姜黄素对肝星状细胞增殖和凋亡的影响及其机制研究

Effect and mechanism of curcumin on proliferation and apoptosis in hepatic stellate cells

目的探讨姜黄素是否通过调控白细胞介素-6(IL-6)/信号转导和转录活化因子3(IL-6/STAT3)信号通路而调控肝纤维化鼠肝星状细胞((HSC))增殖、凋亡。方法(1)5个不同浓度(0,10,20,40,80μmol·L^-1)的姜黄素处理HSC,分别作为空白组和4个浓度姜黄素组。(2)将细胞分为4组:空白组(未处理细胞)、JSI-124组(10μmol·L^-1抑制剂JSI-124)、中浓度姜黄素组(姜黄素40μmol·L^-1),联合组(姜黄素40μmol·L^-1和IL-6100 ng·mL^-1共同处理)。以CCK-8法检测HSC细胞增殖水平,流式细胞术检测HSC细胞凋亡情况。分别加入IL-6/STAT3信号通路激活剂与抑制剂,观察其对HSC细胞增殖和凋亡的影响。以蛋白质印迹法检测IL-6和磷酸化STAT3(p-STAT3)蛋白表达量。结果(1)于72 h,空白组和很低、低、中、高4个浓度姜黄素组的细胞凋亡率分别为(4.56±0.98)%,(8.33±1.65)%,(16.61±2.07)%,(22.44±1.97)%和(24.36±2.68)%;这5组的IL-6蛋白相对表达量分别为0.79±0.09,0.62±0.07,0.51±0.05,0.34±0.03和0.30±0.02;这5组的p-STAT3蛋白相对表达量分别为0.81±0.08,0.67±0.05,0.50±0.04,0.32±0.03和0.27±0.02;4个浓度姜黄素组与空白组相比,细胞凋亡率明显升高,而IL-6和p-STAT3蛋白相对表达量明显降低,差异均有统计学意义(均P<0.05)。(2)空白组、JSI-124组、中浓度姜黄素组与联合组的细胞增殖抑制率分别为(0.19±0.05)%,(35.26±5.69)%,(43.26±6.19)%和(13.22±2.18)%;这4组的细胞凋亡率分别为(4.91±0.64)%,(21.69±2.25),(23.61±3.94)%和(6.59±1.22)%;上述指标:JSI-124组与空白组比较,差异均有统计学意义(均P<0.05);联合组与中浓度姜黄素组比较,差异均有统计学意义(均P<0.05)。结论姜黄素可通过抑制IL-6/STAT3信号通路而抑制肝星状细胞增殖,促进细胞凋亡。

Objective To investigate whether curcumin regulates proli-feration and apoptosis of hepatic stellate cells(HSC)in hepatic fibrosis rats by regulating interleukin-6(IL-6)/signaling and transcriptional activator 3(IL-6/STAT3)signaling pathway.Methods(1)HSC were treated with five concentrations of curcumin(0,10,20,40,80μmol·L^-1),as blank group and four concentrations of curcumin groups.(2)The cells were divided into four groups:blank group(untreated cells),JSI-124 group(10μmol·L^-1 inhibitor JSI-124),curcumin-M group(40μmol·L^-1 curcumin),combined group(40μmol·L^-1 curcumin+100 ng·mL^-1 IL-6).The proliferation of HSC cells was detected by CCK-8 method.Flow cytometry was used to detect the effect of curcumin on proliferation in hibition and apoptosis of HSC cells.IL-6/STAT3 signaling pathway activators and inhibitors were added,to observe the effects of HS-6/STAT3 signaling pathway on proliferation and apoptosis of HSC cells.Western blot was used to detect the expression of IL-6 and phosphorylated STAT3(p-STAT3)proteins.Results(1)At 72 h,the apoptosis rates in blank group,and curcumin-VL,-L,-M,-H groups were(4.56±0.98)%,(8.33±1.65)%,(16.61±2.07)%,(22.44±1.97)%,(24.36±2.68)%,respectively;the relative expression of IL-6 protein in the five groups were 0.79±0.09,0.62±0.07,0.51±0.05,0.34±0.03,0.30±0.02,respectively;the relative expression levels of p-STAT3 protein in the five groups were 0.81±0.08,0.67±0.05,0.50±0.04,0.32±0.03,0.27±0.02,respectively;the cell proliferation inhibition rate and apoptosis rate were significantly increased in four concentrations of curcumin compared with the blank group(all P<0.05);the relative expression of IL-6,p-STAT3 protein was significantly reduced,and the difference of the factors were statistically significant(all P<0.05).(2)The cell proliferation inhibition rate in blank group,JSI-124 group,curcumin-M group,and combined group were(0.19±0.05)%,(35.26±5.69)%,(43.26±6.19)% and(13.22±2.18)%;the apoptosis rates in the four groups were(4.91±0.64)%,(21.69±2.25),(23.61±3.94)% and(6.59±1.22)%;there was significant difference of the indicators between the JSI-124 group and the blank group(all P<0.05);comparison between combined group and curcumin-M group,the differences of the indicators were statistically significant(all P<0.05).Conclusion Curcumin can inhibit hepatic stellate cell proliferation and promote apoptosis by inhibiting IL-6/STAT3 signaling pathway.