鼻滴吡非尼酮对百草枯致小鼠肺纤维化的治疗作用  

Intranasal administration of pirfenidone attenuates paraquat-induced pulmonary fibrosis in mice

作　　者：肖振宇 孙阳阳 杨呓 马莹凯 王永安 骆媛 XIAO Zhenyu;SUN Yangyang;YANG Yi;MA Yingkai;WANG Yongan;LUO Yuan(State Key Laboratory of National Security Specially Needed Medicines,Academy of Military Medical Sciences,Beijing 100850,China)

机构地区：[1]军事医学研究院国家安全特需药品全国重点实验室,北京100850

出　　处：《中国药理学与毒理学杂志》2025年第4期268-276,共9页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的探讨不同时间鼻滴吡非尼酮对百草枯所致小鼠肺纤维化的治疗作用。方法8周龄雄性C57BL/6小鼠随机分为6组,每组8只:正常对照(生理盐水)组、吡非尼酮对照组、百草枯组、百草枯+吡非尼酮治疗15、10和5 d组。除正常对照组和吡非尼酮对照组外,其余各组小鼠均ip给予百草枯(35 mg·kg^(-1))建立肺纤维化模型;百草枯+吡非尼酮治疗15、10和5 d组小鼠分别于百草枯染毒后第1、6和11 d开始滴鼻给予吡非尼酮(20 mg·kg^(-1)),每天1次,至第15 d。百草枯染毒后第15 d进行小鼠肺功能检测、微型计算机断层扫描(CT)成像分析和腹主动脉血气测定;HE染色观察肺组织病理变化;Masson染色观察肺组织胶原纤维沉积;Western印迹法检测肺组织纤维化标志物纤连蛋白(FN)、Ⅰ型胶原蛋白(CollⅠ)、E钙黏蛋白(E-cad)、波形蛋白(Vim)、α平滑肌肌动蛋白(α-SMA)表达水平;ELISA检测小鼠肺组织中炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-8、IL-1β及结缔组织生长因子(CTGF)表达水平。结果与正常对照组相比,百草枯组小鼠呼气时间(TE)显著延长、增强停顿(PENH)显著升高、通气量(TV)显著降低;CT扫描成像可见网状高密度影,伴散在磨玻璃样病变;动脉血氧气分压(PaO_(2))、血氧饱和度(SaO_(2))、氧合血红蛋白(FO_(2)Hb)、中心静脉血氧饱和度(ScvO_(2))均显著降低,二氧化碳分压(PaCO_(2))与还原血红蛋白(FHHb)水平显著升高,提示百草枯染毒后小鼠出现低氧血症与高碳酸血症;肺组织出现明显的间质增厚、肺泡塌陷、大量炎症细胞浸润现象,肺组织边缘区域出现明显纤维化病变,大量胶原纤维沉积;肺纤维化标志物FN、CollⅠ、Vim和α-SMA蛋白表达水平显著提高,E-cad表达水平显著降低;肺组织中炎症因子TNF-α、IL-6、IL-8和IL-1β显著升高,促纤维化细胞因子CTGF显著升高。与百草枯组相比,百草枯+吡非尼酮治疗15、10和5 d组�OBJCTIVE To evaluate the therapeutic efficacy of intranasal administration of pirfeni⁃done in treating paraquat-induced pulmonary fibrosis in mice across treatment durations.METHODS Eight-week-old male C57BL/6 mice were randomly divided into six groups(n=8 per group):the normal control group(saline),pirfenidone control group,paraquat group,and three treatment groups receiving a combination of paraquat and pirfenidone for 15,10 and 5 d,respectively.Except the normal and pirfeni⁃done control groups,all the mice received intraperitoneal injection of paraquat(35 mg·kg^(-1))to induce pulmonary fibrosis.In the treatment groups,pirfenidone(20 mg·kg^(-1))was delivered intranasally once daily,beginning on days 1,6,and 11 post-paraquat exposure,until day 15.Fifteen days after paraquat exposure,pulmonary function tests,micro-CT imaging,and arterial blood gas analysis were performed.Histopathological changes and collagen fiber deposition in lung tissues were examined using HE and Masson staining respectively.The protein expression levels of fibrosis markers,including fibronectin(FN),collagen typeⅠ(CollⅠ),E-cadherin(E-cad),vimentin(Vim),andα-smooth muscle actin(α-SMA),were detected by Western blotting.Additionally,inflammatory and pro-fibrotic cytokines,such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-8,IL-1β,and connective tissue growth factor(CTGF),were quantified using ELISA.RESULTS Compared with the normal control group,mice treated with paraquat exhibited significant respiratory alterations,including prolonged expiratory time(TE),increased enhanced pause(PENH),and reduced tidal volume(TV).CT imaging revealed reticular high-density shadows and ground-glass opacities in paraquat-treated mice.Blood gas analysis showed reduced partial pressure of oxygen(PaO_(2)),oxygen saturation of blood(SaO_(2)),fractional oxygen saturation of hemoglobin(FO_(2)Hb),and central venous oxygen saturation(ScvO_(2)),along with increased partial pressure of carbon dioxide(PaCO_(2))and fractional deoxyhemoglobin saturat

关 键 词：吡非尼酮 百草枯 肺纤维化 鼻滴 炎症因子 

分 类 号：R96[医药卫生—药理学] R974[医药卫生—药学]