胆固醇对体外分离培养的豚鼠胆囊Cajal样间质细胞的影响

Influence of cholesterol on interstitial Cajal-like cells of guinea pig gallbladder isolated and cultured in vitro

目的研究胆固醇对体外分离培养的豚鼠胆囊Cajal样间质细胞(ICLCs)增殖、凋亡的影响。方法体外分离培养豚鼠胆囊ICLCs,用含不同质量浓度[0 mg/L(空白组)及12.5、25、50、100 mg/L(实验组)]的胆固醇培养液作用于细胞24 h后,CCK-8法检测细胞的增殖率,流式细胞术检测细胞的凋亡率,Western blot检测细胞受体酪氨酸激酶(c-kit)蛋白的表达量,实时荧光定量PCR检测细胞c-kit mRNA的表达水平。结果用含25、50、100 mg/L质量浓度胆固醇的培养液培养ICLCs后,与空白组相比,实验组ICLCs的增殖率明显下降(P<0.05);ICLCs的凋亡率明显增加(P<0.05);c-kit蛋白表达及c-kit mRNA表达均明显下降(P<0.05)。结论高浓度的胆固醇能够抑制ICLCs的增殖并促进ICLCs的凋亡,ICLCs增殖率下降可能是SCF/c-kit信号通路受阻所致。

Objective To investigate the effects of cholesterol on proliferation and apoptosis of interstitial Cajal-like cells(ICLCs) isolated from guinea pig gallbladder in vitro.Methods ICLCs of guinea pig gallbladder were isolated and cultured in vitro.After the ICLCs were treated with cholesterol medium containing different concentrations(0,12.5, 25, 50,100 mg/L) for 24 h,cell proliferation was detected by cell-comting kit-8 method,cell apoptosis was detected by flow cytometry,c-kit protein expression was detected by Western blot,and c-kit mRNA expression was detected by real time fluorescence quantitative PCR.Results After the ICLCs were treated with cholesterol medium containing different concentrations(25,50,100 mg/L) for 24 h,the proliferation rate of ICLCs decreased significantly(P<0.05),the apoptotic rate of ICLCs increased significantly(P<0.05),the expression of c-kit protein and c-kit mRNA decreased significantly(P<0.05) in 25、50、100 mg/L cholesterol concentration group compared with 0 mg/L.Conclusions High concentration of cholesterol can inhibit the proliferation of ICLCs and promote the apoptosis of ICLCs.The decrease of the proliferation rate of ICLCs may be due to the blockage of SCF/c-kit signaling pathway.