| 【Background】Depression is a class of persistent and obvious state of mind,slow thinking,will decline in activity,cognitive impairment as the main clinical manifestations of mood disorders.World Health Organization(WHO)global coping rate survey shows that depression has become the world’s fourth largest disease.The use of antidepressants is currently the primary means of treating various depressive disorders.According to the annual report of the US Poison Control System’s National Toxic Data System from 2006 to 2014,the incidence of exposure to antidepressants is on the rise,and antidepressant exposure has become the fifth most common poisoning-related event,AMI exposure and its death were all at the first place in all antidepressant drugs.TCAs is still as the first choice for the treatment of depression drugs in a lot of areas of China.Amitriptyline(AMI)and its active metabolite decamethrin have complex pharmacological effects,including inhibition of 5-HT reuptake,inhibition of NE reuptake,inhibition of DA reuptake,anticholinergic effect,antagonize histamine effect and so on.In addition to treating various types of depression,AMI also used to relieve chronic pain and so on.AMI showed a variety of adverse reactions,of which the most common and most serious cardiovascular response,especially arrhythmia(tachycardia)and low blood pressure is a common cause of death.It has been suggested that cardiotoxicity of AMI is associated with sodium channel,potassium channel and calcium channel ion channel,but these studies focus on clinical manifestations(ECG changes)and single channels and their influencing factors,and the conclusions are relatively scattered and difficult to Comparison and evaluation,it is difficult to fully and systematically explain the mechanism of AMI cardiotoxicity.【Objectives】In this study,we established the rat model of AMI acute poisoning and screeninged of myocardial protein.Review the literature,we focused on the abnormal expression of different myocardial proteins in AMI poisoning models and try to explore the possible mechanism of cardiovascular damage caused by AMI.(1)To establish the acute AMI poisoning model of rats,observe the effect of AMI acute poisoning on the behavior of rats and the morphological changes of multiple organs in rats;(2)To determine the LD50 of AMI acute poisoning in rat models;(3)Screening differentially expressed proteins of myocardial injury induced by AMI acute poisoning in rats;(4)To determine the function of differentially expressed proteins and to explore the molecular toxicological mechanism of myocardial injury induced by AMI acute poisoning in rats;(5)To find possible signs of acute AMI poisoning myocardial injury.【Materials and Methods】(1)Establishment of rat AMI acute poisoning model: 55 SD rats were randomly divided into 5 AMI poisoning groups and 1 blank control group.The 5 AMI poisoning groups were injected intraperitoneally by using 15mg/mL AMI normal saline solution in 65mg/kg,78mg/kg,94mg/kg,113mg/kg and 136mg/kg respectively.The rats in the control group were sacrificed by intraperitoneal injection of 2mL normal saline.Infected group and the control group were killed after 24 h.Take the hearts,rinse and weighed,cut the apex of the ventricular tissue fluid nitrogen frozen-80℃ preservation preparation,the remaining myocardial tissue in the paraformaldehyde solution fixed parallel histopathological examination.(2)LD50: According to the mortality rate data obtained from the above 5 groups(10 rats per group),the Sun’s modified Kouge method(point oblique method)and the Bliss method were used to calculate the acute AMI toxicity of LD50 in rat.(3)Two different groups of ventricular myocytes were extracted and extracted from the myocardium.The proteins were screened by 8-labeled iTRAQ combined with LC-MS/MS.(4)To explore the molecular toxicological mechanism of myocardial injury induced by AMI acute poisoning in rats,and to find possible markers of acute AMI poisoning myocardial injury.【Results】(1)Behavioral changes: AMI poisoning groups of rats were two kinds of stable symptoms,paralysis and convulsions.After the exposure,such as movement disorders and mouth breathing,respiratory rate changes and other symptoms,but its frequency were low.(2)Histopathological changes: there were no obvious pathology changes in macroscopic observation of rat hearts and there were only dotted sarcoplasm condensation in myocardial fibers.There were no obvious pathology changes in other organs.There were some liver necrosis in some rats of high dose groups.(3)LD50: two methods were used to calculate the LD50 of AMI acute poisoning,and we get LD50 relatively close about 96 mg/kg;(4)Screening the different proteins: AMI acute poisoning rat myocardial protein for iTRAQ technology combined with liquid chromatography tandem mass spectrometry(LCMS/MS)detection,screening out the statistically significant(p<0.05)in myocardial differentially expressed proteins more than 2200 kinds,among them up to the standard of screening protein has 352 kinds.【Conclusion】(1)By using a single 15 mg/mL AMI of saline solution to intraperitoneal injection can establish AMI rats model of acute poisoning.Canister to stay lie less rapidly happened after the move,paralysis,and cramps,convulsions can be used as a most successful performance;(2)By using a single 15 mg/mL AMI of saline solution to intraperitoneal injection,calculate LD50 SD rats about 96 mg/kg;(3)The iTRAQ technology combined with LC-MS/MS can be used for AMI acute poisoning rat myocardial differences in protein detection;(4)Through to the AMI acute poisoning rat myocardial differences in protein appraisal and analysis of the Ca2+ channel and its related proteins,myocardial contraction related proteins and protein involved in mitochondrial respiratory chain are more obvious differences in expression,the causes may be associated with AMI infected. |
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