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目的探讨携带编码幽门螺杆菌UreB基因质粒(pLUB)的重组耻垢分枝杆菌(Mycobacterium smegmatis,M.s)经灌胃免疫在小鼠体内的分布和安全性。方法将携带绿色荧光蛋白(GFP)基因质粒(pLUB)的重组耻垢分枝杆菌灌胃BALB/c小鼠后8周,处死小鼠,观察胃、心、肝、脾、肺和肾组织中荧光蛋白的分布,测定小鼠肌酐及ALT;处死小鼠,分离出肺、肾、肝并分别测定其重量及HE染色。结果在BALB/c小鼠胃、脾组织中均检测到绿色荧光蛋白的表达,其他组织及对照组未检测到绿色荧光;小鼠体质量、各脏器质量、各血清肌酐和ALT含量与对照组无差异(P>0.05)。结论携带GFP的重组耻垢分枝杆菌可在胃和脾组织分布,对小鼠无系统毒性作用。
Objective To investigate the distribution and safety of recombinant Mycobacterium smegmatis (M.s.) harboring the gene encoding UreB gene of Helicobacter pylori in mice by intragastric administration. Methods BALB / c mice were inoculated with recombinant M. smegmatis harboring green fluorescent protein (GFP) gene plasmid (pLUB) for 8 weeks. Mice were sacrificed to observe the changes in stomach, heart, liver, spleen, lung and kidney Fluorescent protein distribution, determination of creatinine and ALT in mice; mice were sacrificed and lung, kidney and liver were isolated and their weight and HE staining were determined respectively. Results The expression of green fluorescent protein (GFP) was detected in the stomach and spleen of BALB / c mice, and no fluorescence was detected in other tissues and controls. The body weight, organ quality, serum creatinine and ALT levels in mice were compared with those in control No difference (P> 0.05). Conclusion Recombinant Mycobacterium smegmatis carrying GFP can be distributed in stomach and spleen tissues and has no systemic toxicity to mice.