糜蛋白酶抑制剂保护糖尿病仓鼠肾功能的机制研究
Protective effect of chymase inhibitor on renal function in diabetic hamsters
目的 观察糜蛋白酶抑制剂对糖尿病仓鼠肾脏的保护作用，进而探讨其保护肾脏损伤的机理。 方法 链脲佐菌素腹腔注射法制备仓鼠糖尿病模型。通过分子生物学技术、免疫组织化学技术、免疫印记技术检测各组动物糜蛋白酶，RAS成分：ACE、肾素、ANG-I、ANG-II；氧化应激水平：8-OhdG，NOX-4，p22phox，以及TGF-β1的表达情况。 结果 糜蛋白酶抑制剂抑制了糖尿病仓鼠肾内ANG-II升高，同时明显降低了8-OHdG 分泌水平。NOX-4和p22phox染色表明糖尿病组血管球和肾小管区域氧化产物显著增高，而糜蛋白酶抑制剂抑制氧化产物的升高(P<0.01)。Western blot 证实了糖尿病组血管球NOX-4和p22phox蛋白的水平较对照组高，糜蛋白酶抑制剂显著降低了其表达。血管球TGF-β1表达糖尿病组高于对照组，同样糜蛋白酶抑制剂治疗后大大降低了TGF-β1水平。 结论 糜蛋白酶抑制剂能保护糖尿病仓鼠的肾功能，其机制是通过降低肾内ANG-II水平和氧化应激水平来保护糖尿病肾脏损伤。
Objective To observe the protective effect of chymotrypsin inhibitors on diabetic hamster kidney, and to explore mechanism underlying the protection. Methods Diabetic hamsters was established by intraperitoneal injection of STZ. The animal chymotrypsin, RAS components: ACE, renin, ANG-I, ANG-II; oxidative stress levels: 8-OhdG, NOX-4, p22phox, and the expression of TGF-β1 were determined by molecular biologic techniques, immunohistochemistry, western blotting technique. Results Chymotrypsin inhibitors inhibited the increase of ANG-II in diabetic hamster kidney, but significantly reduced the level of 8-OHdG secretion. NOX-4 and p22phox staining indicated significantly increased products of oxidation products in diabetic glomerulus and tubules, which could be significantly inhibited by chymotrypsin inhibitors (P<0.01). Western blot confirmed that the level of NOX-4 and p22phox increased in the diabetic group than in the control group, and chymotrypsin inhibitor could significantly reduce its expression. Glomerular TGF-β1 expression in the diabetic group was higher than that in the control group, and the chymotrypsin inhibitor therapy could significantly reduce the levels of TGF-β1. Conclusions Chymotrypsin inhibitors can protect diabetic hamster kidney function, and the underlying mechanism is through reducing renal ANG-II levels and oxidative stress levels to protect diabetic kidney damage.
Chymase inhibitor / Oxidative stress / Kidney / Diabetic hamsters
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国家自然科学基金（31140072）；辽宁省科技厅计划项目（2011225015）；辽宁省自然科学基金（20102140）
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