摘要
目的基于“肺-肠轴”理论,从肠道微生态角度研究泻白散对脂多糖(LPS)诱导的肺损伤幼龄大鼠的抗炎作用机制。方法将60只幼龄雄性SD大鼠随机分为空白组、模型组、阿奇霉素组及泻白散低、中、高剂量组,每组10只。每日于早晚固定时间雾化吸入LPS水溶液(0.5 mg·mL^(-1))15 mL,每次30 min,连续3 d,复制肺损伤(肺炎)大鼠模型。泻白散低、中、高剂量分别为3.4、6.7、13.4 g·kg^(-1);阿奇霉素第1天给药剂量为1.1 mg·kg^(-1),第2~7天给药剂量为0.55 mg·kg^(-1)。模型复制期间,各给药组大鼠同时灌胃相应药物,每日1次,连续7 d。采用HE染色法观察肺组织病理变化;ELISA法测定肺泡灌洗液中的肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、白细胞介素1β(IL-1β)水平;免疫组化法测定肺组织中的IL-6、IL-1β水平;ELSIA法测定肺、结肠组织中的P物质(SP)、血管活性肠肽(VIP)水平;16S rDNA法分析肠道菌群;斯皮尔曼(Spearman)法分析生化指标与肠道菌群的相关性。结果(1)与空白组比较,模型组大鼠的部分肺泡壁明显增厚,肺泡数量有所下降,部分肺泡结构破坏,出现炎性细胞浸润现象;肺泡灌洗液中的TNF-α、IL-6、IL-1β水平显著升高(P<0.05,P<0.001);肺组织中的IL-6水平显著升高(P<0.001),IL-1β水平升高但差异无统计学意义(P>0.05);肺、肠组织中的SP水平显著升高(P<0.05,P<0.001),VIP水平明显降低(P<0.05)。与模型组比较,各给药组大鼠肺组织炎症病变均出现不同程度好转;阿奇霉素组及泻白散低、高剂量组大鼠肺泡灌洗液中的TNF-α、IL-6水平显著降低(P<0.05,P<0.001),各给药组大鼠肺泡灌洗液中的IL-1β水平均显著降低(P<0.01,P<0.001);阿奇霉素组及泻白散高剂量组大鼠肺组织中的IL-6水平显著降低(P<0.05,P<0.001),各给药组大鼠肺组织中的IL-1β水平均显著降低(P<0.001);泻白散中剂量组大鼠肺、肠组织中的SP水平显著降低(P<0.01,P<0.001),VIP水平显著升高(P<0.05,P<0.01)。(2)与空白组比较,模型组大鼠肠道菌群的Shannon、Simpson Alpha多样性指数均无明显变化(P>0.05);与空白组或模型组比较,泻白散中剂量组大鼠肠道菌群的Shannon、Simpson指数均明显降低(P<0.05)。泻白散对LPS诱导的肺损伤大鼠的肠道菌群多样性、肠道微生物群落构成、肠道菌群标志物等均具有调节作用。泻白散对LPS诱导的肺损伤大鼠肠道菌群的影响与其对肺、肠组织中炎性因子IL-6、IL-1β及SP、VIP等神经肽的调节存在潜在相互作用关系有关。结论泻白散对LPS诱导的肺损伤幼龄大鼠肺部炎症具有明显改善作用,该作用与调节肠道菌群及免疫反应之间的相互作用有关。
Objective To investigate the anti-inflammatory mechanism of Xiebai San(XBS)in lipopolysaccharide(LPS)-induced lung injury in juvenile rats from the perspective of gut microbiota based on the"gut-lung axis"theory.Methods Sixty SD rats were randomly divided into six groups(n=10 per group):control,model,Azithromycin,and XBS low-(3.4 g·kg^(-1)),medium-(6.7 g·kg^(-1)),and high-dose(13.4 mg·kg^(-1))groups.The lung injury(pneumonia)model was established by intratracheal aerosol inhalation of LPS solution(0.5 mg·mL^(-1),15 mL per session,30 minutes per session,twice daily for 3 consecutive days).Drug treatments were administered once daily for 7 days:Azithromycin(1.1 mg·kg^(-1)on day 1,0.55 mg·kg^(-1)on days 2-7)and XBS at respective doses via oral gavage.Pathological changes in lung tissues were assessed by HE staining.Levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in bronchoalveolar lavage fluid(BALF)were measured by ELISA.Immunohistochemistry was used to quantify IL-6 and IL-1β in lung tissues.Substance P(SP)and vasoactive intestinal peptide(VIP)levels in lung and colon tissues were determined by ELISA.Gut microbiota composition was analyzed via 16S rDNA sequencing,and Spearman correlation analysis was performed to assess relationships between biochemical indices and microbial taxa.Results(1)Compared with the blank group,the model group showed significantly thickened alveolar walls,reduced alveolar numbers,structural damage to some alveoli,and