摘要
【目的】探讨清郁和降汤对反流性食管炎(RE)大鼠的治疗作用及机制。【方法】将48只雄性SD大鼠随机分为假手术组,模型组,中药低、中、高剂量组及西药组,每组8只。除假手术组,其余5组大鼠采用前胃结扎联合外置幽门部分结扎术建立反流性食管炎模型。成功造模后,中药低、中、高剂量组分别对应给予清郁和降汤7.9、15.8、31.6 g·kg^(-1)·d^(-1)灌胃,西药组给予泮托拉唑蒸馏水混悬液8.4 mg·kg^(-1)·d^(-1)、莫沙比利蒸馏水混悬液0.19 mg·kg^(-1)·d^(-1)灌胃,假手术组与模型组给予等体积蒸馏水灌胃。连续灌胃14 d后,将所有大鼠处死取材。采用苏木素-伊红(HE)染色法观察食管组织病理学改变,采用酶联免疫吸附分析(ELISA)检测血清中5-羟色胺(5-HT)、色氨酸羟化酶1(TPH1)、P物质(SP)、降钙素基因相关肽(CGRP)的含量,分别采用实时定量聚合酶链反应(qRT-PCR)法、蛋白免疫印迹(Western Blot)法对应检测食管组织G蛋白偶联胆汁酸受体5(TGR5)、瞬时受体电位锚蛋白1(TRPA1)、瞬时受体电位香草酸亚型1(TRPV1)、磷脂酶C(PLC)、蛋白酶激活受体2(PAR2)mRNA或蛋白表达水平。【结果】与假手术组比较,模型组大鼠可见食管黏膜出现较为明显的损伤及炎细胞浸润,血清中5-HT、TPH1、SP、CGRP含量显著增高(P<0.01),食管组织中TRG5、TRPA1、TRPV1、PLC、PAR2 mRNA及蛋白表达水平显著增高(P<0.01);与模型组比较,中药低、中、高剂量组及西药组大鼠食管黏膜病理损伤得到明显改善,血清中5-HT、TPH1、SP、CGRP含量显著降低(P<0.01),食管组织中TRG5、TRPA1、TRPV1、PLC、PAR2 mRNA及蛋白表达水平显著降低(P<0.05)。【结论】清郁和降汤可有效改善大鼠反流性食管炎,其机制可能与抑制TGR5/TRPA1通路,减少5-HT释放,进而降低食管敏感性有关。
Objective To investigate the therapeutic effect and mechanism of Qingyu Hejiang Decoction on rats with reflux esophagitis(RE).Methods Forty-eight male SD rats were randomly divided into sham-operation group,model group,low-,medium-and high-dose groups of Chinese medicine and western medicine group,with 8 rats in each group.In addition to the sham-operation group,the remaining five groups of rats were used to establish a reflux esophagitis model by combining anterior gastric ligation with external pyloric partial ligation.After successful modeling,the low-,medium-and high-dose groups of Chinese medicine were given 7.9,15.8,31.6 g·kg^(-1)·d^(-1)gavage respectively,the western medicine group was given 8.4 mg·kg^(-1)·d^(-1)of Pantoprazole in distilled water suspension and 0.19 mg·kg^(-1)·d^(-1)of Mosapride in distilled water suspension,and the shamoperation group and the model group were given equal volume of distilled water by gavage.After 14 days of continuous gavage,all rats were killed and the materials were taken.The histopathological changes of the oesophagus were observed by hematoxylin-eosin(HE)staining,and the serum levels of 5-hydroxytryptamine(5-HT),tryptophan hydroxylase 1(TPH1),substance P(SP),and calcitonin gene-related peptide(CGRP)were detected by enzyme-linked immunosorbent assay(ELISA).Real-time quantitative polymerase chain reaction(qRT-PCR)and protein immunoblotting(Western Blot)methods were used to detect the mRNA or protein expression levels of G protein-coupled bile acid receptor 5(TGR5),transient receptor potential anchoring protein 1(TRPA1),transient receptor potential vanilloid isoform 1(TRPV1),phospholipase C(PLC),and proteaseactivated receptor 2(PAR2)in esophageal tissue,respectively.Results Compared with the sham-operation group,rats in the model group showed more obvious damage and inflammatory cell infiltration in the oesophageal mucosa,with significantly higher serum levels of 5-HT,TPH1,SP and CGRP(P<0.01),and significantly higher levels of mRNA and protein expressions of TRG5,TR
作者
周易
黄雨晴
叶松
ZHOU Yi;HUANG Yu-Qing;YE Song(Hubei University of Chinese Medicine,Wuhan 430065 Hubei,China;Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430061 Hubei,China)
出处
《广州中医药大学学报》
CAS
2023年第12期3115-3122,共8页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
叶松全国名老中医专家传承工作室建设项目(国中医药人教函[2022]75号)
湖北省科技厅省自然科学基金创新发展联合基金项目(编号:2022CFD141)。
