利用端粒和端粒酶PCR芯片筛选易发/抗肺纤维化小鼠肺组织差异基因研究  

Screening of discrepant genes in lung tissues of pulmonary fibrosis susceptible/resistant mice by telomere and telomerase PCR microarray

作　　者：贾兆乾 张潇 邢源 闫华 赵欣悦 李小宇 王勇懿 叶雨萌 郝延辉 左红艳 李杨 JIA Zhao-qian;ZHANG Xiao;XING Yuan;YAN Hua;ZHAO Xin-yue;LI Xiao-yu;WANG Yong-yi;YE Yu-meng;HAO Yan-hui;ZUO Hong-yan;LI Yang(Institute of Radiation Medicine,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;Academy of Medical Sciences,Zhengzhou University,Zhengzhou 450001,China;The First Affiliated Hospital of Hebei North University,Zhangjiakou,Hebei 075000,China;The Sixth Medical Center of Chinese PLA General Hospital,Beijing 100048,China)

机构地区：[1]军事科学院军事医学研究院辐射医学研究所,北京100850 [2]郑州大学医学科学院,郑州450001 [3]河北北方学院附属第一医院,河北张家口075000 [4]解放军总医院第六医学中心,北京100048

出　　处：《军事医学》2023年第1期26-32,共7页Military Medical Sciences

基　　金：北京市自然科学基金(7132149)。

摘　　要：目的通过建立易发/抗放射性肺纤维化(RIPF)小鼠模型,利用端粒和端粒酶实时荧光定量PCR(RT-qPCR)芯片筛选易发/抗RIPF小鼠照射前后肺组织差异基因并初步验证。方法采用20 Gy^(60)Coγ射线照射C57BL/6J和C3H/HeN小鼠胸部;通过HE、天狼星红染色,羟脯氨酸、转化生长因子β(TGF-β)和结缔组织生长因子(CTGF)含量检测等,比较这两种小鼠RIPF进程的差异;采用端粒和端粒酶RT-qPCR芯片,分别比较照射前、照射后3个月两种小鼠肺组织端粒和端粒酶相关基因的表达差异,并采用RT-qPCR对差异基因进行初步验证。结果照射后3个月,C57BL/6J小鼠肺组织纤维化性病变较C3H/HeN小鼠更为明显;C57BL/6J小鼠肺组织中羟脯氨酸含量显著高于照射前和C3H/HeN小鼠(P<0.05),且其肺泡壁Ⅰ型胶原沉积显著多于C3H/HeN小鼠;C57BL/6J小鼠肺组织促纤维化细胞因子TGF-β和CTGF含量明显高于C3H/HeN小鼠(P<0.05)。端粒和端粒酶RT-qPCR芯片筛选发现,两种对照组小鼠肺组织差异基因48个;照射后3个月两种小鼠肺组织差异基因32个;照射后3个月,C57BL/6J小鼠肺组织与照射前相比有8个差异基因,C3H/HeN小鼠肺组织与照射前相比有4个差异基因。对差异基因端锚聚合酶2(Tnks2)基因进行验证,照射前及照射后3个月C3H/HeN小鼠肺组织中Tnks2的表达均高于C57BL/6J小鼠(P<0.05)。结论^(60)Coγ射线照射前后易发/抗肺纤维化小鼠肺组织存在多个端粒及端粒酶相关差异基因。Objective To establish a model of pulmonary fibrosis-prone and fibrosis-resistant mice and screen the differential genes in lung tissues before and after irradiation by telomere and telomerase real-time fluorescence quantitative PCR(RT-qPCR)microarray.Methods The chests of C57BL/6J and C3H/HeN mice were irradiated with 20 Gy^(60)Co gamma rays.The differences in radiation-induced pulmonary fibrosis(RIPF)progression between them were studied via HE staining,sirius red staining,hydroxyproline content,TGF-βand CTGF detection.Telomere and telomerase RT-qPCR microarray was used to compare the expressions of telomere and telomerase related genes in their lung tissues before irradiation and 3 months after irradiation.The differential genes were verified by RT-qPCR.Results The fibrotic lesions of C57BL/6J mice were more obvious than those of C3H/HeN mice 3 months after irradiation.The content of hydroxyproline in lung tissues of C57BL/6J mice was significantly higher than before irradiation and in C3H/HeN mice(P<0.05),and much more collagen typeⅠwas deposited in the alveolar wall of C57BL/6J mice than in that of C3H/HeN mice.The contents of TGF-βand CTGF in lung tissues of C57BL/6J mice were significantly higher than in those of C3H/HeN mice(P<0.05).There were 48 differential genes in telomere and telomerase related pathways in lung tissues of control C57BL/6J and C3H/HeN mice and 32 differential genes were found in the lung tissues of the two types of mice 3 months after irradiation.Moreover,8 differential genes were discovered in the lung tissues of C57BL/6J mice 3 months after irradiation and 4 discrepant genes were detected in the lung tissues of C3H/HeN mice after irradiation.The differential gene Tnks2 was verified.The expression of Tnks2 in lung tissues of C3H/HeN mice 3 months after irradiation was higher than that of C57BL/6J mice and before irradiation(P<0.05).Conclusion Differential genes in telomere and telomerase related pathways in lung tissues of fibrosis-susceptible and fibrosis-resistant mice are foun

关 键 词：小鼠 放射性肺纤维化 端粒 端粒酶 实时荧光定量PCR芯片 端锚聚合酶2 基因表达 

分 类 号：R563[医药卫生—呼吸系统]