基于网络药理学和实验验证探讨大黄牡丹汤改善大肠湿热型急性胰腺炎大鼠胰腺损伤的机制  

Mechanism of Dahuang Mudantang in Improving Pancreatic Injury in Rats with Acute Pancreatitis of Dampness-heat in Large Intestine Syndrome Based on Network Pharmacology and Experimental Verification

作　　者：王琼[1] 汪永锋[1] 张延英[1,2] 宋冰 郭超 刘馨鸿[3] 白敏 汪湛东 文林林 赵泓彰[1] WANG Qiong;WANG Yongfeng;ZHANG Yanying;SONG Bing;GUO Chao;LIU Xinhong;BAI Min;WANG Zhandong;WEN Linlin;ZHAO Hongzhang(Gansu University of Chinese Medicine,Lanzhou 730000,China;Technology Center of Laboratory Animal Industry,Lanzhou 730000,China;Gansu Agricultural University,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学,兰州730000 [2]甘肃省实验动物行业技术中心,兰州730000 [3]甘肃农业大学,兰州730000

出　　处：《中国实验方剂学杂志》2023年第20期61-68,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82160871);甘肃省自然科学基金项目(22JR5RA591);甘肃省中医药管理局重点项目(GZKZ-2021-10);甘肃省中医药研究中心课题(ZYZX-2020-ZX22)。

摘　　要：目的:揭示大黄牡丹汤对大肠湿热型急性胰腺炎大鼠胰腺损伤的干预作用,并结合网络药理学探讨其可能机制。方法:随机将96只SPF级Wistar大鼠分为6组,空白组、模型组、大黄牡丹汤低、中、高剂量组(3.5、7、14 g·kg^(-1))、清胰利胆颗粒组(3 g·kg^(-1))每组16只。除空白组外,其余各组大鼠采用“高温高湿环境+高糖高脂饮食+5%牛磺胆酸钠逆行胰胆管注射法”建立大肠湿热型急性胰腺炎(AP)大鼠模型。空白组、模型组大鼠给予等体积蒸馏水灌胃,治疗组于造模前1 h、造模后12、24 h分别给药,末次给药1 h后采集样本。观察大鼠一般情况;评价大肠湿热型模型;采用生化法检测血清淀粉酶(AMS)、C反应蛋白(CRP)含量情况;苏木素-伊红(HE)染色观察胰腺组织病理形态;采用网络药理学预测大黄牡丹汤干预急性胰腺炎的可能靶点,并采用分子生物学技术验证相关靶点。结果:与空白组比较,模型组大鼠精神萎靡、毛发杂乱且无光泽、粪便软黏、黄而恶臭、肛温升高并伴有弓背扭体反应,血清中AMS、CRP含量明显升高(P<0.05),光镜下可见胰管走形异常、小叶间隙紊乱及炎症细胞浸润,腺泡细胞水肿、充血、坏死等病理改变;与模型组比较,各治疗组大鼠一般生存状况有不同程度改善、扭体反应减轻、粪便软黏、黄而恶臭明显改善,胰腺组织水肿、坏死程度减轻,血清中AMS、CRP含量降低(P<0.05),以大黄牡丹汤高剂量组效果最明显(P<0.05)。网络药理学预测结果表明常春藤皂苷元、β-谷甾醇、槲皮素是与疾病靶点连接最为广泛的活性化合物,而蛋白质-蛋白质相互作用(PPI)发现蛋白激酶B(Akt)、肿瘤蛋白P53(TP53)、肿瘤坏死因子(TNF)、白细胞介素-6(IL-6)、转录激活因子(JUN)、血管内皮生长因子α(VEGFα)、白细胞介素-1β(IL-1β)、血管内皮细胞黏附分子1(VCAM1)是“药物-疾病”中较为关键靶点,京都基因与基�Objective:To reveal the intervention effect of Dahuang Mudantang on pancreatic injury in rats with acute pancreatitis(AP)of dampness-heat in large intestine syndrome and explore its possible mechanism based on network pharmacology.Method:Ninety-six SPF-grade Wistar rats were randomly divided into the following six groups:a blank group,a model group,low-,medium-,and high-dose Dahuang Mudantang groups(3.5,7,and 14 g·kg^(-1)),and a Qingyi Lidan granules group(3 g·kg^(-1)),with 16 rats in each group.The AP model of dampness-heat in large intestine syndrome was induced in rats except for those in the blank group by"high-temperature and high-humidity environment+high-sugar and high-fat diet+retrograde injection of 5%sodium taurocholate into the pancreaticobiliary duct".The blank and model groups received equal volumes of distilled water by gavage,while the treatment groups were administered Dahuang Mudantang or Qingyi Lidan granules 1 hour before modeling,and 12 and 24 hours after modeling.Samples were collected 1 hour after the last administration.The general conditions of the rats were observed.The AP model of dampnessheat in large intestine syndrome was evaluated.Serum amylase(AMS)and C-reactive protein(CRP)levels were determined using biochemical methods.Pancreatic tissue morphology was observed using hematoxylineosin(HE)staining.Network pharmacology was employed to predict potential targets of Dahuang Mudantang in the intervention in AP,and molecular biology technique was used to verify relevant targets.Result:Compared with the blank group,the model group exhibited lethargy,unkempt fur,loose and foul-smelling stools,elevated anal temperature with arching and twisting reactions,significantly increased serum levels of AMS and CRP(P<0.05),abnormal pancreatic ductules,disordered interlobular spaces,and inflammatory cell infiltration in histopathological examination,as well as pathological changes including pancreatic acinar cell swelling,congestion,and necrosis.Compared with the model group,the treatment groups sh

关 键 词：急性胰腺炎 大肠湿热型 大黄牡丹汤 丝裂原活化蛋白激酶P38 蛋白激酶-2 人组织抗原R 

分 类 号：R2-0[医药卫生—中医学] R33R318.14R576R365.4