Satoh K, Wakui H, Komatsuda A, Nakamoto Y, Miura A B, Itoh H, Tashima Y
Third Department of Internal Medicine, Akita University School of Medicine, Japan.
Ren Fail. 1994;16(3):313-23. doi: 10.3109/08860229409044872.
We purified 90-kDa heat-shock protein (HSP90) from murine brains and produced a specific antibody against the protein in a rabbit. This antibody cross-reacted with rat renal HSP90 on immunoblot analysis. Using the antibody, we observed serial immunohistochemical localizations of HSP90 in rat kidneys with cisplatin-induced acute renal failure. In normal kidneys, HSP90 was mainly localized in the cytoplasm of distal tubules and collecting ducts. Twenty-four hours after the cisplatin exposure, a rapid expression of HSP90 was observed in the cytoplasm of epithelial cells in the Henle's loops (especially at the luminal side), although there was little change in these cells on light microscopy. Degenerative changes of epithelial cells appeared in the S3 segment of proximal tubules on day 3, and epithelial cell regeneration in this portion was found from day 5. On day 5, HSP90 was markedly expressed in both the cytoplasm and the nucleus of epithelial cells in the S3 segment with a granular pattern. The induced HSP90 was then accumulated in the cytoplasm of these cells on day 7 and disappeared on day 14. Immunoblot analysis of isotonic buffer-extractable renal fractions showed that there was a rapid induction of HSP90 from day 1, and that the maximum induction of HSP90 in the extract at day 5 was 6-fold that of a control. These results suggest that HSP90 plays some role related to functional abnormalities of the Henle's loops at the luminal side, and in the regeneration of damaged cells in the S3 segment of proximal tubules, during the course of cisplatin-induced acute tubular injury.
我们从小鼠大脑中纯化了90-kDa热休克蛋白（HSP90），并在兔体内制备了针对该蛋白的特异性抗体。在免疫印迹分析中，该抗体与大鼠肾脏HSP90发生交叉反应。利用该抗体，我们观察了顺铂诱导的大鼠急性肾衰竭肾脏中HSP90的系列免疫组织化学定位。在正常肾脏中，HSP90主要定位于远端小管和集合管的细胞质中。顺铂暴露24小时后，在亨氏袢上皮细胞的细胞质中观察到HSP90的快速表达（尤其是在管腔侧），尽管在光学显微镜下这些细胞几乎没有变化。第3天近端小管S3段出现上皮细胞退行性改变，第5天发现该部位上皮细胞再生。第5天，S3段上皮细胞的细胞质和细胞核中均有明显的HSP90颗粒状表达。诱导的HSP90在第7天积聚在这些细胞的细胞质中，并在第14天消失。等渗缓冲液可提取的肾脏组分的免疫印迹分析表明，从第1天开始HSP90迅速诱导，第5天提取物中HSP90的最大诱导量是对照的6倍。这些结果表明，在顺铂诱导的急性肾小管损伤过程中，HSP90在亨氏袢管腔侧功能异常以及近端小管S3段受损细胞的再生中发挥了一定作用。
