新生大鼠肾脏缺血性损伤致肾小管上皮细胞ADF与G-ACTIN的改变

Alternations of ADF and G-actin in Renal Tubular Epithelial Cell of Neonatal Rats During Kidney Ischemic Injury.

目的了解ADF和G-ACTIN在新生大鼠肾脏缺血损伤中的变化情况,探讨ADF对肌动蛋白细胞骨架的影响机理及其在损伤发病机制中所起的作用。方法新生大鼠分为缺血10MIN组、缺血30MIN组和对照组,每组各8只。通过钳夹肾蒂不同时段,建立新生大鼠肾脏缺血动物模型。同一张肾脏组织切片上,同时使用兔抗鸡ADF抗血清和鼠G-ACTIN单克隆抗体,通过FITC和TRITC标记的不同二抗检测实验组和对照组ADF和G-ACTIN在肾小管上皮细胞的表达情况。结果对照组ADF和G-ACTIN染色均匀地分散在肾小管上皮细胞胞浆内。缺血损伤后,ADF和G-ACTIN的分布发生明显变化,出现在TEC顶侧区的聚集,同时肾小管腔颗粒中也出现两者的染色。ADF的表达在缺血前后差异无统计学意义(P>0.05),G-ACTIN的表达在缺血后出现增加,差异有统计学意义(P>0.05)。结论生理情况下,ADF和G-ACTIN在肾小管上皮细胞内呈弥散分布。缺血损伤使ADF激活并向顶侧区聚集,解聚微绒毛轴心的F-ACTIN束,出现该处G-ACTIN的浓集,造成微绒毛肌动蛋白骨架的破坏,出现微绒毛的断裂、脱落等病理变化。

Objective To study the changes of ADF and G-actin in renal tubular epithelial cell of neonatal rats during kindney ischemic injury. To explore the interactions between ADF and G-actln and their relationships with actin cytoskeleton. Methods Twenty-four neonatal rats were used to the experiments due to have the weights from 30 to 40 g respectively. Renal ischemia was induced by clamping the left renal pedicle for different periods of time that was in 10 min or 30 min. The kidneys of normal neonatal rats were functioned as the control group. Two kinds of primary antibodies, which one was rabbit anti-chick ADF antiserum and another was mouse monoclonal G-actin antibody, were used on one same section of kidney tissue. FITC-labeled and TRITC-labeled secondary antibodies were used to identify ADF and G-actin respectively. These samples were examined by using a fluorescene microscope. Results In control group, ADF and G-actin showed to be diffused in cytoplasm of renal tubular epithelial cell. In ischemic injury group, the distributions of ADF and G-actin were changed significantly to apical region of tubular epithelial cells and the renal tubula lumen. Regardless of the ADF expression to not change but G-actin was increased after kidney ischemia. Conclusion Under the physiolgical condition, ADF and G-actin got a diffuse distribution in cytoplasm of renal tubular epitbelial cell. However, when the kidney got in ischemia, the distributions of ADF and G-actin were changed significantly to apica region of tubular epithelial cell or the renal tubular lumen.