Notch信号通路在妊娠期DN小鼠中的表达意义

Expression significance of Notch signaling pathway in mice with gestational DN

目的探究Notch信号通路在妊娠期糖尿病肾病(DN)小鼠肾组织中的表达意义。方法采用链脲佐菌素(STZ)建立DN小鼠模型,DN小鼠(腹腔注射STZ溶液)和对照组小鼠(腹腔注射等量枸椽酸盐缓冲液),每组各6只,2、4、8周时分别收集小鼠血液检测生化指标,8周末处死小鼠取其肾脏组织,HE染色观察两组小鼠肾组织形态,免疫组织化学染色观察两组小鼠肾组织Notch1表达情况,Western blot检测Notch信号通路中Notch1、Jagged1、Hes1、Hey1蛋白表达水平,Real-time PCR检测Notch信号通路中的Notch1、Jagged1、Hes1和Hey1 mRNA相对表达量。结果2周、4周、8周时,DN组小鼠血糖(Glu)、糖化血红蛋白(HbA1c)、血肌酐(Scr)、尿素氮(BUN)及尿酸(UA)水平均显著高于对照组小鼠,且HbA1c、Scr、BUN及UA水平随着DN病程延长而升高(DN组2周vs.对照组:t值分别为11.804、7.481、2.362、5.621、2.557;DN组4周vs.对照组:t值分别为14.431、9.936、4.449、7.329、3.977;DN组8周vs.对照组:t值分别为12.415、13.593、7.619、9.654、6.981,P<0.05)。DN组小鼠Notch1、Jagged1、Hes1和Hey1蛋白表达水平均显著高于对照组小鼠(t值分别为3.781、9.738、12.231、6.684,P<0.05)。DN组小鼠Notch1、Jagged1、Hes1和Hey1 mRNA相对表达量均显著高于对照组小鼠(t值分别为6.411、9.510、15.490、9.498,P<0.05)。结论DN小鼠肾组织中Notch信号通路蛋白表达明显增加,Notch信号通路可能在DN肾脏损害的发生和发展中发挥着重要作用。

Objective To explore the expression and significance of Notch signaling pathway in kidney tissues of mice with gestational diabetic nephropathy(DN).Methods Streptozotocin(STZ)was used to establish the DN mouse model.6 DN mice(intraperitoneal injection of STZ solution)and 6 control mice(intraperitoneal injection of equal amount of citrate buffer)were enrolled.At 2,4 and 8weeks,blood was collected from mice to detect biochemical indicators.At the end of the 8th week,the mice were sacrificed to take their kidney tissues.The kidney tissues of the two groups were observed by hematoxylin eosin(HE)staining.The expressions of Notch1 in the kidney tissues of the two groups of mice were observed by immunohistochemical staining.Western blot was used to detect the protein expression levels of Notch1,Jagged1,Hes1 and Hey1 in the Notch signaling pathway.Real-time polymerase chain reaction(PCR)was used to detect the relative expression levels of Notch1,Jagged1,Hes1 and Hey1 mRNA in the Notch signaling pathway.Results At 2,4,and 8 weeks,the levels of blood glucose(Glu),glycosylated hemoglobin(HbA1c),serum creatinine(Scr),blood urea nitrogen(BUN)and uric acid(UA)in the DN group were significantly higher than those in the control group,and the levels of HbA1c,Scr,BUN and UA increased with the prolonged course of DN(control group vs DN group at 2 weeks,t=11.804,7.481,2.362,5.621 and 2.557,respectively;control group vs DN group at 4 weeks,t=14.431,9.936,4.449,7.329 and 3.977 respectively;control group vs DN group at 8 weeks,t=12.415,13.593,7.619,9.654 and 6.918,respectively,P<0.05).The protein expression levels of Notch1,Jagged1,Hes1 and Hey1 were significantly higher in the DN group than those in the control group(t=3.781,9.738,12.231 and 6.684,respectively,P<0.05).The mRNA relative expression levels of Notch1,Jagged1,Hes1 and Hey1 in the DN group were all significantly higher compared with those in the control group(t=6.411,9.510,15.490 and 9.498,respectively,P<0.05).Conclusion The expression of Notch signaling pathway protein in kidney tissue of DN mice is significantly increased.Notch signaling pathway may play an important role in the occurrence and development of DN kidney damage.