摘要
目的探讨维生素D(VD)对甲状腺功能减退(HT)肾损伤幼鼠肾小管上皮细胞间充质转化(EMT)的影响,以及其对肿瘤坏死因子受体相关因子6(Traf6)/转化生长因子-β活化激酶1(TAK1)通路的调控机制。方法通过丙基硫尿嘧啶(PTU)灌胃构建幼鼠HT模型,以过表达TAK1(pc DNA3.1-TAK1)作功能挽救实验;50只SPF级雄性SD大鼠分为正常组、HT组、VD低剂量(HT+VD-L)组、VD高剂量(HT+VD-H)组、HT+VD-H+pc DNA3.1-TAK1(HT+VD-H+pc)组,每组10只。全自动生化仪检测各组大鼠血清血肌酐(Scr)和血尿素氮(BUN)的含量;脱氧核糖核苷酸末端转移酶介导的缺口末端标记法试剂盒(TUNEL)检测肾组织中的细胞凋亡;免疫组化检测肾组织中转化生长因子β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)和上皮钙黏蛋白(E-cadherin)的表达;Western blot法检测肾组织中B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、Traf6、TAK1和磷酸化TAK1(p-TAK1)的表达。结果VD能明显降低HT幼鼠血清中Scr和BUN的含量,下调肾组织中的细胞凋亡率,降低肾组织中TGF-β1和α-SMA的表达,上调E-cadherin的表达;抑制肾组织中Traf6、p-TAK1和Bax的表达,升高肾组织中Bcl-2的表达,差异均具有统计学意义(均P<0.05)。结论维生素D能抑制HT幼鼠肾小管上皮细胞的EMT,降低肾组织中的细胞凋亡率,减轻肾组织的病理损伤,改善其肾功能,这与抑制Traf6/TAK1信号的激活有关。
Objective To investigate the effect of vitamin D(VD)on epithelial mesenchymal transition(EMT)of renal tubular epithelial cells in infant rats with hypothyroidism(HT)and its regulatory mechanism on tumor necrosis factor receptor related factor 6(Traf6)/transforming growth factorβactivated kinase 1(TAK1)pathway.Methods Propylthiouracil(PTU)was used to establish the HT model of infant rat by intragastric administration.Overexpression of TAK1(pc DNA3.1-TAK1)was used to perform the functional salvage experiment.The animals were divided into normal group,HT group,VD low dose(HT+VD-L)group,VD high dose(HT+VD-H)group and HT+VD-H+pc DNA3.1-TAK1(HT+VD-H+pc)group with 10 rats in each group.Automatic biochemical analyzer was used to detect the contents of creatinine(Scr)and blood urea nitrogen(BUN)in serum.Terminal-deoxynucleoitidyl transferase-mediated nick end labeling was used to detect the apoptosis in renal tissue.Immunohistochemistry was used to detect the expressions of transforming growth factor-β1(TGF-β1),α-smooth muscle actin(α-SMA)and E-cadherin(E-cadherin)in renal tissue.Western blot was used to detect the expressions of B cell lymphoma-2(Bcl-2),Bcl2-associated x protein(Bax),Traf6,TAK1 and phosphorylated TAK1(p-TAK1)in renal tissue.Results VD could significantly decrease the contents of Scr and BUN in serum,down-regulate the rate of apoptosis in renal tissue,decrease the expression of TGF-β1 andα-SMA in renal tissue,up-regulate the expression of E-cadherin,inhibit the expression of Traf6,p-TAK1 and Bax in renal tissue,and increase the expression of Bcl-2 in renal tissue.And the differences were statistically significant(all P<0.05).Conclusion Vitamin D can inhibit the EMT of renal tubular epithelial cells,reduce the rate of apoptosis in renal tissue,reduce the pathological injury of renal tissue and improve renal function in infant rats with HT,which is related to the inhibition of the activation of Traf6/TAK1 signaling pathway.
作者
李鸿燕
张丽敏
冀娟
刘旭颖
LI Hongyan;ZHANG Limin;JI Juan;LIU Xuying(Department of Endocrinology,The Affiliated Hospital of Hebei University,Baoding 071000,Hebei,China;Department of Nephrology,The Affiliated Hospital of Hebei University,Baoding 071000,Hebei,China)
出处
《西部医学》
2024年第8期1115-1122,共8页
Medical Journal of West China
基金
保定市科技计划项目(2241ZF303)。
