摘要
目的:细胞极性是肾小管上皮细胞发挥生理功能的结构基础。本研究的目的是明确成体大鼠肾小管坏死后修复过程中与胚胎大鼠肾小管发育过程中细胞极性形成过程的一致性。方法:通过皮下注射硫酸庆大霉素制作出生后8周雄性Wistar大鼠急性肾小管坏死后自然修复的动物模型,分别在注射庆大霉素后第7天(用药5d,停药2d)、第14天和第28天采集肾脏标本;另外采集妊娠20d大鼠的胚胎肾脏标本。观察成体大鼠肾小管坏死后修复再生过程中与胚胎大鼠肾小管发育过程中肾小管的形态学变化;以Na+-K+-ATP酶作为细胞极性的标志物,采用免疫荧光染色技术对比观察成体大鼠肾小管坏死后修复再生过程中与胚胎大鼠肾小管发育过程中细胞极性形成的过程。结果:(1)成体大鼠注射庆大霉素后第7天,肾小管上皮细胞坏死、脱落,肾小管基底膜裸露,管腔内有大量脱落的上皮细胞碎片;第14天,肾小管基底膜出现新再生的肾小管上皮细胞;第28天,大多数肾小管结构基本恢复正常。(2)成体大鼠注射庆大霉素后第7天,裸露的肾小管基底膜没有Na+-K+-ATP酶的表达;第14天,新再生的肾小管上皮细胞有Na+-K+-ATP酶的表达,但此时Na+-K+-ATP酶没有明显的极性分布,胞膜和胞浆均有表达;第28天,Na+-K+-ATP酶呈极性分布,主要表达在肾小管上皮细胞的侧基底膜。(3)胚胎大鼠肾小管发育过程是从S形体发育成为不成熟的肾小管,再发育成为成熟的肾小管。(4)胚胎大鼠肾小管发育过程中,Na+-K+-ATP酶的表达从没有极性到有极性分布,即Na+-K+-ATP酶从肾小管上皮细胞的胞膜和胞浆均有表达到只局限在侧基底膜表达。结论:成体大鼠肾小管坏死后修复过程中细胞极性形成的过程与胚胎大鼠肾小管发育过程中细胞极性形成的过程是一致的,提示大鼠肾小管坏死后修复再生的过程类似肾小管胚胎发育的过程。
Objective： Cell polarity is the structural basis that plays an important role in the physiological function of the renal tubular epithelial cells. Our study aimed to clear the consistency of the cell polarity formation during the repair/regeneration after renal tubular necrosis in adult rat and the renal tubular development in embryonic rat. nethods：A natural repair and regeneration model after acute tubular necrosis was made in eight - week - old male Wistar rat by injecting gentamicin subcutaneously for five days. The rats were sacrificed on days 7, 14 and 28 after the first gentamicin injection respectively and the bilateral kidneys of these rats were rapidly removed. In addition, the embryonic kidneys of rats were removed on the 20th day of pregnancy. The morphological changes of the renal tubules during the repair and regeneration after renal tubular necrosis and the development in rat were observed. Through using Na ^＋ - K ^＋ - ATPase as the marker of cell polarity, the immunofluoreseence staining technique was used to observe comparatively the formation of cell polarity in these two processes. Results： （ 1 ） On day 7 after the first administration of gentamicin, necrosis and detachment of renal tubules epithelial cell were found, the tubular basement membrane tended to be exposed, and a large number of epithelial cell debris was observed in the lumen; on day 14, regenerative renal tubular epithelial cells could be observed along the tubular basement membrane; on day 28, most of the tubular structure basically returned to normal. （2） On day 7 after the first administration, Na ^＋ - K ^＋ - ATPase didn＇ t express in the exposed basement membrane; on day 14, Na ^＋ - K^＋ - ATPase expressed in cytomembrane and cytoplasm of the renal regenerative tubular epithelial cells, and its expression wasn＇t polar contributed; on day 28, the expression of Na ^＋ - K ^＋ - ATPase was polar contributed, and mainly in the lateral - basal plasma membrane. （3） The process of development of rat renal tubules was from S - shaped body to immature tubules to mature tubules finally. （4） In the process of devel- opment of rat renal tubules, the expression of Na ^＋ -K^＋ -ATPase was from non polar contribution to polar contribution, as is from cytomembrane and cytoplasm to only in the lateral - basal plasma membrane. Conclusion： The process of cell polarity formation during repair/regeneration after renal tubular necrosis in adult rat is consist with the process of cell polarity formation during renal tubular de- velopment in embryonic rat, suggesting the repair processes after renal tubular necrosis in adult rat are similar as the processes of renal tubular development in embryonic rat.
出处
《中国中西医结合肾病杂志》
2013年第5期381-383,I0001,共4页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
国家重大科学研究计划项目(No.2011CB944004)
国家自然科学基金面上项目(No.30971377)
