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中国癌症防治杂志 ›› 2018, Vol. 10 ›› Issue (2): 95-99.doi: 10.3969/j.issn.1674-5671.2018.02.05

• 基础研究 • 上一篇    下一篇

通过新生树鼩乙型肝炎病毒感染模型动态观察宿主体内TLR-4和IL-6的表达

  

  1. 广西医科大医学附属肿瘤医院实验研究部;广西医科大学研究生院
  • 出版日期:2018-04-25 发布日期:2018-06-13
  • 通讯作者: 杨春 E-mail:cxl_yang@163.com
  • 基金资助:

    国家自然科学基金资助项目(81460418)

Detecting changes in TLR-4 and IL-6 during hepatitis B virus infection in a tree shrew model

  • Online:2018-04-25 Published:2018-06-13

摘要:

目的 通过观察接种乙型肝炎病毒(hepatitis B virus,HBV)后新生树鼩体内细胞因子TLR-4和IL-6的变化,初步探讨HBV感染过程中宿主体内的免疫反应。方法 将31只树鼩幼崽分为实验组(n=25)和对照组(n=6)。实验组接种人HBV,接种后第8周至第24周定期检测树鼩外周血中乙肝表面抗原(HBsAg)和HBV- DNA,根据检测结果将实验组分为慢性感染组7只(血清HBsAg或HBV-DNA持续阳性时间达到24周)和疑似慢性感染组5只(血清HBsAg或HBV-DNA不连续出现2次或2次以上阳性),其余13只HbsAg和HBV-DNA均阴性,不纳入研究;对照组不做任何处理。采用双抗体夹心法检测树鼩外周血TLR-4、IL-6蛋白表达水平,RT-PCR法检测树鼩肝组织中TLR-4、IL-6 mRNA的表达水平。 结果 观察期间内,慢性感染组和疑似慢性感染组外周血TLR-4与IL-6蛋白的表达均高于对照组(P<0.05);肝组织中IL-6的mRNA表达水平高于对照组(P<0.05);慢性感染组TLR-4 mRNA表达水平高于对照组(P<0.05)。结论 TLR-4、IL-6在树鼩HBV感染模型中表达升高,可能在HBV感染过程中起重要作用。

关键词: 乙型肝炎病毒, TLR-4, IL-6, 树鼩, 免疫反应

Abstract:

Objective To detect changes in TLR-4 and IL-6 in tree shrews infected by hepatitis B virus. Methods A total of 25 tree shrews were inoculated with human hepatitis B virus (HBV) and then divided into an HBV-infected group,suspected HBV-infected group and control group based on detection of the HBV marker HBsAg or HBV DNA using a photochemical method or fluorescence- probe PCR. HBV-infected animals showed persistent positive results for HBsAg or HBV DNA,while the suspected HBV-infected group showed two or more positive results for HBsAg or HBV DNA. Levels of TLR-4 and IL-6 in peripheral blood and liver tissues were measured using,respectively,enzyme-linked immunosorbent assay (ELISA) and hematoxylin- eosin staining. Levels of TLR-4 and IL-6 mRNA in liver tissue of all three groups were measured using real-time fluorescence quantitative PCR. All assays were performed between post-natal weeks 8 and 24. Results Seven animals were assigned to the HBV-infected group,while five were assigned to the suspected HBV-infected group; all control animals showed negative results. Levels of TLR-4 in the peripheral blood of infected and suspected infected animals were significantly higher than in control animals (P<0.05)at weeks of 8,12,16,20,and 24. Levels were not significantly different between the infected and suspected infected groups. Similar results were observed for levels of IL-6 in peripheral blood. Levels of TLR-4 and IL-6 mRNA in liver tissue were significantly higher in the infected and suspected infected groups than in the control group(P<0.05). Conclusion Levels of TLR-4 and IL-6 are elevated in tree shrews following infection with HBV virus,and these signaling molecules may play an important role in the infection process.

Key words: Hepatitis B virus, TLR-4, IL-6, Tree shrew, Immune response