大黄附子汤对重症急性胰腺炎早期大鼠结肠黏液层作用的研究  

Effect of Da Huang Fu Zi Tang on colonic mucus layer in early stage of severe acute pancreatitis in rats

作　　者：宋轶[1] 唐远莉 路晓光[1] 战丽彬[3] 陈海铭[4] 路童 芮庆林[5] 范治伟[1] Song Yi;Tang Yuanli;Lu Xiaoguang;Zhan Libin;Chen Haiming;Lu Tong;Rui Qinglin;Fan Zhiwei(Department of Emergency Medicine,the Affiliated Zhongshan Hospital of Dalian University,Dalian 116001,China;Graduate School of Dalian University,Dalian 116622,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,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,大连116001 [2]大连大学研究生院,大连116622 [3]辽宁中医药大学中医药创新工程技术中心,沈阳110847 [4]辽宁中医药大学附属医院急诊医学科,沈阳110847 [5]江苏省中医院急诊医学科,南京210004

出　　处：《中华实验外科杂志》2023年第1期35-39,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(81673801、81473512)。

摘　　要：目的探讨大黄附子汤对重症急性胰腺炎(SAP)早期大鼠结肠黏液层的作用机制。方法18只健康雄性Sprague-Dawley大鼠,随机数字表法分为3组(每组6只):对照组(Sham组)、重症急性胰腺炎组(SAP组)、重症急性胰腺炎-大黄附子汤组(DHFZT-SAP组)。逆行胰胆管注射4%牛磺胆酸钠(1 ml/kg)构建SAP模型,对照组开腹翻动胰腺组织后关腹。Sham组和SAP组造模后6、12 h分别给予1.5 ml生理盐水(37℃)灌胃;DHFZT-SAP组对应给予1.5 ml大黄附子汤(37℃)灌胃。于造模24 h后,经腹主动脉取血,酶联免疫吸附试验(ELISA)检测血清淀粉酶、内毒素、TNF-α和IL-6的含量;取胰腺、肺和结肠组织行病理学观察并评分;采用过碘酸希夫/阿利新蓝(PAS/AB)染色法观察结肠黏液层及杯状细胞的变化;采用免疫组织化学法检测结肠组织黏蛋白-2(MUC2)及结肠上皮细胞间Occludin蛋白的表达。组间比较用单因素方差分析。结果SAP组胰腺、肺、结肠组织可见水肿淤血及部分坏死,组织病理评分高于Sham组(9.33±1.86比0.17±0.41,F=130.761,P<0.05;4.50±1.22比0.33±0.52,F=58.962,P<0.05;9.0±1.67比0.17±0.41,F=157.809,P<0.05);而DHFZT-SAP组胰腺、肺、结肠呈轻度水肿淤血,病理评分低于SAP组(5.50±1.05比9.33±1.86,F=19.307,P<0.05;2.67±1.03比4.50±1.22,F=7.857,P<0.05;6.0±1.41比9.0±1.67,F=11.250,P<0.05)。SAP组血清淀粉酶、内毒素、TNF-α和IL-6含量高于Sham组(2254.89±197.29比224.66±42.82,F=606.760,P<0.05;68.90±8.55比23.01±4.92,F=129.806,P<0.05;292.03±20.33比56.0±11.60,F=610.122,P<0.05;198.14±17.36比30.86±2.63,F=544.778,P<0.05);DHFZT-SAP组血清淀粉酶、内毒素、TNF-α和IL-6含量低于SAP组(1353.93±54.23比2254.89±197.29,F=116.332,P<0.05;34.0±4.70比68.90±8.55,F=76.712,P<0.05;152.35±14.43比292.03±20.33,F=188.416,P<0.05;102.18±7.01比198.14±17.36,F=167.720,P<0.05)。SAP组大鼠结肠黏液层完整性被破坏,厚度和隐窝中杯状细胞数量低于Sham组(4.90±0.45比44.82±1.69,F=31Objective To study the effect of Da Huang Fu Zi Tang(DHFZT)on colonic mucus layer in rats with severe acute pancreatitis(SAP)in the early stage.Methods Totally,18 healthy male Sprague-Dawley rats with SPF grade were randomly divided into 3 groups(n=6):Sham group,SAP group,DHFZT-SAP group.The SAP and DHFZT-SAP groups were injected with 4% sodium taurocholate(1 ml/kg)retrograde pancreatic bile duct to construct SAP model.In the Sham group,the pancreatic tissue was turned over several times and then the abdomen closed.The Sham group and SAP group were given 1.5 ml normal saline(37℃)6 h and 12 h after operation by gavage,respectively.DHFZT-SAP group was given 1.5 ml DHFZT correspondingly.At 24 h after operation,blood samples were extracted from the abdominal aorta and the contents of serum amylase,endotoxin(LPS),TNF-αand IL-6 were detected by enzyme linked immunosorbent assay(ELISA).Pancreas,lung and colon tissues were taken for pathological examination and score.Periodic acid schiff/alcian blue(PAS/AB)staining was used to observe the changes of colonic mucus layer and goblet cells.The immunohistochemistry was used to detect the expression of mucin-2(MUC2)in colon tissue and occludin between intestinal epithelial cells.Single factor analysis of variance was used for comparison between groups.Results(1)The pancreas,lung and colon tissues of SAP group were edema,congestion and partial necrosis,and the pathological scores were higher than those of Sham group(9.33±1.86,0.17±0.41,F=130.761,P<0.05;4.50±1.22,0.33±0.52,F=58.962,P<0.05;9.0±1.67,0.17±0.41,F=157.809,P<0.05).However,DHFZT-SAP group showed mild edema and congestion,and the pathological scores were lower than those in SAP group(5.50±1.05,9.33±1.86,F=19.307,P<0.05;2.67±1.03,4.50±1.22,F=7.857,P<0.05;6.0±1.41,9.0±1.67,F=11.250,P<0.05).(2)Compared with Sham group,the contents of serum amylase,LPS,TNF-αand IL-6 in SAP group were significantly increased(2254.89±197.29,224.66±42.82,F=606.760,P<0.05;68.90±8.55,23.01±4.92,F=129.806,P<0.05;292.03±20.33,

关 键 词：重症急性胰腺炎 结肠 杯状细胞 大黄附子汤 

分 类 号：R285.5[医药卫生—中药学]