Effects of grape seed proanthocyanidin B2 pretreatment on oxidative stress and renal tubular epithelial cell apoptosis after renal ischemia reperfusion in mice

Abstract Purpose To investigate the effects of grape seed proanthocyanidin B2 (GSPB2) preconditioning on oxidative stress and apoptosis of renal tubular epithelial cells in mice after renal ischemia-reperfusion (RIR). Methods Forty male ICR mice were randomly divided into 4 groups: Group A: mice were treated with right nephrectomy. Group B: right kidney was resected and the left renal vessel was clamped for 45 minutes. Group C: mice were intraperitoneally injected with GSPB2 before RIR established. Group D: mice were intraperitoneally injected with GSPB2 plus brusatol before RIR established. Creatinine and urea nitrogen of mice were determined. Pathological and morphological changes of kidney were checked. Expressions of Nrf-2, HO-1, cleaved-caspase3 were detected by Western-blot. Results Compared to Group B, morphology and pathological damages of renal tissue were less serious in Group C. Western-blot showed that expressions of Nrf-2 and HO-1 in Group C were obviously higher than those in Group B. The expression of cleaved-caspase3 in Group C was significantly lower than that in Group B. Conclusion GSPB2 preconditioning could attenuate renal oxidative stress injury and renal tubular epithelial cell apoptosis by up-regulating expressions of Nrf-2 and HO-1 and down-regulating the expression of cleaved-caspase-3, but the protective effect could be reversed by brusatol.

摘要
研究目的：探讨葡萄籽原花青素B2（grape seed proanthocyanidin B2，GSPB2）预处理对小鼠肾缺血再灌注（renal ischemia-reperfusion，RIR）后肾小管上皮细胞氧化应激与细胞凋亡的影响。
实验方法：将40只雄性ICR小鼠随机分为4组：A组仅行右侧肾切除术；B组切除右侧肾脏，并夹闭左侧肾蒂45分钟以构建肾缺血再灌注模型；C组在构建肾缺血再灌注模型前，对小鼠腹腔注射GSPB2；D组在构建肾缺血再灌注模型前，对小鼠腹腔注射GSPB2联合鸦胆子苦醇（brusatol）。检测各组小鼠血清肌酐与尿素氮水平，观察肾脏组织的病理形态学变化，并采用蛋白质印迹法（Western-blot）检测核因子E2相关因子2（Nrf-2）、血红素氧合酶1（HO-1）及活化半胱氨酸天冬氨酸蛋白酶3（cleaved-caspase3）的表达水平。
实验结果：与B组相比，C组小鼠肾脏组织的形态学与病理损伤程度更轻。蛋白质印迹法检测结果显示，C组小鼠肾脏组织中Nrf-2与HO-1的表达水平显著高于B组，cleaved-caspase3的表达水平则显著低于B组。
研究结论：GSPB2预处理可通过上调Nrf-2与HO-1的表达、下调cleaved-caspase3的表达，减轻小鼠肾缺血再灌注后的肾脏氧化应激损伤与肾小管上皮细胞凋亡，但该保护作用可被鸦胆子苦醇逆转。