棉酚对2型糖尿病大鼠肾脏SGK1、Col－Ⅳ基因表达的影响  

Effects of gossypol on expression of SGK1 and Col-IV in nephridial tissue of diabetic rats

作　　者：张虎祥[1] 万丽[1] 肖艳[1] 吴晓烨[1] 王蓉蓉[1] 陈筱菲[2] 方周溪[3] 陈国荣[1] 

机构地区：[1]温州医学院附属第一医院病理科,325000 [2]温州医学院附属第一医院检验科,325000 [3]温州医学院附属第一医院电镜室,325000

出　　处：《浙江医学》2012年第24期1951-1954,1957,共5页Zhejiang Medical Journal

基　　金：浙江省科技攻关项目（2007C34003）;温州市科技局（Y2111190041）

摘　　要：目的观察棉酚对2型糖尿病大鼠肾脏组织形态的影响。探讨糖皮质激素诱导的蛋白激酶1（SGKl）、IV型胶原（Col-IV）在糖尿病肾病发病中的作用及棉酚干预的机制。方法雄性SD大鼠30只随机分成正常对照组、2型糖尿病组、棉酚干预组（各10只）。后两组给以高脂饮食加小剂量（30mg／kg）链脲佐菌素（STZ）诱导2型糖尿病大鼠模型。此后，棉酚干预组按15mg·kg-1·d-1剂量棉酚（溶于2％羧甲基纤维素）灌胃4周。从第5周起，按15mg／kg剂量棉酚每周给予维持剂量一次至第12周末。检测大鼠生化指标：血糖、血尿素氮（BUN）、尿蛋白；称量大鼠体重（Bw）、肾重（Kw）并计算肾指数（K／Bw）；用光镜及电镜观察大鼠肾脏的组织形态学改变；逆转录-PCR法检测肾脏SGKl、Col-IVmRNA含量。结果2型糖尿病组大鼠血糖[（2748±1．94）mmol／L]、BUN[（14．89±482）mm01／L]、24h尿蛋白[（37．64±14．36）mg／24h]、K／Bw（00112±00015）较正常组[（6．31±042）mm01／L、（6．79±032）mmol／L、（739±2．15）mg／24h、（00055±0．0003）1明显升高（P〈0．01）。光镜下，2型糖尿病组大鼠肾小球体积普遍增大，个别肾小球出现萎缩。PAS染色阳性基质增多，主要集中在肾小球团的中心；电镜下，2型糖尿病组大鼠肾小球毛细血管基底膜不规则增厚，足细胞突起肿胀，可见足突融合。与正常对照组比较，2型糖尿病组大鼠肾Col-IV、SGKl基因mRNA表达显著升高。经棉酚治疗后，’肾脏生化指标明显好转，组织病理改变减轻，Col-IV基因mRNA表达水平降低，SGKl基因mRNA表达有降低趋势。结论棉酚干预组大鼠肾脏组织形态改变有减轻趋势，可能与降低2型糖尿病大鼠肾脏Col-IVmRNA表达及影响SGKImRNA表达有关。Objective To investigate the effects of gossypol on diabetic nephropathy and the possible mechanism. Methods Thirty male Sprague-Dawley rats were divided randomly into three groups： normal control group, type 2 diabetic group and gossypol-treated group. After being fed with high-fat feeding for 4 weeks, the latter two groups were injected with low dosage streptozotocin （30mg/Kg）intraperitoneally to induce type 2 diabetes. Diabetic rats in gossypol-treated group were given gossypol at the dosage of 15mg/kg.d for 4 weeks by gavage, afterwards once per week for 8 week. The rats were sacrificed after fasting for 12h and 24-h urine samples were collected. The concentrations of blood glucose, blood urea nitrogen（ BUN ）and 24-h urine protein were measured. The morphological changes of the kidney were studied by light microscopy and transmission elec- tron microscopy. The mRNA expression of SGK1, Col-IV kidney tissue was assayed by semi-quantity RT-PCR. Results Com- pared with normal control group, the level of glucose, BUN and the 24-h urine protein in diabetic group were significantly in- creased （P〈0.01）. The volume of glomerulus was enlarged, the deposition of PAS positive substance in the glomerular intersti- tium increased under light microscopy, and the glomerular basal membrane was thickened significantly under electron mi- croscopy in diabetic group, but they were ameliorated in gossypol-treated group. The mRNA expression of SGK （P〈0.05）and Col-IV （P〈0.01） was increased significantly compared with normal control group. After the treatment of gossypol, the mRNA ex- pression of Col-IV was decreased compared with type 2 diabetic group （P〈0.05）. Conclusion Gossypol may delay the progress of diabetic nephropathy by down-regulating mRNA expression level of Col-IVin kidney of type 2 diabetic rats.

关 键 词：糖尿病肾病 棉酚 IV型胶原 基因表达 逆转录多聚酶链反应 

分 类 号：R151.2[医药卫生—营养与食品卫生学]