摘要
目的:探讨外感清热解毒方对脂多糖(LPS)诱导急性肺损伤(ALI)大鼠的影响及作用机制。方法:20只雄性Wistar大鼠随机分为对照组、模型组、外感清热解毒方组、miR-495抑制剂组。气管滴注LSP(5 mg/kg)成功复制ALI大鼠模型,给予相应干预。定量PCR检测大鼠肺组织miR-495水平;苏木精-伊红(HE)染色观察肺组织病理改变;酶联免疫吸附试验检测肺泡灌洗液(BALF)中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、IL-1β、IL-18、IL-10炎症介质和髓过氧化物酶(MPO)活性;蛋白质免疫印迹法检测肺组织核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、凋亡相关微粒蛋白(ASC)、胱天蛋白酶-1(Caspase-1)、GSDMD蛋白表达。结果:与对照组比较,模型组大鼠肺组织miR-495表达下调(P<0.05),出现病理损伤,BALF中IL-6、TNF-α、IL-1β、IL-18含量升高,IL-10含量下降,MPO活性增强;肺组织NLRP3、ASC、Caspase-1蛋白表达增加(均P<0.05)。与模型组和miR-495抑制剂组比较,外感清热解毒方组大鼠肺组织miR-495表达上调(P<0.05),病理损伤减轻,BALF中IL-6、TNF-α、IL-1β、IL-18含量下降,IL-10含量升高,MPO活性减弱;NLRP3、ASC表达下调(均P<0.05),Caspase-1表达差异无统计学意义(P>0.05)。除对照组外,均可见GSDMD蛋白切割,外感清热解毒方组蛋白切割表达水平明显低于模型组和miR-495抑制剂组。结论:外感清热解毒方对ALI大鼠具有保护作用,其机制可能与调控miR-495/NLRP3/GSDMD信号通路,减轻炎症反应,抑制细胞焦亡有关。
Objective:This paper aims to investigate the effect and mechanism of Waigan Qingre Jiedu Formula on lipopolysaccharide(LSP)-induced acute lung injury(ALI)in rats.Methods:Twenty male Wistar rats were randomly divided into a control group,a model group,a Waigan Qingre Jiedu Formula group,and a miR-495 inhibitor group.The rat model of ALI was successfully induced by intratracheal instillation of LSP(5 mg/kg),and corresponding interventions were performed.qPCR was used to detect the level of miR-495 in rat lung tissue.HE staining was used to observe the pathological changes in lung tissue.The levels of IL-6,TNF-α,IL-1β,IL-18,and IL-10 inflammatory cytokines and myeloperoxidase(MPO)activity in bronchoalveolar lavage fluid(BALF)were detected by ELISA.Western blot was used to detect the expression of nucleotide binding oligomerization domain-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein(ASC),cysteine aspartate protease-1(Caspase-1),and gasdermin D(GSDMD)proteins in lung tissue.Results:Compared with the control group,the expression of miR-495 in the lung tissue of rats in the model group was down-regulated(P<0.05),and pathological injury occurred.The content of IL-6,TNF-α,IL-1β,and IL-18 in BALF increased,and that of IL-10 decreased.MPO activity increased.NLRP3,ASC,and Caspase-1 protein expression increased in lung tissue(all P<0.05).Compared with the model group and inhibitor group,the expression of miR-495 in lung tissue of the Waigan Qingre Jiedu Formula group was up-regulated(P<0.05),and pathological injury was alleviated.The content of IL-6,TNF-α,IL-1β,and IL-18 in BALF was decreased,and that of IL-10 was increased.MPO activity was decreased.NLRP3 and ASC expression were down-regulated(all P<0.05),and there was no significant difference in Caspase-1 expression(P>0.05).Except for the control group,GSDMD protein cleavage was observed in all groups,and the expression level of protein cleavage in the Waigan Qingre Jiedu Formula group was significantly lower than that of the model group and
作者
刘玉婷
王浩艺
麦静愔
何青青
何东仪
陈建杰
成扬
LIU Yuting;WANG Haoyi;MAI Jingying;HE Qingqing;HE Dongyi;CHEN Jianjie;CHENG Yang(Shanghai Guanghua Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai 200052,China;Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)
出处
《世界中医药》
CAS
2023年第23期3328-3333,3341,共7页
World Chinese Medicine
基金
国家自然科学基金项目(81774098)
第五批全国中医临床优秀人才研修项目
上海中医药大学预算内科研项目(2020LK84)
上海市长宁区创新团队项目(2020100100061)。
