自然感染OPA的羔羊肺组织中自噬相关因子的表达分析

Expression analysis of autophagy-related factors in lung tissues of lambs naturally infected with OPA

为研究细胞自噬现象在绵羊肺腺瘤(ovine pulmonary adenocarcinoma,OPA)发生发展过程中的作用,本试验以自然感染OPA的羔羊肺组织为材料,以健康羔羊肺组织为对照组,利用免疫组织化学(IHC)染色、实时荧光定量PCR(qPCR)和Western blot等方法综合检测了自噬基因MAP1LC3B、BECN1、SQSTM1及其对应的自噬蛋白LC3、Beclin1、P62的表达情况。结果显示:在对照组中,LC3、Beclin1、P62主要表达于细支气管上皮细胞和肺泡上皮细胞中;在OPA肺组织中,LC3、Beclin1、P62主要表达于肿瘤化增生的细支气管上皮细胞、增生的Ⅱ型肺泡上皮细胞和浸润的巨噬细胞中。与对照组相比,OPA肺组织中MAP1LC3B基因表达量极显著升高(P<0.01),BECN1基因表达量显著升高(P<0.05),而SQSTM1基因表达量则极显著降低(P<0.01)。在蛋白水平上,OPA肺组织中LC3和Beclin1表达量均显著高于对照组(P<0.05),而P62表达量则极显著低于对照组(P<0.01)。结果表明自然感染OPA的羔羊肺组织中自噬水平明显高于健康羔羊肺组织,提示当病毒入侵机体时机体为了抵抗病毒带来的伤害并维持内环境的稳态,细胞自噬水平升高。本试验为深入探讨自噬在OPA发生发展过程中的作用提供了依据。

In order to study the role of autophagy in the tumorigenic mechanism of ovine pulmonary adenocarcinoma(OPA),in this study,the lung tissues of lambs naturally infected with OPA were used as the test material,and the lung tissues of healthy lambs were used as the control group,immunohistochemical(IHC) staining,quantitative real-time PCR(qPCR) and Western blot assays were used to detect the expression of autophagic genes MAP1 LC3 B,BECN1,SQSTM1 and their corresponding proteins LC3,Beclin1,P62.The results showed that LC3,Beclin1,P62 were mainly expressed in bronchiolar epithelial cells and alveolar epithelial cells in the control group and in tumor-proliferating bronchiole epithelial cells,proliferating type Ⅱ alveolar epithelial cells and infiltrating macrophages in the OPA lung tissues.In addition,compared with the control group,the expression of MAP1 LC3 B gene was extremely significantly increased(P<0.01),the expression of BECN1 gene was significantly increased(P<0.05),while the expression of SQSTM1 gene was extremely significantly decreased(P<0.01) in the OPA lung tissues.At the protein level,the expression of LC3 and Beclin1 in the OPA lung tissues was significantly higher than that in the control group(P<0.05),while the expression of P62 in the OPA lung tissues was extremely significantly lower than that in the control group(P<0.01).The results indicated that the autophagy level in the naturally OPA-infected lambs lung tissues was significantly higher than that in healthy lambs,suggesting that the level of autophagy was increased in order to resist the damage caused by the virus and maintain the stability of the internal environment when the virus invaded the body.This study provides basic informations for an in-depth study of the role of autophagy in the development of OPA.