Effective Part And Mechanism Of Ligusticum Chuanxiong Hort. On The Treatment Of Acute Myocardial Ischemia

Objective:In this study,this topic by adopting the method of Traditional Chinese Medicine（TCM）chemical preparation of Ligusticum chuanxiong Hort.different chemical parts,including Ligusticum chuanxiong Hort.water extract（LCW）,Ligusticum chuanxiong Hort.alcohol extract（LCA）,Ligusticum chuanxiong Hort.volatile oil（LCO）,Ligusticum chuanxiong Hort.total alkaloids（LCK）,Ligusticum chuanxiong Hort.total phenolic acids（LCP）on the application of rhizoma ligustici wallichii promoting blood circulation to remove blood stasis efficacy for the treatment of myocardial ischemia,discuss the role of active substance and the possible mechanism.Methods:1.Rat models of acute myocardial ischemia induced by isoproterenol（ISO）and acute myocardial ischemia induced by pituitrin（Pit）were established respectively.The rats were randomly divided into control group,model group,anolol group,LCW group,LCA group,LCO group,LCK group and LCP group.In the Ligusticum chuanxiong group,the different extracts of Ligusticum chuanxiong Hort.were given according to the amount of raw medicine,the dosage was 3.33g/kg,and the anti pain group was given 9mg/kg,all by 1 time/d,and lasted for 5 days.Then ISO 40mg/kg was injected into the abdominal cavity or Pit 0.5U/kg was injected under sublingual vein.The electrocardiogram changes of rats before and after the model were recorded by electrocardiogram.The content or activity of AST,CK,LDH and SOD and MDA related to oxidative damage were measured by the kits,and the content or activity of cTnT in the myocardium was measured.The cardiac tissues of 3 rats were randomly selected from each group.Western blot analysis of the expression of apoptosis-related proteins Bax and Bcl-2 in myocardial tissue.In addition,in the model experiment of acute myocardial ischemia induced by Pit,8 rats were taken from each group,and the model was duplicated with the same method.HE staining was used to observe the morphological changes of the myocardium.2.Further studies on the mechanism of volatile oil from effective parts were carried out.Rats were randomly divided into control group,model group,isosorbide dinitrate group,Ligusticum chuanxiong volatile oil Low,medium and high doses group and high dose+LY group.After 5 days of low,medium and high dose of Ligusticum chuanxiong and the anti pain group was given,the model of acute myocardial ischemia induced by Pit was produced by the same method.10 minutes before modeling,the high dose group+LY of Chuanxiong volatile oil was intraperitoneally injected with LY294002（PI3K/Akt pathway specific blocker）0.3mg/kg.The contents or activities of SOD and MDA related to AST,CK,LDH and oxidative damage were detected by the changes of electrocardiogram in rats.The cTnT,Ca²⁺content,Na⁺-K⁺-ATPase,Ca²⁺-ATPase and Mg²⁺-ATPase activity were measured by kits.The cardiac tissues of 3 rats were collected and the protein expressions of Akt,p-Akt,Bax and Bcl-2 in myocardium were measured by Western blot.8 rats were taken from each group and the acute myocardial ischemia model induced by Pit was replicated by the same method.HE staining was used to observe the morphological changes of myocardial ischemia and the detection of cardiomyocyte apoptosis.Results:1.Screening results of effective fraction of Ligusticum chuanxiong Hort for acute myocardial ischemia（1）Effect of Ligusticum chuanxiong Hort.on acute ischemic injury induced by ISO in SD rats.In the experiment of acute ischemic injury of SD rats induced by ISO,the activity of LDH and CK in the serum of the model group increased,the activity of SOD decreased,the content of MDA increased,and the content of cTnT in the myocardium increased,and there was a significant difference compared with the control group（P<0.05）.At the same time,the abnormal elevation or inversion of T wave in electrocardiogram indicated that the model was successful.Compared with the model group,the different Ligusticum chuanxiong Hort.parts（LCW、LCA、LCO、LCK、LCP）could improve the abnormal T wave elevation or inversion in the electrocardiogram,and significantly reduce the serum LDH,CK,MDA and cTnT content in the myocardium,and the SOD viability was significantly enhanced（P<0.05）.After acute ischemia,the expression of Bax in the model group increased and the Bcl-2 expression decreased.The different parts of isosorbide dioxone and Chuanxiong could decrease the expression of Bax and increase the expression of Bcl-2,which significantly reduced Bax/Bcl-.2 proportions,the difference was significant（P<0.05）.