inflammatory cell infiltration;levels of TNF-α,IL-6,and IL-1β in bronchoalveolar lavage fluid(BALF)were significantly increased(P<0.05,P<0.001);IL-6 levels in lung tissue were markedly elevated(P<0.001)while IL-1β showed no significant change(P>0.05);substance P(SP)levels in lung and intestinal tissues were significantly increased(P<0.05,P<0.001),whereas vasoactive intestinal peptide(VIP)levels were significantly decreased(P<0.05).Compared with the model group,all treatment groups exhibited varying degrees of improvement in pulmonary inflammation:the Azithromycin group and low-/high-dose XBS groups showed significantly reduced TNF-α and IL-6 levels in BALF(P<0.05,P<0.001),while all treatment groups demonstrated significantly decreased IL-1β levels(P<0.01,P<0.001);the Azithromycin group and high-dose XBS group had significantly lower IL-6 levels in lung tissue(P<0.05,P<0.001),and all treatment groups showed significantly reduced IL-1β levels(P<0.001);the medium-dose XBS group exhibited significantly decreased SP levels(P<0.01,P<0.001)and increased VIP levels(P<0.05,P<0.01)in lung and intestinal tissues.(2)Compared with the blank group,the model group showed no significant changes in Shannon or Simpson Alpha diversity indices of gut microbiota(P>0.05);however,the mediumdose XBS group demonstrated significantly lower Shannon and Simpson indices compared with either the blank or model group(P<0.05).XBS modulated gut microbiota diversity,microbial community composition,and microbial biomarkers in LPS-induced lung injury rats,and these effects were potentially associated with its regulatory actions on inflammatory factors(IL-6,IL-1β)and neuropeptides(SP,VIP)in lung and intestinal tissues.Conclusion XBS alleviates LPS-induced lung inflammation in juvenile rats,likely via gut microbiota-immune crosstalk.
作者
姚素媛
张庆玲
王坤然
邓凡莹
王瑞麒
YAO Suyuan;ZHANG Qingling;WANG Kunran;DENG Fanying;WANG Ruiqi(Institute of Adult Education,Heilongjiang University of Chinese Medicine,Harbin 150040 Heilongjiang,China;Institute of the First Clinical College of Heilongjiang University of Chinese Medicine,Harbin 150040 Heilongjiang,China;Institute of Basic Medicine,Heilongjiang University of Chinese Medicine,Harbin 150040 Heilongjiang,China;College of Pharmacy,Heilongjiang University of Chinese Medicine,Harbin 150040 Heilongjiang,China;Institute of Pharmaceutical&Food Engineering,Shanxi University of Chinese Medicine,Jinzhong 030619 Shanxi,China)
出处
《中药新药与临床药理》
北大核心
2025年第10期1658-1669,共12页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
黑龙江省中医药科研项目(20230830225018)
黑龙江省卫生健康委科技计划项目(20221313050712)。
关键词
泻白散
肺损伤
肺炎
肺-肠轴
肠道菌群
抗炎作用
免疫反应
幼龄大鼠
Xiebai San
lung injury
pneumonia
gut-lung axis
gut microbiota
anti-inflammatory effect
immune response
juvenile rats
