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目的:研究清肺理痰方各有效成分组对急性肺损伤大鼠的影响及可能的作用机制。方法:SD大鼠随机分为正常对照组、模型组、地塞米松组、清肺理痰方水提取物组、乙醇提取物组、石油醚萃取物组、乙酸乙酯萃取物组、正丁醇萃取物组、残余水层组;用以上中药复方各有效成分组预防性给药,采用气管内注入脂多糖致大鼠急性肺损伤;观察造模前后大鼠的一般情况;采用酶联免疫吸附试验(ELISA)检测血清中白介素(IL)-1β、肿瘤坏死因子(TNF)-α、白介素(IL)-6的水平;光镜下观察肺组织病理变化;免疫组织化学技术观察大鼠肺组织中核因子κB(NF-κB)p65蛋白表达情况。结果:与正常对照组比较,模型组大鼠肺组织呈现典型的炎症病理特征,肺组织损伤明显,肺湿/干重比、血清中IL-1β,TNF-α,IL-6含量及肺组织中NF-κB p65蛋白表达明显升高(P<0.01)。与模型组比较,清肺理痰方各有效成分组、地塞米松组肺组织病理损伤有不同程度地减轻,肺湿/干重比、血清中IL-1β,TNF-α,IL-6含量及肺组织中NF-κB p65蛋白表达出现不同程度的降低,其中以水提取物组和乙醇提取物组降低最明显(P<0.01或P<0.05),与地塞米松组疗效相当。结论:中药复方清肺理痰方各有效成分组可不同程度地减轻内毒素致急性肺损伤大鼠的病理损害,其机制可能通过抑制NF-κB信号转导通路,进而减少细胞因子(如IL-1β,TNF-α,IL-6)分泌以减轻脏器组织损伤,其中以水提取物组和乙醇提取物组疗效最明显。
Objective: To study the effect and its possible mechanism of each active ingredient of Qingfei Li Tan Decoction on acute lung injury in rats. Methods: SD rats were randomly divided into normal control group, model group, dexamethasone group, Qingfei Liutan Fang water extract group, ethanol extract group, petroleum ether extract group, ethyl acetate extract group, Alcohol extract group and residual water layer group. Using the above active ingredients of the compound prescription of Chinese medicine for prophylactic administration, tracheal injection of lipopolysaccharide induced acute lung injury in rats. The general situation of rats before and after modeling was observed. Enzyme-linked immunosorbent assay The levels of IL-1β, TNF-α and IL-6 in serum were detected by ELISA. The pathological changes of lung tissue were observed under light microscope. The expression of IL-1β was detected by immunohistochemistry The expression of nuclear factor kappa B (NF-κB) p65 protein in tissues. Results: Compared with the normal control group, the lung tissue of the model group showed typical pathological features of inflammation, lung injury, lung wet / dry weight ratio, serum IL-1β, TNF-α, IL-6 content and lung tissue NF-κB p65 protein expression was significantly increased (P <0.01). Compared with the model group, the pathological changes of the lungs in the active ingredients of Qingfei Li Tan Decoction group and the dexamethasone group were alleviated to varying extents, the wet / dry weight ratio of lungs, the contents of IL-1β, TNF-α and IL-6 (P <0.01 or P <0.05), which was comparable to that of dexamethasone group. The levels of NF-κB p65 protein in lung tissue were significantly lower than those in dexamethasone group. Conclusion: The active constituents of Qingfeishutan recipe can relieve the pathological damage induced by LPS-induced acute lung injury in rats to a certain extent. The mechanism may be through the inhibition of NF-κB signal transduction pathway and the decrease of cytokines (such as IL -1β, TNF-α, IL-6) secretion to reduce organ tissue injury, of which the water extract group and ethanol extract group the most obvious effect.