Objective To explore the effects of pirfenidone(PFD)on the proliferation,activation and glycolytic pathway of human hepatic stellate cell line LX2.Methods LX2 cells were treated with 10 µg/L transforming growth factor-β1(TGF-β1)for activation.LX2 cells were divided into normal group(0.1％DMSO),control group(10 μg/L TGF-β1+0.1％DMSO)and experimental groups(10 μg/L TGF-β1+2,4,6,8 mmol/L PFD,respectively).LX2 cell proliferation ability was evaluated by CCK-8 assay and colony formation assay.Glucose and lactic acid levels were detected in cell culture supernatants by the corresponding kits.Western blot was used to detect protein expression of α-smooth muscle actin(α-SMA),collagen type Ⅰ Alpha 1(COL1A1),glucose transporter 1(Glut1),hexokinase 2(HK2),phosphofructokinase platelet-type(PFKP),pyruvate kinase M2(PKM2),lactate dehydrogenase A(LDHA),and monocarboxylate transporter 1(MCT1).Quantitative Real-time PCR was applied to measure mRNA expression levels of α-SMA,COL1A1,Glut1,HK2,PFKP,PKM2,LDHA,and MCT1.Results When compared with normal group,LX2 cells in control group exhibited enhanced proliferation ability,increased protein and mRNA expression levels of α-SMA and COL1A1(P＜0.05),increased glucose consumption and accumulation of extracellular lactate,elevated protein expression levels of Glut1,HK2,PFKP,PKM2,LDHA,and MCT1 as well as elevated mRNA expression levels of Glut1,HK2,PKM2,and LDHA.When compared with control group,LX2 cells in experimental groups displayed inhibited proliferation,reduced protein expression levels of α-SMA and COL1A1,decreased glucose consumption and accumulation of extracellular lactate,reduced protein expression levels of Glut1,HK2,PFKP,PKM2,LDHA,and MCT1 as well as reduced mRNA expression levels of Glut1,HK2,PKM2,and LDHA(P＜0.05).Conclusion PFD can inhibit the LX2 cell proliferation and activation,reduce the secretion of extracellular matrix.Its anti-fibrotic ability may be related to the downregulation of glycolysis levels and disruption of cellular energy metabolism.
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