不同剂量的重组腺相关病毒载体介导的人血管抑素对糖尿病肾病大鼠肾功能的影响

Effect of different doses of rAAV-AS on the renal function in rats with diabetic nephropathy

目的探讨不同剂量的重组腺相关病毒载体介导的人血管抑素(recombinant adeno-associated viruscarrying human angiostatin gene,rAAV-AS)对糖尿病肾病(diabetic nephropathy,DN)大鼠肾功能的影响。方法60只清洁级雄性SD大鼠随机抽取10只作为正常对照组(NC组),其余造模为糖尿病大鼠。大鼠空腹12 h腹腔注射链尿佐菌素(STZ)60 mg/kg,72 h后尾静脉测血糖大于16.7 mmol/L记为造模成功,适应性喂养1周后,把糖尿病大鼠随机分为DN组(n=10),rAAV-0组(n=10),AS1组(n=10,1×1012VG/kg),AS2组(n=10,2×1012VG/kg),AS3组(n=10,4×1012VG/kg)。AS1组、AS2组和AS3组3组大鼠尾静脉注射rAAV-AS,NC组和DN组分别注射同等剂量的生理盐水,rAAV-0组注射同等剂量rAAV-0空载体;从第2周起,整个实验过程共持续8周。8周后检测各组大鼠的体重(body weight,BW)、肾重(kidney weight,KW),计算肾指数(kidney in-dex,KI),检测血糖(blood glucose,BG)、糖化血红蛋白(hemoglobin A1c,HbA1c)、尿素氮(blood urea nitrogen,BUN)、血肌酐(serum creatinine,SCr)、尿微量白蛋白(miscroalbunminuria,MAU)。结果与NC组相比,DN、rAAV-0、AS1组的KI、MAU、BUN、SCr、BG、HbA1c明显增高(P<0.05);与DN组相比,AS2组和AS3组KI、MAU、BUN、SCr明显降低(P<0.05);与rAAV-0组相比,AS2组和AS3组KI、MAU、BUN、SCr降低(P<0.05);与AS1组相比,AS2组和AS3组KI、MAU、BUN、SCr降低(P<0.05)。结论中等剂量和大剂量rAAV-AS对DN大鼠肾脏具有保护作用,且不依赖血糖降低而起作用。

Objective To investigate the effect of different doses of recombinant adeno - associated virus carrying human angiostatin gene （rAAV -AS） on the renal function in rats with diabetic nephropathy. Methods 10 out of 60 male SD rats of clean grade were randomly allotted as the normal control group, while the others were used to establish models of diabetic rats. Streptozotocin （STZ） 60 mg/kg was intraperitoneally injected in rats at the end of 12 h of fasting to induce diabetic model. 72 h later, the blood glucose level of caudal vein was above 16.7 mmol/L, and the diabetic model was marked as successful. After a week of adaptation, diabetic rats were randomized into 5 groups （n = 10） ： group DN, group rAAV-0, group AS1 （1 × 10^12 VG/kg）, group AS2 （2 × 10^12 VG/kg）, group AS3 （4× 10^12 VG/ kg）. Different doses of rAAV - AS were injected into groups AS1, AS2 and AS3 from the caudal vein, and the corresponding doses of normal saline were injected in group NC and corresponding doses of rAAV - 0 idler injected in group DN. At the end of the 8th weeks, their body weight （BW） and kidney weight （KW） were measured to calculate kidney index （KI）. The blood glucose （BG）, glycosylated hemoglobin （HbAlc）, miscroalbunminuria （MAU）, blood urea nitrogen （BUN） and serum creatinine （SCr） were determined. Results Compared with group NC, the levels of KI, BG, HbAlc, MAU, BUN and SCr increased significantly in DN, rAAV - 0 and AS1 groups （ P 〈 0.05）. Compared with group DN, the levels of KI, MAU, BUN and SCr significantly decreased in groups AS2 and AS3 （P 〈0. 05）. Compared with group rAAV - 0, the levels of KI, MAU, BUN and SCr significantly decreased in groups AS2 and AS3 （P 〈 0.05 ）. Compared with group AS1, the levels of KI, MAU, BUN and SCr significantly decreased in groups AS2 and AS3 （P 〈 0.05 ）. Conclusions rAAV - AS of medium and large doses has protective effect on the kidneys in diabetic rats, independent of the decrease of blood glucose level.