胆固醇对兔胆管成纤维细胞增生的影响  

Effects of cholesterol on proliferation of cultured fibroblasts of bile duct in rabbit

作　　者：陈宝莹[1] 魏经国[1] 王耀程[1] 杨向新[1] 钱济先[2] 于军[3] 陈志南[4] 徐静[4] 吴道澄[5] 

机构地区：[1]中国人民解放军第四军医大学唐都医院放射科,陕西省西安市710038 [2]中国人民解放军第四军医大学唐都医院骨科,陕西省西安市710038 [3]中国人民解放军第四军医大学生理学教研室,陕西省西安市710032 [4]中国人民解放军第四军医大学细胞工程中心,陕西省西安市710032 [5]中国人民解放军第四军医大学化学教研室,陕西省西安市710032

出　　处：《世界华人消化杂志》2002年第5期566-570,共5页World Chinese Journal of Digestology

摘　　要：目的:研究胆固醇(CL)对兔胆管成纤维细胞增生的影响,并对其机制进行探讨。 方法:分离、纯化、培养兔胆管成纤维细胞,完全随机分为两组:对照组、中等浓度胆固醇组(0.6g/L)和高浓度胆固醇组(1.2g/L)。以MTT法测定各组细胞的增生情况、并以流式细胞仪(FCM)测定细胞周期。进而采用免疫组化法定量分析各组细胞内增生细胞核抗原(PCNA)的表达情况。 结果:中等浓度CL(0.4～0.8g/L)可以使培养的兔胆管成纤维细胞MTT A_(490nm)值显著升高,CL(0.4g/L)A_(490nm值为0.422±0.015,较对照组增加51.4％(P<0.05);CL(0.6g/L)A_(490nm)值达到最大为0.486±0.019,较对照组增加95.2％(P<0.01);0.8g/L CL组A_(490nm)值为0.472±0.017较对照组增加89.6％(P<0.01);高浓度CL(1.2g/L)明显抑制细胞生长,A_(490nm)值为0.163±0.018,较对照组下降34.5％(P<0.05)。流式细胞仪分析显示,与正常组相比,中等浓度CL(0.6g/L)显著增加G_2/M期细胞比例,从9.8％上升为13.0％,S期细胞比例从11.7％上升为33.4％,而显著减少进入G_0/G_1期的细胞比例,从78.5％下降为53.6％。高浓度CL(1.2g/L)显著减少G_2/M期细胞比例,从9.8％降低为6.7％,S期细胞比例从11.7％下降为5.9％,而显著增加进入G_0/G_1期的细胞比例,从78.5％上升为87.4％。AIM: To investigate how cholesterol can affect the proliferation of bile duct fibroblasts cultured from pure-bred rabbits.METHODS: The cultured bile duct fibroblasts were divided randomly into three groups: control, moderate and high concentration. The growth of bile duct fibroblasts was measured by the means of MTT and flow cytometer (FCM) respectively after treated by water-soluble cholesterol. The effect of cholesterol on the intracellular level of PCNA was measured by immunohistochemistry staining.RESULTS: There was significant increaseof the opticalabsorbance value at 490 nm (A490nm) in bile duct fibroblasts in moderate concentration group(0.4 - 0.8 g/L). The value (A490nm) of CL (0.6 g/L)increased up to the maximum (0.486 ± 0.019) and that of CL( 1.2 g/L)decreased to 0.163 ± 0.018, compared to control group (0.249± 0.025) (P<0.01). Flow cytometric analysis indicated that moderate concentration group (0. 6 g/L) increased in the number of bile duct fibroblasts in S-phase (from 11.7 % to 33.4 %). There was a high expression of PCNA in bile duct fibroblasts of moderate concentration group(0.6 g/L) and the staining intensity of moderate concentration group (126.3 ± 10.2 % AU) was significantly higher than that of control group (88.3 ± 11. 6 % AU). High concentration group (1. 2 g/L) decreased the expression of PCNA and the staining intensity was 96.3 ± 11.4 % AU ( P < 0,05, vs middle concentration group).CONCLUSION: Moderate concentration group of CL can promote the proliferation of bile duct fibroblasts, which is associated with the over-expression of PCNA.

关 键 词：胆固醇 胆管成纤维细胞增生 影响 胆系疾病 

分 类 号：R575.6[医药卫生—消化系统]