Wang Ai-wu, Liu Yan, Hu Kong-xin, Cheng Qian
Department of Stomatology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2012 Dec;47(12):725-9. doi: 10.3760/cma.j.issn.1002-0098.2012.12.007.
To investigate an effective purification method for removing endotoxin from Prevotella intermedia.
The main protein ingredients of bacteria prepared from ammonium sulfate precipitation were further treated with octyl phenyl polyoxyethylene ether (Triton X-114), and then processed at 4°C, 37°C and 25°C. The obtained aqueous phase after at least two more cycle repeated operations was assayed for endotoxin by Western blotting, LAL-clotting method, in vitro cell stimulation and in vivo animal experiments.
Western blotting and LAL-clotting method demonstrated that the reduction in endotoxin level was greater than 99.99% and recovery of the proteins after endotoxin removal was greater than 90% with Triton X-114 treatment for 3 cycles. The cytokines expression level was lower in both in vitro cell stimulation and in vivo animal experiments than in untreated group (P < 0.05).
The extraction method provides a new choice for endotoxin removal from large volumes of the oral Prevotella intermedia.
研究一种从中间普氏菌中去除内毒素的有效纯化方法。
用硫酸铵沉淀法制备细菌的主要蛋白质成分，再用辛基苯基聚氧乙烯醚（吐温X - 114）进一步处理，然后在4℃、37℃和25℃下进行处理。至少再重复两个循环操作后，通过蛋白质免疫印迹法、鲎试剂凝胶法、体外细胞刺激和体内动物实验对内毒素进行检测。
蛋白质免疫印迹法和鲎试剂凝胶法表明，经吐温X - 114处理3个循环后，内毒素水平降低超过99.99%，去除内毒素后蛋白质回收率超过90%。在体外细胞刺激和体内动物实验中，细胞因子表达水平均低于未处理组（P < 0.05）。
该提取方法为从大量口腔中间普氏菌中去除内毒素提供了新的选择。
