摘要
目的 研究多聚肌苷-多聚胞苷酸钠盐(Poly I∶C)对裸鼹鼠自噬水平的影响.方法 成年裸鼹鼠以及C57BL/6小鼠分别随机分成Poly I∶C给药组和对照组,分别腹腔注射Poly I∶C和生理盐水.Poly I∶C给药12h后,组织切片HE染色观察小肠组织的病理变化,电镜观察细胞结构的变化,实时定量PCR检测小肠组织中炎症相关因子IL-1、ATG5的mRNA表达情况,蛋白印迹检测小肠组织中自噬相关蛋白LC3B、Beclin 1表达情况.结果 Poly I∶C给药后,裸鼹鼠小肠组织未出现显著的病理变化,但C57BL/6小鼠小肠组织腺体部出现广泛性出血;电镜观察发现,裸鼹鼠小肠组织细胞中线粒体增多、变长,自噬体数量显著增加,而小鼠组织细胞线粒体有大量的空泡化,滑面内质网扩张;实时定量PCR检测未发现组织中IL-1、ATG5 mRNA表达有显著变化;蛋白印迹检测发现,给药后裸鼹鼠组织中LC3B蛋白表达显著上调(P<0.05),而C57BL/6小鼠则下调(P<0.05).结论 Poly I∶C给药引起裸鼹鼠自噬水平上调,同时,裸鼹鼠自噬水平的上调亦可以提高机体的适应能力,进而起到抵抗Poly I∶C损伤的作用.
Objective To study the effect of polyinosinic-polycytidylic acid(Poly I: C) on the autophagy in naked mole rat. Methods The adult naked mole rats and C57BL/6 mice were randomly divided into Poly I: C treatment group (ip Poly I: C) and control group (ip saline). Twelve hours after Poly I: C administered, the differences in the intestinal pathological changes and subcellular organelles between naked mole rats and C57BL / 6 mice were observed by light and electron microscopy respectively. The mRNA expression of inflammation-related factor IL-1 and autophagy-related gene ATG5 in intestinal tissue were detected by quantitative real-time PCR. Detection of autophagy-related proteins LC3B and Beclin 1 was conducted by Western blot. Results Naked mole rats did not show significant pathological changes after Poly I: C treatment, but intestinal tissue glands of C57BL/6 mouse show serious hemorrhage. Further observation by electron microscope found that the mitochondria's size was elongagted, number increased and more autophagy in naked mole rats' intestinal tissue cells. Meanwhile, a lot of vacuolatation of mitochondria, dilation of smooth endoplasmic reticulum was observed in mouse intestinal tissues. Significant difference was not found in mRNA expression of IL-1, ATG5 by real-time quantitative PCR. In Western blot detection, LC3B protein expression was significantly increased (P〈0.05) in naked mole rats, after administration, but that of C57BL/6 mice were reduced (P〈0.05). Conclusion Poly I: C administration induced upregulation of autophagy in naked mole rat. While, autophagy may regulate the inflammatory response in turn, which played an important role in naked mole rats' anti-Poly I:C injury.
出处
《实验动物与比较医学》
CAS
2014年第5期411-416,共6页
Laboratory Animal and Comparative Medicine
基金
上海市科技发展基金项目(No.12140900200)、(No.13140900400)和(NO.31402028)联合资助