摘要
目的 采用单侧肾切除联合链脲佐菌素(STZ)诱导构建糖尿病肾病(DKD)大鼠模型,并探讨Apelin-13对DKD纤维化的影响。方法 健康SD雄性大鼠28只,随机分为4组:假手术组、单侧肾切组、DKD组和Apelin-13组。DKD组单侧肾切后,一次性给予腹腔注射STZ,4、6、8周检测各组大鼠体重、随机血糖变化。8周后收集大鼠尿液、血液,检测大鼠尿微量白蛋白/肌酐比值,酶联免疫试剂盒检测大鼠血清中Apelin-13水平的表达变化。Apelin-13组在DKD成模后,给予腹腔注射Apelin-13 20μg/kg,苏木精-伊红(HE)和马松(MASSON)染色检测肾脏组织的病理学变化,并用实时荧光定量PCR和Western blot法检测纤维化标志物α-平滑肌肌动蛋白(SMA)mRNA和蛋白表达情况。结果 与假手术组和单侧肾切组相比,DKD组大鼠体重增长缓慢或出现负增长趋势,随机血糖明显升高,平均值为26.6mmol/L,尿微量白蛋白/肌酐比值升高。血清Apelin-13水平在单侧肾切组和DKD组均下降,且DKD组下降趋势更明显(P<0.05)。HE染色显示DKD组大鼠肾脏肾小管萎缩、肾小管上皮细胞变性、胶质纤维增生。MASSON染色显示Apelin-13组胶质纤维减少。与假手术组和单侧肾切组相比,α-SMA mRNA和蛋白在DKD组升高(P<0.05),给予Apelin-13注射后,α-SMA mRNA和蛋白水平均下降(P<0.05)。结论 Apelin-13可能在DKD纤维化发展中起着重要作用。
Objective To establish the rat model of diabetic kidney disease(DKD)induced by unilateral nephrectomy combined with streptozotocin(STZ),and study the effect of Apelin-13 on fibrosis of DKD.Methods Twenty-eight healthy male SD rats were randomized to the sham operation group,the unilateral nephrectomy group,DKD group(with unilateral nephrectomy combined with intraperitoneal injection of streptozotocin)and Apelin-13 group.The DKD group received one-time intraperitoneal of STZ,the changes of body weight and random blood glucose were monitored in each group at 4,6 and 8 weeks.Urine and blood were harvested after 8 weeks,and the serum Apelin-13 levels and microalbumin/creatinine ratio were detected.The Apelin-13 group was given intraperitoneal injection of 20μg/kg Apelin-13 after successful DKD modeling.HE and MASSON staining were used to detect pathological changes of renal tissue.Real-time PCR and Western blot were used to detect mRNA and protein expression ofα-smooth muscle actin(α-SMA),a marker of fibrosis.Results Compared to the sham operation group and the unilateral nephrectomy group,the weight of DKD group increased slowly or showed a negative trend(P<0.05).The random blood glucose was significantly increased with an average of 26.6mmol/L,elevated the urinary microalbumin/creatinine ratio(P<0.05).The level of serum Apelin-13 decreased in the unilateral nephrectomy group and the DKD group,and it was the lowest in the DKD group(P<0.05).Renal tubule atrophy,renal tubule epithelial cell degeneration and fiber hyperplasia were determined by pathological staining in DKD group.The glial fibers decreased in the Apelin-13 group.Compared with the sham operation group and the unilateral nephrotomy group,the mRNA and protein ofα-SMA were increased in DKD group(P<0.05),and the level ofα-SMA was decreased after intraperitoneal injection of Apelin-13(P<0.05).Conclusion Apelin-13 may play an important role in the development of fibrosis in DKD.
作者
王琦
刘旭静
胡玉娟
宋德刚
田桂珍
刘玉林
Wang Qi;Liu Xujing;Hu Yujuan(Department of Pharmacy,the Fifth People's Hospital of Jinan,Jinan 250022)
出处
《中国现代医药杂志》
2023年第10期15-19,共5页
Modern Medicine Journal of China
基金
济南市科技发展计划(临床医学科技创新计划)项目(编号:202019083)。
