6 cells and suppress the mRNA expression levels of PDGF-BB and PDGFR-β.In addition,melatonin could suppress the protein levels ofα-SMA,ERK1/2 and p-ERK1/2 in PDGF-BB-induced HSC-T6 cells.Conclusion These findings show that melatonin can inhibit the proliferation and activation of HSC-T6 cells.Its mechanism may be related to the inhibition of PDGF-BB,PDGFR-βexpression and the suppression of ERK1/2 signaling pathway.