（2）Effect of Ligusticum chuanxiong Hort.on acute ischemic injury induced by Pit in SD ratsIn the experiment of acute ischemic injury of SD rats induced by Pit,the activity of LDH and CK in the serum of the model group increased,the activity of SOD decreased,the content of MDA increased,and the content of cTnT in the myocardium increased,which was significantly different from that of the control group（P<0.05）.At the same time,ECG T wave was abnormal elevation or inversion,myocardial tissue HE staining observation,myocardial fiber rupture,inflammatory infiltration phenomenon,suggesting that the success of modeling.Compared with the model group,the different parts can improve the abnormal T wave elevation or inversion in the ECG after ischemia,and significantly reduce the serum LDH,CK,MDA and cTnT content in the myocardium,but the SOD activity is significantly enhanced,the difference is significant（P<0.05）.After acute myocardial ischemia,compared with the normal group,the expression of Bax in the myocardial tissue of the model group was increased,while the expression of Bcl-2 was decreased.The different parts of isosorbide dioxone and Chuanxiong could decrease the expression of Bax in the myocardial tissue and increase the expression of Bcl-2.,Significantly reduce the proportion of Bax/Bcl-2,the difference was significant（P<0.05）.The pathological staining of HE showed that the myocardium in the model group had inflammatory infiltration,disorder of myocardial fiber arrangement,myocardial fibrosis,and some phenomena of nucleus contraction and necrosis of the nucleus of the myocardium.Different parts of Ligusticum chuanxiong can reduce the disorder of myocardial fiber arrangement and fibrosis,reduce the infiltration and necrosis of myocarditis after ischemia.2.The effective mechanism of Ligusticum chuanxiong Hort.on acute myocardial ischemiaIn the experiment of acute ischemic injury of SD rats induced by Pit,the activity of LDH and CK in the serum of model rats increased,the activity of SOD decreased,the content of MDA increased,the content of cTnT in the myocardium increased,the activity of ATP ase decreased,and the Ca²⁺concentration increased significantly,and there was a significant difference compared with the control group（P<0.05）.At the same time,electrocardiogram T wave abnormal elevation or inversion,myocardial tissue HE staining observation,myocardial fiber rupture,inflammatory infiltration and other phenomena,myocardial tissue HE staining observation,myocardial fiber rupture,inflammatory infiltration and other phenomena,suggesting the success of the model.Compared with the model group,each dose group（low,middle and high）of sac and Ligusticum chuanxiong Hort.volatile oil（low,medium and high）can improve the abnormal T wave elevation or inversion in the ECG after ischemia,obviously reduce the serum LDH,CK,MDA and cTnT content in the myocardium,but the SOD activity is obviously enhanced,and the ATP active ase in the myocardium group is enhanced(Na⁺-K⁺-ATP ase,Ca²⁺-ATP ase and Mg²⁺-ATP ase),Ca²⁺concentration decreased significantly,the difference was significant（P<0.05）.After acute ischemia,the expression of p-Akt decreased,the expression of Bax increased,and the expression of Bcl-2 decreased in the model group.The expression of p-Akt increased,the ratio of p-Akt/Akt increased,the expression of Bax decreased,the expression of Bcl-2 increased,and Bax/Bcl-2 decreased significantly in each dose group（low,medium,and high）.The results showed a significant difference（P<0.05）.However,the high dose+LY group of Ligusticum chuanxiong Hort.volatile oil has a weak effect on the improvement of ischemic myocardium related indexes.There is no significant difference in myocardial ase spectrum,oxidative damage related ase,Ca²⁺concentration and ATP ase activity compared with the model group（P>0.05）.Myocardial morphological changes in pathological injury and cardiomyocyte apoptosis were also more serious than those in other groups of Ligusticum chuanxiong Hort.There was no significant difference in the expression level of p-Akt,Bax and Bcl-2between cardiac tissue and model group（P>0.05）.Conclusions:In this paper,the methods of ISO/Pit induced acute myocardial ischemia in rats were used to study the therapeutic effect of different parts of Ligusticum chuanxiong on ischemic rats,and the mechanism of its action was discussed.The main conclusions are as follows:1.Different parts of Ligusticum chuanxiong Hort.（water extract,alcohol extract,volatile oil,alkaloid,total phenolic acid）can reduce the expression of myocardial damage markers,improve the expression of ases in oxidative damage related ases,regulate apoptosis related proteins,decrease the expression of Bax and increase the expression of Bcl-2,reduce myocardial ischemia injury,and have certain protection.Protective effect.When the amount of Chuanxiong was 3.33g/kg,the effect of Chuanxiong volatile oil was better.2.Ligusticum chuanxiong Hort.volatile oil is the effective part of Ligusticum chuanxiong Hort in the treatment of acute myocardial ischemia in rats.Its effect is mainly antioxidant injury,alleviating myocardial cell injury and apoptosis.The mechanism of its action is closely related to the activation of PI3K/Akt signaling pathway;when the signal pathway is suppressed,the effect of Ligusticum Chuanxiong against myocardial blood is significantly reduced.