Liu Fang, Liu TianTian, Sun Min, Zhou JingMin, Xue Feng, Chen ShuangHui, Chen Jia, Zhang Lei
Department of Pediatrics, Pudong New Area People's Hospital, Shanghai, 201200, People's Republic of China.
Department of Pediatrics, East Hospital Affiliated to Tongji University, Shanghai, 200123, People's Republic of China.
Infect Drug Resist. 2021 Mar 3;14:859-867. doi: 10.2147/IDR.S292413. eCollection 2021.
is a predominant cause of community-acquired respiratory infections. We recently discovered the clinical efficacy of Maxing shigan decoction (MXSG) in infection and designed a study to explore the mechanism of action.
Serum IL-1β, IL-18, and TNF-α, and transcript expression of the NLR Family, Pyrin Domain Containing Protein 3 (NLRP3) were measured in the peripheral blood mononuclear cells (PBMCs) of 30 children with infection and 30 healthy donors. An in vitro model of infection in A549 cell culture was used to explore the curative effects and mechanisms of MXSG. Pyroptosis was measured by flow cytometry with activated caspase-1 and propidium iodide staining. IL-1β, IL-18, and TNF-α, and NLRP3 transcript expression were measured by qRT-PCR. Protein expression of NLRP3, Caspase-1, pro-caspase-1, IL-1β, pro-IL-1β, and GSDMD-N was determined by Western blotting. Experimental confirmation was performed in NLRP3-overexpressing A549 cells and in the presence of an NLRP3 inhibitor, INF39.
infection-induced IL-1β, IL-18, TNF-α, and mRNA expression of NLRP3 in PBMCs and promoted pyroptosis in A549 cells. It also induced IL-1β, IL-18, TNF-α, and up-regulated NLRP3, ro-IL-1β, Caspase-1, Pro-Caspase-1, and GSDMD-N in culture. Similar to the NLRP3 inhibitor INF39, MXSG (0.1, 0.2, and 0.4 mg/mL) suppressed pyroptosis induced by infection and decreased IL-1β ( < 0.001), IL-18, TNF-α in culture. MXSG down-regulated NLRP3, pro-IL-1β, Caspase-1, pro-Caspase-1, and GSDMD-N in infected cultures and mitigated NLRP3 overexpression-induced pyroptosis.
MXSG mitigates -induced pyroptosis in A549 cells via the NLRP3 inflammasome.
是社区获得性呼吸道感染的主要原因。我们最近发现了麻杏石甘汤（MXSG）在感染中的临床疗效，并设计了一项研究来探索其作用机制。
检测了30例感染患儿和30例健康供体外周血单个核细胞（PBMCs）中血清白细胞介素-1β（IL-1β）、白细胞介素-18（IL-18）、肿瘤坏死因子-α（TNF-α）以及含吡啉结构域的NLR家族蛋白3（NLRP3）的转录表达。采用A549细胞培养中的感染体外模型来探究MXSG的疗效和机制。通过活化的半胱天冬酶-1和碘化丙啶染色，利用流式细胞术检测细胞焦亡。采用qRT-PCR检测IL-1β、IL-18、TNF-α以及NLRP3转录表达。通过蛋白质印迹法测定NLRP3、半胱天冬酶-1（Caspase-1）、前半胱天冬酶-1、IL-1β、前白细胞介素-1β（pro-IL-1β）和Gasdermin D-N（GSDMD-N）的蛋白表达。在过表达NLRP3的A549细胞中以及存在NLRP3抑制剂INF39的情况下进行实验验证。
感染诱导PBMCs中IL-1β、IL-18、TNF-α以及NLRP3的mRNA表达，并促进A549细胞中的细胞焦亡。它还在培养物中诱导IL-1β、IL-18、TNF-α，并上调NLRP3、前白细胞介素-1β、Caspase-1、前半胱天冬酶-1和GSDMD-N。与NLRP3抑制剂INF39相似，MXSG（0.1、0.2和0.4mg/mL）抑制感染诱导的细胞焦亡，并降低培养物中IL-1β（P<0.001）、IL-18、TNF-α的水平。MXSG下调感染培养物中的NLRP3、前白细胞介素-1β、Caspase-1、前半胱天冬酶-1和GSDMD-N，并减轻NLRP3过表达诱导的细胞焦亡。
MXSG通过NLRP3炎性小体减轻感染诱导的A549细胞焦亡。
