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目的:探讨STZ诱导的糖尿病小鼠肾脏发生上皮-间质转分化(EMT)的情况。方法:将80只C57BL小鼠随机分为正常对照组(NC组)和糖尿病组(DM组),每组40只。DM组小鼠用1%STZ(streptozotocin,链脲佐菌素)溶液按60mg/kg体质量的剂量进行腹腔注射,每天1次,连续6天。NC组小鼠平行腹腔注射同等体积0.1mol/L的柠檬酸钠缓冲液。再将成模小鼠随机分为A、B批次,A批次用于动态观察生存率、小鼠体质量及随机血糖的监测;B批次用于在造模后第4、8、12周末观察肾组织的病理变化,并用Western blot、免疫荧光染色的方法观察肾组织中EMT标志蛋白α-SMA和E-cadherin的表达。结果:STZ诱导的糖尿病模型小鼠出现糖尿病典型症状如多饮、多尿等,血糖持续在高水平状态,体质量增长缓慢。在造模后12周末,DM组小鼠较NC组小鼠累积生存率显著降低,两组比较差异具有统计学意义(P<0.001)。在造模后的第8周末,DM组小鼠肾脏出现明显的病理改变,到第12周末时,绝大部分肾小管上皮细胞被梭形的肌成纤维细胞取代,肾小球空泡,基底膜增厚。造模后的第4、8、12周末时,DM组小鼠E-cadherin表达量均显著低于NC组小鼠(P=0.004,0.026,0.004);而在第8和12周末时,DM组α-SMA表达量显著升高(P=0.009,0.015)。在第12周末,肾组织冰冻切片E-cadherin和α-SMA、免疫荧光染色结果与上述结果一致。结论:STZ诱导的糖尿病模型小鼠有较典型的糖尿病临床改变,且肾组织发生了EMT。
Objective: To investigate the occurrence of epithelial-mesenchymal transition (EMT) in kidney of STZ-induced diabetic mice. Methods: Eighty C57BL mice were randomly divided into normal control group (NC group) and diabetic group (DM group), 40 rats in each group. DM mice were injected intraperitoneally with 1% STZ (streptozotocin) solution at a dose of 60 mg / kg body weight once daily for 6 days. The mice in NC group were injected intraperitoneally with the same volume of 0.1mol / L sodium citrate buffer. Then the model mice were randomly divided into A, B batches, A batch for the dynamic observation of survival rate, body weight and random blood glucose monitoring; Batch B was used in the modeling after 4,8,12 weekend The pathological changes of renal tissues were observed. The expressions of EMT marker α-SMA and E-cadherin in renal tissues were observed by Western blot and immunofluorescence staining. Results: The mice with diabetes mellitus (STZ) induced typical symptoms of diabetes, such as polydipsia, polyuria and so on. The blood glucose level kept at a high level and the body weight increased slowly. At the 12th week after modeling, the cumulative survival rate of mice in DM group was significantly lower than that in NC group. There was significant difference between the two groups (P <0.001). At the end of the 8th week after model establishment, the pathological changes in the kidneys of DM mice were obvious. By the end of the 12th week, most of the renal tubular epithelial cells were replaced by fusiform myofibroblasts, glomerular vacuolization, Membrane thickening. At the end of the 4th, 8th and 12th week after modeling, the expression of E-cadherin in DM group was significantly lower than that in NC group (P = 0.004,0.026,0.004); at the end of 8th and 12th week, DM Group α-SMA expression was significantly increased (P = 0.009,0.015). At the end of the 12th week, the frozen section of renal tissue E-cadherin and α-SMA, immunofluorescence staining results consistent with the above results. CONCLUSIONS: STZ-induced diabetic mice have more clinical changes of typical diabetes and EMT occurs in renal tissues.