贻贝粘蛋白对痔病大鼠模型治疗作用的实验研究  

Therapeutic effect of mussel adhesive protein on hemorrhoids disease in rats

作　　者：田茂生 高记华 戚文月 刘朝阳 吴琳琳 高策 顾铭[5] 康瑞娟[5] 李利霞 TIAN Maosheng;GAO Jihua;QI Wenyue;LIU Zhaoyang;WU Linlin;GAO Ce;GU Ming;KANG Ruijuan;LI Lixia(Hebei University of Chinese Medicine,Shijiazhuang 050091,China.;the First Affiliated Hospital of Hebei University of Chinese Medicine,Shijiazhuang 050011;Key Laboratory of Gastroenterology Research of Integrated Traditional Chinese and Western Medicine of Hebei Province,Shijiazhuang 050011;Shanxi Provincial People’s Hospital,Taiyuan 030002;Institute of Process Engineering,Chinese Academy of Sciences,Beijing 100190)

机构地区：[1]河北中医学院,石家庄050091 [2]河北中医学院第一附属医院,石家庄050011 [3]河北省中西医结合胃肠病研究重点实验室,石家庄050011 [4]山西省人民医院,太原030002 [5]中国科学院过程工程研究所,北京100190

出　　处：《中国实验动物学报》2022年第6期742-750,共9页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金面上项目(82174381);国家重点研发计划项目(2016YFC1103200);中国博士后科学基金项目(2021M701035);河北中医学院科研能力提升重点项目(KTZ2019033);河北中医学院研究生创新能力培养资助项目(XCXZZBS2021019)。

摘　　要：目的本研究基于直肠局部用药要求建立巴豆油诱导的痔病大鼠模型,探究贻贝粘蛋白(MAP)的抗痔疗效和黏附性,并探讨其作用机制。方法以巴豆油为致炎剂建立痔病大鼠模型,将40只SPF级SD大鼠按体重随机分为4组,造模后1 h各组大鼠予以相应治疗。D4取材,处死前30 min将1%伊文思蓝(EBD)注入大鼠尾静脉,计算肛肠系数,评价血管通透性,Western Blot法检测黏蛋白Muc2、Muc4的蛋白相对表达量,ELISA法检测大鼠血清TNF-α、IL-6、MDA表达量。通过在体实验和离体实验比较异硫氰酸荧光素(FITC)标记的MAP在大鼠直肠的荧光成像,观察比较MAP在直肠的黏附性。结果MAP可减少大鼠肛门直肠肿胀、分泌物和渗出情况,肛肠系数结果提示MAP可降低肛门直肠肿胀程度(P<0.01),EBD含量提示MAP可降低局部血管通透性(P<0.01),增加黏蛋白Muc2、Muc4的相对表达量(P<0.05),降低血清TNF-α、IL-6、MDA表达量(P<0.01),荧光成像结果显示MAP在直肠环境内表现出良好的黏附性和稳定性。结论海洋生物材料贻贝粘蛋白具有良好黏附性,能够减轻大鼠肛门直肠黏膜肿胀、增加黏膜黏蛋白表达、抑制炎症、修复黏膜屏障、促进黏膜愈合、改善痔病大鼠模型症状,可为痔病药物的临床前研究提供可靠依据。Objective The anti-hemorrhoid efficacy and adhesion of mussel adhesive protein(MAP)was studied and its mechanism evaluated.Methods Croton oil was used to stimulate the anorectal mucosa to establish the model.Forty specific-pathogen-free grade Sprague-Dawley rats were divided into four experimental groups.One hour after model creation,different treatments were applied to each group by rectal administration under anesthesia.General observation was performed on the fourth day.A 1%solution of EBD was injected into the tail vein 30 min before rats were sacrificed.Approximately 8 mm of rectum was cut off and weighed to calculate the anorectal coefficient.Vascular permeability was evaluated by measuring EBD concentrations.Western Blot was used to detect relative protein expression levels of Muc2 and Muc4,and ELISA was used to detect expression of serum tumor necrosis factor(TNF-α),interleukin(IL-6),and malonaldehyde(MDA)in rats.Fluorescence imaging of FITC-labeled MAP at different times was compared and observed by fluorescence microscopy.Results General observations revealed that MAP could reduce the redness,edema,mucosal secretion,and exudation in the rectal area of rats.Anorectal coefficient result suggested that MAP could reduce the degree of rectal swelling(P<0.01),and EBD contents suggested that MAP could reduce local vascular permeability(P<0.01).Relative expression levels of mucins Muc2 and Muc4 were increased(P<0.05),whereas expression levels of serum TNF-α,IL-6,and MDA were decreased(P<0.01).Fluorescence imaging indicated that FITC-MAP could maintain good adhesion and stability in the rectum.Conclusions MAP could increase the expression of mucosal mucin,inhibit inflammation,repair the mucosal barrier,promote mucosal healing,and improve the symptoms of hemorrhoids in a rat model,which can provide a reliable basis for preclinical research of hemorrhoids drugs.

关 键 词：痔病 巴豆油模型 贻贝粘蛋白 黏膜屏障 黏附性 

分 类 号：Q95-33[生物学—动物学]