Induction and subcellular localization of protein kinase C isozymes following renal ischemia
Induction and subcellular localization of protein kinase C isozymes following renal ischemia
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DOI：
10.1046/j.1523-1755.2001.0590051789.x
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发表时间：
2001-05-01
影响因子：
19.6
通讯作者：
Padanilam, BJ
中科院分区：
文献类型：
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作者：
Padanilam, BJ
背景我们以前曾报道过，活化C激酶受体（RACK 1）的表达诱导缺血后/预灌注损伤的肾脏，蛋白激酶C（PKC）的活化保护肾细胞免受缺氧损伤。本研究旨在确定RACK 1的诱导表达是否伴随着PKC同工酶表达水平和亚细胞分布的变化。采用双侧肾动脉夹闭60 min的方法，造成大鼠急性肾功能衰竭的缺血/再灌注损伤模型。通过免疫印迹分析证明了全肾匀浆中可溶性和颗粒组分之间各种PKC同工酶的表达水平和易位。采用免疫组化法检测损伤后肾组织中PKC同工酶的表达。诱导PKC α、β II和ζ从可溶性部分移位到损伤后的颗粒部分。免疫定位显示，在缺血/再灌注损伤（IRI）后0至30分钟，近端小管上皮细胞（PTEC）中诱导PKC α，β III和zeta表达。在一天伤后，α同工酶易位到质膜的未受损的PTEC，而它易位到细胞核在受损的PTEC。PKC β II表达沿着未损伤的PTEC的基底和侧面，而在损伤的PTEC中，其分布于脱落细胞的胞浆中。一天后，PKC ζ表达沿着受损PTEC的顶侧。在损伤后7天，α和zeta同工酶的表达定位于再生的PTEC的质膜，PKC β II同工酶的表达定位于某些间质鳗鱼。诱导表达。易位，和细胞内的空间分布的酶表明，他们可能介导的IRI过程中的多个过程。
Background. We have previously reported that the expression of the receptor for activated C kinase (RACK1) is induced post-ischemia/preperfusion injury to the kidney, and activation of protein kinase C (PKC) protects renal cells from hypoxic injury. This study was done to determine whether the induced expression of RACK1 is accompanied by changes in the level of expression and subcellular distribution of PKC isozymes.Methods. Ischemia/reperfusion injury resulting in acute renal failure was induced by 60 minutes of bilateral renal artery clamping in rats. The expression levels and translocation of various PKC isozymes between soluble and particulate fractions in whole kidney homogenates were demonstrated by immunoblot analysis. The expression pattern of the various PKC isozymes in the kidney postinjury was performed by immunohistochemistry.Results. PKC alpha, beta II, and zeta were induced and translocated from the soluble fraction to the particulate fraction post-injury. Immunolocalization showed PKC alpha, beta III and zeta expression to be induced in the proximal tubule epithelial cell (PTEC) at 0 to 30 minutes post-ischemia/reperfusion injury (IRI). At one-day postinjury, the alpha isozyme was translocated to the plasma membrane of the undamaged PTEC, while it was translocated to the nucleus in damaged PTEC. PKC beta II expression was along the basal and lateral side of the undamaged PTEC, while it was distributed in the cytoplasm of sloughed cells in the damaged PTEC. PKC zeta expression at one day was along the apical side of the damaged PTEC. At seven-days postinjury, the expressions of the alpha and zeta isozymes were localized to the plasma membrane of the regenerating PTEC and the expression of PKC beta II isozyme to certain interstitial eels.Conclusion. The induced expression. translocation, and the intracellular spatial distributions of the enzymes suggest that they may mediate multiple processes during IRI.