1、文章编号:1673-2995(2023)04-0255-05论著清肺排毒汤对脂多糖诱导急性肺损伤模型小鼠的作用及机制邱爱珠1，徐晔青2，欧阳翌国1，张斌1，曾丹1，文芳1(1 湖南中医药高等专科学校，湖南 株洲412012;2 株洲市中心医院普外科，湖南 株洲412001)摘要:目的观察清肺排毒汤对脂多糖诱导的急性肺损伤的作用，并探索其机制。方法采用脂多糖(5 mg/kg)经气道滴注建立小鼠急性肺损伤模型，将模型小鼠随机分成3 组:空白对照组、模型组和治疗组。造模后2 h，空白对照组、模型组给予0 9%氯化钠注射液 0 3 mL 灌胃，治疗组给予清肺排毒汤0 3 mL(含生药2 g/mL)灌胃
2、治疗，每隔4 h 一次，共3 次。末次给药12 h 后处死小鼠，取小鼠肺组织，HE 染色观察肺组织病理变化情况并检测肺湿/干质量(wet/dry mass，W/D)比值，收集各组小鼠肺泡灌洗液，ELISA 法检测白介素 1(interleukin-1，IL-1)、IL-10、肿瘤坏死因子(tumor necrosis factor-，TNF-)的含量，定量聚合酶链反应(quantitative PC，qPC)检测肺组织中 NLP3 炎性小体表达情况。结果HE 染色显示，与空白对照组相比，模型组肺组织结构破坏严重，肺泡间隔变宽，肺泡萎缩，模型构建成功;与模型组相比，治疗组肺组织结构破坏程度减轻，
3、肺泡间隔厚度变小，充血水肿有所缓解。与模型组相比，治疗组肺组织 W/D 明显降低(P 0 05)。ELISA 结果显示，与空白对照组相比，模型组 IL-1、TNF-显著上升，IL-10 的表达显著下降(P 0 05);与模型组相比，治疗组 IL-1、TNF-显著下降，IL-10 的表达显著上升(P 0 05)。qPC 结果显示，与空白对照组相比，NLP3 在模型组中高表达，而治疗组中 NLP3 的表达量较模型组显著降低(P 0 05)。结论清肺排毒汤可能通过抑制 NLP3 炎性信号通路影响相关炎性因子的表达，减轻炎性反应，对脂多糖诱导的急性肺损伤起到保护作用。关键词:清肺排毒汤;脂多糖;急性肺
4、损伤中图分类号:259文献标志码:AEffect and mechanism of Qingfei Paidu decoction on LPS induced acute lung inju-ry in miceQIU Aizhu1，XU Yeqing2，OUYANG Yiguo1，ZHANG Bin1，ZENG Dan1，WEN Fang1(1 Hunan Traditional Chinese Med-icine College，Zhuzhou，Hunan Province，412012;2 Department of General Surgery，Zhuzhou Central Ho
5、spital，Zhuzhou，Hunan Province，412001，China)Abstract:ObjectiveTo observe the effect of Qingfei Paidu decoction on lipopolysaccharide induced acute lung injury and explore itsmechanism MethodsThe acute lung injury model of mice was established by intratracheal instillation of lipopolysaccharide(5 mg/k
6、g)The model mice were randomly divided into three groups:blank control group，model group and treatment group Two hours after mod-eling，the blank control group and the model group were given 0 3 mL of 0 9%sodium chloride injection by gavage，while the treatmentgroup was given 03 mL of Qingfei Paidu De
7、coction(containing 2 g/mL of crude drug)by gavage，once every four hours，a total of threetimes The mice were killed 12 hours after the last administration The lung tissue of the mice was taken，and the pathological changes ofthe lung tissue were observed by HE staining，and the ratio of wet/dry mass(W/
8、D)of the lung was measured The alveolar lavage fluid ofthe mice in each group was collected，and the content of interleukin-1(IL-1)，IL-10，tumor necrosis factor-(TNF-)was measured byELISA，Quantitative polymerase chain reaction(qPC)was used to detect the expression of NLP3 inflammatory bodies in lung t
