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中华妇幼临床医学杂志(电子版) ›› 2005, Vol. 01 ›› Issue (02) : 110 -封一. doi: 10.3877/cma.j.issn.1673-5250.2005.02.113

论著

缺血/再灌注损伤对乳鼠肾小管上皮细胞辅肌动蛋白表达和分布变化的影响
姚裕家, 余波, 杨凡, 陈大鹏   
  1. 四川大学华西第二医院儿科(成都,610041)
  • 出版日期:2005-08-25
  • 基金资助:
    本研究为四川大学华西医学中心CMB基金资助项目(00-722)

The changes of α-actinin in renal tubular epithelial cells during kidney ischemia/reperfusion injury in neonatal rats

Yu-jia YAO, Bo YU, Fan YANG, Da-peng CHEN   

  1. Department of Pediatrics, the Second University Hospital, Sichuan University, Chengdu, 610041, China.
  • Published:2005-08-25
引用本文:

姚裕家, 余波, 杨凡, 陈大鹏. 缺血/再灌注损伤对乳鼠肾小管上皮细胞辅肌动蛋白表达和分布变化的影响[J]. 中华妇幼临床医学杂志(电子版), 2005, 01(02): 110-封一.

Yu-jia YAO, Bo YU, Fan YANG, Da-peng CHEN. The changes of α-actinin in renal tubular epithelial cells during kidney ischemia/reperfusion injury in neonatal rats[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2005, 01(02): 110-封一.

目的

通过研究缺血/再灌注不同时段辅肌动蛋白(α-actinin)在乳鼠肾小管上皮细胞(RETs)中分布变化,探讨α-actinin在肾脏缺血/再灌注损伤中所起作用。

方法

取出生14d的SD大鼠,采用先钳夹单侧肾动脉再松开的方式,建立不同时段肾脏缺血/再灌注模型,并设立正常对照组。应用免疫荧光法检测α-actinin在RETs上的分布和表达,并分析、计算各组α-actinin的阳性表达量。然后,对所得数据使用SPSS统计软件做方差分析。

结果

正常对照组α-actinin主要集中分布在RETs基底膜附近和管腔面附近的胞浆中。缺血0. 5h组,管腔面附近的α-actinin明显减少,基底膜附近可见α-actinin染色。再灌注2h组,管腔面附近的α-actinin消失,基底膜附近的α-actinin也减少,其连续性遭到破坏。再灌注120h组,α-actinin在基底膜附近的分布与正常对照组比较差异无显著意义,在管腔面附近仍有分布和对照组比较,差异仍然无显著意义。在基底侧,再灌注2h组α-actinin的表达量明显低于对照组,差异有显著意义(P<0. 05)。缺血0.5h组和再灌注120h组α-actinin的表达量和对照组相比,差异无显著性意义(P>0. 05)。在管腔侧,缺血0. 5h组、再灌注2h组和再灌注120h组α-actinin的表达量,均明显低于对照组,差异有显著意义(P<0.05)。

结论

α-actinin正常情况下,分布于RETs基底侧和管腔侧。缺血/再灌注损伤使其表达和分布减少,这可能导致微绒毛的破坏和整合素的去极化而影响肾功能。

Objective

To study the role of α-actinin in kidney ischemia/reperfusion injury by investigating the changes of α-actinin expression and its distribution in renal tubular epithelial cells (RETs) at different ischemia/reperfusion time stages.

Methods

The unilateral renal artery of neonatal SD rats was clamped for 0. 5h, then the kidneys were reperfused by removing the clamp for different time intervals. Immunofluorescent staining was used to show the distribution and expression of α-actinin in renal tubular epithelial cells. The quantities of α-actinin were counted by ImagePlus-Pro system. All data were analyzed with SPSS software.

Results

In the control, α-actinin located mainly in the plasma near the basal and apical membrane. After ischemia for 0. 5h, the distribution of α-actinin around the apical membrane decreased significantly, and some α-actinin could be detected in the lumen. After reperfusion for 2h, no α-actinin could be seen near the apex and those near the basal membrane also decreased, showing a damaged continuity. After reperfusion for 120h, compared with the control the distribution of α-actinin showed no difference in the basal region but reduced in the apical region. After 2h of reperfusion, the expression of α-actinin in the basal region was significantly lower than the control. After 2h and 120h of reperfusion, the expression of α-actinin in the apical region was significantly lower than the control.

Conclusion

Normally α-actinin is located in the plasma near the basal and apical membrane of RETs. Under ischemia/reperfusion injury, its expression and distribution drop down, which may induce the damage of microvilli and the depolarization of integrin of RETs, and thus affects the renal function.

图1 对照组α-actinin的分布 ×400
图2 缺血0.5h组α-actinin的分布× 400
图3 再灌注2h组α-actinin的分布 ×400
图4 再灌注120h组α-actinin的分布×400
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