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目的:本实验研究了以非致死剂量脂多糖(Lipopolysaccharide,LPS)攻击半乳糖葡胺(D-galactosamine,D-GalN)增敏小鼠的死亡是否伴有重要器官的凋亡特征。方法:实验组小鼠以LPS(0.05μg)与D-GalN(20mg)直接攻击,而对照组小鼠仅仅以LPS或D-GalN,在规定实验时间经乙醚麻醉后,取出脑、肾、心、脾、肺、肝,并立即匀浆和抽提DNA,对所抽提的各器官的DNA以常规琼脂糖凝胶电泳分析是否存在DNA梯度现象;并测定血浆中乳酸脱氢酶和谷氨酸草酰乙酸转移酶,同时将DNA片段化现象与小鼠致死作用的关联性在实验组和对照组间做比较。结果:LPS攻击B-GalN增敏的小鼠4h时,在所检测的器官中,仅仅肝脏可见到凋亡特征DNA片段化现象,至7h时小鼠死亡。而仅用LPS或D-GalN攻击的对照组小鼠,不仅全部存活而且肝脏DNA分子完整。结论:LPS攻击B-GalN增敏的小鼠,肝脏是唯一出现DNA片段化的靶器官,由细菌LPS引发小鼠的死亡,肝脏的损伤可能是关键的原因。
AIM: To investigate whether the death of mice sensitized with D-galactosamine (D-GalN), a non-lethal dose of lipopolysaccharide (LPS), is associated with apoptosis of vital organs. Methods: The mice in the experimental group were challenged directly with LPS (0.05μg) and D-GalN (20mg), while the control mice were anesthetized with LPS or D-GalN only after being anesthetized with diethyl ether for brain, kidney and heart , Spleen, lungs and liver, and immediately homogenized and extracted DNA, the extracted DNA of each organ was analyzed by conventional agarose gel electrophoresis for the presence of DNA gradient phenomenon; and determination of plasma lactate dehydrogenase and glutamine Acid oxaloacetate transferase, and the association between DNA fragmentation and lethal effects in mice was compared between experimental and control groups. Results: LPS challenged B-GalN sensitized mice 4h, the apoptotic DNA fragmentation was found only in the liver in the organs tested, and the mice died at 7h. Control mice challenged with LPS or D-GalN not only all survived but also had intact liver DNA molecules. Conclusion: LPS challenge B-GalN sensitized mice, the liver is the only target organ for DNA fragmentation. The death of mice induced by bacterial LPS and liver damage may be the key reasons.