| Objective:To explore pathophysiology of Contraction of heart collaterals and Heart vessel blockage stasis,and give Tongluo intervention to both.Methods:1.Contraction of heart collaterals:① The animal model of coronary artery spasm was established in rats by single high-dose intravenous injection of pituitrin.The changes of rat’s electrocardiogram,myocardial microcirculation and myocardial blood flow were observed in order to evaluate the model.②TXL was given by gavage once a day for 7 days before,and the animal model was established 1 hour after the last.The levels of plasma ET-1,NO,TXB2 and 6-keto-PGF1α in rats were detected.The histomorphology changes of myocardium were observed by HE staining,Nagar-Olsen staining and transmission electron microscopy.The apoptotic index and the expression of Bax and Bcl-2 protein of myocardium were respectively determined by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling(TUNEL)assays and immunohistochemical staining.2.Heart vessel blockage stasis:①TXLU was given by gavage once a day for 7 days before,and the AMI animal model was induced by ligations of left anterior descending 1 hour after the last.6 hours later,the changes of rat’s electrocardiogram and histomorphology were observed.The levels of plasma ET-1,Ang II,TXB2 and 6-keto-PGF1α in rats were detected by radioimmunoassay.The apoptotic index and the expression of Bax and Bcl-2 protein of myocardium were respectively determined by terminal deoxynucleotide transferase mediated dUTP nick-end labeling(TUNEL)assays and immunohistochemical staining.② 24 hours later,the area of myocardial infarction was measured by TTC staining.The protein expression of HIF-la and VEGF were detected by Western blotting.the expression of CD34 and PDGF-B were determined by immunohistochemical staining.The mRNA expression of eNOS、ET-1、ICAM-1 and VCAM-1 were detected by Real-time quantitative PCR.Results:1.①Compared with normal group,ST segment was elevated,myocardial arterioles were constricted,and myocardial blood flow was decreased in the model group(P<0.05 or P＜0.01).②Compared with normal group,the levels of plasma ET-1,NO,TXB2 and TXB2/6-Keto-PGF1α were significantly increased(P<0.05 or P<0.01)while the level of 6-keto-PGF1α was decreased significantly(P<0.05).The area of myocardial ischemia was significantly increased(P<0.05).Compared with model group,the levels of plasma ET-1,TXB2 and TXB2/6-Keto-PGF1αwere significantly decreased(P<0.05 or P<0.01)while the level of 6-keto-PGF1α and NO were increased significantly in tongluo group(P<0.05).The area of myocardial ischemia was significantly decreased(P<0.05).③Karyopyknosis and margination of the chromatin in some myocardial cells were observed under transmission electron microscopy.The mitochondria of some myocardial cells were congregated and swollen with some ruptured mitochondrial membrane and disrupted mitochondrial ridge in model group.The mitochondria was mild swollen and mitochondrial ridge was complete.There was little karyopyknosis and mitochondria congregated in tongluo group.The apoptosis index,the expression of Bax and the ratio of Bax/Bcl-2 in model group were significantly increased(P<0.01).Compared with model group,the apoptosis index,the expression of Bax and the ratio of Bax/Bcl-2 were decreased(P<0.01).2.①Compared with sham group,the levels of plasma ET-1,TXB2 and TXB2/6-Keto-PGF1α were significantly increased(P<0.05 or P<0.01)while the level of 6-keto-PGF1α was decreased signif icantly(P<0.05).Compared with model group,the levels of plasma ET-1,TXB2 and TXB2/6-Keto-PGF1α were significantly decreased(P<0.05 or P<0.01)while the level of 6-keto-PGF1α was increased significantly in tongluo group(P<0.05).②The apoptosis index,the expression of Bax and the rat io of Bax/Bc1-2 in model group were significantly increased in model group(P<0.05 or P<0.01).Compared with model group,the apoptosis index,the expression of Bax and the ratio of Bax/Bcl-2 were decreased while the expression of Bcl-2 was increased(P<0.05 or P<0.01).③The protein expression of HIF-1 a,VEGF and PDGF-B were increased significantly in model group(C<0.05 or P<0.01).Compared with model group,The MVC and.the protein expression of HIF-1 a,VEGF and PDGF-B were increased significantly while the area of myocardial infarction was decreased in tongluo group.④The mRNA expression of eNOS、ET-1、ICAM-1 and VCAM-1 were increased significantly in model group(P<0.05 or P<0.01).Compared with model group,The mRNA expression of eNOS was increased significantly while mRNA expression of ET-1、ICAM-1 and VCAM-1 were decreased in tongluo group.Conclusion:1.Contraction of heart collaterals:①The CAS animal model was established successfully which lasts for more than 30mins.②Endothelial dysfunction participates in pathophysiology of CAS,and TXL can rectify endothelial dysfunction in order to anti-spasm,inhibit thrombosis and improve myocardial blood circulation.③Apoptosis caused by Bax/Bcl-2 participates in pathophysiology of myocardial injury,which may be related to ischemia reperfusion injury.Tongxinluo can protect ischemic myocardium by means of Bax and Bax/Bcl-2 down-regulation.2.Heart vessel blockage stasis:①Endothelial dysfunction participates in pathophysiology of AMI,and tongxinluo can rectify endothelial dysfunction so as to mitigate myocardial injury.②Apoptosis and necrocytosis participate jointly in myocardial injury In AMI early stages.The imbalance of expression of Bax and Bcl-2 regulate and control myocardium apoptosis.Tongxinluo can protect ischemic myocardium by means of Bcl-2 up-regulation and Bax down-regulation.③The pathways of HIF-1α-VEGF/PDGF-B and VEGF-eNOS-NO participate jointly in the process of angiogenesis.Tongxinluo can promote angiogenesis by means of HIF-1α,VEGF,PDGF-B and eNOS up-regulation.④Inflammatory reaction and cellular cytotoxicity caused by ICAM-1 and VCAM-1 participate in the pathological process of AMI.Tongxinluo can decrease the expression of ICAM-1 and VCAM-1 in myocardium which can alleviate the inflammation reaction and cellular cytotoxicity,reduce the infarct sie and protect ischemic myocardium. |
PDF Full Text Request