摘要
AIM:To investigate the effect of cholesterol (Ch) on the growth and functional protein expression of rabbit bile duct fibroblasts.METHODS:The cultured bile duct fibroblasts were divided randomly into two groups:the control group and the experiment group (fibroblasts were incubated respectively with 0.6g/L Ch for 12, 24, 36 and 48 h).The growth and DNA synthesis of bile duct fibroblasts were measured by the means of ^3H-TdR incorporation.The total protein content of fibroblast was measured by BSA protein assay reagent kit,then the expression of α-actin was analyzed semiquantitatively by Western blot.RESULTS:After treatment with 0.6g/L Ch for 12, 24,36 and 48h, the values of ^3H-TdR incorporation of bile duct fibroblasts were respectively 3.1±0.39, 3.8±0.37,4.6±0.48 and 5.2±0.56mBq/cell, and the values of the corresponding control groups were 3.0±0.33, 3.2±0.39,3.7±0.49 and 4.3±0.43mBq/cell. After comparing the values of experiment groups and their corresponding control groups,it was found that the 3H-TdR incorporation of bile duct fibroblasts after treatment with 0.6g/L Ch for 24, 36 and 48h were significantly increased (P<0.05, P<0.01, P<0.01),while the ^3H-TdR incorporation of 12-h group was not different statistically from its control group.Ch had no obvious effect on the total protein content of fibroblasts.After incubated with 0.6g/L Ch for 12, 24, 36 and 48h,the total protein content of each experiment group was not altered markedly compared with its corresponding control group.The values of experiment groups were 0.246±0.051,0.280±0.049, 0.263±0.044 and 0.275±0.056ng/cell, and those of corresponding control groups were 0.253±0.048,0.270±0.042,0.258±0.050 and 0.270±0.045 ng/cell.Western blot analysis revealed that the α-actin expression in fibroblasts affected by Ch for 12 and 24 h was not markedly changed compared with their corresponding control groups (P>0.05),the values of total gray scale of 12- and 24-h groups were 1 748±185 and 1 756±173,respectively. But after stimulation with Ch for 36 h, the total gray scale of fibroblasts (1923±204) was significantly higher than that of control group (1734±197).When the time of Ch treatment was lengthened to 48h, the α-actin expression was markedly elevated, the total gray scale was 2189±231 (P<0.01 vs control group).CONCLUSION:Moderately concentrated Ch can promote the proliferation of bile duct fibroblasts at early stage, With the prolongation of Ch treatment, the α-actin expression of fibroblasts was also increased,but the hypertrophy of fibroblasts was not observed,
AIM:To investigate the effect of cholesterol(Ch)on the growth and functional protein expression of rabbit bile duct fibroblasts. METHODS:The cultured bile duct fibroblasts were divided randomly into two groups:the control group and the experiment group(fibroblasts were incubated respectively with 0.6 g/L Ch for 12,24,36 and 48 h).The growth and DNA synthesis of bile duct fibroblasts were measured by the means of ~3H-TdR incorporation.The total protein content of fibroblast was measured by BSA protein assay reagent kit,then the expression of α-actin was analyzed semi- quantitatively by Western blot. RESULTS:After treatment with 0.6 g/L Ch for 12,24, 36 and 48 h,the values of ~3H-TdR incorporation of bile duct fibroblasts were respectively 3.1±0.39,3.8±0.37, 4.6±0.48 and 5.2±0.56 mBq/cell,and the values of the corresponding control groups were 3.0±0.33,3.2±0.39, 3.7±0.49 and 4.3±0.43 mBq/cell.After comparing the values of experiment groups and their corresponding control groups,it was found that the ~3H-TdR incorporation of bile duct fibroblasts after treatment with 0.6 g/L Ch for 24,36 and 48 h were significantly increased(P<0.05,P<0.01,P<0.01), while the ~3H-TdR incorporation of 12-h group was not different statistically from its control group.Ch had no obvious effect on the total protein content of fibroblasts.After incubated with 0.6 g/L Ch for 12,24,36 and 48 h,the total protein content of each experiment group was not altered markedly compared with its corresponding control group. The values of experiment groups were 0.246±0.051, 0.280±0.049,0.263±0.044 and 0.275±0.056 ng/cell,and those of corresponding control groups were 0.253±0.048, 0.270±0.042,0.258±0.050 and 0.270±0.045 ng/cell. Western blot analysis revealed that the α-actin expression in fibroblasts affected by Ch for 12 and 24 h was not markedly changed compared with their corresponding control groups (P>0.05),the values of total gray scale of 12-and 24-h groups were 1 748±185 and 1 756±173,respectively.But after stimulation with Ch for 36 h,the total gray scale of fibroblasts(1 923±204)was significantly higher than that of control group(1 734±197).When the time of Ch treatment was lengthened to 48 h,the α-actin expression was markedly elevated,the total gray scale was 2 189±231(P<O.01 vs control group). CONCLUSION:Moderately concentrated Ch can promote the proliferation of bile duct fibroblasts at early stage.With the prolongation of Ch treatment,the α-actin expression of fibroblasts was also increased,but the hypertrophy of fibroblasts was not observed.
