α2-巨球蛋白对X线照射后人皮肤成纤维细胞的抗氧化及抗纤维化的作用  

Effect of Alpha2-macroglobulin on Anti-Oxidation and Anti-Fibrosis in Human Skin Fibroblasts after X-ray Irradiation

作　　者：孔祥波[1] 刘仰 陈雪英[2] 李浣洋 李洁 葛雅平[2] 房思炼[2] 

机构地区：[1]中山大学附属孙逸仙纪念医院口腔科,广东广州510120 [2]中山大学附属第六医院口腔颌面外科,广东广州510655

出　　处：《中山大学学报（医学版）》2018年第1期48-53,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金(2015A030313064);广东省科技计划项目(2014A020212127)

摘　　要：【目的】研究α2-巨球蛋白(α2M)对X线照射后人皮肤成纤维细胞(HSF)超氧阴离子(.O_2^-)含量、超氧化物歧化酶(SOD)活性和HSF细胞向肌成纤维细胞转化过程中的影响。【方法】分别使用0、5、10、15、20 Gy X线照射HSF细胞,在照射后第1天检测细胞培养基上清液中。O_2^-含量、SOD活性的变化,在照射后第5天采用Western blot检测α-平滑肌肌动蛋白(α-SMA)的蛋白表达变化,以选定合适的照射剂量。用以上合适剂量照射HSF细胞,分别于照射前1 h、照射后1 h实验组细胞培养基中加入终浓度为0.25 mg/m L,0.5 mg/mL的α2M,检测并比较不同实验组细胞培养基上清液中.O_2^-含量、SOD活性和α-SMA蛋白表达的变化情况。【结果】5~20 Gy X线照射HSF细胞后,.O_2^-含量逐渐增多,SOD活性逐渐降低,α-SMA蛋白表达量逐渐升高,差异具有统计学意义(P<0.05)。10 Gy X线照射后1 h加入α2M,可明显减少照射后HSF细胞。O_2^-含量,提高SOD活性,下调放射后HSF细胞α-SMA的蛋白表达,差异具有统计学意义(P<0.05),照射前1 h给药则无明显变化。【结论】X线照射后的HSF细胞明显增加.O_2^-释放量,同时SOD活性降低,并有向肌成纤维细胞转化的趋势;而X线照射后加入α2M可明显减少受照射HSF细胞的.O_2^-释放量,提高其SOD活性,并抑制向肌成纤维细胞转化的趋势,说明α2M可通过抗氧化和抗纤维化起到放射保护作用。【Objective】To explore the effect of alpha2-macroglobulin(α2M)on superoxide anion(.O_2^-)content,superoxide dismutase(SOD)activity and the process of cell-to-myofibroblast transformation in human skin fibroblasts(HSF)after X-ray irradiation.【Methods】HSF cells were irradiated with 0,5,10,15 and 20 Gy X-ray. The change of.O_2^- content and SOD activity in the supernatant of cell culture medium were measured on the first day after irradiation.The protein expression of alpha-smooth muscle actin(α-SMA)was detected by Western blot on the fifth day after irradiation. The most sensitive radiation dose is selected. HSF cells were irradiated with the above sensitive dose. Respectively,1 h before irradiation,1 h after irradiation,the experimental group cell culture medium was added to a final concentration of 0.25 mg/mL,0.5 mg/mL of α2M. The change of.O_2^-content,SOD activity and the protein expression of α-SMA were detected.【Results】HSF cells were irradiated with 5~20 Gy doses of X-ray..O_2^-content increased,SOD activity decreased and α-SMA protein expression increased gradually(P<0.05). The addition of α2M at 1 h after 10 Gy X-ray irradiation reduced the.O_2^-content,increased the SOD activity and downregulated the protein expression of α-SMA in HSF cells(P<0.05). There was no significant change in the administration at 1 h before irradiation. 【Conclusion】HSF cells increased.O_2^- content significantly,while SOD activity decreased,and the tendency to transform myofibroblasts after Xray irradiation. α2M can reduce the.O_2^-content,increase the SOD activity in HSF cells and inhibit the transformation of fibroblasts into myofibroblasts after irradiation. Indicating that α2M can play a role in radiation protection by anti-oxidation and anti-fibrosis.

关 键 词：Α2-巨球蛋白 放射损伤 人皮肤成纤维细胞 抗氧化 抗纤维化 

分 类 号：R782.3[医药卫生—口腔医学]