Experimental Study On The Effect Of Thoracic Duct Drainage And Salvianolic Acids On Severe Acute Pancreatitis Associate Intestinal Barrier Damage

Objectives:Severe Acute Pancreatitis(SAP),a common surgical disease,is one of the acute abdomen. It’s a dangerous clinical process which has thecharacteristics of many complications, difficult therapy and high mortality.Intestines as a central stress reaction organ that the integrity of intestinalbarrier function is closely related to the severity of SAP. During SAP,ischemia-reperfusion, impaired microcirculation, inflammation, excessiveproduction of cytokines, intestinal motility disorders, excessive apoptosis ofintestinal epithelial cells, intestinal flora disorders, growth factor deficiencyand intestinal epithelial cell excessive apoptosis and other reasons which led tothe intestinal barrier function disturbance (IBFD). The combined effects of theimpaired intestinal barrier, weak intestinal motility, immune suppression andother reasons give rise to intestinal bacteria and endotoxin translocation.Dislocated bacteria and endotoxin are closely relevant to the secondaryinfection of pancreas and the necrotic organization around the pancreas,systemic sepsis and （Multiple organ dysfunction syndrome，MODS）.Therefore IBFD is called the "engine" of SAP. Thus, preventing IBFD,blocking the translocation of bacterial toxins or other intestinal harmfulsubstances, improving intestinal, pancreatic and other visceralmicrocirculation disturbance are significant to prevent clinical process andimprove the prognosis of patients. In this study, one part of the animal modelin rats give Salvianolic Acids, the other part establish an animal model ofthoracic duct drainage on the basis of SAP. By discussing mechanism of theCervical thoracic duct drainage and Salvianolic Acids protective effect on SAPrats, we provide new ideas and experimental basis being used to guide clinical treatment of intestinal barrier dysfunction with SAP.Methods:One hundred and forty-four clean SD healthy male rats, weighing300g-380g were randomly divided into6groups: sham operation group (J),the control group of Cervical thoracic duct drainage(Sham), SAP group (S),Cervical thoracic duct drainage+SAP group(L), Salvianolic Acids+SAPgroup(D) and Cervical thoracic duct drainage+Salvianolic Acids+SAPgroup(LD). Each group then randomly divided into2h group,6h group and12h group, with8in each. sham operation group: found left jugular trunk thenclosed the incision+closed the incision after turning off the pancreas+tail-vein fluid infusion; the control group of Cervical thoracic duct drainage:Cervical thoracic duct drainage+closed the incision after turning off thepancreas; SAP group:Rat model was induced by retrograde injection of5%sodium taurocholate into the biliopancreatic duct(0.11ml/100g weight)+tail-vein fluid infusion; Cervical thoracic duct drainage+S AP group: madeSAP rat model after achieving Cervical thoracic duct drainage；SalvianolicAcids+SAP group：injected Salvianolic Acids after making SAP model；Cervical thoracic duct drainage+Salvianolic Acids+SAP group：InjectedSalvianolic Acids after achieving Cervical thoracic duct drainage and makingSAP model. Each teams undergo laparotomy on the basis of the regulationtime, visual inspection ascitic colour and pancreatic change. Collected ascites,blood， thoracic duct lymph in each hour,intestinal and pancreatic tissuesamples. The amylase (AMY) level of ascites, lymph and serum weremeasured by biochemical technique. Used UV spectroscopy to detect catalase(CAT) and glutathione-peroxidase (GSH-PX) level in ascites, lymph andserum. The level of Endotoxin in lymph and plasma were determined by theEndpoint chromogenic method. Histopathological sections of pancreas anddistal ileum tissue were observed to analyzed pathology score based onApplication HemateinEosin.Results：1Compared with J group and Sham group at the corresponding time point: the pathological changes of pancreas and intestine, as well as Thevolume of ascitic fluid,the level of serum amylase, ascitic fluid amylase,palsma endotoxin in SAP group,L group,D group,LD group were increasedsignificantly on all time point（P<0.05or P<0.01）. The level of serumGSH-PX and CAT, GSH-PX and CAT in Pancreas and intestine tissuehomogenates in SAP group,L group,D group,LD groupare were decreasedsignificantly on all time point（P<0.05or