摘要
目的探讨抗炎通腑方对脓毒症急性肺损伤(ALI)大鼠的影响及作用机制。方法将35只大鼠随机分为正常组、模型组、地塞米松(DEX)组、中药(TCM)组和TCM+DEX组,每组7只。分别给予相应药物灌胃、造模。酶联免疫吸附实验(ELISA)法检测白细胞介素(IL)-6、IL-1β、IL-18、肿瘤坏死因子-α(TNF-α)含量及血清中巨噬细胞计数;采用实时荧光定量多聚核苷酸链式反应(RT-qPCR)和蛋白免疫印迹(Western blot)检测肺组织中干扰素基因刺激因子(STING)、磷酸化干扰素基因刺激因子(P-STING)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白信使核糖核酸(ASC mRNA)和相关蛋白的表达水平。测定肺组织湿/干比(W/D),观察肺组织病理学改变。检测肺组织中巨噬细胞表达情况。结果与正常组比较,各组指标显著升高;与模型组比较,DEX组、TCM组及TCM+DEX组各指标均显著降低。与模型组比较,TCM+DEX组改变最为明显,肺组织损伤改善,W/D值(4.77±0.29 vs.3.78±0.48)及巨噬细胞计数(13.39±2.06 vs.7.09±1.42)均降低(P<0.05),IL-6(152.51±22.27 vs.58.92±13.53)、IL-1β(126.19±10.02 vs.45.69±6.67)、IL-18(59.12±6.31 vs.31.75±4.23)及TNF-α(126.57±8.25 vs.49.59±8.12)水平均降低(P均<0.01),肺组织中线粒体DNA(mtDNA)损伤及释放、P-STING(0.32±0.03 vs.0.16±0.03)、STING mRNA(19.24±2.70 vs.0.32±0.16)、NLRP3 mRNA(20.03±5.06 vs.1.20±0.04)、ASC mRNA(16.96±4.31 vs.3.41±2.52)和相关蛋白的表达均降低(P<0.01)。结论抗炎通腑方可能通过调控mtDNA-STING-NLRP3信号通路减轻脓毒症ALI大鼠的炎症。
Objective To observe the effects of anti-inflammatory Tongfu formula on sepsis-induced acute lung injury(ALI)in rats and to explore its mechanism.Methods The 35 rats were randomly divided into normal group,model group,dexamethasone(DEX)group,traditional Chinese medicine(TCM)group and TCM+DEX group,with 7 rats in each group.The corresponding drugs were given by intragastric administration and the model was made.The contents of interleukin(IL)-6,IL-1β,IL-18,tumor necrosis factor-α(TNF-α)and serum macrophage count were detected by enzyme linked immunosorbent assay(ELISA).The expression levels of stimulator of interferon genes(STING),phosphorylated(P)-STING,NOD-like receptor thermal protein domain associated protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC)mRNA and related proteins in lung tissues were detected by reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR)and Western blot.The wet/dry ratio(W/D)of lung tissue was measured and the pathological changes of lung tissue were observed.The expression of macrophages in lung tissues was detected.Results Compared with the normal group,the indicators of other groups significantly increased.Compared with model group,the indicators in DEX group,TCM group and TCM+DEX group significantly decreased.The further comparison between the model group and the TCM+DEX group showed,lung tissue injury was improved,W/D value(4.77±0.29 vs.3.78±0.48)and macrophage count(13.39±2.06 vs.7.09±1.42)were decreased(P<0.05),IL-6(152.51±22.27 vs.58.92±13.53),IL-1β(126.19±10.02 vs.45.69±6.67),IL-18(59.12±6.31 vs.31.75±4.23)and TNF-α(126.57±8.25 vs.49.59±8.12)were decreased in the TCM+DEX group(P<0.01);The damage and release of mitochondrial DNA(mtDNA)in lung tissues,the expressions of P-STING(0.32±0.03 vs.0.16±0.03),STING mRNA(19.24±2.70 vs.0.32±0.16),NLRP3 mRNA(20.03±5.06 vs.1.20±0.04),ASC mRNA(16.96±4.31 vs.3.41±2.52)and related proteins in lung tissue were decreased in the TCM+DEX group(P<0.01).Conclusions Anti-inflammatory Tongfu formula may reduce inflammation in sepsis-induced acute lung injury rats by regulating mtDNA-STING-NLRP3 signaling pathway.
作者
霍雁
岳迪
Huo Yan;Yue Di(Department of Traditional Chinese Medicine,Shanghai Tongren Hospital Affiliated to Shanghai Jiaotong University School of Medicine,Shanghai 200336,China)
出处
《中国急救医学》
CAS
CSCD
2024年第4期323-329,共7页
Chinese Journal of Critical Care Medicine
基金
上海交通大学医学院附属同仁医院一般学科课题(tr2023xk35)。
关键词
抗炎通腑方
STING-NLRP3信号通路
脓毒症
急性肺损伤(ALI)
炎症
地塞米松
巨噬细胞计数
Anti-inflammatory Tongfu formula
Stimulator of interferon genes(STING)-NOD-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathway
Sepsis
Acute lung injury(ALI)
Inflammation
Dexamethasone
Macrophages count
