抗增殖蛋白在db／db小鼠肾损伤中的变化及干预研究

The change of prohibitin in the kidney injury of db/db mice and its intervention

目的研究抗增殖蛋白在糖尿病肾病中的分子机制和葡萄籽原花青素B2（GSPB2）的保护作用。方法以雄性db／m小鼠为健康对照组、db／db小鼠为2型糖尿病模型组（糖尿病组）、GSPB2干预组，各8只，每日给予GSPB2（30mg·kg-1·d-1）灌胃。于18周末留取血清测定空腹血糖（FBG）、糖基化终末产物（AGE。）和单核细胞趋化蛋白-1（MCP-1）的含量，留取肾脏组织行苏木素伊红（HE）染色观察组织病理变化，透射电镜观察肌浆网超微结构变化，应用免疫印迹分析测定抗增殖蛋白的蛋白表达。结果糖尿病组小鼠在第8、12、16、18周体质量较健康对照组增加（均P〈0．01），在第12、16、18周，GSPB2能抑制糖尿病组体质量增加（P〈0．05）。糖尿病组FBG、血清AGEs和MCP-1含量较健康对照组均升高（P〈0．01），GSPB2能抑制FBG、AGEs和MCP-1（P〈0．01）。可见db／db小鼠肾小球体积增大，系膜细胞增生，细胞外基质增多，GSPB2干预组肾小球体积增大及系膜细胞增生情况较糖尿病组减轻；超微结构发现糖尿病组系膜细胞插入，基底膜增厚，足细胞数量减少，足突细胞融合，GSPB2干预组的基底膜增厚及足细胞损伤均较糖尿病模型鼠改善。糖尿病组抗增殖蛋白的蛋白表达较健康对照组降低（P〈0．01），而GSPB2能够增加抗增殖蛋白的蛋白表达（P〈0．01）。结论GSPB2对db／db小鼠。肾脏损伤起到一定的保护作用，其机制可能与抗糖基化、抗炎和增加抗增殖蛋白的表达有关。

Objective To study the molecular mechanisms of prohibitin in the diabetic nephropathy and its effect. Methods C57BLKS/J db/db mice were selected randomly as diabetic model group （DM group, n=8） and the db/m mice were selected as control group （CC, n=8） , and C57BLKS/J db/db mice lavage treated with GSPB2 for 10 weeks were considered as treatment group （30mg/kg body weight/day, DMT, n：8）. At the end of 18 weeks, fasting blood glucose （FBG）, advanced glycation end products （AGEs） and monocyte chemotactic protein 1 （MCP1） were determined. Morphological changes of the kidney tissue were examined by light microscopy and transmission electron microscope. The expression o{ prohibitin in the kidney tissue was determined by Western blot. Results The body weight was higher in DM groups than in control group at 8, 12, 16, 18 weeks （all P〈0.01）. The body weight was significantly inhibited at 12, 16, 18 weeks after GSPB2 administration in the DMT group as compared to the DM group （all P〈0.05）. Serum FBG, AGEs and MCP-1 levels were significantly higher in DM group than in control group （all P〈0.01）. GSPB2 significantly decreased the serum levels of FBG, AGEs and MCP-1 in db/db mice （all P〈 0.01）. The glomerular volume, mesangial cell proliferation, extracellular matrix in DM group was significantly increased as compared with CC group by light microscopy. GSPB2 significantly reduced the glomerular volume, mesangial cell proliferation in db/db mice. The result of ultrastructural microscopy showed that in DM group, mesangial cell was inserted into the sub-endothelium, basement membrane became thicken, the number of podocytes was decreased and the podocytes were fused, and the thicken basement membrane and podocytes injure were improved in the DMT group. Moreover, the protein expression of prohibitin was higher in DM group than in control group （P〈0.01）, while GSPB2 could decrease the protein expression of prohibitin （P〈 0.01 ）. Conclusions The mechanism of protective effects of GSPB2in diabetic nephropathy can be attributed to its anti glycation, anti-inflammatory and the increase of prohibitin expression, which will provide a theoretical basis for the prevention and treatment of diabetic nephropathy.