褪黑素对肝星状细胞中PI3K/AKT信号通路的影响

Effect of melatonin on PI3K/AKT signaling pathway in hepatic stellate cells

目的探究褪黑素对转化生长因子-1(TGF-C1)诱导的肝星状细胞系(HSC-T6)中PI3K/AKT信号通路的影响。方法将HSC-T6细胞分成5组:对照组、模型组、褪黑素低浓度组(1 nml/L)、褪黑素中浓度组(1 imoUL)、褪黑素高浓度组(0.1 mmoUL)。培养箱培养24 h后,用无血清培养基培养,除对照组外,其余各组均加入TGF-C1,在三组不同浓度的褪黑素组加入对应浓度的褪黑素,药物作用48 h&MTT检测褪黑素对HSC-T6细胞增殖的影响;细胞免疫组化检测褪黑素对HSCT6细胞中P-OKT蛋白表达的影响;Western blot检测褪黑素对HSC-T6细胞中PI3K和P-AKT蛋白表达的影响;qPCR检测褪黑素对HSC-T6细胞中$平滑肌肌动蛋白(a-SMA)mRNA表达的影响。结果与对照组相比,TGF-C1能显著诱导HSC-T6细胞增殖(2<0.01),增强PI3K和PTKT蛋白的表达(2<0.01),增强$SMA mR-NA的(2<0.01);型相,褪黑素度著抑制HSCT6细胞增殖(2<0.01),褪黑素能明显抑制PI3K和P-OKT蛋白的表达较模型组(2<0.01或2<0.05),褪黑素能明显抑制aSMA mRNA的表达较模型组(2<0.01)。结论褪黑素能显著抑制HSC-T6细胞增殖,其作用机制可能与抑制PI3K/AKT信号通路有关.

Objective To investigate the efect of melatonin on PI3K/AKT signaling pathway in hepatie sellate cell line(HSC-T6)induced by transforming growth factor-B1(TGF-β1).Methods The HSC-T6 cells were divided into 5 groups:control group,model group,melatonin low concentration group(1 nmol/L),melatonin medium concen-tration group(1 umol/L),melatonin high concentration group(0.1 mmol/L).Afer being cultured for 24 h,they were replaced with FBS-free medium and treated with TGF-β1 excepted the control group,three groups of different concentrations of melatonin were added with corresponding concentrations of melatonin.After drugs incubation for 48 h,the effect of melatonin on the proliferation of HSC-T6 cells was detected by MTT assay.The effect of melato-nin on the protein expression of P-AKT was detected by immunohistochemistry.The efect of melatonin on the pro-teins expression of PI3K and P-AKT were detected by Wester blot.The effect of melatonin on the mRNA expres-sion of a SMA was detected by qPCR.Results Compard with the control roup,TGF-β1 could significantly in-duce the proliferation of HSC-T6 cells(P<0.01),elevate the proteins expression of PI3K and P-AKT,and ele-vate the mRNA expression of x-SMA(P<0.01).Compared with the model group,the high concentration of me-latonin could significantly inhibit the proliferation of HSC-T6 cells(P<0.01),melatonin could significantly inhibit the proteins expression of PI3K and P-AKT compare with the model group(P<0.01 or P<0.05),melatonin could significanly inhibit the mRNA expression of a-SMA compare with the model group(P<0.01).Conclusion Me-latonin can inhibit the proliferation of HSC-T6 cells,which may be related to the inhibition of PI3K/AKT signaling pathway.