摘要
目的:探究胰岛素样生长因子1受体(insulin-like growth factor 1 receptor,IGF-1R)抑制剂抑制糖尿病肾病小鼠的肾脏纤维化。方法:选取90只8周龄C57/BL6雄鼠,依据随机数字表法分为5组,每组18只。依次为A组(糖尿病肾病组)、B组(IGF-1R组)、C组(血管紧张素转换酶抑制剂组)、D组(胰岛素组)和对照组。A组、B组、C组和D组雄鼠腹腔注射100 mg/kg链脲霉素,48 h随机血糖≥16.7 mmol/L,尿糖(+~4+)定性为糖尿病。造模8周后,A组使用生理盐水灌胃,B组48 h/次使用30 mg/kg IGF-1R抑制剂灌胃,C组48 h/次使用30 mg/kg IGF-1R抑制剂灌胃,D组皮下注射1~2 U/kg胰岛素。常规检测各组尿肌酐、血糖、24 h蛋白排泄量。16周龄时对各组小鼠进行麻醉处死,收集其肾组织样本,行常规石蜡包埋后进行组织病理学检测。结果:对照组小鼠血糖正常,A组、B组和C组小鼠血糖均高于16.7 mmol/L,D组小鼠血糖低于16.7 mmol/L,差异具有统计学意义(P<0.05)。A组小鼠尿蛋白排泄率高于对照组小鼠,B组和D组小鼠尿蛋白排泄率低于A组,差异具有统计学意义(P<0.05)。A组小鼠尿蛋白肌酐比高于对照组小鼠,B组小鼠尿蛋白肌酐比低于A组,D组小鼠尿蛋白肌酐比低于A组,差异具有统计学意义(P<0.05)。组织原位杂交检测,糖尿病肾病时肾组织中Snail1和IGF-1表达上调,A组小鼠IGF-1 mRNA表达变化无统计意义(P>0.05),Snail1 mRNA表达明显降低,差异具有统计学意义(P<0.05)。C组和D组小鼠肾组织中Snail1和IGF-1表达无明显变化(P>0.05)。免疫组织化学染色,A组小鼠肾组织中Snail1和IGF-1表达高于对照组,差异具有统计学意义(P<0.05)。B组小鼠IGF-1蛋白表达无明显变化,Snail1蛋白表达降低,差异具有统计学意义(P<0.05)。C组和D组小鼠较A组小鼠Snail1和IGF-1蛋白表达无明显变化。结论:抑制IGF-1R或沉默IGF-1可抑制肾小管上皮细胞间质转分化,缓解肾纤维化。
Objective:To investigate the possible mechanism of insulin-like growth factor 1 receptor(IGF-1R)inhibitor inhibiting renal fibrosis in mice with diabetic nephropathy.Methods:Ninety 8-week-old C57/BL6 male rats were equally and randomly divided into five groups:group A(diabetic nephropathy group),group B(IGF-1R group),group C(angiotensin-converting-enzyme inhibitor group),group D(insulin group),and control group.Groups A-D were intraperitoneally injected with 100 mg/kg streptozotocin to establish a diabetes model,which was confirmed with 48 h random blood glucose(≥16.7 mmol/L)and urine glucose(+~4+).After 8 weeks of modeling,group A was given normal saline by gavage,group B was given 30 mg/kg IGF-1R inhibitor by gavage once every 48 h,group C was given 30 mg/kg angiotensin-converting-enzyme inhibitor once every 48 h,and group D was injected subcutaneously with 1-2 U/kg insulin.The urine creatinine,blood glucose,a nd 24-hour protein excretion in each group were routinely tested.At the age of 16 weeks,the mice in each group were anesthetized and sacrificed.The renal tissue samples were collected and routinely paraffinembedded for histopathological examination.Results:The blood glucose in the control group was normal.The blood glucose levels in groups A-C were higher than 16.7 mmol/L,and that in group D was lower than 16.7 mmol/L.There was a significant difference in blood glucose between the groups(P<0.05).The urinary protein excretion rate in group A was significantly higher than that in the control group(P<0.05);the urinary protein excretion rates in group B and D were significantly lower than that in group A(P<0.05).The urinary protein/creatinine ratio in group A was significantly higher than that in the control group(P<0.05);compared with group A,groups B and D had a significantly lower urine protein/creatinine ratio(both P<0.05).In situ hybridization assay showed that the expression of Snail1 and IGF-1 in the renal tissue was up-regulated in mice with diabetic nephropathy.Compared with the control group,group A had no significant change in the mRNA expression of IGF-1(P>0.05),but had significantly lower mRNA expression of Snail1(P<0.05);no significant changes in the mRNA expression of Snail1 and IGF-1 in the renal tissue were observed in groups C and D(P>0.05).Immunohistochemical staining results showed that compared with the control group,group A had significantly higher protein expression of Snail1 and IGF-1 in the renal tissue(P<0.05);group B had no significant change in the protein expression of IGF-1(P>0.05),but had significantly lower protein expression of Snail1(P<0.05).No significant changes in the protein expression of Snail1 and IGF-1 were observed in groups C and D compared with group A(P>0.05).Conclusion:Inhibiting IGF-1R or silencing IGF-1 can inhibit renal tubular epithelial-mesenchymal transition and relieve renal fibrosis.
作者
张晓利
胡威利
Zhang Xiaoli;Hu Weili(Department of Nephropathy and Rheumatism,The Third Affiliated Hospital of Xinxiang Medical University)
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2020年第3期363-367,共5页
Journal of Chongqing Medical University
作者简介
张晓利,Email:zhangxiaoli329@163.com,研究方向:肾小球疾病的基础与临床研究。
