摘要
目的 研究暴发性肝衰竭小鼠肾脏I型1,4,5-三磷酸肌醇受体（IP3R）表达的变化。方法 采用D-氨基半乳糖和内毒素联合腹腔注射制备暴发性肝衰竭小鼠动物模型。动物随机分为4组,即等渗盐水对照组、LPS对照组、D-氨基半乳糖对照组、暴发性肝衰竭组（2、6、9 h）。应用免疫组织化学、Western blot及RT-PCR技术检测暴发性肝衰竭进程中,小鼠肾脏IP3R蛋白质的定位、表达及mRNA表达的变化。结果 Ⅰ型IP3R主要分布于肾小球系膜细胞和血管平滑肌细胞的胞浆内。在暴发性肝衰竭组小鼠,6 h时I型IP3R免疫组织化学染色阳性细胞开始增加,9 h时更为明显（6 h：x^2=7．11,P〈0．01,9 h： x^2=9．15,P〈0.01）。Western blot结果与免疫组织化学结果一致,6 h时开始增加（t=3．16,P〈0．05）,9 h达到最高值（t=5．43,P〈0．01）,与等渗盐水对照组相比差异有统计学意义。RT PCR结果显示暴发性肝衰竭小鼠2 h时I型IP3R mRNA即开始增加,6 h时达到最高值,9 h时有所恢复,但与等渗盐水对照组相比差异均有统计学意义（2 h：t=2．47,P〈0．05,6 h：t=4．42,P〈0.01,9 h：t=2．16,P〈0．05）。结论 在暴发性肝衰竭进程中,肾脏I型1,4,5-三磷酸肌醇受体表达增强,其mRNA也呈上调趋势。
Objective To study the changes of expression of type Ⅰ inositol 1,4,5 triphosphate receptors （IP3RI） in the kidneys of mice with fulminant hepatic failure （FHF） in order to understand the role renal vasoconstriction plays in the development of hepatorenal syndrome （HRS）. Methods One hundred twenty male Balb/c mice were divided into 4 groups. In the fourth group （60 mice） lipopolysaccharide with Dgalactosamine was injected intraperitoneally to induce acute liver necrosis. The first-third groups （20 mice in each group） served as controls and those mice were given NS, LPS or GalN intraperitioneally. At the end of 2 h, 6 h, and 9 h, mice were sacrificed and their livers and kidneys were removed and examined histologically. Immunohistochemistry, Western blot and reverse transcription PCR （RT-PCR） were used to detect the distribution and expression of IP3RI in the kidneys. Results IP3RI protein was localized in the cytoplasma of glomerular mesangial cells and vascular smooth muscle cells in the kidneys. In the kidney tissues from mice with FHF at 6 h and 9 h, IP3RI-positive staining cells increased significantly （6 h： x^2 = 7.11, P 〈 0.01; 9 h： x^2 = 9.15, P 〈 0.01）. Western blot demonstrated a consistent and significant increase of IP3RI expression in mice with FHF at 6 h and 9 h （6 h： t = 3.16, P 〈 0.05; 9 h： t = 5.43, P 〈 0.01）. Using RT-PCR we observed that IP3RI mRNA in FHF samples at 2 h, 6 h and 9 h was markedly up-regulated in comparsion to that of the controls （2 h：t = 2.47, P 〈 0.05; 6 h： t = 4.42, P 〈 0.01; 9 h： t = 2.16, P 〈 0.05）. Conclusion The expression of IP3RI protein increased in glomerular mesangial cells and renal vascular smooth muscle cells of FHF mice. Perhaps this was caused by IP3RI mRNA up-regulation.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2007年第6期403-407,共5页
Chinese Journal of Hepatology
基金
国家自然科学基金（30270607）
作者简介
闻颖 女，34岁，副主任医师。
通讯作者：刘沛，Email：syliupei2003@yahoo．com.cn
