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目的:通过体内外实验探讨清肺理痰方对克雷伯菌诱导的细菌性肺炎的作用机制。方法:将60只C57BL/6小鼠随机分为正常对照组、模型对照组、头孢克肟52 mg/kg组、清肺理痰方2.2、4.5、9 g/kg组，通过气管滴注克雷伯菌悬浮液建立细菌性肺炎的动物模型。造模8 h后，各组灌胃给予相应药物或等体积生理盐水，连续灌胃3 d。检测小鼠肺组织载菌量，计算肺指数；HE染色观察肺组织病理损伤；IHC染色观察肺组织JAK/STAT信号通路蛋白的表达。体外实验：用热灭活的克雷伯菌诱导小鼠肺泡巨噬细胞(MH-S)建立细胞炎症模型。将肺泡巨噬细胞分为空白对照组、模型对照组、清肺理痰方2.5%、5%、10%含药血清组，流式细胞术检测肺泡巨噬细胞吞噬荧光微球能力；qRT-PCR法检测细胞中白细胞介素6(Il6)、Il1b、肿瘤坏死因子(Tnfα)的表达；Western blot法检测细胞JAK/STAT相关通路蛋白的表达。结果:与正常对照组相比，模型对照组小鼠肺组织载菌量显著增加，肺指数显著升高(P<0.01),肺组织炎症细胞浸润增加，肺组织p-JAK1、p-JAK2、p-STAT3、p-STAT6蛋白的表达明显上调(P<0.05或P<0.01);与模型对照组相比，清肺理痰各组小鼠肺组织载菌量显著减少，肺指数显著降低(P<0.01),肺组织炎症细胞浸润减少，肺组织p-JAK1、p-JAK2、p-STAT3、p-STAT6蛋白表达明显下调(P<0.05)。与空白对照组相比，模型对照组细胞吞噬率升高，MH-S细胞Il6、Il1β、Tnfα的表达显著上调(P<0.01)、p-JAK1/JAK1、p-JAK2/JAK2、p-STAT3/STAT3、p-STAT6/STAT6的比值明显升高(P<0.05或P<0.01);与模型对照组相比，清肺理痰含药血清各组能明显增强肺泡巨噬细胞吞噬荧光微球的能力(P<0.01);MH-S细胞Il6、Il1β、Tnfα的表达明显下调(P<0.05或P<0.01),p-JAK1/JAK1、p-JAK2/JAK2、p-STAT3/STAT3、p-STAT6/STAT6的比值明显降低(P<0.05或P<0.01)。结论:清肺理痰方通过调控JAK/STAT通路，改善肺炎克雷伯菌所致细菌性肺炎。
Abstract：Objective:To investigate the mechanism by which Qingfei Litan Formula(清肺理痰方,QFLTF) ameliorates Klebsiella pneumoniae-induced bacterial pneumonia. Methods:In vivo experiments: Sixty C57BL/6 mice were randomly divided into normal control, model control, cefixime(52 mg/kg),and QFLTF groups(2.2,4.5,and 9 g/kg). Bacterial pneumonia was induced by intratracheal instillation of K. pneumoniae suspension. Eight hours after infection, mice received corresponding treatments or saline via oral gavage for three consecutive days. Pulmonary bacterial load and lung index were assessed. Lung histopathology was evaluated by hematoxylin and eosin(HE) staining, and JAK/STAT pathway protein expression in lung tissue was detected by immunohistochemistry(IHC). In vitro experiments: MH-S alveolar macrophages were stimulated with heat-inactivated K. pneumoniae(iKp) to establish an inflammatory model. Cells were divided into control, model, and QFLTF-containing serum groups(2.5%,5%,10%). Phagocytic capacity of alveolar macrophages was assessed by flow cytometry using fluorescent microspheres. Expression levels of interleukin 6(Il6),interleukin 1β(Il1β),and tumor necrosis factor α(Tnfα) were determined using qRT-PCR,and JAK/STAT pathway protein expression was analyzed by Western blotting(WB). Results:In in vivo experiments, compared with the normal control group, the model group showed significantly increased pulmonary bacterial load and lung index(P<0.01),severe inflammatory cell infiltration, and markedly upregulated p-JAK1,p-JAK2,p-STAT3,and p-STAT6 expression(P<0.05 or P<0.01). Compared with the model control group, QFLTF treatment significantly reduced bacterial load and lung index(P<0.01),alleviated inflammatory infiltration, and downregulated p-JAK1,p-JAK2,p-STAT3,and p-STAT6 expression(P<0.05). In in vitro experiments, compared to the fluorescent microsphere group, QFLTF-containing serum significantly enhanced the phagocytic capacity of MH-S cells(P<0.01). Compared with the normal control group, the model control group exhibited significantly upregulated Il6,Il1β,and Tnfα expression(P<0.01) and increased p-JAK1/JAK1,p-JAK2/JAK2,p-STAT3/STAT3,and p-STAT6/STAT6 ratios(P<0.05 or P<0.01). Compared with the model control group, QFLTF-containing serum significantly downregulated Il6,Il1β,and Tnfα expression(P<0.05 or P<0.01) and reduced p-JAK1/JAK1,p-JAK2/JAK2,p-STAT3/STAT3,and p-STAT6/STAT6 ratios(P<0.05 or P<0.01). Conclusion:QFLTF ameliorates K. pneumoniae-induced bacterial pneumonia by modulating the JAK/STAT signaling pathway.
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基本信息:
DOI：10.13412/j.cnki.zyyl.20250929.018
中图分类号:R285.5
引用信息:
[1]魏若君,史渊源,丁奇.清肺理痰方通过调控JAK/STAT通路改善克雷伯菌所致细菌性肺炎[J].中药药理与临床,2026,42(06):18-24.DOI:10.13412/j.cnki.zyyl.20250929.018.
基金信息:
国家自然科学基金青年科学基金项目(编号：82405053); 深圳市基础研究专项自然科学基金计划面上项目(编号：JCYJ20230807150505010)
2025-09-30
2025-09-30
2025-09-30