射干麻黄汤通过调节SIRT1-NF-κB信号通路干预哮喘大鼠气道炎症的机制  

Mechanism of Shegan Mahuang Decoction(射干麻黄汤)on Airway Inflammation in Asthmatic Rats by Regulating SIRT1-NF-κB Signaling Pathway

作　　者：彭帅 方向明[1] 叶卫东 袁亚美[1] 高鹏飞 马忍 蔡旻 PENG Shuai;FANG Xiangming;YE Weidong;YUAN Yamei;GAO Pengfei;MA Ren;CAI Min(Anhui University of Chinese Medicine,Hefei 230038,Anhui,China)

机构地区：[1]安徽中医药大学,安徽合肥230038

出　　处：《辽宁中医药大学学报》2023年第9期18-23,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074401);安徽省高校优秀人才支持计划项目(gxyq2021186);安徽省质量工程项目(2020kcszyjxm142);安徽中医药大学自然科学研究重点项目(2021zrzd18)。

摘　　要：目的探究射干麻黄汤对卵清蛋白(OVA)诱导的哮喘大鼠气道炎症的防治作用及机制。方法将105只SPF雄性SD大鼠随机分为正常组,模型组,地塞米松组,桂龙咳喘宁组,射干麻黄汤高、中、低剂量组,每组15只。以卵清蛋白联合氢氧化铝复制大鼠哮喘模型,造模21 d后,正常组、模型组每天给予等体积的0.9%氯化钠溶液灌服,地塞米松组按0.405 mg/(kg·d)灌胃给药,桂龙咳喘宁按0.405 g/(kg·d)灌胃给药,射干麻黄汤高、中、低剂量组按12.96、6.48、3.24 g/(kg·d)灌胃给药。灌胃4周后,在末次OVA激发后2 h内解剖大鼠并取出肺组织、支气管肺泡灌洗液(BALF)。苏木素-伊红(HE)染色观察肺组织的形态学变化及炎症细胞浸润;酶联免疫吸附剂测定(ELISA)法测量BALF中白细胞介素(IL)-4、IL-5和肿瘤坏死因子(TNF)-α的含量以及OVA特异性免疫球蛋白E(IgE)的浓度;蛋白免疫印迹法(Western blot)检测大鼠肺组织中沉默信息调节因子1(SIRT1)、核因子kappa B(NF-κB)p65和蛋白表达水平;反转录聚合酶链反应(RT-qPCR)检测SIRT1、NF-κB p65mRNA的表达水平。结果HE染色发现各治疗组支气管壁损伤明显改善,炎性细胞浸润减少;ELISA结果显示,与正常组比较,模型组的各项炎性因子IgE、IL-4、IL-5和TNF-α均显著升高(P<0.01);与模型组比较,各治疗组(射干麻黄汤组、地塞米松组、桂龙咳喘宁组)的IgE、IL-4、IL-5和TNF-α均明显降低(P<0.01);RT-qPCR结果显示,与正常组比较,模型组的SIRT1mRNA相对表达水平显著下降(P<0.01),而NF-κB p65mRNA相对表达则显著升高(P<0.01);与模型组比较,各治疗组的SIRT1mRNA均显著升高(P<0.01),而治疗组的NF-κB p65mRNA均显著下降(P<0.01);Western blot结果显示,与正常组比较,模型组的SIRT1相对表达水平显著下降(P<0.01),而NF-κB p65相对表达则显著升高(P<0.01);与模型组相比,各治疗组的SIRT1均显著升高(P<0.01),而NF-κB p65均显著下降(P<0.01)。结论射干麻�Objective To explore the preventive and therapeutic effect of Shegan Mahuang Decoction(射干麻黄汤)on airway inflammation in asthmatic rats induced by ovalbumin(OVA)and its mechanism.Methods 105 SPF male SD rats were randomly divided into normal group,model group,dexamethasone group,Guilong Kechuanning group and high,medium,low dose Shegan Mahuang Decoction groups,with 15 rats in each group.The asthma model of rats was made with ovalbumin and aluminum hydroxide.After 21 days of modeling,the normal group and model group were given the same volume of normal saline every day,the dexamethasone group was given 0.405 mg/(kg*d)dexamethasone by gavage,the Guilong Kechuanning group was given 0.405 g/(kg·d)Guilong Kechuanning(桂龙咳喘宁)by gavage,and the high,middle and low dose groups of Shegan Mahuang decoction were given 12.96,6.48 and 3.24 g/(kg:d)Shegan Mahuang Decoction by gavage.After 4 weeks of intragastric administration,the rats were dissected and the lung tissue and bronchoalveolar lavage fluid(BALF)were taken out within 2 hours after the last OVA challenge.Hematoxylin eosin(HE)staining was used to observe the morphological changes of lung tissue and inflammatory cell infiltration;To determine of interleukin-4(IL-4),interleukin-5(IL-5)and tumor necrosis factor(TNF)in BALF by enzyme-linked immunosorbent assay-α(ELISA-α)and the concentration of OVA specific immunoglobulin E(IgE);To determine of silent information regulator 1(SIRT1)and nuclear factor kappa B(NF-κB)p65 in rat lung tissues by Western blot and protein expression level;To determine of SIRT1 and NF-κB p65 by reverse transcription polymerase chain reaction(RT qPCR)mRNA expression level.Results HE staining showed that the damage of bronchial wall was significantly improved and the infiltration of inflammatory cells was reduced in each treatment group;The results of ELISA showed that the inflammatory factors IgE,IL-4,IL-5 and TNF-αin the model group were higher than those in the normal group(P<0.01);Compared with the model group,IgE,IL-4,IL-

关 键 词：射干麻黄汤 炎症 沉默信息调节因子1/核因子kappa B p65 支气管哮喘 

分 类 号：R289.5[医药卫生—方剂学] R562.25[医药卫生—中药学]