Construction Of Acute Pancreatitis Model In Dogs And Octreotide Treatment

Background:Acute pancreatitis is a common inflammatory disease,which can lead to multiple organ failure and high mortality during acute attack.Fluid resuscitation,analgesia and nutrition are the most basic clinical treatments.At present,in the research of AP therapeutic drugs,rats,mice and other experimental animals are mostly used to construct acute pancreatitis animal models,but there is a lack of related research on establishing AP models through dogs and primates.Beagle dogs with less difference with human physiology are used to complete the modeling,in order to provide reference for the construction of stable acute pancreatitis model.As a somatostatin analogue,octreotide has the function of inhibiting the secretion of polypeptide hormones in the gastrointestinal and pancreatic system and regulating gastrointestinal activity,so it is widely used in the clinical treatment of many diseases.In this study,the preparation technology of octreotide acetate freeze-dried powder injection was optimized,the preparation prescription was screened and the product stability was explored.The therapeutic effect of octreotide acetate freeze-dried powder on canine acute pancreatitis model was explored,in order to provide a new idea for the efficient diagnosis and treatment of AP.Objective:To optimize the prescription process of aseptic freeze-dried octreotide acetate preparation,to determine the content of octreotide acetate aseptic freeze-dried preparation by SERS in order to provide a basis for quality standard evaluation,and to explore the construction method of canine acute pancreatitis model and the efficacy of octreotide freeze-dried powder injection preparation on acute pancreatitis model dogs Method:1.According to the quality standard of freeze-dried powder injection in Chinese Pharmacopoeia,several different prescriptions were screened,samples were prepared by vacuum freeze-drying,and the stability of octreotide freeze-dried preparation under different conditions was investigated.2.The stretching vibration peak of mono-substituted phenyl ring of octreotide alanine was selected as the characteristic peak,the quantitative analysis method of Raman spectrum of octreotide acetate aqueous solution was established,and the related chromatographic conditions were optimized3.Four healthy beagle dogs were selected and operated by retrograde injection of cholangiopancreatic duct.The canine acute pancreatitis model was established by injection of sodium taurocholate and trypsin.4.Ten healthy beagle dogs were randomly divided into three groups: control group（n=4）,routine treatment group（n=4）and octreotide group（n=4）.The AP model was established by operation,and the control group was injected with normal saline after operation.the routine treatment group and octreotide treatment group were treated after operation.the lipase level,amylase level and blood routine of the dogs in each group were monitored before operation and 1-7 days after operation,and the curative effect of octreotide was evaluated.Results:1.The specification of octreotide freeze-dried powder injection was determined as0.05 mg,and the preparation prescription was selected as follows: 0.05 mg octreotide acetate,15 mg dextran 40 and 15 mg mannitol,1ml water for injection,the p H range was4.5-6.5.2.The SERS method for evaluating the quality standard of octreotide was determined: the stretching vibration peak of the mono-substituted phenyl ring of octreotide alanine was selected as the characteristic peak and the Raman shift 1000cm-1 was selected.The Raman spectrometer sets the parameters: the range is 400 cm to1500cm,the wavelength output of the Hemuri ne laser is 532 nm,the multiple of the objective lens is adjusted to 50 times,the laser attenuation parameter is 10%,the resolution is set to 1cm-1,the exposure time is set for 30 s,the integral is 3 times,and15 to 30 points are detected randomly.3.The postoperative monitoring results showed that the specific protein was positive;the levels of lipase and amylase were significantly higher than those before operation;ultrasound images showed that the pancreas was enlarged and the echo was abnormal,which was consistent with the characteristics of acute pancreatitis.Canine acute pancreatitis model was established successfully.4.The results of postoperative monitoring showed that the levels of lipase and amylase in the control group,routine treatment group and octreotide treatment group were significantly higher than those before operation,the detection of specific protein was positive,and the method of model construction was stable and repeatable.The levels of amylase and lipase in octreotide group were significantly lower than those in octreotide group（P < 0.05）.After operation,the levels of amylase and lipase in octreotide group were significantly lower than those in octreotide group（P < 0.05）.The results of ultrasound were good.Conclusion:1.The optimized preparation prescription of octreotide was selected as follows:0.05 mg octreotide acetate,15 mg dextran 40 and 15 mg mannitol,1ml water for injection,the p H range was 4.5-6.5.the experiment proved that the method was stable.2.The Raman spectrometer sets parameters: the range is 400 cm to 1500 cm,the wavelength output of the Hemuri ne laser is 532 nm,the objective lens multiple is adjusted to 50 times,the laser attenuation parameter is 10%,the resolution is set to 1cm-1,the exposure time is set for 30 s,and the integral is 3 times.Under this condition,the octreotide is detected,and the method is effective.3.Retrograde injection of sodium taurocholate and trypsin into biliopancreatic duct can stably establish canine acute pancreatitis model.4.Octreotide is effective in the treatment of acute pancreatitis in beagle dogs and reduces the occurrence of complications.