Luciano Melissa N, da Silva Paulo H, Chaim Olga M, dos Santos Vera Lucia P, Franco Célia Regina C, Soares Maria Fernanda S, Zanata Silvio M, Mangili Oldemir C, Gremski Waldemiro, Veiga Silvio S
Department of Cell Biology, Federal University of Paraná, Brazil.
J Histochem Cytochem. 2004 Apr;52(4):455-67. doi: 10.1177/002215540405200404.
Brown spider (Loxosceles genus) venom causes necrotic lesions often accompanied by fever, hemolysis, thrombocytopenia, and acute renal failure. Using mice exposed to Loxosceles intermedia venom, we aimed to show whether the venom directly induces renal damage. The experimental groups were composed of 50 mice as controls and 50 mice that received the venom. Light microscopic analysis of renal biopsy specimens showed alterations including hyalinization of proximal and distal tubules, erythrocytes in Bowman's space, glomerular collapse, tubule epithelial cell blebs and vacuoles, interstitial edema, and deposition of eosinophilic material in the tubule lumen. Electron microscopic findings indicated changes including glomerular epithelial and endothelial cell cytotoxicity as well as disorders of the basement membrane. Tubule alterations include epithelial cell cytotoxicity with cytoplasmic membrane blebs, mitochondrial changes, increase in smooth endoplasmic reticulum, presence of autophagosomes, and deposits of amorphous material in the tubules. We also found that the venom caused azotemia with elevation of blood urea levels but did not decrease C3 complement concentration or cause hemolysis in vivo. Confocal microscopy with antibodies against venom proteins showed direct binding of toxins to renal structures, confirmed by competition assays. Double-staining immunofluorescence reactions with antibodies against type IV collagen or laminin, antibodies to venom toxins, and fluorescent cytochemistry with DAPI revealed deposition of toxins in glomerular and tubule epithelial cells and in renal basement membranes. Two-dimensional electrophoresis showed venom rich in low molecular mass and cationic toxins. By immunoblotting with antibodies to venom toxins on renal extracts from venom-treated mice, we detected a renal binding toxin at 30 kD. The data provide experimental evidence that L. intermedia venom is directly involved in nephrotoxicity.
棕色蜘蛛（洛氏蛛属）毒液会导致坏死性损伤，常伴有发热、溶血、血小板减少和急性肾衰竭。我们以暴露于中间洛氏蛛毒液的小鼠为研究对象，旨在探究该毒液是否直接引发肾损伤。实验组由50只作为对照的小鼠和50只接受毒液的小鼠组成。对肾活检标本进行的光学显微镜分析显示出多种改变，包括近端和远端肾小管玻璃样变、鲍曼囊内出现红细胞、肾小球塌陷、肾小管上皮细胞出现泡状突起和空泡、间质水肿以及嗜酸性物质在肾小管腔内沉积。电子显微镜检查结果表明存在多种变化，包括肾小球上皮细胞和内皮细胞的细胞毒性以及基底膜紊乱。肾小管改变包括上皮细胞的细胞毒性，伴有细胞质膜泡状突起、线粒体变化、滑面内质网增多、自噬体出现以及无定形物质在肾小管内沉积。我们还发现该毒液会导致氮质血症，血尿素水平升高，但在体内不会降低C3补体浓度或引发溶血。用针对毒液蛋白的抗体进行共聚焦显微镜检查显示毒素与肾脏结构直接结合，竞争试验证实了这一点。用抗IV型胶原蛋白或层粘连蛋白的抗体、抗毒液毒素的抗体进行双重染色免疫荧光反应，以及用DAPI进行荧光细胞化学分析，结果显示毒素沉积在肾小球和肾小管上皮细胞以及肾基底膜中。二维电泳显示毒液富含低分子量和阳离子毒素。通过用抗毒液毒素的抗体对经毒液处理的小鼠肾脏提取物进行免疫印迹分析，我们检测到一种30 kD的肾脏结合毒素。这些数据提供了实验证据，表明中间洛氏蛛毒液直接参与肾毒性作用。
