活血清解汤对重症急性胰腺炎大鼠肠道免疫屏障功能的影响及机制  

Effect of Huoxueqingjie decoction on intestinal immune barrier function in rats with severe acute pancreatitis and the mechanism

作　　者：梁超[1] 汪扬 刘新博 李善宝 LIANG Chao;WANG Yang;LIU Xinbo;LI Shanbao(Department of Hepatobiliary and Pancreatic Surgery,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Department of General Surgery,Shanghai General Hospital,School of Medicine,Shanghai Jiaotong University,200080 Shanghai,China)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院肝胆胰外科,上海200437 [2]上海交通大学医学院附属上海市第一人民医院普通外科,上海200080

出　　处：《中国普通外科杂志》2022年第9期1182-1193,共12页China Journal of General Surgery

基　　金：上海中医药大学附属岳阳中西医结合医院青年孵育计划基金资助项目(2019YYQ03)。

摘　　要：背景与目的:重症急性胰腺炎(SAP)易并发肠道免疫屏障功能障碍,笔者前期发现活血清解汤可缓解SAP,但其作用是否与肠道免疫屏障功能有关尚不清楚。因此,本研究探讨活血清解汤对SAP大鼠肠道免疫屏障功能的作用及机制。方法:将SD大鼠随机分成假手术组、SAP模型组(模型组)、SAP模型+活血清解汤治疗组(治疗组),SAP模型采用5%牛磺胆酸钠胰胆管逆行注射法诱导,治疗组造模后给予活血清解汤灌胃,假手术组与模型组给予等体积生理盐水代替。分别在术后6、12 h处死动物,ELISA检测各组血清中的促炎因子TNF-α、抗炎因子IL-4、血淀粉酶及脂肪酶的指标变化;HE染色检测胰腺及小肠组织的病理变化;qRT-PCR及Western blot检测小肠组织巨噬细胞M1型标志物iNOS及M2型标志物Arg-1的表达。将大鼠小肠巨噬细胞分别用牛磺胆酸钠、活血清解汤、牛磺胆酸钠+活血清解汤处理,以无处理的大鼠小肠巨噬细胞为空白对照,用qRT-PCR及Western blot检测各组细胞NF-κB、iNOS、TNF-α、IL-4、Arg-1、TIPE2和PPAR-γ的表达。结果:体内实验中,与假手术组比较,模型组与治疗组术后的TNF-α、IL-4、血淀粉酶、脂肪酶水平均有不同程度升高(部分P<0.05),但治疗组TNF-α、血淀粉酶、脂肪酶升高的程度明显低于模型组,而IL-4升高的水平明显大于模型组(均P<0.05);模型组与治疗组术后胰腺与小肠组织均出现明显的病理损伤,但治疗组的损伤程度小于模型组,且随时间延长,模型组的损伤程度加重,治疗组则减轻,模型组与治疗组胰腺与小肠组织病理评分差异均有统计学意义(均P<0.05);iNOS蛋白与mRNA的表达在模型组小肠组织中表达明显升高,而在治疗组的小肠组织中明显降低(均P<0.05),Arg-1蛋白与mRNA的表达模型组小肠组织后期(12 h)明显升高,而在治疗组的早期(6 h)与后期(12 h)均明显升高,且升高程度大于模型组(均P<0.05)。体外实验Background and Aims:Severe acute pancreatitis(SAP) is always associated with intestinal immune barrier dysfunction.The authors previously found that Huoxueqingjie(HXQJD) decoction can alleviate SAP damage,but whether this action is involved in the intestinal immune barrier function is unclear.Therefore,this study was conducted to investigate the effect of HXQJD decoction on the intestinal immune barrier function in rats with SAP,and the mechanism.Methods:The SD rats were randomly divided into sham operation group,SAP model group(SAP group),and SAP model plus HXQJD decoction treatment group(treatment group).The SAP model was established by retrograde biliopancreatic duct infusion of 5% sodium taurocholate(Na T).After the operation,HXQJD decoction was administrated by gavage in the treatment group,and the sham operation group and model group were treated with normal saline in the same fashion.Rats were sacrificed at 6 and 12 h after the operation.The serum pro-inflammatory factor TNF-α,antiinflammatory factor IL-4,as well as blood amylase and lipase levels were detected by ELISA assay.The pathological changes in the pancreas and small intestine were observed by HE staining.The expressions of M1 macrophage phenotype marker i NOS and M2 macrophage phenotype marker Arg-1 in small intestine tissue were determined by q RT-PCR and Western blot.Rat intestinal macrophages were treated with Na T,HXQJD decoction,or Na T plus HXQJD decoction,respectively,using the untreated rat intestinal macrophages as blank control.The expressions of NF-κB,i NOS,TNF-α,Arg-1,IL-4,TIPE2,and PPAR-γ were examined by q RT-PCR and Western blot,respectively.Results:In invivo study,the levels of TNF-α,IL-4,and serum amylase and lipase were increased in different degrees after operation in model group and treatment group compared with sham operation group(partial P0.05) in the rat intestinal macrophages after Na T treatment alone;the protein and m RNA expression levels of Arg-1,IL-4,TIPE2,and PPAR-γ were significantly increased(all P<0.05) whi

关 键 词：胰腺炎 急性坏死性 活血清解汤 巨噬细胞 肠 大鼠 

分 类 号：R657.5[医药卫生—外科学]