摘要
目的探讨转化生长因子β_1(TGF-β_1)促进 BALB/c 313成纤维细胞向肌成纤维细胞转化及对早期生长反应基因1(Egr-1)表达的影响。方法实验组用 TGF-β_1 10 ng/ml 处理 BALB/c3T3成纤维细胞,对照组不给予 TGF-β_1。采用流式细胞仪检测孵育24、48、72 h 的α平滑肌肌动蛋白(α-SMA)阳性细胞百分比,用免疫细胞化学方法及 Western 免疫印迹法检测 TGF-β_1处理15、30、60、90、120、180、240 min 的 Egr-1表达情况。结果用 TGF-β_1 10 ng/ml 培养24 h,实验组和对照组的α-SMA 阳性细胞百分率分别为(6.65±0.48)%和(5.53±0.62)%;继续培养至48和72 h,实验组的α-SMA 阳性细胞百分率[(28.38±3.60)%和(36.04±O.73)%]显著高于对照组[(9.49±0.21)%和(15.23±0.33)%]。免疫细胞化学染色检测 Egr-1的结果显示,培养90 min 时实验组细胞中出现棕黄色阳性染色颗粒,而对照组阳性染色细胞较少。Western 免疫印迹法检测 Egr-1的结果显示,实验组培养15 min 时的灰度水平(55±26)和对照组(38±18)无明显差别;随着给予 TGF-β_1时间延长,实验组灰度水平逐渐增加,60和90 min 时(115±14和129±22)显著高于对照组,之后实验组灰度水平逐渐降低,在240 min 时达最低点(39±7)。结论 TGF-β_1能诱导 BALB/c 3T3成纤维细胞向肌成纤维细胞分化,并且在此过程的早期 Egr-1表达增加。
Objective To investigate the effects of transforming growth factor （TGF）-β1 on fibroblasts differentiation into myofibroblasts, and the change of Egr-1 expression in the course of differentiation in BALB/e 3T3 fibroblasts. Methods BALB/e 3T3 fibroblasts of the experiment group were treated with 10 ng/ml TGF-β1 and those of the control group were treated without TGF-β1. The expression of a-smooth muscle aetin （SMA） was detected by flow eytometer after incubation for 24, 48, and 72 h. The expression of Egr-1 was detected by immunocytechemistry and Western blot method after 10 ng/ml TGF-β1 incubation for 15, 30, 60, 90, 120, 180, and 240 rain. Results The percentage of α-SMA positive cells was （6. 65 ± 0. 48）% when cells were treated with 10 ng/ml TGF-β1 for 24 h, and the control group was （5.53 ±0.62）% ,which showed no statistical significance. However the percentage was up to （28.38 ± 3.60） % after treated with TGF-β1 for 48 h, and the control group was （ 9.49 ± 0. 21 ） % ; At 72 h, the percentage was （ 36. 04 ± 0.73 ） % and the control group was （ 15. 23 ± 0. 33 ） % ; The difference was statistically significant. Immunocytochemistry showed that Egr-1 protein positive signal was brown in color. The color was light in the control group. The gray-level of the control group was 38 ± 18 measured by Western blot when cells were treated without TGF-β1. After treated with 10 ng/ml TGF-β1 for 15 min, the level was up to 55 ± 26, which had no statistical significance. The level was increased with longer incubation. It was 115 ± 14 at 60 rain and reached its peak level 129 ± 22 at 90 rain,which was significantly higher than the control group. The level was decreased after 90 min and reached neap 39 ± 7 at 240 rain. Conclusion TGF-β1 increased the expression of α-SMA in BALB/e 3T3 fibroblasts and affected the Egr-1 protein expression during the early course of fibroblasts differentiation into myofibroblasts.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
2007年第4期269-272,共4页
Chinese Journal of Tuberculosis and Respiratory Diseases
