The Mechanism Of Action Of MIRI Based On The Induction Of Inflammatory Response By Mast Cell Degranulation

Purpose With the help of rat myocardial ischemia-reperfusion injury model,mast cell related inflammatory reaction was the breakthrough point.To study the protective effect of Tongxinluo preadministration on ischemia-reperfusion injury myocardium and the mechanism of inflammation induced by mast cell degranulation,and to explore the protective effect of Tongxinluo on ischemia-reperfusion injury myocardium.Methods Thirty-six healthy adult male SD rats were randomly divided into sham-operation group,model group and Tongxinluo group.The rats in the Tongxinluo group were given Tongxinluo medicine,and the sham operation group and the model group were given the same amount of normal saline for gastric perfusion.The model was made for 7 days.Myocardial ischemia-reperfusion injury model was established by reversible left anterior descending coronary artery ligation(1 h ischemia,2 h reperfusion),In the sham operation group,only the puncture thread was not ligated.Electrocardiogram(ECG)was measured 10 minutes after modeling and 2 hours after reperfusion.Blood samples from abdominal aorta were collected and blood routine was detected.After centrifugation,calcium ion,creatine kinase,lactate dehydrogenase,trypsin and c-fos immunoreactive double labeling,spleen tyrosine kinase were detected by Elisa.The phosphorylation level of extracellular signal-regulated kinases is 5-hydroxytryptamine;The left ventricular tissue of the rat myocardium was stained with hematoxylin and eosin and the histopathologic changes were observed under optical microscope.The level of beta aminohexosidase and chymotrypsin was detected by ELISA method,and the apoptosis index of myocardial cells was detected by DNA fracture in situ end labeling method.The expression of PKC epsilon and PKC Delta mRNA in myocardial tissue was detected by real-time fluorescent quantitative polynucleotide chain reaction;Western blot assay was used to detect the expression of myocardial tissue.The expression level of protein kinase was measured.Results The results of electrocardiogram after reperfusion:the T wave in the model group was significantly higher than that of the sham operation group.The T wave in the Tongxinluo group was significantly lower than that of the model group,and the decline was about 0.06mm in the model group.Blood:the model group compared with the sham group of white blood cells(WBC)and elevated(P<0.01),Tongxinluo group rats WBC,platelet(PLT)level lower than the model group(P<0.05);the number of red blood cells of rats and hemoglobin values were not significantly different(P>0.05);Ca2+,CK and LDH:the Ca2+,CK and LDH increased in the model group than those in the sham operation group(P<0.01 or P<0.05),and the levels of Ca2+,CK and LDH in Tongxinluo group were lower than those in the model group(P<0.01 or P<0.05).TPS,c-fos,Syk,5-HT and histamine in serum index:the model group compared with the sham operation group(P<0.01 or P<0.05),decreased in Tongxinluo group compared with model group(P<0.01 or P<0.05);serum ERK:model group decreased significantly compared to the sham operation(P<0.01).Tongxinluo group was significantly higher than the model group(P<0.01).HE staining:the inflammatory cells infiltration in the sham operation group was accompanied by necrosis of some myocardial cells.The infiltration of inflammatory cells in the model group was most obvious,accompanied by the coagulation and necrosis of cardiac muscle cells,and the infiltration of inflammatory cells in Tongxinluo group was not obvious.Beta hexosaminidase and chymase:the model group compared with the sham operation group(P<0.05),decreased in Tongxinluo group compared with model group(P<0.05);myocardial apoptosis index:the model group compared with the sham operation group(P<0.01),decreased in Tongxinluo group than in the model group(P<0.01);PKC epsilon and PKC delta mRNA:The model group compared with the sham group was significantly increased(P<0.01),Tongxinluo group was significantly lower than the model group,the difference was statistically significant(P<0.01);PKC ?:the model group compared with the sham operated group was significantly increased(P<0.01),Tongxinluo group was significantly lower than the model group,the difference was statistically significant(P<0.01).Conclusion Tongxinluo preconditioning can reduce myocardial ischemia reperfusion injury,and its mechanism is related to the inflammatory response to the inhibition of mast cell degranulation.The protective effect of Tongxinluo on cardiomyocytes is related to the up regulation of anti apoptotic gene expression,self repair of cells and inhibition of cardiomyocyte apoptosis.Tongxinluo drugs may reduce the expression of protective factor ERK and inhibit the inflammatory reaction related to mast cell degranulation and reduce myocardial injury by decreasing the inflammatory index of TPS,c-fos,Syk and 5-HT in serum.The protective effect of Tongxinluo on myocardial reperfusion may be related to reducing the expression of beta aminoglucosidase,chymotrypsin and PKC,and inhibiting the degranulation of mast cells.