促血管生成素重组腺病毒对糖尿病鼠肾血管新生的影响

Effect of recombinant adenovirus angiopoietin-1 on renal angiogenesis in diabetic rats

摘要:

目的  通过SD大鼠糖尿病肾病（DN）模型，观察经腺病毒外源性表达Ang-1对肾脏血管新生的作用，以期为DN的临床治疗提供参考。方法  SD大鼠DN造模选用链脲佐菌素（STZ），设计两个大的组别，即正常对照组和DN模型组，并将DN模型组进一步划分为3个亚组，即糖尿病组、空白载体组、Ang-1腺病毒组。STZ造模成功后的第8周开始，从尾静脉注入Ang-1腺病毒载体以及空白载体，在8、12、20和28周，检测各组大鼠的尿蛋白水平，实时定量聚合酶链法分析肾脏Ang-2 mRNA水平。结果  ①DN模型组大鼠的尿蛋白水平较正常对照组升高（P <0.01），且只有在20周前，空白载体组及糖尿病组的尿蛋白水平才低于Ang-1腺病毒组（P <0.01）；②在正常对照组大鼠肾组织中未检测到尿蛋白和Ang-2 mRNA表达，而其他DN模型组（糖尿病、空白载体、Ang-1腺病毒组）大鼠12周后的尿蛋白和Ang-2 mRNA表达升高，特别是Ang-1腺病毒组升高（峰值在20周）更为明显（P <0.05），在28周时降低，糖尿病、空白载体和Ang-1腺病毒组的Ang-2 mRNA表达比较差异有统计学意义（P <0.05）；③Ang-2与尿蛋白排泄量呈负相关（r =-0.601，P <0.05）。结论  外源性给予Ang-1对糖尿病肾脏新生微血管生成有保护作用。

Abstract:

Objective To establish diabetic nephropathy rat model and observe diabetic kidney exogenous expression of angiopoietin-1 (Ang-1) by adenovirus and to study its effect on generation of new blood vessels in the kidneys, in order to provide reference for clinical treatment of diabetic nephropathy. Methods Streptozotocin (STZ) was chosen for diabetic nephropathy modeling of SD rats. The model rats were divided into two groups, namely normal control group and diabetic nephropathy group; and diabetic nephropathy model group was further divided into three subgroups as diabetic group, empty vector group and Ang-1 adenovirus group. Since the eighth week of successful rat modeling, Ang-1 adenovirus vector and empty vector were injected through tail vein. At different time, such as the 8th week, 12th week, 20th week and 28th week, the level of urine protein content of each group was detected, and kidney level of Ang-2 mRNA was analyzed using real-time quantitative PCR. Results Urinary proteins of the diabetic nephropathy model group were significantly higher than those in the healthy rats (P < 0.01), and only in the first 20 weeks, urine protein levels of the empty vector group and the diabetic group were significantly lower than those in the Ang-1 treatment group (P < 0.01). Urine protein content and Ang-2 mRNA were not detected in healthy kidney tissue of the control group; while those in the diabetes group, empty vector group and Ang-1 adenovirus group were significantly elevated after the 12th week, especially those of the Ang-1 adenovirus group with the peak values at the 20th week (P < 0.05); and reduced in the 28th week, Ang-2 mRNA levels of the diabetic group, empty vector group and Ang-1adenovirus group were statistically different (P < 0.05). Ang-2 and urinary protein excretion showed a negative correlation (r = -0.601, P < 0.05). Conclusions The exogenous administration of Ang-1 in diabetic kidney has a protective effect on angiogenesis.