四氢生物蝶呤在2型糖尿病肾病小鼠肾脏NO生成中的作用  

Effects of tetrahydrobiopterin on NO production in the kidney of type 2 diabetic nephropathy mice

作　　者：汪建云[1] 王栋栋[1] 陆昭磊 朱闯[1] 张帆[1] 郭昊[1] 印晓星[1] 

机构地区：[1]徐州医学院新药与临床应用重点实验室,徐州江苏221004

出　　处：《中国药理学通报》2014年第4期514-519,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81173104);江苏省高校自然科学基金资助项目(No 09KJD310011);徐州市科技计划项目(No XZZD1156);徐州医学院院长专项人才基金资助项目(No 2011KJZ19);江苏省大学生创新计划(No 201310313028)

摘　　要：目的研究四氢生物蝶呤(tetrahydrobiopterin,BH4)对2型糖尿病肾病(diabetic nephropathy,DN)小鼠肾脏中NO生成的影响,为2型DN的防治寻找新的靶点提供实验依据。方法 12周龄发展为糖尿病肾病的db/db小鼠分为2组:DAHP组(150 mg·kg-1)、DN组(5%DMSO生理盐水)。同周龄db/m小鼠作NS组(5%DMSO生理盐水)。各组小鼠腹腔注射给药7 d,处死。化学比色法检测小鼠空腹血糖、24 h尿蛋白、血肌酐及iNOS酶活性;高效液相色谱法检测BH4水平;免疫组化法及Western bolt法检测iNOS蛋白;Griess法检测NO水平。结果 DN组血糖、血肌酐、24h尿量及尿蛋白、BH4水平、iNOS蛋白及酶活性、NO水平均明显高于NS组;DAHP组24 h尿量及尿蛋白、BH4水平、iNOS蛋白及酶活性、NO水平均明显低于DN组。结论在自发性2型DN小鼠肾脏中,BH4含量明显增加,促使iNOS蛋白表达及酶活性增加,从而导致NO生成增多,引起多尿及蛋白尿。Aim To observe the effects biopterin （ BH4 ） on nitric oxide （NO） of tetrahydro- production in the kidney of type 2 diabetic nephropathy （DN） mice, and to find a new target for the treatment of type 2 DN. Methods The 12 week-old db/db mice developed in- to DN phase were divided into 2 groups ： DAHP group,subjected to intraperitoneal injection of 150 mg ~ kg-1 DAHP （n = 8） ; DN group, subjected to intraperitone- al injection of same dose of normal saline containing 5% DMSO （n = 6）. The age-matched db/m mice （NS group） were subjected to intraperitoneal injection of same dose of normal saline containing 5 % DMSO （ n = 6）. Three groups of mice were treated for 7 days. Then the fasting blood-glucose, serum creatinine, u- rine protein and activity of iNOS were determined by chemical colorimetry. And the iNOS protein in renal cortex was determined by immunohistochemisty and western blot, respectively. BH4 was measured by HPLC method. NO level was determined by Griess method. Results The levels of fasting blood-glucose, serum creatinine, 2dh urine volume, 24h urine pro- tein, BH4, iNOS and NO in DN group were signifi-cantly higher than those in NS group;The levels of ser- um creatinine, urine volume, urine protein, BH4, iN- OS and NO in DAHP group were significantly lower than those in DN group. Conclusion In the kidney of type 2 DN mice, the increased BH4 contributes to over- production of NO by the increased iNOS expression, and resultes in the increase of urine volume and urine protein.

关 键 词：四氢生物蝶呤 INOS NO 2型糖尿病肾病 DB db小鼠 蛋白尿 

分 类 号：R332[医药卫生—人体生理学] R322.61[医药卫生—基础医学]