摘要
中医肺肠合治法治疗急性肺损伤疗效显著,该研究以体现肺肠合治的麻黄汤和大承气汤联合应用,探究肺肠合治法经核转录因子-κB(nuclear factor kappaB,NF-κB)/NOD样受体家族3(nucleotide binding oligomerization domain-like receptors-3,NLRP3)通路及肺泡巨噬细胞活化对急性肺损伤大鼠肺组织炎症的影响,揭示肺肠合治法治疗急性肺损伤的作用机制。采用脂多糖腹腔注射制备急性肺损伤大鼠模型,给予肺肠合治法高(10 g·kg^(-1))、中(7.5 g·kg^(-1))、低(5 g·kg^(-1))剂量灌胃,每8 h给药1次,共3次。苏木素-伊红(haematoxylin-eosin,HE)染色法观察肺组织病理组织学变化,进行炎症评分;免疫组织化学法检测肺泡巨噬细胞活化,酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)法检测血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α),白细胞介素-18(interleukin-18,IL-18)含量;蛋白免疫印迹(Western blot)法检测肺组织中核转录因子-κB p65(nuclear factor kappaB p65,NF-κB p65)、磷酸化核转录因子-κB p65(phosphorylated-nuclear factor kappaB p65,p-NF-κB p65)、核因子抑制蛋白α(inhibitor kappaB,IκBα)、磷酸化核因子抑制蛋白α(phosphorylated-inhibitor kappaB alpha,p-IκBα)、NLRP3蛋白表达;实时荧光定量PCR(qRT-PCR)法检测肺组织中TNF-α、IL-18、NLRP3、NF-κB p65 mRNA表达。结果显示,肺肠合治法各组大鼠肺组织损伤明显减轻,炎症评分降低,肺泡巨噬细胞活化程度降低;血清炎症因子TNF-α、IL-18含量显著降低;肺组织p-NF-κB p65/NF-κB p65、p-IκBα/IκBα、NLRP3蛋白相对表达降低;肺组织TNF-α、IL-18、NLRP3、NF-κB p65 mRNA表达降低。综上结果,肺肠合治法对急性肺损伤有明显的治疗作用,其作用机制可能是通过抑制NF-κB/NLRP3信号通路,抑制巨噬细胞活化,减少炎症因子的激活和释放,进而抑制炎症反应。
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-αand 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.
作者
寇育乐
王文霸
闫曙光
李京涛
史捷
惠毅
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,Shanxi University of Chinese Medicine,Xianyang 712046,China;Department of Infectious Disease,Affiliated Hospital of Shannci University of Chinese Medicine,Xianyang 712000,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2022年第1期151-158,共8页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81703974)
陕西省自然科学基础研究计划面上项目(2021JM-473)
陕西中医药大学学科创新团队建设项目(2019-YL05)。
关键词
肺肠合治
麻黄汤
大承气汤
急性肺损伤
NF-κB/NLRP3信号通路
炎症反应
lung and intestine combination therapy
Mahuang Decoction
Dachengqi Decoction
acute lung injury
NF-κB/NLRP3 signaling pathway
inflammatory response
