摘要
目的探讨糖尿病大鼠肾组织丝裂原活化蛋白激酶磷酸酶-1(MKP-1)的表达特征及在细胞外基质重构中的作用。方法雄性Wister大鼠60只,腹腔一次性注射链脲佐菌素(STZ,65 mg/kg)制备糖尿病大鼠模型,48 h后血糖≥16.7 mmol/L,尿糖3+~4+者入选糖尿病组(DM),DM组造模成功后1、2、4、8、12周各组宰取10只大鼠,另取10只作为对照组。免疫组化检测肾组织MKP-1和磷酸化p38丝裂原活化蛋白激酶(p38 MAPK)层粘连蛋白(LN)的表达。Western印迹检测肾组织MKP-1和磷酸化p38 MAPK的水平,RT-PCR检测MKP-1和纤维粘连蛋白(FN)mRNA的表达。结果与对照组相比,糖尿病组大鼠肾组织1~8周MKP-1蛋白及mRNA的表达降低(P<0.01),磷酸化p38 MAPK水平在第1、2、4、8周表达升高(P<0.01),在第12周差异无统计学意义(P>0.05);LN 4~12周均高于对照组(P<0.01),FN mRNA 2~12周均高于对照组(P<0.01)。结论 MKP-1在糖尿病肾组织中表达下降,在细胞外基质重构中发挥一定作用,可能与p38 MAPK的去磷酸化有关。
Objective To investigate expression characteristics of mitogen-activated protein kinase phosphatase-1（ MKP-1） in renal tissues of rats with diabetic mellitus（ DM） and its effect on extracellular matrix（ ECM） reconstruction. Methods A total of 60 male Wister rats were used to establish diabetic models by injecting Streptozotocin（ STZ,65 mg/kg） intraperitoneally for one time. At 48 h after injection,rats with blood glucose equal or larger than16. 7 mmol/L and 3 ＋-4 ＋ harnzucker were selected as DM group,and then rats were respectively sacrificed in the 1 st,2 nd,4 th,8 thand 12 thweeks after successful model establishment for 10 rats in one time,and other 10 rats were selected as control group. Immunohistochemistry test was used to detect expressions of MKP-1,p38 mitogen-activated protein kinase（ p38 MAPK） and laminin（ LN） in renal tissues. Western blot method was used to detect expressions of MKP-1 and phosphorylation p-p38 MAPK,and reverse transcription-polymerase chain reaction（ RT-PCR） was used to detect expressions of MKP-1 and fibronectin（ FN） mRNA. Results Compared with those in control group,expressions of MKP-1 protein and mRNA renal tissues of rats with diabetic mellitus in the 1 st-8 thweeks after successful model establishment were significantly decreased（ P〈0. 01）; phosphorylation p-p38 MAPK expressions in the 2 nd-8 thweeks after successful model establishment were significantly increased（ P〈0. 01）,while the phosphorylation p-p38 MAPK expression in the 12 th weeks after successful model establishment showed no significant difference（ P〈0. 05）; LN expressions were gradually increased in the 4 th-12 thweeks compared with those in control group（ P〈0. 01）; FN mRNA expressions were significantly increased from the 2 ndto 12 thweeks（ P〈0. 01）. Conclusion MKP-1 expression can be found decreased in renal tissues of rats with diabetic mellitus,and it plays a certain role in pathogenesis of extracellular matrix reconstruction,which may be related to dephosphorylation of p38 MAPK pathway.
作者
王丽晖
吴广礼
林静
黄旭东
杨新军
汪晶华
陈云爽
赵维
WANG Li-hui;WU Guang-li;LIN Jing;HUANG Xu-dong;YANG Xin-jun;WANG Jing-hua;CHEN Yun-shuang;ZHAO Wei(Departmet of Nephrology,Bethune International Peace Hospital of PLA,Shijiazhuang 050082,Chin)
出处
《解放军医药杂志》
CAS
2018年第5期9-14,共6页
Medical & Pharmaceutical Journal of Chinese People’s Liberation Army
基金
河北省自然科学基金资助项目(H2016206578)
