牛月黄胆酸钠诱导大鼠急性胰腺炎肺肠损伤模型制备方法的研究  

Study on the preparation method of the rat model for sodium taurocholate induced acute pancreatitis-associated lung and intestinal injury

作　　者：文林林 刘馨鸿 张延英 汪湛东 宋冰 康万荣 白敏 郭超 赵泓彰 王琼 杨润泽 刘亮亮 汪永锋 WEN Lin-lin;LIU Xin-hong;ZHANG Yan-ying;WANG Zhan-dong;SONG Bing;KANG Wan-rong;BAI Min;GUO Chao;ZHAO Hong-zhang;WANG Qiong;YANG Run-ze;LIU Liang-liang;WANG Yong-feng(School of Basic Medicine,Gansu Universityof,TraditionalChinese Medicine,Lanzhou 730000,Gansu Province,China;Chinese Medicine Department,Zhuanglang County Peoples Hospital,Pingliang 744600,Gansu Province,China;School of Animal Medicine,Gansu Agricultural University,Gansu 730070,Lanzhou Province,China;Gansu Provincial Laboratory Animal Industry Technology Center,Lanzhou 730000,Gansu Province,China;Chinese Medicine Department,Nanhu Central Health Center,Pingliang 744600,Gansu Province,China)

机构地区：[1]甘肃中医药大学基础医学院,甘肃兰州730000 [2]庄浪县人民医院中医科,甘肃平凉744600 [3]甘肃农业大学动物医学院,甘肃兰州730070 [4]甘肃省实验动物行业技术中心,甘肃兰州730000 [5]南湖中心卫生院中医科,甘肃平凉744600

出　　处：《中国临床药理学杂志》2023年第6期873-877,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(82160871);甘肃省自然科学基金资助项目(22JR5RA591,20JR5RA186);甘肃省中医药管理局重点基金资助项目(GZKZ-2021-10)。

摘　　要：目的建立大鼠急性胰腺炎(AP)肺肠损伤动物模型,通过比较不同时间点大鼠胰腺、肺、结肠病理改变、血清生化及炎症细胞因子含量,探讨AP大鼠肺肠损伤模型复建的最佳时间。方法用经胰胆管逆行注射5%牛[月黄]胆酸钠的方法建立AP大鼠肺肠损伤模型,将模型大鼠分为模型12 h组、模型24 h组、模型48 h组,每组20只。另取20只正常大鼠作为空白组,空白组大鼠经胰胆管逆行注射0.9%生理盐水。分别于造模后12、24和48 h采集相对应组大鼠的全血及胰腺、结肠、肺组织。用生化法检测血清中的淀粉酶(AMS)、C反应蛋白(CRP)含量;用免疫组化法检测胰腺、肺、结肠组织中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和诱导型一氧化氮合酶(INOS)表达水平。结果空白组、模型12 h组、模型24 h组和模型48 h组全血血清中AMS含量分别为(8.15±4.30)、(24.11±3.70)、(35.83±7.12)和(32.69±10.17)U·L^(-1);CRP含量分别为(8.54±4.93)、(23.31±11.04)、(37.56±5.98)和(32.09±14.61)mg·L^(-1);胰腺组织中TNF-α表达含量分别为1.65±0.18、4.70±0.49、16.39±1.38和12.91±0.56;IL-6表达含量分别为2.17±0.25、5.23±0.92、17.18±2.86和10.71±1.91;INOS表达含量分别为2.46±1.26、7.81±0.77、19.60±1.73和16.04±0.75;肺组织中TNF-α表达含量分别为1.91±0.21、4.57±0.27、19.13±2.22和15.90±1.27;IL-6表达含量分别为1.25±0.24、6.18±0.66、17.27±0.32和13.96±6.61;INOS表达含量分别为2.38±0.25、8.48±3.08、17.47±2.27和10.72±6.30;结肠组织中TNF-α表达含量分别为1.94±0.14、8.61±0.74、18.71±2.23和14.60±2.13;IL-6表达含量分别1.98±0.61、7.07±1.58、14.47±0.91和12.63±1.00;INOS表达含量分别为2.52±0.79、8.31±0.86、16.22±1.50和13.66±1.13。模型组的上述指标与空白组比较,其中模型24 h组的差异最明显,差异均有统计学意义(均P<0.05)。结论AP肺肠损伤模型以24 h为最佳成模时间点。Objective To establish an animal model of lung and intestinal injury in rats with acute pancreatitis(AP)and to investigate the best time to reconstruct the lung and intestinal injury model in rats with AP by comparing the pathological changes in the pancreas,lung and colon,serum biochemistry and inflammatory cytokine levels at different time points.Methods SPF grade Wistar rats were used to replicate the AP rat lung intestinal injury model by retrograde injection of 5%Sodium taurocholate via the pancreaticobiliary duct,and the model mice were divided into model 12 h,24 h and 48 h groups,with 20 mice in each group.Another 20 normal mice were taken as the blank group,and the blank group rats were retrogradely injected with 0.9%saline via pancreaticobiliary duct.Whole blood and tissues of pancreas,colon and lung were collected from each group at 12 h,24 h and 48 h after modeling,respectively.The levels of amylase(AMS)and C-reactive protein(CRP)in whole blood serum were measured by biochemical methods;the expression levels of tumor necrosis factorα(TNF-α),interleukin 6(IL-6)and inducible nitric oxide synthase(INOS)in pancreas,lung and colon tissues were measured by immunohistochemistry,respectively.Results In the blank,model 12 h,model 24 h and model 48 h groups the AMS levels in whole blood serum were(8.15±4.30),(24.11±3.70),(35.83±7.12)and(32.69±10.17)U·L^(-1),respectively;CRP levels were(8.54±4.93),(23.31±11.04),(37.56±5.98)and(32.09±14.61)mg·L^(-1),respectively.In pancreatic tissues:TNF-αexpression levels were 1.65±0.18,4.70±0.49,16.39±1.38 and 12.91±0.56,respectively;IL-6 expression levels were 2.17±0.25,5.23±0.92,17.18±2.86 and 10.71±1.91,respectively;INOS expression levels were 2.46±1.26,7.81±0.77,19.60±1.73 and 16.04±0.75,respectively.In lung tissues:TNF-αexpression levels were 1.91±0.21,4.57±0.27,19.13±2.22 and 15.90±1.27,respectively;IL-6 expression levels were 1.25±0.24,6.18±0.66,17.27±0.32 and 13.96±6.61,respectively;INOS expression levels were 2.38±0.25,8.48±3.08,17.47

关 键 词：急性胰腺炎 肺损伤 肠损伤 牛[月黄]胆酸钠 模型 

分 类 号：R97[医药卫生—药品]