肾蕨对乙酸致小鼠肛门溃疡的作用及NF-κB,TNF-α,IL-1β和COX-2表达的影响  

Effect of Nephrolepis cordifolia on Acetic Acid-induced Anal Ulceration in Mice and Levels of NF-κB,TNF-α,IL-1β and COX-2

作　　者：王小青[1] 高杨[1] 马帅[1] 商亚珍[1] WANG Xiao-qing;GAO Yang;MA Shuai;SHANG Ya-zhen(Hebei Key Laboratory of Research and Development for Traditional Chinese Medicine, Chengde Medical College, Chengde 067000, Chin)

机构地区：[1]承德医学院河北省中药研究与开发重点实验室

出　　处：《中国实验方剂学杂志》2018年第8期122-127,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省中医药管理局项目(2017191);河北省高校重点学科建设项目(冀教高[2013]4号);承德医学院课题项目(201529);承德医学院大学生创业训练计划项目(201717)

摘　　要：目的：研究肾蕨叶水提取物（NLD）对小鼠肛门溃疡模型的作用及机制。方法：以20%乙酸（HAc）0.5 m L涂抹小鼠肛门,制备肛门溃疡模型,将50只昆明小鼠随机分为正常组、模型组、NLD低、中、高剂量（0.45,0.9,1.8 g·kg^-1）组,每组10只。各组小鼠自外涂给NLD计时,第10天处死。采集肛门、直肠组织,实时荧光定量逆转录聚合酶链式反应（Real-time PCR）检测各组小鼠肛门、直肠组织中核转录因子（NF）-κB,肿瘤坏死因子（TNF）-α,白细胞介素（IL）-1β和环氧合酶（COX）-2mRNA表达;蛋白免疫印迹法（Western blot）检测各组小鼠肛门、直肠组织中NF-κB,TNF-α,IL-1β和COX-2蛋白表达;苏木素-伊红（HE）染色观察小鼠肛门、直肠组织形态学变化。结果：NLD可恢复小鼠体质量增长趋势,降低小鼠肛门脏器指数,下调小鼠肛门、直肠组织中NF-κB,TNF-α,IL-1β和COX-2 mRNA（P〈0.01）,降低小鼠肛门、直肠组织中NF-κB,TNF-α,IL-1β和COX-2蛋白表达（P〈0.01）;NLD 0.9,1.8 g·kg^-1剂量组可抑制炎性细胞的浸润及黏膜下层血管扩张,改善相应炎症反应。结论：NLD对肛门溃疡模型小鼠的肛门溃疡有明显的改善作用,其机制可能与下调NF-κB mRNA和蛋白的表达,进而下调其下游因子TNF-α,IL-1β与COX-2的基因表达有关。Objective： To investigate the therapeutic effect of Nephrolepis cordifolia leaf decoction（NLD）on anal ulcer model in mice and explore its mechanism. Method： The anal ulcer model was prepared by applying0. 5 m L of 20% acetic acid（HAc） into the anus of mice. The 50 Kunming mice were randomly divided into normal group,model group,low,middle and high dose NLD（0. 45,0. 9 and 1. 8 g·kg^-1） groups,10 mice in each group. All the mice were killed on the 10 thday after NLD application. Their anal and rectal tissues were collected and the mRNA expression levels of nuclear transcription factor-κB（NF-κB）,tumor necrosis factor-α（TNF-α）,interleukin-1β（IL-1β） and cyclooxygenase-2（COX-2） in the anal and rectal tissues of each group were detected by Real-time fluorescence quantitative reverse transcription polymerase chain reaction（Real-time PCR）. Western blot was used to detect the protein expression levels of NF-κB,TNF-α,IL-1β and COX-2 in anal and rectal tissues of mice in each group. Morphological changes of anal and rectal tissues in mice were observed by hematoxylin-eosin（HE） staining. Result： NLD could restore the growth of mice body mass,reduce the rectoanal coefficient and down-regulate NF-κB,TNF-α,IL-1β and COX-2 mRNA expression in the anal and rectal tissues of mice（P〈0. 01）,inhibit the activity of NF-κB,TNF-α,IL-1β and COX-2 protein expression in the anal and rectal tissues of mice（P〈0. 01）. NLD 0. 9 and 1. 8 g·kg^-1 could inhibit the infiltration of inflammatory cells and submucosa vasodilation and improve the corresponding inflammatory response. Conclusion： NLD has protective effect on the inflammatory response of acetic acid-induced anal ulcer in mice,and the mechanism may be related to inhibiting NF-κB mRNA and protein expression and down-regulating the gene expression of downstream inflammatory factors TNF-α,IL-1β and COX-2.

关 键 词：肾蕨叶水提取物 溃疡 核转录因子-ΚB P65 肿瘤坏死因子-α 白细胞介素-1β 环氧合酶-2 

分 类 号：R285.5[医药卫生—中药学] R22[医药卫生—中医学]