鼻内滴注与气管滴注PM2.5建立大鼠急性肺损伤模型的比较研究  

Comparison of intranasal and tracheal instillation of PM2.5 suspension in construction of rat acute lung injury model

作　　者：张行行 赵麓 孙欠欠 赵安东 周甜雨 王川[1] 刘继平[1] 王斌[1] ZHANG Hang-hang;ZHAO Lu;SUN Qian-qian;ZHAO An-dong;ZHOU Tian-yu;WANG Chuan;LIU Ji-ping;WANG Bin(Shaanxi University of Chinese Medicine,Key Laboratory of Pharmacodynamic Mechanism and Material Basis of Traditional Chinese Medicine,Shaanxi Provincial Administration of Traditional Chinese Medicine,Xianyang 712046,China)

机构地区：[1]陕西中医药大学,陕西省中医药管理局中药药效机制与物质基础重点研究室,陕西咸阳712046

出　　处：《中国病理生理杂志》2022年第12期2298-2304,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81473385);陕西中医药大学学科创新项目(No.2019-YL13)。

摘　　要：目的:通过比较鼻内滴注与非暴露式气管滴注细颗粒物(PM2.5)混悬液构建急性肺损伤(ALI)大鼠模型的差异,为研究PM2.5气道染毒选择合理的动物模型提供参考。方法:以200μL/kg的PM2.5混悬液(10mg/kg)鼻腔滴注或气管滴注10次,每3 d一次,构建ALI模型。通过HE染色观察肺组织结构及中性粒细胞浸润情况;用试剂盒检测支气管肺泡灌洗液(BALF)中酸性磷酸酶(ACP)、碱性磷酸酶(AKP)和乳酸脱氢酶(LDH)活性;ELISA检测血清中白细胞介素6(IL-6)和转化生长因子β(TGF-β)含量;免疫组化检测肺组织中TGF-β和IL-17的蛋白表达;RT-qPCR检测肺组织中IL-17和叉头框蛋白P3(Foxp3)的mRNA表达。结果:与对照组相比,鼻内滴注组和气管滴注组大鼠肺组织均出现显著的病理损伤;血清中IL-6水平显著升高,TGF-β水平显著下降(P<0.05);BALF中ACP、AKP和LDH活性均显著升高(P<0.05);肺组织中IL-17的蛋白和mRNA表达,以及Th17/Treg比值显著升高(P<0.05),TGF-β蛋白和Foxp3 mRNA表达量显著下降(P<0.05)。两组不同部位滴注组相比,气管滴注组肺损伤较鼻内滴注组严重,血清中IL-6水平和BALF中ACP活性显著升高(P<0.05);肺组织Th17/Treg值显著升高,TGF-β蛋白表达显著下降(P<0.05);其他指标无显著差异。结论:两种方式滴注PM2.5均能造成不同程度的肺部损伤,增加促炎细胞因子的产生,加强Th2应答;两种不同部位滴注相比,气管滴注造成的肺组织损伤和炎症反应较严重。AIM:To compare intranasal and tracheal instillation of particulate matter 2.5(PM2.5)suspension in the construction of rat acute lung injury(ALI)model.METHODS:The rat ALI model was established with 200μL/kg of PM2.5 suspension(10 mg/kg)through intranasal or tracheal instillation once per 3 days for 10 times.The histological structure and neutrophil infiltration in lung tissues were observed by HE staining.The activity of acid phosphatase(ACP),alkaline phosphatase(AKP)and lactate dehydrogenase(LDH)in bronchoalveolar lavage fluid(BALF)was detected using commercially available kits.The content of interleukin-6(IL-6)and transforming growth factor-β(TGF-β)in serum was detected by ELISA.The protein expression of TGF-βand IL-17 in lung tissues was detected by immunohistochemistry,and the mRNA expression of IL-17 and forkhead box protein P3(Foxp3)in lung tissues was detected by RTqPCR.RESULTS:Compared with control group,the obvious lung injury and significant increase in cytotoxicity indexes(ACP,AKP and LDH)in BALF were found after intranasal and tracheal instillation of PM2.5 suspension(P<0.05).Serum IL-6 level was increased,while TGF-βwas decreased after instillation.The expression level of IL-17 and the ratio of Th17/Treg were increased,while the expression levels of TGF-βand Foxp3 were decreased in lung tissues after instillation.The lung injury was more serious after tracheal instillation compared with intranasal instillation.Serum IL-6 level,ACP activity in BALF and the ratio of Th17/Treg in lung tissues were increased after tracheal instillation compared with intranasal instillation,while the expression of TGF-βprotein in lung tissues was decreased.CONCLUSION:Intranasal and tracheal instillation of PM2.5 suspension induces lung injury in rats,and enhances the production of proinflammatory cytokines and the Th2 response.Compared with intranasal instillation,tracheal instillation exacerbates lung injury.

关 键 词：鼻内滴注 气管滴注 细颗粒物 急性肺损伤 

分 类 号：R563[医药卫生—呼吸系统] R363.2[医药卫生—内科学]