ADAMTSL3促大鼠糖尿病肾病心肌纤维化和SGLT-2抑制剂的疗效观察

Effect of ADAMTSL3 on myocardial fibrosis and SGLT-2 inhibitor in diabetic nephropathy in rats

目的 探讨ADAMTSL3(Adisintegrin-like and metalloprotease domain with thrombospondin type I motifs-like-3)在大鼠糖尿病肾病心肌纤维化中的作用机制和SGLT-2抑制剂(Sodium-glucose co-transporter 2)的应用效果。方法 SPF级雄性Wistar大鼠28只经高脂高糖饲料喂养联合腹腔注射链脲佐菌素(SMZ)50 mg/kg进行糖尿病肾病造模,造模成功后分为模型组和治疗组,另设正常对照组大鼠,治疗组给予SGLT-2抑制剂(卡格列净10 mg/kg)灌胃。3组大鼠进行心脏B超动态分析心脏结构改变。对大鼠心肌组织进行病理染色、qRT-PCR、Western Blot检测ADAMTSL3基因表达水平,同时观察使用SGLT-2抑制剂后大鼠心肌的病理改变。结果 与正常对照组相比,模型组在左心室重量(LVM)、左心室收缩末壁厚度(LVPWs)、左心室短轴缩短率(LVFS%)、左射血分数(LVEF%)上的差异均有统计学意义(P<0.05)。与模型组相比,治疗组大鼠糖尿病肾病诱导心肌组织纤维化相对面积降低,以及在大鼠心肌纤维化中的Ⅰ型胶原蛋白(CollagenⅠ)、纤维连接蛋白(Fibronectin)表达降低(P<0.05)。与正常对照组相比,模型组大鼠心肌组织的ADAMTSL3蛋白表达显著增加;与模型组相比,治疗组大鼠心肌组织的ADAMTSL3蛋白表达显著降低(P<0.05)。与正常对照组相比,模型组中衰老标志物p-p53、p21、p16表达水平升高;与模型组相比,治疗组p-p53、p16表达水平均降低,差异有统计学意义(P<0.05)。结论 ADAMTSL3表达与纤维化相关,因此ADAMTSL3参与促进糖尿病肾病大鼠心肌纤维化,SGLT-2抑制剂治疗可逆转其心肌纤维化病变。

Objective To investigate the mechanism of ADAMTSL3(Adisintegrin-like and metalloprotease domain with thrombospondin type I motifs-like-3) in myocardial fibrosis in rats with diabetic nephropathy and the effect of SGLT-2(Sodium-glucoseco-transporter2). Methods 28 SPF male Wistar rats were fed with high-fat and high-sugar diet combined with intraperitoneal injection of streptozotocin( SMZ) 50 mg/kg to establish diabetic nephropathy model. After successful establishment of the model, the rats were divided into the model group and the treatment group, and the normal control rats were set up also. Rats in the treatment group were given SGLT-2 inhibitor(10mg/kg) intragastric administration. The changes of cardiac structure were dynamically analyzed by B-ultrasound in the three groups. The myocardial pathological staining, qRT-PCR and Western Blot were performed to detect the expression of ADAMTSL3 gene, and the pathological changes of rat myocardium were observed after the use of SGLT-2 inhibitor. Results Compared with the normal control group, the model group had statistically significant differences in left ventricular weight(LVM), left ventricular end systolic wall thickness(LVPWs), left ventricular short axis shortening rate(LVFS%) and left ejection fraction(LVEF%)(P<0. 05). Compared with the model group, the relative area of myocardial fibrosis induced by diabetic nephropathy in the treatment group was decreased, and the expressions of Collagen Ⅰ and Fibronectin were decreased in the treatment group(P<0. 05). Compared with normal control group, ADAMTSL3 protein expression in myocardium of model group was significantly increased. Compared with model group, ADAMTSL3 protein expression in myocardial tissue of the rats in the treatment group was significantly decreased(P<0. 05). Compared with the normal control group, the expression levels of aging markers pp53, p21 and p16 were increased in the model group. Compared with model group, the expression levels of P-p53and p16 in the treatment group were decreased, and the difference was statistically significant(P<0. 05). Conclusion The expression of ADAMTSL3 is related to fibrosis, so ADAMTSL3 is involved in promoting myocardial fibrosis in diabetic nephropathy rats, and SGLT-2 inhibitor therapy can reverse the myocardial fibrosis.