大黄附子汤对重症急性胰腺炎小鼠肠道微皱褶细胞变化的影响  

Effect of Da-Huang-Fu-Zi-Tang on changes of intestinal microfold cells in mice with SAP

作　　者：宋轶[1] 赵伟彤 路晓光[1] 战丽彬[3] 陈海铭[4] 路童 康新 芮庆林[6] Song Yi;Zhao Weitong;Lu Xiaoguang;Zhan Libin;Chen Haiming;Lu Tong;Kang Xin;Rui Qinglin(Department of Emergency Medicine,the Affiliated Zhongshan Hospital of Dalian University,Dalian 116001,China;Graduate School of Zunyi Medical University,Zunyi 563003,China;Innovation Engineering and Technology Center of Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Department of Emergency Medicine,the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Department of Emergency Medicine,the Fifth Affiliated Hospital of Southern Medical University,Guangzhou 510900,China;Department of Emergency Medicine,Jiangsu Hospital of Traditional Chinese Medicine,Nanjing 210004,China)

机构地区：[1]大连大学附属中山医院急诊医学科,辽宁大连116001 [2]遵义医科大学研究生院,贵州遵义563003 [3]辽宁中医药大学中医药创新工程技术中心,辽宁沈阳110847 [4]辽宁中医药大学附属医院急诊医学科,辽宁沈阳110847 [5]南方医科大学第五附属医院急诊医学科,广东广州510900 [6]江苏省中医院急诊医学科,江苏南京210004

出　　处：《中华卫生应急电子杂志》2022年第3期167-173,共7页Chinese Journal of Hygiene Rescue(Electronic Edition)

基　　金：国家自然科学基金(81673801,81473512)。

摘　　要：目的:研究大黄附子汤对重症急性胰腺炎(SAP)小鼠肠道微皱褶细胞(M细胞)变化的影响,为大黄附子汤临床防治SAP提供理论基础。方法:选取40只健康SPF级C57BL/6J雄性小鼠,随机分为4组(每组10只):空白对照组、大黄附子汤对照组、SAP组、大黄附子汤治疗组,其中SAP组和大黄附子汤治疗组分别以3.5 g/kg剂量腹腔注射20%L-精氨酸,空白对照组和大黄附子汤对照组注射等剂量的等渗盐水;于造模后12、24、36 h,空白对照组和SAP组给予等渗盐水0.2 mL,大黄附子汤对照组和治疗组给予大黄附子汤0.2 mL灌肠,均于造模完成48 h后处死取材,使用ELISA检测血清淀粉酶、内毒素、IL-1β及TNF-α含量,取回肠及胰腺组织行HE染色、评分,免疫组化染色检测M细胞特异性蛋白GP2并评分。结果:(1)一般情况:对照组腹腔注射后小鼠一般情况好,精神状态良好,能正常进水,四肢活动自如,行动未受影响;SAP组小鼠腹腔注射后一般情况差,精神萎靡,反应迟钝,行动迟缓,呼吸急促,拱背收腹,饮水减少。(2)SAP组淀粉酶、内毒素、IL-1β及TNF-α水平较对照组明显升高(P<0.05);与SAP组进行对比,大黄附子汤治疗组血清淀粉酶、内毒素、IL-1β及TNF-α水平出现显著下降(P<0.05)。(3)HE染色:SAP组胰腺及回肠组织坏死严重,可见大量白细胞浸润。大黄附子汤治疗组胰腺及回肠组织轻度坏死,可见少量中性粒细胞等白细胞浸润。(4)免疫组化染色:SAP组与对照组相比GP2表达降低(P<0.05);相较于SAP组,大黄附子汤治疗组GP2的表达水平升高(P<0.05)。结论:大黄附子汤治疗可改善作为肠道免疫应答起始的M细胞数量与功能,增强肠道免疫应答,减轻肠免疫屏障损伤,减少内毒素入血,改善SAP症状。Objective To study the effect of Da-Huang-Fu-Zi-Tang on the changes of intestinal microfold cells(M cells)in mice with severe acute pancreatitis(SAP),so as to provide a theoretical basis for the clinical prevention of SAP.Methods Forty healthy SPF C57BL/6J male mice were selected and randomly divided into 4 groups(n=10):blank group,Da-Huang-Fu-Zi-Tang control group,SAP group,Da-Huang-Fu-Zi-Tang treatment group;the experimental group were injected with 20%PH 7.0 L-arginine into the abdominal cavity at a dose of 3.5 g/kg,and the control group were injected with the same amount of normal saline into the abdominal cavity,blank control Group and SAP group were given saline 0.2 mL by enema at 12,24 and 36 hours after modeling,and the Da-Huang-Fu-Zi-Tang control group and treatment group were given Da-Huang-Fu-Zi-Tang 0.2 mL by enema correspondingly.At 48 hours after modeling,mice were executed and selected.Serum amylase,endotoxin,IL-1t,and TNF-nlasecuted and selected.er modelingnd treatment group were given Da-Huang-Fu-Zi-Tang,and the pathology of the pancreas was analyzed using the modified Schmidt scoring,Chiu’s intestinal mucosal injury scoring criteria were used to histologically score the ileum,and the ileum was taken for immunohistochemical staining to detect GP2 changes,which is a specific protein of M cells.Results The blank control group and the Da-Huang-Fu-Zi-Tang control group were generally in good condition after intraperitoneal injection,with good mental state,normal water intake,free movement of limbs,and unaffected actions.After intraperitoneal injection,the mice in the experimental group were generally in poor condition,mentally depressed,unresponsive,sluggish,shortness of breath,abdomen arched back,and reduced drinking water.The levels of amylase,endotoxin,IL-1toxins of amylasewater.l group of Da-Huang-Fu-Zi-Tang were not significantly changed compared with the blank control group(P<0.05).Compared with SAP group,serum amylase,endotoxin,IL-1toxinlaseth SAP groupr.l group of Da-Huang-Fu-Zi-Tang were n

关 键 词：重症急性胰腺炎 M细胞 肠黏膜免疫屏障 大黄附子汤 

分 类 号：R576[医药卫生—消化系统]