六味小青龙汤抑制p38MAPK/p22phox通路减轻哮喘大鼠慢性炎症作用机制研究  

Mechanism of Liuwei Xiaoqinglong Decoction(六味小青龙汤)Inhibiting p38MAPK/p22phox Pathway and Alleviating Chronic Inflammation in Asthmatic Rats

作　　者：陈秀峰[1] 肖臻[1] 姜之炎[1] 姜永红[1] 李文[1] 刘秀秀[1] 王明晶 CHEN Xiufeng;XIAO Zhen;JIANG Zhiyan;JIANG Yonghong;LI Wen;LIU Xiuxiu;WANG Mingjing(Longhua Hospital Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院,上海200032

出　　处：《辽宁中医药大学学报》2023年第2期41-48,共8页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(82104924);上海市临床重点专科建设项目(shslczdzk04102);上海市进一步加快中医药传承创新发展三年行动计划项目(ZY<2021-2023>-0206-01)。

摘　　要：目的探讨六味小青龙汤抑制p38MAPK/p22phox哮减轻喘大鼠慢性炎症的作用机制。方法120只SD大鼠分为空白组,模型组,六味小青龙汤低、中、高剂量组,地塞米松组6组,酶联免疫吸附测定法(ELISA)检测各组大鼠干预后血清,支气管肺泡灌洗液(BALF)白细胞介素-4(IL-4)、白细胞介素-5(IL-5),肿瘤坏死因子-α(TNF-α)水平;苏木精-伊红(HE)染色观察各组肺组织病理改变,比较肺组织炎症细胞浸润;实时荧光定量聚合酶链式反应(Real-time PCR)检测各组肺组织p38MAPKmRNA、p22phoxmRNA表达;蛋白免疫印迹法(Western blot)检测各组DUOX1、DUOX2、p38MAPK及HO-1蛋白表达。结果模型组BALF IL-4、IL-5、TNF-α水平均高于空白组(P<0.05),地塞米松及六味小青龙汤能降低哮喘大鼠BALF中IL-4、IL-5含量(P<0.05)。p22phoxmRNA、p38MAPKmRNA在模型组中表达升高(P<0.05),中剂量六味小青龙汤能降低p22phoxmRNA表达(P<0.05),低、中、高剂量六味小青龙汤组均能使p38MAPKmRNA表达降低(P<0.05),地塞米松对p22phoxmRNA、p38MAPKmRNA表达的影响不明显(P>0.05)。DUOX1在初期模型组中表达较空白组升高,六味小青龙汤高剂量能降低DUOX1的表达(P<0.05)。DUOX2在模型组中较空白组表达升高,六味小青龙汤中、高剂量能降低DUOX2的表达(P<0.05)。HO-1在模型组中较空白组表达升高,高剂量六味小青龙汤以及地塞米松能降低HO-1的表达(P<0.05)。p38MAPK蛋白在模型组中较空白组表达升高,六味小青龙汤中、高剂量以及地塞米松均能降低p38MAPK蛋白表达(P<0.05)。六味小青龙汤及地塞米松组肺组织炎症细胞浸润均较模型组改善。结论六味小青龙汤可抑制p38MAPK/p22phox通路减轻哮喘大鼠肺组织病变和慢性炎症,下调p38MAPKmRNA、22phoxmRNA与DUOX1、DUOX2、HO-1、p38MAPK蛋白表达,降低IL-4、IL-5、TNF-α水平。Objective To explore the mechanism of Liuwei Xiaoqinglong Decoction(六味小青龙汤)inhibiting p38MAPK/p22phox asthma and alleviating chronic inflammation in asthmatic rats.Methods 120 SD rats were divided into blank group,model group,low-dose,medium-dose,and highdose groups of Liuwei Xiaoqinglong Decoction and dexamethasone group.The interleukin-4(IL-4),interleukin-5(IL-5)and tumor necrosis factor-α(TNF-α)level in bronchoalveolar lavage fluid(BALF)were detected by enzyme-linked immunosorbent assay(ELISA).Hematoxylin eosin(HE)staining was used to observe the pathological changes of lung tissue and compare the infiltration of inflammatory cells in lung tissue.The expressions of p38MAPK mRNA and p22phox mRNA in lung tissues of each group were detected by real-time fluorescence quantitative polymerase chain reaction(real-time PCR).The protein expressions of duox1,DUOX2,p38MAPK and HO-1 were detected by Western blot.Results IL-4,IL-5 and TNF-αin the model group were higher than those in the blank group(P<0.05).Dexamethasone and Liuwei Xiaoqinglong Decoction could reduce the contents of IL-4 and IL-5 in BALF of asthmatic rats(P<0.05).The expression of p22phox mRNA and p38MAPK mRNA increased in the model group(P<0.05).Medium-dose Liuwei Xiaoqinglong Decoction could reduce the expression of p22phox mRNA(P<0.05),and low-dose,medium-dose and high-dose Liuwei Xiaoqinglong Decoction could reduce the expression of p38MAPK mRNA(P<0.05).Dexamethasone had no significant effect on the expression of p22phoxmRNA and p38MAPK mRNA(P>0.05).The expression of DUOX1 in the initial model group was higher than that in the blank group.High-dose Liuwei Xiaoqinglong Decoction could reduce the expression of DUOX1(P<0.05).The expression of DUOX2 in the model group was higher than that in the blank group.Medium-dose and high-doses of Liuwei Xiaoqinglong Decoction could reduce the expression of DUOX2(P<0.05).The expression of HO-1 in the model group was higher than that in the blank group.High-dose Liuwei Xiaoqinglong Decoction and dexamet

关 键 词：六味小青龙汤 p38MAPK/p22phox通路 哮喘 慢性炎症 

分 类 号：R285.5[医药卫生—中药学]