Cruz Anselmo Hernández, Mendonça Ronaldo Z, Petricevich Vera L
Facultad de Medicina, Universidad Autónoma de Estado de Morelos, Mexico.
Mediators Inflamm. 2005 Dec 14;2005(6):349-59. doi: 10.1155/MI.2005.349.
Crotalus durissus terrificus venom (Cdt) is toxic for a variety of eukaryotic cells, especially at high concentrations. However its effects on host immune cells are not well known. The purpose of this study was to determine the effect of Cdt on functional status and the mediators production in peritoneal macrophages. The effects of Cdt were analyzed in vitro and were detected using functional status of macrophages as determined by the H(2)O(2) release, spreading percentage, phagocytic index, vacuole formation, and mediators production. Several functional bioassays were employed: cytotoxicity was determined by taking the lyses percentage and the presence of hydrogen peroxide (H(2)O(2)) in macrophages, using the horseradish peroxidase-dependent oxidation of phenol red and nitric oxide (NO) in the supernatants of macrophages by the Griess reaction. The tumor necrosis factor (TNF) activity was detected by measuring its cytotoxic activity on L929 cells, and the production the level of other cytokines was assayed using enzyme-linked immunosorbent assay. In vitro studies revealed that Cdt produced (a) a discrete increase in the release of H(2)O(2) and vacuole formation; (b) a decrease in spreading percentage and in the phagocytic index; and (c) an increment in the mediators production. More pronounced increments of IL-6 and TNF were observed after 24 and 48 hours, respectively. Maximum levels of IFN-gamma and NO were observed after 96 hours. Interestingly, levels of all mediators presented a discreet decrease, as the amount of Cdt was increased. In contrast, the IL-10 levels observed for all doses studied here did not alter. The IL-6/IL-10 ratio may possibly reflect the balance of pro- and anti-inflammatory cytokines in macrophages, which may be manifested in the inflammatory status during the envenoming processes. Taken together, these data indicate that Cdt have a differential effect on macrophage activation and that this venom is a potent inhibitor of anti-inflammatory response.
巴西矛头蝮蛇毒（Cdt）对多种真核细胞具有毒性，尤其是在高浓度时。然而，其对宿主免疫细胞的影响尚不清楚。本研究的目的是确定Cdt对腹膜巨噬细胞功能状态和介质产生的影响。在体外分析了Cdt的作用，并通过巨噬细胞的功能状态进行检测，这些功能状态由H₂O₂释放、铺展百分比、吞噬指数、液泡形成和介质产生来确定。采用了几种功能性生物测定法：通过测定巨噬细胞中的裂解百分比和过氧化氢（H₂O₂）的存在来确定细胞毒性，利用辣根过氧化物酶依赖的酚红氧化反应以及通过Griess反应测定巨噬细胞上清液中的一氧化氮（NO）。通过测量其对L929细胞的细胞毒性活性来检测肿瘤坏死因子（TNF）活性，并使用酶联免疫吸附测定法测定其他细胞因子的产生水平。体外研究表明，Cdt产生了以下影响：（a）H₂O₂释放和液泡形成略有增加；（b）铺展百分比和吞噬指数降低；（c）介质产生增加。分别在24小时和48小时后观察到IL-6和TNF有更明显的增加。在96小时后观察到IFN-γ和NO的最高水平。有趣的是，随着Cdt量的增加，所有介质的水平都有轻微下降。相比之下，此处研究的所有剂量的IL-10水平均未改变。IL-6/IL-10比值可能反映了巨噬细胞中促炎和抗炎细胞因子的平衡，这可能在蛇咬伤过程中的炎症状态中表现出来。综上所述，这些数据表明Cdt对巨噬细胞激活有不同的影响，并且这种毒液是抗炎反应的有效抑制剂。
