WTAP对博来霉素诱导的肺纤维化胶原沉积的影响  

Effect of WTAP on collagen deposition in bleomycin-induced pulmonary fibrosis

作　　者：张云森 刘震宇 刘芷言 林丽婵 沙纪名[3] 陶辉[1,2] 陈齐 Zhang Yunsen;Liu Zhenyu;Liu Zhiyan;Lin Lichan;Sha Jiming;Tao Hui;Chen Qi(Dept of Anesthesiology and Perioperative Medicine,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601;Key Laboratory of Anesthesiology and Perioperative Medicine of Anhui Higher Education Institutes,Hefei 230601;Dept of Thoracic Surgery,The Second Affiliated Hospital of Anhui Medical University,Hefei 230601)

机构地区：[1]安徽医科大学第二附属医院麻醉与围术期医学科,合肥230601 [2]麻醉与围术期医学安徽普通高校重点实验室,合肥230601 [3]安徽医科大学第二附属医院胸外科,合肥230601

出　　处：《安徽医科大学学报》2025年第2期266-271,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省高校自然科学研究项目(编号:2023AH040376)。

摘　　要：目的探讨Wilms肿瘤1相关蛋白(WTAP)对博来霉素所致的肺纤维化组织胶原沉积的影响。方法60只小鼠随机分为4组:对照组(Control组)、博来霉素诱导肺纤维化组(BLM组)、肺纤维化慢病毒空载体对照组(BLM+LV-NC组)、肺纤维化WTAP慢病毒组(BLM+LV-WTAP组)。采用腹部皮下注射博来霉素(35 mg/kg)建立实验性肺纤维化小鼠模型,每周2次,共8次。造模结束后,使用Western blot检测纤维化相关标志物α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(CollagenⅠ)、纤维连接蛋白(Fibronectin)的蛋白表达情况以及WTAP蛋白表达情况;使用Masson染色和Sirius Red染色检测胶原沉积情况;采用RT-qPCR检测WTAP mRNA表达情况、WTAP慢病毒侵染效果及CollagenⅠmRNA表达情况。结果与Control组比较,BLM组肺组织α-SMA(P<0.001)、CollagenⅠ(P<0.001)、Fibronectin(P<0.01)蛋白表达均升高;Masson染色(P<0.001)和Sirius Red染色(P<0.001)证实BLM组肺组织出现明显胶原沉积;且BLM组肺组织中WTAP蛋白表达升高(P<0.01)。与Control组比较,BLM组肺组织中WTAP mRNA表达升高(P<0.001);与BLM+LV-NC组比较,BLM+LV-WTAP组肺组织中WTAP mRNA表达下降(P<0.001),证明病毒侵染有效。WTAP慢病毒侵染后,BLM+LV-WTAP组胶原纤维沉积减少(P<0.001)、CollagenⅠmRNA水平降低(P<0.001)及蛋白表达下降(P<0.001)。结论敲低WTAP表达可减少博来霉素所致的小鼠肺纤维化组织的胶原沉积,改善实验性肺纤维化。Objective To explore the effect of Wilms′tumor 1-associated protein(WTAP)on tissue collagen deposition in pulmonary fibrosis caused by bleomycin.Methods 60 mice were randomly divided into four groups:control group(Control group),Bleomycin-induced pulmonary fibrosis group(BLM group),pulmonary fibrosis lentivirus empty vector control group(BLM+LV-NC group),pulmonary fibrosis WTAP lentivirus group virus group(BLM+LV-WTAP group).Experimental pulmonary fibrosis mouse model was established by subcutaneous injection of bleomycin(35 mg/kg)into the abdomen,twice a week for a total of 8 times.After modeling,Western Blot was used to detect the protein expression of fibrosis-related markersα-smooth muscle actin(α-SMA),type I collagen(CollagenⅠ),fibronectin(Fibronectin),and WTAP protein.Masson staining and Sirius Red staining were used to detect collagen deposition.RT-qPCR was used to detect WTAP mRNA expression,WTAP lentivirus infection effect,and CollagenⅠmRNA expression.Results Compared with the Control group,the expression of pulmonary fibrosis markersα-SMA(P<0.001),CollagenⅠ(P<0.001),and Fibronectin(P<0.01)protein in the BLM group all increased.Masson staining(P<0.001)and Sirius Red staining(P<0.001)confirmed that significant collagen deposition occurred in the lung tissue of the BLM group.In addition,the expression of WTAP protein in the lung tissue of the BLM group increased(P<0.01).Compared with the Control group,the expression of WTAP mRNA in the BLM group increased(P<0.001).Compared with the BLM+LV-NC group,the expression of WTAP mRNA in the tissues of the BLM+LV-WTAP group decreased(P<0.001),proving that virus infection is effective.After infection with WTAP lentivirus,collagen fiber deposition decreased(P<0.001),CollagenⅠmRNA(P<0.001)level decreased,and protein(P<0.001)expression decreased in the BLM+LV-WTAP group.Conclusion Knocking down of WTAP can reduce collagen deposition in bleomycin-induced pulmonary fibrosis tissue in mice and improve experimental pulmonary fibrosis.

关 键 词：Wilms肿瘤1相关蛋白 肺纤维化 博来霉素 表观转录组 胶原 N6-甲基腺苷 

分 类 号：R332[医药卫生—人体生理学] R563.9[医药卫生—基础医学] R977.6