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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 effect of TGF-β 1 on the transformation of BALB / c 313 fibroblasts into myofibroblasts and the expression of Egr-1. Methods In experimental group, BALB / c3T3 fibroblasts were treated with TGF-β 1 10 ng / ml, while the control group was not given TGF-β 1. The percentage of α-smooth muscle actin (α-SMA) positive cells incubated for 24, 48, and 72 h were detected by flow cytometry. The expressions of α-SMA positive cells were detected by immunocytochemistry and Western immunoblotting. , 120,180,240 min Egr-1 expression. Results The percentage of α-SMA positive cells in the experimental group and the control group was (6.65 ± 0.48)% and (5.53 ± 0.62)%, respectively, after cultured with TGF-β 1 10 ng / ml for 24 h. (28.38 ± 3.60)% and (36.04 ± 0.73)%] in the control group [(9.49 ± 0.21)% and (15.23 ± 0.33)%], respectively. Immunocytochemical staining of Egr-1 results showed that 90 min after culture cells in the experimental group brown positive staining particles, while the control group less positive staining cells. The results of Western blotting analysis of Egr-1 showed that there was no significant difference in gray level (55 ± 26) and control group (38 ± 18) at 15 min in experimental group. With the prolongation of TGF-β 1, the experimental group The grayscale gradually increased at 60 and 90 min (115 ± 14 and 129 ± 22), which was significantly lower than that in the control group. The gray level of the experimental group decreased gradually after reaching 90 min (39 ± 7). Conclusion TGF-β 1 induces the differentiation of BALB / c 3T3 fibroblasts into myofibroblasts and increases the expression of Egr-1 in the early stage of the process.