Mechanism Research Based On Acupoint Proteomics Of Acupuncture At Different Acupoints Improving Cardiac Function In AMI Rats

Objective:To explore the effect differences of acupuncture at different acupoints in the treatment of AMI rats,and study the local common and unique key proteins and signal pathways of acupoints after acupuncture at different acupoints by proteomics technology,so as to provide experimental basis for revealing the acupoint initiation mechanism of acupuncture effect differences.Method:1 Comparative study on the effect of acupuncture at different acupoints on cardiac function in AMI ratsThe wister rats were randomly divided into a sham operation group(n=9)and a model group(n=36).The rats in the model group were ligated the left anterior descending coronary artery to establish the rat AMI model.The sham operation group only had thoracotomy without ligation of the left anterior descending coronary artery.The rat AMI model was further randomly divided into model group,acupuncture PC6 group,acupuncture BL15 group,and acupuncture ST36 group,with 9 rats in each group.Among them,the PC6 acupuncture group,the acupuncture BL15 group,and the acupuncture ST36 group were respectively given hand acupuncture treatment at the corresponding points on both sides,using the twisting technique,the frequency was 120 times/min,the stimulation was 2 minutes,the needles were retained for8 minutes,and then repeated Acupuncture was performed once for 20 minutes for 7 consecutive days.The sham operation group and the model group were not given acupuncture treatment,but were given the same restraint and fixation measures as the acupuncture group for 20 minutes.The left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS)were detected by small animal echocardiography,and the myocardial infarction area was observed by TTC staining in the rat heart,and compared and analyzed.Therapeutic effect of acupuncture at different points on AMI rats.2.Screening of local key differential proteins and signaling pathways after acupuncture at different acupointsOn the premise of clarifying the differences in the therapeutic effects of acupuncture at different acupoints,the 4D-LFQ proteomic technology was used to analyze and identify the local tissue proteins of acupoints.First,the types of protein components and differential protein screening analysis were performed to screen the proteins whose differential expression changes in each group were up-regulated or down-regulated by 1.5 times,and had statistical differences;these differential proteins were further applied Gene Ontology(GO)function,KEGG pathway,Protein cluster analysis and protein interaction network analysis.By comparing the similarities and differences of local proteins of acupuncture at different acupoints and their relationship with acupuncture effect,the key differential proteins and signal pathways that acupuncture may improve cardiac function in AMI were screened.Result:1.Effects of acupuncture at different acupoints on cardiac function and myocardial infarction size in AMI ratsAfter the AMI model was created,the thickness of the left ventricular wall of the rats was significantly reduced,and the range of motion was weakened;compared with the shamoperated group,the LVEF and LVFS values were significantly decreased(P<0.01).Compared with the model group,the LVEF and LVFS values of the acupuncture PC6 group were significantly increased(P<0.05),the acupuncture BL15 group showed an upward trend,and the acupuncture ST36 group had no significant effect.At the same time,the AMI model rats had obvious myocardial infarction area,and acupuncture could improve the myocardial infarction area of AMI rats to different degrees,and the improvement effect was the most obvious in the PC6 group.2.Local key differential proteins and signaling pathways of acupuncture at different acupoints(1)The differential proteins between the model group and the sham operation group were analyzed.It was found that there was a common differential protein among PC6,BL15 and ST36,and there are 13 common differential proteins between PC6 and BL15 points;The results of differential protein interaction network analysis showed that there were no shared differential proteins among the three acupoints of PC6,BL15 and ST36,and 6 shared differential proteins between PC6 and BL15;KEGG pathway enrichment results show that there is no common KEGG pathway between PC6,BL15 and ST3,and there is a common KEGG pathway between PC6 and BL1,namely the pentose phosphate pathway,and the following Mainly adjusted,indicating that after the AMI model was created,the protein changes of PC6 and BL15 were convergent,meaning the local energy metabolism was reduced.(2)Differential analysis of the differential proteins between the acupuncture group and the model group showed that there were 176 common differential proteins between PC6,BL15 and ST36,267 common differential proteins between PC6 and BL15,491 common differential proteins between PC6 and ST36,and only 186 common differential proteins between BL15 and ST36.There are 38 common differential proteins between PC6,BL15 and ST36,69 unique differential proteins in PC6,83 unique differential proteins in BL15,and 59 unique differential proteins in ST36;The KEGG pathway enrichment results showed that there were two shared KEGG pathways between PC6,BL15 and ST36,namely the neural ligand-receptor interaction pathway and the carbon metabolism pathway.The action pathway is mainly up-regulated,and the carbon metabolism pathway is mainly down-regulated;PC6 has 4 own KEGG pathways,namely glucagon pathway,myocardial contraction pathway and other pathways,and the downregulated pathway is the main one,and BL15 has 15 KEGG pathways.,namely the inflammatory mediator regulation pathway of TRP channel and the actin cytoskeleton regulation pathway,etc.,and down-regulation is the main one.There are 5 self-owned KEGG pathways in ST36,namely synaptic vesicle cycle,etc.,and Mainly the above tone;The results of MCODE analysis of differentially expressed proteins showed that the differential protein functions of PC6 mainly involved oxidative phosphorylation,endocytosis,and complement and coagulation cascades,,in which endocytosis and complement and coagulation cascade related proteins are mainly up-regulated,oxidative phosphorylation related proteins are mainly down regulated,The differential protein functions of BL15 mainly involve complement and coagulation cascade and endoplasmic reticulum-Golgi transport,and all of them are mainly up-regulated.The protein interaction network of ST36 mainly involves oxidative phosphoric acid and ribosomes,in which ribosome-related proteins are mainly upregulated,and oxidative phosphorylation-related proteins are mainly down-regulated.Conclusion:1.The effect of acupuncture at PC6 on improving cardiac function in AMI rats is better than that at BL15 and ST36.2.In the state of acupuncture,PC6,BL15 and ST36 all activated the local neuroactive ligand-receptor interaction pathway in the acupoint area,but it cannot be considered to be related to the synergistic effect of the three acupoints in the treatment of AMI;the specific effects of acupoints The signaling pathway of PC6 may be related to the endocytosis pathway,BL15 may be related to the endoplasmic reticulum-Golgi transport process,and ST36 may be related to the protein synthesis process,but it is not clear that these activation pathways are related to the acupuncture effect of three points.3.In the state of AMI model,the local protein of PC6 and BL15 has a convergent change,which shows that the function of pentose phosphate pathway is reduced,but there is no similar change at ST36,which may be a manifestation of the specific correlation between Zang Fu organs and meridians.