摘要
目的探讨鹅去氧胆酸(CDCA)对重症急性胰腺炎(SAP)大鼠胰腺及肠道损伤的影响及意义。方法将45只SPF级雄性SD大鼠随机分为SAP组、CDCA组和假手术(Sham)组,每组15只。SAP组大鼠先普食喂养一周后采用胰胆管逆行注射5%牛磺胆酸钠的方法建立SAP模型,CDCA组大鼠先喂养含0.5%CDCA的同质饲料一周,再进行造模,建模后24 h收集组织标本,Sham组大鼠喂养方法同SAP组,麻醉开腹仅拨动十二指肠降部后关腹。首先,使用HE染色法观察小肠和胰腺组织的病理学改变;其次,使用ELISA法检测大鼠血清淀粉酶、脂肪酶、二胺氧化酶(DAO)、肠型脂肪酸结合蛋白(IFABP)、D-乳酸(D-Lac)、IL-1β、IL-6、IL-18和TNF-α水平;采用TUNEL法检测肠黏膜上皮细胞凋亡情况;最后,使用Western blot测小肠组织TGR5以及NLRP3蛋白表达。结果与SAP组相比,CDCA干预后的大鼠胰腺和小肠组织病理损伤减轻,病理评分下降(P<0.05);ELISA法检测得出血清炎性因子(IL-1β、IL-6、IL-18、TNF-α)、淀粉酶、脂肪酶以及血清DAO、IFABP和D-Lac的水平也明显降低(P<0.05)。TUNEL检测结果显示Sham组肠黏膜上皮细胞的凋亡较SAP组也有明显缓解(P<0.05);经过CDCA干预后,小肠黏膜细胞凋亡指数及凋亡率较SAP组有明显下降(P<0.05)。Western blot结果显示,SAP造模后大鼠的小肠组织中TGR5蛋白表达较Sham组明显增多(P<0.05);同时,小肠组织的NLRP3在进行SAP造模后也明显增多,而经CDCA干预后,小肠组织NLRP3蛋白表达也降低。结论CDCA能显著降低SAP大鼠的胰腺及肠道损伤,发挥一定的保护作用,其作用机制可能与激活肠道的TGR5受体来抑制NLRP3的表达有关。
Objective To investigate the effects and significance of chenodeoxycholic acid(CDCA)on pancreatic and intestinal injury in rats with severe acute pancreatitis(SAP).Methods Forty-five male SD rats were randomly divided into three groups(15 rats of each group),namely SAP group,CDCA group and sham-operated(Sham)group.Rats in SAP group were first fed with universal diet for one week and then modeled by retrograde injection of 5%sodium taurocholate into the pancreaticobiliary duct.CDCA group were first fed with homogenous diet containing 0.5%CDCA for one week and then modeled,and tissue specimens were collected 24 hours after modeling.Tissue and serum specimens were collected.Firstly,the pathological changes of small intestine and pancreas tissues were observed by HE staining method;secondly,the levels of serum amylase,lipase,DAO,IFABP,D-Lac,IL-1β,IL-6,IL-18 and TNF-αwere detected by ELISA method.The apoptosis of intestinal mucosal epithelial cells was detected by TUNEL assay.Finally,the small intestine tissues were detected by WB method.The expression of TGR5 protein and inflammatory vesicles NLRP3 in small intestine tissue was detected by WB method.Results Compared with the SAP group,the histopathological damage of the pancreas and small intestine in the rats after CDCA intervention was reduced and the pathological score decreased(P<0.05),and the levels of serum inflammatory factors(IL-1β,IL-6,IL-18,TNF-α),amylase,lipase,and serum DAO,IFABP and D-Lac were also significantly reduced by ELISA(P<0.05).TUNEL assay results showed that apoptosis of intestinal mucosal epithelial cells was also significantly alleviated compared with the SAP group(P<0.05);finally,the expression of TGR5 protein as well as inflammatory vesicles NLRP3 in small intestinal tissues was also reduced after CDCA intervention.Conclusion CDCA significantly reduced pancreatic and intestinal injury in SAP rats and exerted certain protective effects,and this effect may be achieved by activating the TGR5 receptor in the intestine to inhibit the NLRP3 inflammasome.
作者
张帆
汤礼军
黄竹
吴俊
黄尚卿
ZHANG Fan;TANG Lijun;HUANG Zhu;WU Jun;HUANG Shangqing(School of Clinical Medical Sciences,Southwest Medical University,Luzhou 646000,Sichuan,China;PLA Center of General Surgery,General Hospital of Western Theater Command,Chengdu 610083,China;College of Medicine,Southwest Jiaotong University,Chengdu 610031,China)
出处
《西部医学》
2022年第12期1723-1728,1742,共7页
Medical Journal of West China
基金
国家临床重点专科项目(41732113)
国家自然科学基金(81772001)。
关键词
鹅去氧胆酸
重症急性胰腺炎
肠道损伤
Chenodeoxycholic acid
Severe acute pancreatitis
Intestinal injury
