肾缺血再灌注损伤对细胞凋亡的影响

EFFECT OF RENAL ISCHEMIA-REPERFUSION INJURY ON APOPTOSIS

目的探讨肾缺血再灌注(I-R)损伤对细胞凋亡的影响。方法健康雄性Wistar大鼠96只,随机分为假手术组(分离肾包膜)、I-R组(建立大鼠肾I-R损伤模型)与α-硫辛酸组(I-R前20 min腹腔注射α-硫辛酸100mg/kg)。分别在缺血再灌注0、3、6、24h时处死大鼠,取血和肾组织。全自动生化仪检测各组血清尿素氮(BUN)、肌酐(Scr)水平。硫代巴比妥酸法、比色法分别测定肾脏组织中丙二醛(MDA)、谷胱甘肽(GSH)含量。免疫组化法检测肾脏组织中肿瘤坏死因子相关受体6(TRAF6)、天冬氨酸特异性半胱氨酸蛋白酶3(Caspase-3)蛋白的表达。结果 I-R组3、6、24h时BUN、Scr、GSH、MDA、TRAF6、Caspase-3水平与假手术组、α-硫辛酸组相比,差异有显著性(F=13.875~2 933.93,q=4.30~116.79,P<0.05);α-硫辛酸组再灌注3、6、24h时BUN、Scr、GSH、MDA、TRAF6、Caspase-3水平与假手术组相比,差异亦有显著性(q=3.00~49.52,P<0.05)。结论肾I-R损伤时氧自由基增多,TRAF6表达增多,最终导致细胞凋亡。

Objective To investigate the influence of renal ischemia-reperfusion （I-R） injury on apoptosis. Methods Ninety-six healthy male Wistar rats were divided into three groups in random： sham-operated group （renal capsule dissociated）~ I- R group （I-R rat model created） ~ a-lipoic acid group （a dose of a-lipoic acid 100 mg/kg was administered by intrapertonel injection 20 min before I-R）. The rats were sacrificed at different time points of I-R （0 h, 3 h, 6 h and 24 h） for tissue and blood collection. Serum creatinine （Cr） and urea nitrogen （BUN） were determined by automatic biochemical meter, the GSH and MDA content in renal tissue were deteced by penthiobarbital method and colorimetry, respectively, the expressions of tumor-necrosis factor recep- tor-associated factor 6 （TRAF6） and Caspase-3 were deteced by immunohistochemistry technique. Results Compared with sham-operation group and a-lipoic acid group, the contents of MDA, TRAF6, Caspase-3 and GSH in I-R group at different time points （3 h, 6 h and 24 h） were statistically different （F： 13.875--2 933.93,q^4.30-- 116.79,P^0.05）. Compared with sham- operation group, the contents of MDA, TRAF6, Caspase-3 and GSH in the a-lipoic acid group at different points （3,6 and 24 h） were statistically different （q=3.00-49.52,P〈0.05）. Conclusion The increase of oxygen radical and TRAF6 in renal ischemi- a-reperfusion injury can finally result in cell apoptosis.