2-甲氧基雌二醇对瘢痕疙瘩成纤维细胞活性及Bcl-2和Bax表达水平的影响

摘要

Objective To explore the effect of 2-methoxyestradiol (2ME2) on the expression of Bcl-2, Bax and cell activity of keloid fibroblasts. Method 6 cases of chest keloid were sampled for primary keloid fibroblast cell culture. The 3rd generation of fibroblasts were extracted as control group (K group), DMSO control group (CTL group) and study group (2ME2 group), and the cells were cultured with routine culture media, media containing 0.7‰DMSO, or containing 6.975μmmol/L 2ME2+0.7‰DMSO, respectively, after 24 h, the cell activity was measured by CCK8 kit, and Western Blot was used to evaluate the Bcl-2 and Bax expression in each group. Result After 24 h of cell culturing, the fibroblasts in 2ME2 group showed apoptosis appearance with membrane invagination and decreased cytoplasm content. The cell activity in 2ME2 group was (50.40％±5.59％), significantly lower than that in K group (99.12％±7.39％) and in CTL group (98.67％± 3.78％), and the difference was of statistical significance (P<0.001). Western Blot showed that, in 2ME2 group, the Bcl-2 expression was (0.11±0.04), which was higher than the (0.06±0.02) in K group and (0.07± 0.01) in CTL group, with significant difference observed (P<0.05);while the Bax expression in 2ME2 group was (2.64 ± 3.12), and was significantly higher than that in K group at (0.40 ± 0.47) and in CTL group at (0.54 ± 0.50) (P<0.05); in 2ME2 group, the Bcl-2/Bax ratio was (0.08±0.06), which was significantly lower than that in K group at (0.61±0.62) and in CTL group at (0.53 ± 0.48), with significant difference observed (P<0.001). Conclusion 2ME2 could remarkably inhibit the cell activity of keloid fibroblasts, increase the expression of Bcl-2 and Bax protein and decrease the Bcl-2/Bax ratio, promote cell apoptosis.%目的 探讨2-甲氧基雌二醇(2ME2)对瘢痕疙瘩成纤维细胞活性及细胞凋亡蛋白Bcl-2、Bax表达水平的影响.方法 对6例胸部瘢痕疙瘩患者经手术切除的瘢痕疙瘩组织行成纤维细胞原代培养,取第3代细胞分为对照组(K组)、DMSO对照组(CTL组)及实验组(2ME2组),分别用常规培养基、含有0.7‰DMSO的培养基、含有6.975μmmol/L 2ME2+0.7‰DMSO的培养基进行培养,培养24 h后通过CCK8细胞活性试验和蛋白质印迹(Western Blot)法检测细胞活性及Bcl-2、Bax蛋白的表达水平.结果 细胞培养24 h后,2ME2组可见细胞凋亡形态,细胞膜内陷,细胞质成分减少.2ME2组的细胞活性为(50.40％±5.59％),明显低于K组的(99.12％±7.39％)和CTL组的(98.67％±3.78％),差异均有统计学意义(P﹤0.001).Western Blot结果显示,2ME2组的Bcl-2蛋白表达水平为(0.11±0.04),高于K组的(0.06±0.02)和CTL组的(0.07±0.01),差异均有统计学意义(P﹤0.05);2ME2组的Bax蛋白表达水平为(2.64±3.12),高于K组的(0.40±0.47)和CTL组的(0.54±0.50),差异均有统计学意义(P﹤0.05);2ME2组的Bcl-2/Bax比值为(0.08±0.06),明显低于K组的(0.61±0.62)和CTL组的(0.53±0.48),差异均有统计学意义(P﹤0.001).结论 2ME2能够抑制瘢痕疙瘩成纤维细胞的增殖活性,提高Bcl-2和Bax的表达水平,降低Bcl-2/Bax的比值,促进细胞凋亡.