| Objective: to establish a model of canine acute pancreatitis and treat it with Bone Marrow Mesenchymal Stem cell-derived Bone ulesin(bmc-cm),to explore the clinical therapeutic effect of Bone Marrow Mesenchymal bmc-cm on canine acute pancreatitis,and to provide a basis for clinical application of bmc-cm in the treatment of canine acute pancreatitis.Methods:(1)establishment of dog acute pancreatitis modelTwenty-four healthy Chinese farm dogs were selected and divided into the control group(n=6)and the model group(n=18).In the model group,5% sodium beaver-cholate(0.1ml/kg)and recombinant trypsin(3500U/ kg)were mixed and retrograde injected through the auxiliary pancreatic duct to establish the dog AP model.In the control group,0.5%NACL was injected through the pancreatic duct retrograde with the same dose.After modeling,the levels of White Blood Cell(WBC),c-reactive protein,serum amylase(AMY)and Lipase(LIP)were detected,which were determined by combining with the rapid test plate for pancreatitis in idesdog.(2)grouping treatment and index collectionAfter modeling,the control group was group A,and the model group was randomly divided into groups B,C and D.Group A received routine treatment(tongstatin,5% glucose normal saline + creatinine + coenzyme A+ adenosine triphosphate + vitamin C,normal saline + ampicillin sodium for injection).Group B received conventional treatment(the same treatment as group A),group C added bmc-cm on the basis of conventional symptomatic treatment,and group D added ulinastatin on the basis of conventional symptomatic treatment.At 1,3,5,7,10,15 days after treatment,changes in clinical symptoms were observed and recorded in each group,and changes in concentrations of WBC,CRP,AMY and LIP were detected.B ultrasonic examination,histopathological examination and HE staining were performed on the 15 th day after treatment for histological observation.The obtained data were statistically analyzed using SPSS22.0 software.Test results:(1)results of model establishmentThe accessory pancreatic duct was mixed with 5% sodium bezoar cholate(0.1ml/kg)and recombinant trypsin(3500U/kg)The model was established by retrograde injection.After the injection of the molding agent,the connective tissue membrane of the pancreas immediately filled with edema and began to bleed.After 5min of modeling,the pancreas was enlarged and showed deep purple band bleeding.In group A,0.5%NaCl of the same dose was injected retrograde through the cholangiopancreatic duct.Clinical symptoms including vomiting,diarrhea,loss of appetite,abdominal pain and arched back were found in the model group 12 hours after the operation.12 h postoperatively except CRP in group A mild inflammation in other areas outside the indicators only small amplitude fluctuation are within the normal range,the WBC model group,CRP,AMY,LIP results than the normal range,AMY,LIP up to three times the normal range,with A group of extremely significant difference(P < 0.01),Ed and rapid detection of canine pancreatitis board,group A negative result,model group for positive results.Prompt successful molding.(2)group treatment resultsClinical changes: on the first day of treatment,group A presented abdominal pain and mild diarrhea,palpated abdominal wall was slightly tense,and returned to normal on the third day until the end of the trial.Group B without taking medication 1 day started vomiting,diarrhea and fecal for blood,abdominal pain and palpable abdominal wall more nervous,did not take the drug treatment 3 days have test dogs begin to symptoms of fever,without taking the fifth day of drug therapy has affected puppies appear "prayer" posture,and have a shock occurs,the seventh day shock dogs die a natural death.During this period,most of the experimental dogs in this group were depressed,and some of them got better,but they did not recover naturally after the experiment.On the first day of treatment,the symptoms of group C were not significantly different from those of group B and group C.On the third day of treatment,the abdominal pain was significantly reduced and the appetite was good.Postoperative treatment group D day 1 vomiting,diarrhea,abdominal pain and palpable abdominal wall extremely nervous,depressed with dog spirit appetite waste,treatment 3 days having appetite to recover,but there are still vomiting,diarrhea,etc.,have had symptoms of fever,fifth day get better treatment,treatment 7 days most of the symptoms disappear,abdominal palpation slightly nervous,the rest of the clinical symptoms disappeared.Changes in WBC: the mean time of disappearance of clinical symptoms was 5 days in bmsc-cm group and 7 days in ulinastatin group.The WBC in the bmsc-cm group began to increase after surgery,reached the peak on the 3rd day of treatment,and then returned to the normal range on the 7th day of treatment,with no significant difference compared with the control group(P>0.05).The WBC in the ulinastatin group reached its peak on the 3rd day of treatment,and then began to decrease.The normal range of WBC in the ulinastatin group was