Saadi Samahir, Assarehzadegan Mohammad-Ali, Pipelzadeh Mohammad Hassan, Hadaddezfuli Reza
Department of Biology, East Tehran Branch, Islamic Azad University, Tehran, Iran.
Department of Immunology, Ahvaz Jundishapur University of Medical Sciences, Iran; Department of Immunology, Iran University of Medical Sciences, Tehran, Iran.
Toxicon. 2015 Nov;106:117-21. doi: 10.1016/j.toxicon.2015.09.029. Epub 2015 Sep 28.
The objective of this study was to evaluate the capacity of venom from Androctonus crassicauda to induce expression/production of interleukin (IL)-12 by isolated human monocytes. For this purpose, isolated human monocytes were exposed to different concentrations of the venom (0.16-20 μg/ml) for varying periods (6, 12, and 24 h). Apart from measures of venom cytotoxicity (i.e., lactase dehydrogenase activity [LDH] release), measures of IL-12 p40 mRNA (by Real-time PCR) of IL-12 release (by ELISA) were performed. The results showed that the venom produced significant concentration- and duration of incubation-dependent cytotoxicity. Expression of IL-12 p40 mRNA was significantly increased at all exposure timepoints relative to that in unexposed cells, but was maximal after 6 h of exposure. At that timepoint, the effect from a dose of 2.5 μg venom/ml provided the maximal increase among all doses tested. At the level of the protein itself, IL-12 production remained almost consistently elevated (vs. unexposed control values) across all exposure timepoints, with the greatest formation again occurring after 6 h of incubation at a dose of 2.5 μg venom/ml. The findings from this study demonstrated that venom from the A. crassicauda scorpion contained active constituents that could induce a sustained activation of human monocytes that was manifested, in part, as promotion of the expression/production of IL-12.
本研究的目的是评估厚尾杀人蝎毒液诱导分离的人单核细胞表达/产生白细胞介素(IL)-12的能力。为此,将分离的人单核细胞暴露于不同浓度的毒液(0.16 - 20μg/ml)中不同时间(6、12和24小时)。除了测量毒液的细胞毒性(即乳酸脱氢酶活性[LDH]释放)外,还通过实时定量聚合酶链反应(Real-time PCR)检测了IL-12 p40 mRNA,并通过酶联免疫吸附测定(ELISA)检测了IL-12的释放。结果表明,毒液产生了显著的浓度和孵育时间依赖性细胞毒性。与未暴露细胞相比,在所有暴露时间点IL-12 p40 mRNA的表达均显著增加,但在暴露6小时后达到最大值。在该时间点,2.5μg毒液/ml剂量的效果在所有测试剂量中提供了最大的增加。在蛋白质水平上,在所有暴露时间点,IL-12的产生几乎持续升高(相对于未暴露的对照值),在2.5μg毒液/ml剂量孵育6小时后再次出现最大形成量。本研究结果表明,厚尾杀人蝎毒液含有活性成分,可诱导人单核细胞持续活化,部分表现为促进IL-12的表达/产生。