| Objective:In this study,the model of acute pancreatitis in dogs was established,and the changes of oxidative stress and apoptosis in the treatment of acute pancreatitis in dogs with bone marrow mesenchymal stem cell secretory factor were preliminarily explored,which provided a theoretical basis for the treatment of acute pancreatitis in dogs with bone marrow mesenchymal stem cell secretory factor.Method:(1)Building canine acute pancreatitis model.A total of 48 healthy Chinese pastoral dogs were selected without any medical history The acute pancreatitis(AP)model was established by retrograde injection of sodium bovine cholic acid(0.5 ml/kg)and trypsin(3500 U/kg)into the pancreatic bile duct in 36 of them The remaining 12 were modeled with saline(2)To observe the changes of oxidative stress and apoptosis in canine acute pancreatitis treated with bone marrow mesenchymal stem cells secretory factor.Six normal saline modelling dogs were selected as A group(control group)and 18 successful model-making test dogs were randomly divided into B?C?D three groups,6 in each group.B group was model group,C group was ulinastatin treatment group,and D group was bone marrow mesenchymal stem cell secretory factor(BMSCs-F)treatment group.Superoxide dismutase(SOD),malondialdehyde(MDA),endoplasmic reticulum stress-related protein(CHOP),Caspase-9 in serum were determined by enzyme linked immunosorbent as say-sandwich technique(ELISA)at one day before modeling and that 1,3,5,7,10,15 days after treatment,respectively the corresponding concentration levels,the effects of BMSCs-F treatment AP oxidative stress and necrotizing apoptosis were analyzed(3)Changes of expression of HO-1 in canine acute pancreatitis treated with bone marrow mesenchymal stem cells secretory factor.Six normal saline modelling dogs were selected as the A group(control group),and 18 successful trial dogs were selected,randomly divided into B?C?D three groups,6 in each group.B group was the model group,C group was the ulinastatin treatment group,and D group was the bone marrow mesenchymal stem cell secretory factor(BMSCs-F)treatment group.Two experimental canine pancreatic samples were taken after ld,5d,and 15d of treatment to make paraffin sections.The expression of the HO-1 was observed by immunohistochemistry,the integral optical density(IOD)of the HO-1 was calculated to further analyze the possible mechanism of BMSCs-F treatment AP oxidative stress and apoptosis.Test results:(1)Results of building canine acute pancreatitis model.After modeling 24h,the test dog appeared vomiting,abdominal pain,loss of appetite,and other symptoms,serum amylase,serum lipase more than 5 times before operation.The modeling was successful.(2)Changes of oxidative stress and apoptosis in canine acute pancreatitis treated with bone marrow mesenchymal stem cells secretory factor.The SOD value of the BMSCs-F group began to decrease on the 1st day after treatment,reached the lowest value on the 3rd day,then slowly increased,and did not approach the control group indicator until the 15th day after treatment;The SOD value of the ulinastatin group began to decrease on the 1st day after treatment,increased slowly after reaching the lowest value on the 5th day,and did not approach the control group indicator until the 15th day after treatment;The SOD of the BMSCs-F group compared with that of the model group increased significantly on the 5th day after treatment(P<0.05);The SOD of the ulinastatin group compared with that of the model group showed a highly significant rising trend on the 7th day after treatment(P<0.01);On the 10th day after treatment,the SOD value of the BMSCs-F group was more significantly higher than that of the ulinastatin group(P<0.01).The MDA value of the BMSCs-F group and ulinastatin group began to on the 1st day after treatment,reached a peak on the 5th day,then decreased gradually,and did not approach the control group indicator until the 15th day after treatment;The MDA of the BMSCs-F group compared with that of the model group showed a highly significant downward trend on the 3rd day after treatment(P<0.01);The MDA of the ulinastatin group compared with that of the model group decreased significantly on the 5th day after treatment(P<0.05)and showed a highly significant downward trend on the 7th day(P<0.01);Compared with ulinastatin group,BMSCs-F group showed a highly significant downward trend in MDA value on the 10th daysafter treatment(P<0.01).The CHOP value of the BMSCs-F group began to rise on the 1st day after treatment,reached the highest value on the 3rd day,then decreased slowly,and did not approach the control group indicator until the 15th day after treatment;The CHOP value of the ulinastatin group began to rise on the 1st day after treatment,slowly decreased after reaching the highest value on the 5th day,and did not approach the control group indicatoruntil the 15th day after treatment;The CHOP of the BMSCs-F group compared with that of the model group showed a highly significant downward trend on the 1st day after treatment(P<0.01);The CHOP of the ulinastatin group compared with that of the model group showed a significant downward trend on the 5th day after treatment(P<0.05),and decreased highly significantly on the 7th day after treatment;The CHOP value of the BMSCs-F group compared with that of the ulinastatin group decreased significantly on the 7th day after treatment(P<0.05),and decreased highly significantly on the 10th day(P<0.01).The Caspase-9 value of the BMSCs-F group and ulinastatin group began to rise on the 1st day after treatment,reached the highest value on the 5th days,then gradually decreased,and did not approach the control group indicator until the 15th day after treatment;The Caspase-9 of the BMSCs-F group compared with that of the model group decreased significantly on the 3rd day after treatment(P<0.05),and showed a highly significant downward trend on the 5th day(P<0.01);The Caspase-9 of the ulinastatin group compared with that of the model group appeared significant difference on the 7th day after treatment(P<0.05)and decreased highly significantly on the 15th day(P<0.01);The Caspase-9 value of the BMSCs-F group compared with that of the ulinastatin group decreased significantly on the 5th day after treatment(P<0.05),and decreased highly significantly on the 10th day(P<0.01).(3)Changes of expression of HO-1 in canine acute pancreatitis treated with bone marrow mesenchymal stem cells secretory factor.The IOD in BMSCs-F group and ulinastatin group increased on the 1st day after treatment,reached a peak on the 5th day,then began to decrease,and did not approach the control group indicator until the 15th day after treatment;In BMSCs-F group and ulinastatin group compared with the model group,the IOD increased significantly on the 5th day after treatment(P<0.05).The IOD in BMSCs-F group was significantly higher than that in ulinastatin group on the 5th day after treatment(P<0.05),and very significantly increased on the 15th day(P<0.01).Conclusions:(1)The model of canine acute pancreatitis was successfully established by retrograde accessory pancreatic duct injection of bovine cholic acid sodium(0.5 ml/kg)and trypsin(3500 U/kg).(2)BMSCs-F may be to achieve good therapeutic effect of acute pancreatitis by effectively reducing pancreatic oxidative stress injury and reducing apoptosis.(3)BMSCs-F effectively reduce pancreatic oxidative stress injury and reduce apoptosis,may be related to up-regulation of HO-1 expression. |
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