奥曲肽调控JAK/STAT3/SOCS3信号通路对肝星状细胞相关肝纤维化的影响

Protective effect of octreotide on hepatic stellate cell-related hepatic fibrosis by regulating the JAK/STAT3/SOCS3 signaling pathway

目的:研究奥曲肽(octreotide,OCT)对肝纤维化的保护作用及相关信号转导通路机制。方法:体外培养人肝星状细胞株LX-2,予以不同浓度的奥曲肽(10-3、10-4、10-5、10-6mmol/L)干预不同时间(24 h、48 h、72 h),通过RT-PCR法、Western-blot法检测STAT3、磷酸化STAT3(p-STAT3)、SOCS3在mRNA和蛋白水平的表达。结果:与24 h对照组比较,各OCT组STAT3(F=198.37,P=0.000)、SOCS3(F=173.45,P=0.000)在mRNA水平表达减少,差异具有统计学意义;与48 h对照组比较,各OCT组STAT3(F=105.79,P=0.000)、SOCS3(F=167.38,P <0.05)在mRNA水平明显减少;与72 h对照组比较,各OCT组STAT3(F=87.44,P <0.05)、SOCS3(F=210.66,P=0.000)在mRNA水平显著降低。OCT干预24 h后,各OCT组p-STAT3(F=112.28,P <0.05)、SOCS3(F=146.82,P=0.000)在蛋白水平表达降低;与48 h对照组比较,各OCT组p-STAT3(F=87.34,P <0.05)、SOCS3(F=66.92,P=0.000)在蛋白水平表达减少,差异具有统计学意义;与72 h对照组比较,各OCT组p-STAT3(F=98.05,P <0.05)、SOCS3(F=227.68,P=0.000)在蛋白水平显著下降。相同药物浓度,干预时间越长,SOCS3的表达量越少;相同干预时间,STAT3、p-STAT3的表达量与OCT浓度平行。结论:JAK/STAT3途径及其负调控因子SOCS3是参与OCT调节LX-2细胞增殖及凋亡的机制之一,奥曲肽能够在mRNA及蛋白水平降低STAT3、p-STAT3、SOCS3表达,从而抑制肝纤维化。

Objective:To investigate the protective effect of octreotide(OCT)on hepatic fibrosis and potential mechanism of related signaling pathway.Methods:Human hepatic stellate LX-2 cells were cultured in vitro and were then treated with OCT at different concentrations(10^-3,10^-4,10^-5,and 10^-6 mmol/L)for different durations(24,48,and 72 hours).RT-PCR and Western blot were used to measure the mRNA and protein expression of STAT3,phosphorylated STAT(p-STAT),and SOCS3.Results:Compared with the 24 hour control group,all OCT groups had significant reductions in the mRNA expression of STAT3(F=198.37,P=0.000)and SOCS3(F=173.45,P=0.000).Compared with the 48 hour control group,all OCT groups had significant reductions in the mRNA expression of STAT3(F=105.79,P=0.000)and SOCS3(F=167.38,P<0.05).Compared with the 72 hour control group,all OCT groups had significant reductions in the mRNA expression of STAT3(F=87.44,P<0.05)and SOCS3(F=210.66,P=0.000).After 24 hours of OCT intervention,all OCT groups had significant reductions in the protein expression of p-STAT3(F=112.28,P<0.05)and SOCS3(F=146.82,P=0.000).Compared with the 48 hours control group,all OCT groups had significant reductions in the protein expression of p-STAT3(F=87.34,P<0.05)and SOCS3(F=66.92,P=0.000).Compared with the 72 hour control group,all OCT groups had significant reductions in the protein expression of p-STAT3(F=98.05,P<0.05)and SOCS3(F=227.68,P=0.000).With drug concentration remaining the same,the expression of SOCS3 decreased over the time of intervention.With the intervention time remaining the same,the expression of STAT3 and p-STAT3 increased with the increase in OCT concentration.Conclusion:The JAK/STAT3 pathway and its negative regulatory factor SOCS3 may be involved in the mechanism of OCT in regulating the proliferation and apoptosis of LX-2 cells.OCT can reduce the mRNA and protein expression of STAT3,p-STAT3,and SOCS3 and thus inhibit hepatic fibrosis.