9、issueesultsHE staining showed that compared with the blank control group，the structure of lung tissue in the model group was severelydamaged，the alveolar space was widened，and the alveoli were atrophied;Compared with the model group，the degree of destruction of lungtissue structure in the treatment
10、group was reduced，the thickness of alveolar septa was reduced，and congestion and edema were alleviatedCompared with the model group，the W/D of lung tissue in the treatment group was significantly lower(P 0 05)The results of ELISAshowed that compared with the blank control group，the expression of IL-
11、1 and TNF-in the model group increased significantly，and theexpression of IL-10 decreased significantly(P 0 05);Compared with the model group，IL-1 and TNF-in the treatment group基金项目:湖南中医药高等专科学校新冠肺炎研究专项(3)作者简介:邱爱珠(1984)，女(汉族)，副教授，硕士(E-mail:qiuaizhu2008126 com)552第 44 卷第 4 期2023 年 08 月吉林医药学院学报Journalo
12、fJilinMedicalUniversityVol 44No 4Aug 2023DOI:10.13845/ki.issn1673-2995.2023.04.023decreased significantly，and the expression of IL-10 increased significantly(P 0 05)QPC results showed that compared with the blankcontrol group，NLP3 was highly expressed in the model group，while the expression of NLP3
13、in the treatment group was significantly low-er than that in the model group(P 0 05)ConclusionQingfei Paidu decoction may reduce the inflammatory response by inhibitingthe inflammatory signal pathway of NLP3 and affecting the expression of related inflammatory factors，and play a protective role in l
14、ipopo-lysaccharide induced acute lung injuryKey words:Qingfei Paidu decoction;lipopolysaccharide;acute lung injury急性肺损伤(acute lung injury，ALI)及其严重形式急性呼吸窘迫综合征(acute respiratory distress syn-drome，ADS)是以急性发作、难治性低氧血症、肺水肿及上皮损伤为特征的非心源性呼吸衰竭1，其常见病理改变以弥漫性炎性损伤、呼吸膜和血管等组织结构破坏为主2。目前 ALI/ADS 的临床治疗主要以机械通气为主，缺乏特异的
15、药物治疗3，病死率仍高达 35%46%1，是亟待解决的医学难题之一。清肺排毒汤对 ALI 展示了良好的防治效果，但其机制尚未明确。本研究通过观察清肺排毒汤对脂多糖诱导的 ALI 小鼠的影响，探讨其对 ALI 的作用机制。1材料和方法1 1实验材料药物及试剂:清肺排毒汤(处方:炙甘草、黄芩、枳实、细辛、陈皮6 g，麻黄、杏仁、桂枝、泽泻、猪苓、白术、姜半夏、生姜、紫苑、冬花、射干、藿香 9 g，山药12 g，茯苓 15 g，柴胡 16 g，生石膏 15 30 g，先煎)中药饮片购于上市药品零售企业老百姓大药房(药物浸泡 30 min 后，加水没过药面 5 cm，武火煎至沸腾后改文火，继续煎煮 2
16、0 min，取汁另置;继续加水，煎煮沸腾后，文火续煎20 min，取汁;两次药汁合并后浓縮至每毫升含生药 2 g，并用离心机去除药汁中残渣，4 保存备用)。脂多糖(北京索莱宝科技有限公司，Lot No 111C031，L8880);苏木素染液(北京中衫金桥生物技术有限公司，19120302，ZLI-9610);伊红染色液(北京索莱宝科技有限公司，G1100);小鼠白介素 1(interleukin-1，IL-1)试剂盒(江苏酶免实业有限公司，MM-0040M2);小鼠肿瘤坏死因子(tumornecrosis factor-，TNF-)试剂盒(江苏酶免实业有限公司，MM-0132M2);小鼠 IL-10 试剂盒(江苏酶免实业有限公司，MM-0176M2);超净高级封片胶(珠海贝索生物技术有限公司，YZB，C210703);Scott 蓝化液(北京索莱宝科技有限公司，G1865);Trizon eagent(北京康为世纪生物科技有限公司，CW0580S);超纯NA 提取试剂盒(北京康为世纪生物科技有限公司，CW0581M);HiScript Q T SuperMix for qPC(+gD