摘要
研究在鸡马立克氏病毒(MDV)感染及引发肿瘤的发生、发展过程中肾组织的病变、热休克蛋白60(HSP60)mRNA转录、表达水平的动态变化及HSP60组织细胞内定位,探讨其相关性。通过人工感染建立MD肿瘤模型,利用实时荧光定量RT-PCR,结合病理组织学和免疫组织化学等方法,检测试验鸡肾脏的病理变化,HSP60的组织细胞内定位、蛋白表达量及mRNA转录水平的变化。在注射MDV 21d后肾组织出现明显病理变化;HSP60在肿瘤病灶内的瘤细胞及附近间质巨噬细胞的胞质中呈强阳性表达,在瘤灶外肾小管上皮细胞的胞质内有少量表达;HSP60的表达量,攻毒组始终高于对照组,28～86日龄间差异达极显著水平(P<0.01),28日龄时攻毒组HSP60的表达量分别为空白对照组和疫苗对照组的8.608和12.752倍;7～42日龄间,攻毒组HSP60mR-NA转录水平随病程发展呈开口向下的抛物线样变化,前期(7～21d)转录水平处于上升阶段,后逐渐降低,21d时达转录峰值,分别为空白组和疫苗组的1.222和1.179倍,整个周期内攻毒组HSP60mRNA转录水平均高于空白组,14～28日龄间差异极显著(P<0.01)。MDV引发肿瘤发生发展过程中肿瘤病灶及周围组织内HSP60表达水平升高,HSP60mRNA转录水平相应增加;HSP60表达量同其mRNA转录水平的变化趋势基本相符,但并非完全一致的线性关系,两者与肿瘤的发生、发展密切相关,有望作为鸡MD诊断及发病过程的重要判定指标。
The objective of this study is to investigate the relationship between pathological lesion and transcription, expression and distribution of heat shock protein 60 （HSP60） in kidney of chickens infected by Marek＇s disease virus （MDV）. Tumor animal model was successfully estab- lished by infecting 1 day old chickens with MDV. Real-time RT-PCR, histopathological, immu- nohistochemistrical methods were used to detect the pathological lesion, transcription level, ex- pression level and distribution of HSP60 in the kidney. After 21 days post-infection （PI）, obvi-ous pathological damage appeared in the kidneys of chickens infected by MDV. HSP60 was main- ly distributed in the cytoplasm of the oncocyte and interstitial macrophages in the tumor regions. Within the course of MD, the contents of HSP60 of effeeted group were always higher than blank control group and vaccine control group, which was significantly higher after 28 days, and the mostly was about 8. 608 and 12. 752 times at the age of 28 days. The tendency changes looked like a downward parabola, during 7 to 21-day-old, the level of HSP60 mRNA transcription of effected group was in the rising stage, and then gradually dropped. When 21-day-old the maximum was came on, which was about 1. 222 and 1. 179 times of two control groups. Throughout all stages the level of transcription was higher than blank control group, and during 14 to 28-day-old was significantly higher （P;0.01）. The MDV infection caused resistance to infection and antitumor response in the progress of MD, the stress related kidney damage resulted in the over-expression of HSP60 in tumor lesions and surrounding tissues. A corresponding increase came to the mRNA transcription level of it in kidney tissues. The rapid expression changes of HSP60 was basic con- sistent with its mRNA transportation level, but not completely linear. And they were closely re- lated with the onset and progression of tumors, which might be hallmarks to diagnose and deter- mine the process of tumor caused by
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2013年第4期610-616,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(31101787)
山东省自然科学基金(ZR2010CM035)
