畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (2): 415-421.doi: 10.11843/j.issn.0366-6964.2019.02.020

• 基础兽医 • 上一篇    下一篇

鸡马立克病肝组织肿瘤病变与热休克蛋白转录、表达的相关性研究

段世豪, 路建彪, 李旭勇, 刘成, 张树金, 殷国政, 司振书*, 李玉保*   

  1. 聊城大学农学院, 聊城 252059
  • 收稿日期:2018-07-27 出版日期:2019-02-23 发布日期:2019-02-23
  • 通讯作者: 司振书,主要从事动物疫病流行病学研究,E-mail:sizhenshu@lcu.edu.cn;李玉保,主要从事动物分子病理学研究,E-mail:liyubao@lcu.edu.cn
  • 作者简介:段世豪(1993-),男,山东临沂人,硕士生,主要从事动物分子病理学研究,E-mail:472618792@qq.com
  • 基金资助:

    国家自然科学基金(31101787);山东省自然科学基金(ZR2016CM40;ZR2010CM035)

Relationship between Liver Tissues Pathological Lesion and Expression Level of HSPs in Progress of Marek's Disease

DUAN Shihao, LU Jianbiao, LI Xuyong, LIU Cheng, ZHANG Shujin, YIN Guozheng, SI Zhenshu*, LI Yubao*   

  1. College of Agriculture, Liaocheng University, Liaocheng 252059, China
  • Received:2018-07-27 Online:2019-02-23 Published:2019-02-23

摘要:

研究马立克病毒引发肿瘤的发生、发展过程中,热休克蛋白(heat shock proteins,HSPs)转录表达水平、组织细胞内定位与肝病理组织学损伤之间的相关性,为进一步阐述HSPs的生物学作用奠定基础。通过人工感染建立鸡MD肿瘤模型,定期剖杀试验鸡,采集肝组织,利用组织学、免疫组织化学、荧光定量RT-PCR及ELISA方法,检测肝病理组织学损伤、HSPs组织细胞内定位以及HSPs转录、表达水平的动态变化。结果:在MDV感染14 d后,感染组鸡肝组织可见大小不等的淋巴样瘤细胞浸润和局灶性生长;HSP90、HSP60在淋巴瘤细胞的细胞质内强表达,而HSP27、HSP70主要分布于肿瘤细胞外的肝细胞的细胞质内;感染组HSP70和HSP90 mRNA转录水平始终极显著高于空白对照组和疫苗免疫对照组;感染组HSP90的表达量始终极显著高于空白对照组,HSP70表达量在14和21日龄时,极显著高于空白对照组,在28和35日龄时显著高于空白对照组,42日龄后与空白对照组和疫苗免疫对照组相比差异不显著。在MDV引发鸡肿瘤发生、发展过程中,HSP60、HSP90与HSP70、HSP27之间在肝组织细胞内定位截然不同,且HSP70与HSP90在肝组织内表达量变化趋势差异明显,说明不同HSPs在MD肿瘤发生、发展过程中的生物学作用存在差异。

Abstract:

The experiment was conducted to discuss the biological function of HSPs during the progress of tumor induced by the Marek's disease, the relationship between pathological lesion of liver and distribution, expression level of HSPs were studied. The tumor animal model was established using MD virus. Chickens were timely killed post artificial infection and the livers were sampled. Pathological lesion of liver and distribution, transcription and expression level of HSPs were studied during the progress of tumor by the methods of histopathology, immunohistochemistry, FQ RT-PCR and ELISA. Results were as follows:At 14 days, obvious pathological damage appeared in the livers of chickens attracted by MDV, and unequal-sized lymphoid cells infiltration was found among liver cells. HSP60 and HSP90 were mainly located in the cytoplasm of tumor cell, while HSP27 and HSP70 were mainly located in the hepatocyte around the tumor tissues. Within the course of MD, the transcription level of HSP70 and HSP90 mRNA of challenge group were always significantly higher than those of blank control and vaccine control groups. The expression level of HSP90 of challenge group was always significantly higher than those of blank control group in the test process, while the expression level of HSP70 of challenge group was significantly higher than those of blank controrl group at 14 and 21 d, was significantly higher at 28 and 35 d, and no significant difference after 42 d. The distribution among HSP60, HSP90, HSP27, HSP70 and the trend of expression level between HSP90 and HSP70 are completely different during the progress of tumor induced by MDV, which indicates that different HSPs have different biological functions.

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