基于β2AR/β-arrestin2/NF-κB信号通路的清肠化湿颗粒防治溃疡性结肠炎的作用机制  

Effect of Qingchang Huashi Granule on β2AR/β-arrestin2/NF-κB Signaling Pathway in Ulcerative Colitis

作　　者：戴路明 朱磊 沈洪 DAI Lu-ming;ZHU Lei;SHEN Hong(Affiliated Hospital of Nanjing University of Traditional Chinese Medicine （TCM）, Jiangsu Province Hospital of TCM , Nanjing 210029, China)

机构地区：[1]南京中医药大学附属医院江苏省中医院

出　　处：《中国实验方剂学杂志》2018年第9期86-94,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81673905);国家中医药管理局项目(JDZX2015084);国家中医药行业科研专项(201407001);江苏省临床医学科技专项(BL2014100);江苏高校优势学科建设工程项目(PAPD)

摘　　要：目的：观察清肠化湿颗粒对溃疡性结肠炎（ulcerative colitis,UC）模型大鼠及人结肠癌上皮细胞株HT-29炎症模型β2肾上腺素受体（β2AR）/β-抑制蛋白2（β-arrestin2）/核转录因子-κB（NF-κB）信号通路的干预作用。方法：取8只大鼠作为空白组,其余大鼠采用三硝基苯磺酸（trinitro-benzene-sulfonic acid,TNBS）灌肠法复制UC大鼠模型;待模型建立成功后,随机分为模型组,柳氮磺胺吡啶（SASP,1.0 g·kg（-1））组,清肠化湿颗粒低、中、高剂量（2.8,5.5,11.0 g·kg（-1））组,每组8只,灌胃给药10 d,每日1次;酶联免疫吸附法（enzyme linked immunosorbent assay,ELISA）测定UC大鼠结肠组织中巨噬细胞移动抑制因子（MIF）,基质金属蛋白酶-1（MMP-1）,MMP-2,MMP-9,胰岛素样生长因子-1（IGF-1）,超氧化物歧化酶（SOD）,丙二醛（MDA）,髓过氧化物酶（MPO）,一氧化氮（NO）,一氧化氮合成酶（iNOS）,谷胱甘肽-过氧化物酶（GSH-Px）;免疫组化法检测模型大鼠结肠黏膜β2AR,β-arrestin2,NF-κB定位表达;采用肿瘤坏死因子与脂多糖诱导HT-29细胞炎症模型;噻唑蓝（MTT）比色法检测清肠化湿颗粒对细胞增殖的影响,蛋白免疫印迹法（Western blot）检测细胞中β2AR,β-arrestin2,NF-κB蛋白表达量。结果：造模后,模型组大鼠较空白组大鼠结肠黏膜的结肠损伤分数明显升高;与模型组比较,3个剂量的清肠化湿颗粒给药组结肠黏膜损伤明显下降（P〈0.05,P〈0.01）,大鼠结肠组织IGF-1,SOD,GSH-Px含量与β2AR,β-arrestin2表达明显上升,MIF,MMP-2,MMP-9,MDA,MPO,NO,iNOS含量及NF-κB表达明显下降（P〈0.05,P〈0.01）。结论：清肠化湿颗粒可缓解UC氧化应激反应及肠道炎症,其作用与激活β2AR/β-arrestin2/NF-κB信号通路有关。Objective：To study the effect of Qingchang Huashi granule on β2 adrenergic receptor（β2AR）/β-arrestin2/nuclear transcription factor-κB（NF-κB） signaling pathway in rat and human ulcerative colitis.Method：Eight rats were randomly selected as control group,while the other rats were used to duplicate the ulcerative colitis model induced by trinitrobenzene sulfonic acid; after successful modeling,the rats were randomly divided into model group,salazosulfapyridine（SASP） group（1.0 g·kg^（-1））,and low,medium and high-dose Qingchang Huashi granule groups（2.8,5.5,11.0 g·kg^（-1））,with 8 rats in each group.The drugs were given once a day by gavage for 15 days.The levels of macrophage mobile inhibitory factor（MIF）,matrix metalloproteinase-1（MMP-1）,macrophage migration inhibitory factor（MIF）,matrix metalloproteinase-1（MMP-1）,MMP-2,MMP-9,insulin-like growth factor-1（IGF-1）,superoxide dismutase（SOD）,malondialdehyde（MDA）,myeloperoxidase（MPO）,nitric oxide（NO）,nitric oxide synthase（iNOS）,glutathione peroxidase（GSH-Px） in experimental UC rats were measured by enzyme linked immunosorbent assay method（ELISA）;immunohistochemical method was used to detect the location expressions of β2AR,β-arrestin2,NF-κB in rat models; tumor necrosis factor alpha and lipopolysaccharide were used to induce the HT-29 cell inflammation model; the effect of Qingchang Huashi granule on cell growth was detected by MTT method,the expressions of β2AR,β-arrestin2,NF-κB were measured with Western blot.Result：The injury score of colonic mucosa in model group rats was significantly higher than that of the normal group,expressions of IGF-1,SOD,GSH-Px,β2AR and β-arrestin2 decreased significantly,while expressions of MIF,MMP-2,MMP-9,MDA,MPO,NO,iNOS and NF-κB increased in UC model group（P 〈 0.05,P 〈 0.01）.After intervened with Qingchang Huashi granule,these indicators all recovered to normal.Conclusion：Qingchang Huashi granule can reduce the pathological process o

关 键 词：溃疡性结肠炎 清肠化湿颗粒 Β2肾上腺素受体 β-抑制蛋白2 核转录因子-ΚB 

分 类 号：R22[医药卫生—中医基础理论] R24[医药卫生—中医学]