P<0.01）.2Compared with L group and D group at the corresponding time point:The quantity of ascitic fluid at all time point were increased in SAP group（P<0.01).the pathological changes of pancreas and intestine got higher scoreat12h in SAP group（P<0.01）,The level of serum GSH-PX and CAT at12htime point, GSH-PX and CAT in intestine tissue homogenates at all time point,GSH-PX and CAT in Pancreas tissue homogenates at2h and12h time pointwere all impaired in SAP group（P<0.05or P<0.01). The level of serumamylase, ascitic fluid amylase, plasma endotoxin on all time point wereincreased significantly（P<0.05or P<0.01）.3Compared with L group at the corresponding time point: The quantityof ascitic fluid at all time point were increased in D group（P<0.01).thepathological changes of pancreas and intestine, the level of serum CAT，CATin Pancreas and intestine tissue homogenates，serum GSH-PX and ascitic fluidamylase in D group were not changed significantly on all time poin（tP<0.05）.GSH-PX in intestine tissue homogenates in D group had no obvious differentat12h（P<0.05）. GSH-PX in Pancreas tissue homogenates at all time point,GSH-PX in intestine tissue homogenates at2h and6h were reducedsignificantly in D group（P<0.05）. The level of plasma endotoxin on6h and12h, The level of serum amylase on all time point were higher in D group（P<0.05or P<0.01). While the level of plasma endotoxin on2h wasincreased significantly in L group（P<0.05）.4Compared with LD group at the corresponding time point: The volumeof ascitic fluid at6h and12h time point were increased in D and L group（P<0.01).the pathological score of pancreas and intestine in L group and D group achieved more score at all time point（P<0.01）.The level of serumGSH-PX and CAT as well as GSH-PX in intestine tissue homogenates, CATand GSH-PX in Pancreas tissue homogenates on all time point, the level ofCAT intestine tissue homogenates at2h were dropped obviously in L groupand D group（P<0.05or P<0.01）. The level of CAT and GSH-PX Pancreastissue homogenates in L group and D group are depressed significantly on alltime poin（tP<0.05）. Serum amylase, ascites amylase, plasma endotoxin at alltime point and serum amylase in L6h and L12h in L group and D group wereall increased significantly (P<0.05or P<0.01).5Amylase and endotoxin in Lymph in L group, LD group and Sham:(1)Compared with Sham and LD: the level of Lymph amylase and endotoxin inL group were increased significantly on all time point（P<0.05or P<0.01）.（2）Comparsion among Sham: The level of Lymph amylase at6h and12hand Lymph endotoxin at all time point were much more higher in LD group（P<0.05or P<0.01）.Conclusions:1SAP induces intestinal mucosal pathomorphology damage at earlystage, indicating that this is a successful model to mimic the human SAPcouple with IBFD. Also, the higher level of endotoxin in serum and lymphreveal that SAP damages intestinal mucosal early.2Compare with J and sham group, the level of amylase in thoracic ductlymph and serum in SAP with IBFD increase obviously, especially inthoracic duct lymph. These results show that amylase in lymph may be oneof the major sources of amylase in serum.3The activity of CAT、GSH-PX in serum, lumph, pancreas and smallintestine samples in SAP with IBFD are reduced compare with J and shamgroup, and are further depressed with time going on, revealing thatantioxidant capacity is decreased, the ability of clear oxygen freeradical weakens.4Cervical thoracic duct drainage can improve SAP with IBFD since itcan decrease the transfer of endotoxin and AMY from lymph into bloodstream.5Salvianolic Acids reduce the concentration of endotoxin and AMY inserum, plasma and lymph, lead to attenuate the impairment of pancreas andsmall intestine mucosa.6Cervical thoracic duct drainage restore the activity of CAT、GSH-PX inSAP with IBFD, depress the damage caused by oxidative stress.7Salvianolic Acids restore the activity of CAT、GSH-PX in SAP withIBFD, depress the damage caused by oxidative stress.8Thoracic duct drainage+Salvianolic Acids have more benefit in SAPwith IBFD for Cervical thoracic duct drainage reduce the release ofendotoxin and AMY into bloodstream, improve the activity of CAT、GSH-PX,attenuate the impairment of pancreas and small intestine mucosa. At thesame time, Salvianolic Acids benefit the impaired microcirculation, clearoxygen free radical, inhibit platelet aggregation, thus relieve SAP and IBFD.