糖尿病肾病大鼠肾小动脉骨基质蛋白表达及其与MCP-1的关系  

Relation Between Bone Matrix Proteins and Monocyte Chemoattractant Protein-1 in Renal Small Artery in Diabetic Nephropathy Rats

作　　者：赵安菊[1,2] 黄颂敏[1] 欧三桃[1] 刘芳[1] 陈泽君[1] 胡章学[1] 唐万欣[1] 

机构地区：[1]四川大学华西医院肾脏内科,成都610041 [2]贵州省人民医院干医科,贵阳550002

出　　处：《四川大学学报（医学版）》2012年第3期414-419,共6页Journal of Sichuan University(Medical Sciences)

摘　　要：目的观察糖尿病肾病(DN)大鼠肾实质内小动脉上骨基质蛋白的表达及其变化规律,初步探讨其与炎症因子单核细胞趋化蛋白1(MCP-1)的关系及对糖尿病肾病血管病变的影响。方法设对照组(N组,30只)和糖尿病肾病组(DN组,30只)。4周、12周和24周每组各处死10只大鼠,血生化指标按常规检测;茜素红染色观察肾实质内小动脉周围钙盐沉积,免疫组化方法、原位杂交法检测各时间点肾小动脉中骨基质蛋白核心结合因子α1(cbfα1)、骨形态蛋白2(BMP-2)及MCP-1蛋白和基因表达。以实时荧光定量PCR法作BMP-2及基质Gla蛋白(MGP)的基因定量。结果①DN组各时间点血糖、血尿素氮及24h尿蛋白量均较N组增高(P<0.05),自第12周开始,DN组血肌酐、血磷较N组增高(P<0.05)。②茜素红染色DN组第4周～12周时偶见肾内小动脉少许点状红色钙盐沉积,第24周时可见肾实质内小动脉及周围组织有较多红色点片状沉积,N组茜素红染色为阴性。③免疫组化显示DN组肾小动脉4周时已有cbfα1、BMP-2表达,随时间延长表达逐渐增强,24周表达最强,各时间点均高于N组(P<0.05),原位杂交显示cbfα1、BMP-2仅在肾小动脉中有表达,表达趋势同免疫组化结果;DN组肾实质内小动脉MCP-1蛋白及mRNA在4、12周表达无明显差异,24周表达明显上调,N组肾小动脉无MCP-1mRNA表达。实时荧光定量PCR显示DN组4周时BMP-2mRNA已有表达,随时间延长表达逐渐升高,24周最高(P<0.05);DN组MGP表达量4周最高,随时间延长表达逐渐降低,24周最低(P<0.05)。而N组BMP-2和MGP mRNA各时间点表达量差异均无统计学意义。④DN组cbfα1及BMP-2基因与蛋白均与MCP-1呈正相关。结论①DN大鼠肾内小动脉钙化前即已有骨基质蛋白的表达;②MCP-1可能影响骨基质蛋白cbfα1的表达;③cbfα1、BMP-2、MGP及MCP-1可能参与了DN血管病变。Objective To observe the expressions of bone matrix proteins and monocyte chemoattractant protein-1（（MCP-1） in the renal arteriole of diabetic nephropathy（DN） rats and analyze their correlations and roles in diabetic nephropathy.Methods Adult Sprague-Dawley male rats were used to establish the animal model of diabetic nephropathy induced by peritoneal injection of 55 mg/kg of streptozocin.Calcium deposit around the renal arteriole was observed by alizarin red staining.The protein and mRNA levels of core-bind factor alpha 1（cbfα1）,bone morphogenetic protein 2（BMP-2） and matrix Gla protein（MGP） in renal arteriole of DN rats were detected by immunohistochemistry,in-situ hybridization and real-time PCR.The biochemical indices were detected by routine test.Results ①Blood glucose and Urine protein of 24 h were significantly increased in the renal arteriole of DN rats vs.the control rats（P0.05）,serum creatinine（SCr） and phosphorus were significantly increased from 12 weeks.② Little deposit of calcium salt was observed in the renal arteriole of DN rats at the 4th week and a large amount of deposit was observed at 24th week,but no calcium deposit was observed in control rats.③ Cbfα1 and BMP-2 expressions were significantly increased in the renal arteriole of DN rats from 4 to 24 weeks vs.the control rats.MGP mRNA expression in the renal arteriole of DN rats was significantly decreased from 4 to 24 weeks.MCP-1 expression was obviously upregulated in the renal arteriole of DN rats at 24th week vs.that at 4th and 12th week.No MCP-1 expression was observed in the renal arterioles of control rats.④ MCP-1 were positively correlated with the expression of cbf α1 and BMP-2.Conclusion Bone matrix proteins has already exp ressed in renal arteriole before the formation of vascular calcification.MCP-1 can affect the expression of cbfα1,BMP-2;Cbfα1,BMP-2,MGP and MCP-1 may be involved in the formation of vascular lesions of DN.

关 键 词：糖尿病肾病 血管钙化 炎症因子 骨基质蛋白 

分 类 号：R587.2[医药卫生—内分泌] R692[医药卫生—内科学]