摘要
目的观察还原型谷胱甘肽(GSH)对高糖致大鼠腹膜间皮细胞8-羟基脱氧鸟苷(8-OHd G)及其修复酶8-羟基鸟嘌呤-DNA糖苷酶(r OGG1)表达的影响,探讨其对高糖所致腹膜间皮细胞DNA氧化损伤的保护作用。方法将60只健康雄性SD大鼠按随机数字法分成3组:对照组(n=20,A组):每天1次腹腔注射生理盐水25 ml;模型组(n=20,B组):每天1次腹腔注射4.25%葡萄糖腹透液25 ml+每周1次腹腔注射乳酸红霉素6.25万IU;治疗组(n=20,C组):每天1次腹腔注射4.25%葡萄糖腹透液25 ml+GSH溶液150 mg/(kg·d)+每周1次腹腔注射乳酸红霉素6.25万IU。8周后分别处死每组各20只大鼠,取腹膜组织分别行HE及Masson染色病理学检查、酶联免疫吸附法(ELISA)测定壁层腹膜间皮细胞8-OHd G水平、实时荧光定量PCR检测壁层腹膜间皮细胞r OGG1的表达。结果 1与A组比较,B组腹膜间皮细胞8-OHd G表达上调(0.8843±0.0068,5.0733±2.0068,P<0.05),C组8-OHd G表达低于B组(5.0733±2.0068,1.6867±0.4867,P<0.05);2与A组比较,B组腹膜间皮细胞r OGG1表达减少(0.7576±0.0701,0.2226±0.0565,P<0.05),C组r OGG1表达增高(0.7576±0.0701,1.4793±0.3895,P<0.05);3与A组比较,HE染色时B组和C组可见腹膜间皮细胞变为圆形、柱形,间皮细胞肥大脱落,间皮下结缔组织明显增厚,可见血管生成以及纤维素样物质沉积,还可见成纤维细胞及单核巨噬细胞浸润,在B组表现尤为明显;4与A组比较,Masson染色显示腹膜组织有胶原沉积,B组比C组改变更明显。结论高糖致大鼠腹膜间皮细胞DNA氧化性损伤标记物8-OHd G表达增高,GSH可能通过上调DNA修复酶OGG1m RNA表达,对大鼠腹膜间皮细胞DNA具有保护和修复作用。
[Objeetive] To study the effect of reduced glutathione on the expression of high glucose induced 8- hydroxydeoxyguanosine （8-OHdG） and repaired enzyme 8-hydroxy guanine DNA-glycosidase （rOGGI） in the rat peritoneal mcsothclial cells and therefore to determine the protective effect of reduced glutathione on DNA oxidative damage. [ Methods ] Sixty SD rats were randomly assigned into 3 groups： control, model and treatment groups. Con- trol rats（n =20, A） were injected daily with 25 ml saline, and model rats （n =20, B） daily with 25 ml of 4.25% peri- toneal dialysis solution and weekly 62 500 IU erythromycin, whereas treatment rats were injected （n =20, C） daily with 25 ml of 4.25% peritoneal dialysis solution and 25 ml 150 mg/kg of reduced glutathione and weekly with 62 500 IU of erythromycin. The rats were euthanized 8 weeks later, and the peritoneal tissues were harvested and per- formed for pathological examinations of HE and Masson staining. The expression levels of 8-OHdG and rOGG1 in parietal peritoneal mesothelial cells were measured by ELISA and RT-PCR, respectively. [ Results] The expression level of 8-OHdG in the peritoneal mesothelial cells in the model rats was significantly increased compared with the control rats, whereas that in the treatment rats was more reduced than in the model rats. The expression level of rOGG1 in the peritoneal mesothelial cells in the treatment rats was significantly increased compared to that in the control and model rats. [ Conclusion ] Reduced glutathione reduced the expression of oxidative damage marker 8-0- HdG and increased the expression of DNA repaired enzymes of rOGG1 in rat peritoneal mesothelial cells. Therefore, reduced glutathione may have protective effect on peritoneal mesothelial cells from DNA oxidative damage.
出处
《中国现代医学杂志》
CAS
北大核心
2015年第1期16-20,共5页
China Journal of Modern Medicine