significantly different from that in the bmsc-cm group on the 10 th day of treatment(P<0.01).Changes in CRP: CRP in the bmsc-cm group began to rise 12 h after the operation,reached a peak on the first day of treatment,and then began to decline and returned to normal on the seventh day of treatment,with no significant difference compared with the control group(P>0.05).CRP in ulinastatin group began to rise at 12 h after the operation,reached the peak on the third day of treatment,and then maintained A stable trend of decline.It returned to the normal range on the 10 th day of treatment,but there was still A significant difference compared with group A(P>0.05).CRP in group B increased 12 h after modeling and reached A peak on the 7th day of treatment,and then decreased to 10 days after treatment,which was still above the normal range,showing A very significant difference from group A(P<0.01).AMY changes: AMY in the bmsc-cm group increased to about 3 times the normal value 12 h after the operation,reached the peak on the first day of treatment,and returned to the normal range on the seventh day of treatment.There was no significant difference between the treatment group and group A on the 15 th day of treatment(P>0.05).The peak value of ulinastatin group reached on the 1st day of treatment and there was no significant difference between group C and group C(P>0.05),and it returned to the normal range on the 15 th day of treatment,but there was still A significant difference compared with group A.LIP change: LIP in the bmsc-cm group began to increase postoperatively,reached its peak on the 3rd day after surgery,and then began to decline and returned to the normal range on the 7th day.The peak of ulinastatin group was reached on the 5th day after the operation,which was significantly different from that of bmsc-cm group(P<0.010).Results of ultrasonic scanning: after 15 days of treatment,the duodenal intestinal wall in group A was clearly structured and stratified,the thickness of pancreas was about 0.6cm,the echo was homogeneous,the structure was clear,and the pancreatic capsule was visible.For group B is 12 bowel wall layered blur,intestinal wall appeared fault,tips for gastric ulcer,and visible serous layer thickening,mesenteric echogenicity prompt peritonitis,anechoic around some clues ascites,or bleeding,and the thickness of the pancreas is about 1.35 cm,echo heterogeneity,the capsule is not visible,there are more calcifications and liquid dark space hint for acute hemorrhagic pancreatitis.Visible duodenal stratified group C compared with group B is clear,the thickness of the pancreas is about 0.84 cm,pancreas calcifications is formed,A small amount of visible but no pancreas capsule group A clear,peritoneal normal group D shows the duodenum and abdominal organs form of unofficial stratification is not clear,the pancreas capsule rough invisible,pancreatic tissue increase more calcifications,greater omentum echogenicity calcification prompted to exist.Pathological examination showed that the pancreas in group A returned to normal and was white and white,while in group B the pancreas showed severe hemorrhage and necrosis,and the original structure and color of the pancreas completely disappeared.The structure of the pancreas in group C was intact and slightly thicker than that of the normal pancreas.There was a small amount of calcification in the right lobe of the pancreas.Compared with normal tissues,the pancreatic tissue in group D showed no morphological structure,thickening of the pancreas,formation of a large number of calcification foci,invisible structure,and enlargement and congestion of the spleen.Histopathological sections: on the 15 th day of treatment,histopathological observation was performed.In group A,the pancreas structure was basically normal.Group B pancreatic interstitial edema,broadening,capillary hyperemia,hemorrhage,can see a large number of red blood cells,pancreatic lobule structure damage,glands inherent structure disappears,cytoplasmic vacuoles,acinar cells atrophy,glandular epithelial cell falls off,fracture,a large number of inflammatory cells infiltration,a large number of vacuoles serious formation damage degree than the control group obviously;Group C showed clear glandular structure,mild pancreatic tissue edema,focal necrosis of acinar cells,a small amount of inflammatory cell infiltration and red blood cell overflow.In group D,the pancreatic interstitium was edema and widened,and a small number of red blood cells were also seen in the interstitium,the cytoplasm vacuolized,the inherent structure of the gland disappeared,the glandular epithelial cells were exfoliated,and a small amount of fragmentation,which was less severe than that of the positive control group.Conclusion: the AP model was established by retrograde injection of 5% sodium botal-cholate(0.1ml/kg)and recombinant trypsin(3500U/kg)in the auxiliary pancreatic duct.The therapeutic effect of MSC-CM was better than that of ulinastatin group. |
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