摘要目的 观察急性百草枯(Paraquat,PQ)中毒大鼠肾组织Livin蛋白及Caspase-3的表达及乌司他丁(Ulilnastatin,UTI)的干预作用.方法 将54只SD大鼠随机分为对照组(A组)、PQ组(B组)和UTI组(C组),每组各18只.B,C组以生理盐水稀释PQ 80 mg/kg一次性灌胃;C组灌胃后以UTI 105 U/kg腹腔注射,每天1次;A组等量生理盐水一次性灌胃.于不同处理后24,48,72 h分别用免疫印迹法(Westem-bolt)和免疫组化法测定不同时间点肾组织中Livin蛋白及Caspase-3表达的情况,同时观察大鼠肾组织病理变化.结果 A组肾组织结构清晰;B组肾组织结构清晰度明显下降,染毒24 h可见充血、水肿及空泡变性等病理变化,且随时间延长而加重,重者可见核固缩,细胞结构消失,肾小球、髓质部亦可受累;C组病理变化较B组明显减轻.A组Caspase-3蛋白多不表达;B组染毒24h在皮质部肾小管上皮细胞的胞膜及胞浆中Caspase-3蛋白呈阳性表达;C组亦出现表达,但表达水平较PQ组降低,各组比较差异均有统计学意义(P＜0.01).与A组比较,各时间点B组和C组肾组织Livin蛋白的表达明显增强,尤以C组为甚,差异有统计学意义(P＜0.01).结论 PQ中毒肾损伤主要表现为上皮细胞变性、浊肿、部分坏死.Caspase-3参与了PQ中毒肾损伤的发病过程.UTI对PQ中毒大鼠肾组织有保护作用,可能是通过促进Livin表达、抑制Caspase-3表达起作用,但其调节途径尚需进一步探讨.

abstractsObjective To observe the levels of Livin and Caspase-3 in renal tissue of rats following acute paraquat (PQ) poisoning and the intervention effects of ulinastatin (UTI) . Methods Fifty-four Sprague-Dawley (SD) rats were randomly divided into three experimental groups: control group (group A),PQ poisoning group (group B) and UTI group (group C) (n = 18 in each group) . Rats in group B and group C were administered intragastrically with 80 mg/kg PQ, and rats in group C were treated with 100,000U/kg ulinastatin injected intra-psritoneally once a day; and rats in group A were administered intragastrically with the same volume of saline instead of PQ. At 24, 48, 72 hours after poisoning, the levels of Livin in renal tissue were detected by Westen blotting and the levels of Caspase-3 were detected by immunohistochemistry 24, 48, and 72 hours after poisoning, and the histopathological changes of renal tissue were observed at the same time. Results In the group A, the structure of renal tissue was distinct. In the group B, the distinctness of the structure of renal tissue declined significantly, and swelling, edema and vacuolar degeneration were observed 24 h after poisoning, and pathological changes became more and more obvious keeping pace with time elapsing, and sometimes karyopyknosis appeared and celluar structures disappeared with involvement of renal glomerulus and medulla. These pathological changes were significantly lessened in rats of group C. In the group A, there was little Caspase-3 in renal tissue of rats. Twenty-four hours after poisoning, the caspase-3 in renal tissue of rats of group B was found on the membrane and in the kytoplasm of renal tubular epithelial cells of cortical part. Compared with group B, the level of Caspase-3 in renal tissue of rats of group C decreased significantly to lower level (P ＜0. 01 ) . Compared with group A,the levels of Livin in renal tissue in rats of group B and group C increased significantly at all different intervals (P ＜0. 01 ), and as group B was compared with group C, the difference was statistically significant (P ＜0. 01 ) . Conclusions The main pathologyical changes of renal injury induced by PQ are epithelial swelling, vacuole degenerateion and necrosis. Caspase-3 is involved in the process of renal injury. UTI has a protective effect on the renal tissue of rats following paraquat poisoning through up-regulating the level of Livin and down-regulating the level of Caspase-3, however, the regulation mechanism as well as the pathway is still needed to further study.