藏药十二味翼首散通过抑制中性粒细胞浸润减轻脂多糖诱导急性肺损伤的研究

Tibetan medicine Twelve Flavors Yishou San alleviated lipopolysaccharide-induced acute lung injury through inhibiting neutrophil infiltration

目的探讨藏药十二味翼首散干预急性肺损伤的免疫机制。方法构建脂多糖(lipopolysaccharide,LPS)诱导的急性肺损伤大鼠模型,并以十二味翼首散处理。通过肺部病理切片及血气分析检测明确十二味翼首散对LPS诱导急性肺损伤的干预作用。应用酶联免疫吸附分析(enzyme-linked immunosorbent assay,ELISA)技术检测大鼠肺泡灌洗液促炎因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)及抑炎因子白细胞介素(interleukin,IL)-10的表达情况。流式细胞术检测大鼠肺泡灌洗液、肺、脾及外周血中免疫细胞的变化情况。结果LPS成功诱导大鼠急性肺损伤模型,十二味翼首散可在不同程度上减轻LPS诱导急性肺损伤中炎症细胞的渗出,升高氧分压。大鼠肺泡灌洗液ELISA结果显示,与NC组比较,急性肺损伤大鼠的肺泡灌洗液TNF-α水平升高、IL-10水平降低,而十二味翼首散可缓解该效应。流式细胞术结果表明,十二味翼首散可显著降低大鼠肺泡灌洗液中中性粒细胞浸润,而对巨噬细胞及适应性免疫应答无明显影响。结论十二味翼首散可通过抑制中性粒细胞浸润减轻LPS诱导的急性肺损伤,这为充分认识十二味翼首散治疗作用的效应机制提供实验依据。

Objective To explore the immune mechanism of Tibetan medicine Twelve Flavors Yishou San in intervening with acute lung injury.Methods Construct a lipopolysaccharide(LPS)-induced acute lung injury rat model and treated it with Twelve Flavors Yishou San.The intervention effect of Twelve Flavors Yishou San on LPS induced acute lung injury was determined through lung pathological sections and blood gas analysis.Enzyme linked immunosorbent assay(ELISA)was used to detect the expression of pro-inflammatory cytokine tumor necrosis factor-α(TNF-α)and anti-inflammatory cytokine interleukin(IL)-10 in rat’s bronchoalveolar lavage fluid.Flow cytometry was used to detect changes in immune cells in rat’s bronchoalveolar lavage fluid,lungs,spleen and peripheral blood.Results LPS successfully induced an acute lung injury model in rats,and Twelve Flavors Yishou San could alleviate the exudation of inflammatory cells,elevated oxygen partial pressure in LPS induced acute lung injury to varying degrees.The ELISA results of rat bronchoalveolar lavage fluid showed that compared with normal control group,the TNF-αlevel in the bronchoalveolar lavage fluid of rats with acute lung injury was increased and the IL-10 level was decreased,while the Twelve Flavors Yishou San could alleviate this effect.The results of flow cytometry showed that Twelve Flavors Yishou San could significantly reduce neutrophil infiltration in rat bronchoalveolar lavage fluid,but had no significant effect on macrophages and adaptive immune response.Conclusion Twelve Flavors Yishou San can alleviate LPS induced acute lung injury by inhibiting neutrophil infiltration,providing an important experimental basis for fully understanding the therapeutic mechanism of Twelve Flavors Yishou San.