论文部分内容阅读
目的 :建立血管内皮特异过表达microRNA-155(miR-155)转基因大鼠模型并观察其血压、蛋白尿等表型及妊娠负荷施加后的变化。方法:把外源miR-155基因插入血管内皮细胞特异表达VE-cadherin启动子下游,构建转基因表达载体,将所得载体线性化后,通过受精卵显微注射技术,构建miR-155转基因大鼠,并利用特异引物PCR法鉴定转基因大鼠基因型。通过尾套法、生化检测、组织学检查对未孕和妊娠雌性转基因大鼠的表型进行初步分析。结果:血管内皮特异性过表达miR-155转基因大鼠在胸主动脉、腹主动脉、肾脏、心脏组织miR-155表达较野生型鼠增高2.3、3.3、2.9、3.3倍。未妊娠时miR-155转基因雌性大鼠血压、尿蛋白、血管壁和肾脏结构与非转基因型大鼠比较,差异均无统计学意义。但妊娠后miR-155转基因雌性大鼠血压轻度增高、蛋白尿增加、胎儿生长受限发生率增加,胸/腹主动脉的血管壁结构紊乱和肾小管上皮空泡样变等。结论:血管内皮特异性miR-155转基因大鼠妊娠负荷增加后,表现出子痫前期表型。
OBJECTIVE: To establish a vascular endothelial specific overexpression microRNA-155 (miR-155) transgenic rat model and observe its phenotype such as blood pressure and proteinuria and the changes after pregnancy stress. Methods: The exogenous miR-155 gene was inserted into the downstream of VE-cadherin promoter in vascular endothelial cells to construct the transgene expression vector. After the vector was linearized, the miR-155 transgenic mice were constructed by microinjection of fertilized eggs, The genotypes of transgenic rats were identified by PCR. By tail cuff method, biochemical tests, histological examination of non-pregnant and pregnant female transgenic rats phenotype preliminary analysis. Results: The expression of miR-155 in thoracic aorta, abdominal aorta, kidney and heart tissue of vascular endothelial-specific overexpression miR-155 transgenic mice was 2.3, 3.3, 2.9 and 3.3 times higher than that of wild type mice. There was no significant difference in blood pressure, urinary protein, blood vessel wall and kidney structure between non-transgenic and transgenic rats with miR-155 in nonpregnant rats. However, after pregnancy, miR-155 transgenic female rats slightly increased blood pressure, increased proteinuria, increased fetal growth restriction, thoracic / abdominal aorta vascular wall structure disorder and tubular epithelial vacuolar degeneration. CONCLUSION: Vascular endothelial-specific miR-155 transgenic rats exhibit a phenotype of preeclampsia after increased pregnancy load.