蚓激酶对UUO大鼠肾组织NOX4、FAK、Src的影响

Effects of lumbrokinase on expression of NOX4,FAK,Src in renal interstitial fibrosis of unilateral ureteral obstruction(UUO)rats

目的:观察蚓激酶(lumbrokinase)对单侧输尿管梗阻(UUO)大鼠肾组织NADPH氧化酶4(NOX4)、黏着斑激酶(FAK)、Src激酶表达的影响,并探讨其潜在机制。方法:雄性Sprauge-Dawley(SD)大鼠50只,随机分为5组,每组10只,即假手术组、模型组、蚓激酶低、中、高剂量组(0.9×10^(5)、1.8×10^(5)、3.6×10^(5) U·kg^(-1)),术后第2天开始灌胃给药,假手术组、模型组大鼠给予等体积生理盐水灌胃,连续14 d。免疫组化法检测转化生长因子β1(TGFβ1)、NOX4、FAK、Src蛋白表达变化,Western印迹法和RT-PCR法检测TGFβ1、NOX4、FAK、Src蛋白和mRNA表达变化。结果:与假手术组相比,模型组TGFβ1、NOX4、FAK、Src、α-SMA蛋白和mRNA水平显著升高(均P<0.05)。与模型组比较,免疫组化结果显示,蚓激酶组TGFβ1、NOX4、FAK、Src蛋白水平明显降低(F=10.57、7.086、11.23、7.750,均P<0.05);Western印迹结果显示,蚓激酶组TGFβ1、NOX4、FAK、Src、α-SMA蛋白水平明显降低(F=8.873、11.18、6.902、8.679、5.672,均P<0.05);RT-PCR结果显示,蚓激酶组TGFβ1、NOX4、FAK、Src mRNA水平明显降低(F=5.546、4.285、6.626、3.911、5.914,均P<0.05)。结论:蚓激酶可改善UUO模型大鼠肾间质纤维化(RIF),其机制可能与蚓激酶调控TGFβ1、NOX4、FAK、Src、α-SMA的表达有关。

Objective:To investigate the effects andmechanismof lumbrokinase on expression ofNADPHoxidase 4(NOX4),focal adhesion kinase(FAK)and proto-oncogene tyrosine-protein kinase(Src)in unilateral ureteral obstruction(UUO)-induced renal interstitial fibrosis of rats.Methods:A rat model of UUO was established through ligation of a unilateral ureter.Fifty male Sprauge-Dawley(SD)rats were randomly divided into 5 groups:Sham group,Model group,Low,Medium and High-doses of lumbrokinase treatment groups(0.9×10^(5),1.8×10^(5),3.6×10^(5) U·kg^(-1))with intragastric administration for 14 d.Immunochemistry was used to detect the protein expression of transforming growth factorβ1(TGFβ1),NOX4,FAK,Src,Western blotting and RT-PCR were used to detect the protein and mRNA level of TGFβ1,NOX4,FAK,Src and alpha-smooth muscle actin(α-SMA).Results:Compared with the sham group,the protein and mRNA level of TGFβ1,NOX4,FAK,Src andα-SMA in the model group were increased significantly(all P<0.05).Compared with the UUO group,the protein level of TGFβ1,NOX4,FAK,Src in treatment groups by immunochemistry were decreased significantly(F=10.57,7.086,11.23,7.750,all P<0.05),the protein level of TGFβ1,NOX4,FAK,Src andα-SMA in treatment groups by Western blotting were decreased significantly(F=8.873,11.18,6.902,8.679,5.672,all P<0.05),the mRNA level of TGFβ1,NOX4,FAK,Src andα-SMA in treatment groups byRT-PCRwere decreased significantly(F=5.546,4.285,6.626,3.911,5.914,all P<0.05).Conclusion:Lumbrokinase attenuates renal interstitial fibrosis of UUO rats.Its mechanism is related to the regulation of TGFβ1,FAK,Src,NOX4 andα-SMA expression.