Kou Yu-le, Wang Wen-Ba, Yan Shu-Guang, Li Jing-Tao, Shi Jie, Hui Yi
Department of Respiratory, Affiliated Hospital of Shannxi University of Chinese Medicine Xianyang 712000, China.
College of Basic Medicine, Shannxi University of Chinese Medicine Xianyang 712046, China.
Zhongguo Zhong Yao Za Zhi. 2022 Jan;47(1):151-158. doi: 10.19540/j.cnki.cjcmm.20211027.401.
Lung and intestine combination therapy(LICT) is effective in the treatment of acute lung injury(ALI). In this study, the combination of Mahuang Decoction and Dachengqi Decoction(hereinafter referred to as the combination), a manifestation of LICT, was employed to explore the effect of nuclear factor kappaB(NF-κB)/nucleotide binding oligomerization domain-like receptors-3(NLRP3) pathway and alveolar macrophage activation on the lung inflammation in rats with ALI, for the purpose of elucidating the mechanism of LICT in treating ALI. After the modeling of ALI with limpolysaccharide(LPS, ip), rats were respectively given(ig) the combination at 10, 7.5, and 5 g·kg~(-1)(high-dose, medium-dose, and low-dose LICT groups, separately), once every 8 h for 3 times. Haematoxylin-eosin(HE) staining was used to observe the histopathological changes of lung tissue, followed by the scoring of inflammation. Immunohistochemistry was applied to detect alveolar macrophage activation, enzyme-linked immunosorbent assay(ELISA) was applied to detect the serum content of tumor necrosis factor-α(TNF-α) and interleukin-18(IL-18), Western blot was applied to detect the protein expression of phosphorylated-nuclear factor kappaB p65(p-NF-κB p65), nuclear factor kappaB p65(NF-κB p65), phosphorylated-inhibitor kappaB alpha(p-IκBα), inhibitor kappaB alpha(IκBα), and NLRP3 in lung tissue, and quantitative reverse transcription-PCR(qRT-PCR) was applied to detect the mRNA expression of TNF-α, IL-18, NLRP3, and NF-κB p65 in lung tissue. The results showed that LICT groups demonstrated lung injury relief, decrease in inflammation score, alleviation of alveolar macrophage activation, significant decline in serum content of inflammatory factors TNF-α and IL-18, and decrease of the protein expression of p-NF-κB p65/NF-κB p65, p-IκBα/IκBα, and NLRP3, and mRNA expression of TNF-α, IL-18, NLRP3, and NF-κB p65 in lung tissue. In summary, LICT has definite therapeutic effect on ALI. The mechanism is that it inhibits alveolar macrophage activation by suppressing NF-κB/NLRP3 signaling pathway, thereby reducing the activation and release of inflammatory factors and finally inhibiting inflammation.
肺肠同治疗法（LICT）治疗急性肺损伤（ALI）有效。本研究采用LICT的一种表现形式——麻黄汤与大承气汤联合应用（以下简称联合方），探讨核因子κB（NF-κB）/核苷酸结合寡聚化结构域样受体3（NLRP3）通路及肺泡巨噬细胞活化对ALI大鼠肺炎症的影响，以阐明LICT治疗ALI的机制。采用脂多糖（LPS，腹腔注射）建立ALI模型后，分别给予大鼠（灌胃）10、7.5和5 g·kg⁻¹的联合方（分别为高剂量、中剂量和低剂量LICT组），每8小时1次，共3次。采用苏木精-伊红（HE）染色观察肺组织的组织病理学变化，随后进行炎症评分。应用免疫组织化学检测肺泡巨噬细胞活化情况，应用酶联免疫吸附测定（ELISA）检测血清肿瘤坏死因子-α（TNF-α）和白细胞介素-18（IL-18）含量，应用蛋白质印迹法检测肺组织中磷酸化核因子κB p65（p-NF-κB p65）、核因子κB p65（NF-κB p65）、磷酸化抑制蛋白κBα（p-IκBα）、抑制蛋白κBα（IκBα）和NLRP3的蛋白表达，应用定量逆转录聚合酶链反应（qRT-PCR）检测肺组织中TNF-α、IL-18、NLRP3和NF-κB p65的mRNA表达。结果显示，LICT组肺损伤减轻，炎症评分降低，肺泡巨噬细胞活化减轻，血清炎症因子TNF-α和IL-18含量显著下降，肺组织中p-NF-κB p65/NF-κB p65、p-IκBα/IκBα和NLRP3的蛋白表达以及TNF-α、IL-18、NLRP3和NF-κB p65的mRNA表达均降低。综上所述，LICT对ALI有确切治疗作用。其机制是通过抑制NF-κB/NLRP3信号通路抑制肺泡巨噬细胞活化，从而减少炎症因子的活化和释放，最终抑制炎症。
