TLR4基因敲除对sFlt-1诱导子痫前期小鼠模型临床特征的影响

The Role of TLR4 silence in Adv-sFlt-1 induced mice pre-eclampsia-like phenotype

目的探讨Toll样受体4(Toll-like receptor 4,TLR4)在编码sFlt-1的复制缺陷型重组腺病毒(adenoviral vector containing mouse full-length soluble fms-like tyrosine kinase 1,Adv-sFlt-1)诱导子痫前期小鼠模型临床特征中作用。方法构建Adv-sFlt-1子痫前期临床特征小鼠模型,观察TLR-4基因敲除对Adv-sFlt-1引起小鼠子痫前期临床特征的影响,酶联免疫吸附试验(Enzyme-linked immunosorbent assay,ELISA)测定孕鼠血浆可溶性fms样酪氨酸激-1(soluble fms-like tyrosine kinase 1,sFlt-1)含量,无创尾套法检测小鼠血压、双辛丁酸(bicinchoninic acid,BCA)法检测尿蛋白,肌酐检测试剂盒检测肌酐比值、苏木素-伊红(hematoxylin-eosin staining,HE)染色及透射电镜(transmission electron microscope,TEM)观察肾小球滤膜病理改变。结果 Adv-sFlt-1处理组血压、尿蛋白/肌酐比值明显高于正常对照组、空病毒处理组及TLR-4基因敲除Adv-sFlt-1处理组(P<0.05);Adv-sFlt-1处理组小鼠肾小球滤膜增厚,毛细血管内血栓形成、内皮细胞及足细胞发生病理改变,正常对照组、空病毒处理组及TLR-4基因敲除加Adv-sFlt-1处理组肾小球病理改变不明显。结论成功构建Adv-sFlt-1导致子痫前期临床特征小鼠模型,TLR4基因敲除可逆转AdvsFlt-1导致子痫前期小鼠模型的临床特征。TLR4可能在子痫前期病理过程中起重要作用。

Objective ： We tested whether the female mice lacking TLR4 restored the pre - eclampsia - like phenotype induced by increased sFh - 1. Methods ： The soluble fms - like tyrosine kinase - 1 （ sFh - 1 ） was detected by ELISA. Blood pressure was moni- tored by using a carotid catheter - calibrated eight chamber tail - cuff system. Total urine protein were detected by bicinchoninic acid （BCA） method using a kit. urinary creatinine levels were detected by creatinine Assay Kit. Hematoxylin and eosin （HE） and trans- mission electron microscope （TEM） were used for detecting the histopathological changes of glomeruli of kidney. Results ： The pre - eclampsia - like phenotype of mice was induced by Adv - sFlt - 1. The blood pressure, urine protein/creatinine ratio of Adv - sFh - 1 mice were higher than those of TLR4 - deficient sFlt - 1 mice, Adv - GFP mice and non - treated mice （P 〈 0. 05 ）. The glomerulus shows a ＇ bloodless＇ appearance with a swelling of the capillary endothelial cells , occluded or narrowed lumens, and the thickening of glomerular filtration membrane, microthrombi in capillary lumen in Adv - sFh - 1 mice, but did not see in TLR4 - deficient sFh - 1 mice , Adv - GFP mice and non - treated mice. Conclusion ： Female mice lacking TLR4 reverse the pre - eclampsia - like phenotype induced by increased sFh - 1. TLR4 could play an important role in the pre - eclampsia - like phenotype induced by increased sFh - 1.