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Objective: To explore the effects of electroacupuncture(EA) on metabolic patterns of the prefrontal cortex in rats with acute myocardial ischemia., Methods: Eighteen SD rats were randomly divided into sham group, model group and EA group, with 6 rats in each group. Rats in the model and EA groups were subject to acute myocardial ischemia by ligation of the left anterior descending coronary artery. For the EA group, EA stimulation (1 mA, 2 Hz/15 Hz, 30 min) was applied to "Shenmen"(HT7) -"Tongli"(HT5) once a day for 3 consecutive days. The histopathological changes of myocardial tissue and levels of ischemia modified albumin (IMA) in serum were determined by HE staining and ELISA, respectively. The LC-MS/MS technique was used to characterize the metabolic profiling of the prefrontal cortex. The differentially expressed metabolites were screened by principal component analysis(PCA) and partial least squares-linear discriminant analysis (PLS-LDA), and subsequently Kyoto encyclopedia of genes and genomes (KEGG) metabolic pathway enrichment analysis was performed., Results: Compared with the sham group, the myocardial fibers were disordered and fractured, and content of serum IMA was significantly increased in the model group ( P <0.01), which, however, were significantly decreased in the EA group ( P <0.01). With PCA and PLS-LDA, there were 18 differential metabolites between the model and sham groups. Forty-eight differential metabolites were emerged between the EA and model groups. Three metabolites associated to the sphingolipid metabolism were reversed by EA stimulation, as indicated by KEGG., Conclusion: The molecular mechanism of EA against myocardial ischemia is partially mediated by regulating sphingolipid-related metabolites in the prefrontal cortex.

Objective: To observe the effect of electroacupuncture (EA) on the cardiac function, lymphatic markers, macrophage and inflammatory cytokines in acute myocardial ischemia (AMI) mice, so as to explore its mechanism in improving AMI., Methods: Male C57BL/6 mice were randomly divided into sham operation, model, EA, inhibitor and inhibitor+EA groups, with 10 mice in each group. AMI model was established by occlusion of left anterior descending coronary artery. For mice in the EA group and inhibitor+EA group, EA (1 mA, 2 Hz/15 Hz) was applied to bilateral "Shenmen"(HT7) and "Tongli"(HT5) for 30 min, once daily for consecutive 3 days. Mice in the inhibitor+EA group were given intraperitoneal injection of vascular endothelial growth factor receptor-3 (VEGFR-3) inhibitor SAR131675 30 min before the EA, while mice in the inhibitor group were given intraperitoneal injection of SAR131675 only. The electrocardiogram (ECG) of the neck-thoracic lead was recorded and analyzed by BL-420F biological function experiment system. Histopathologic changes of myocardial tissue were observed after H.E. staining. The contents of lactate dehydrogenase (LDH), cardiac troponin I (cTnI) in serum and interleukin-18 (IL-18) and interleukin-6 (IL-6) in ischemic myocardium were determined by ELISA. The expressions of hyaluronic acid receptor-1 (LYVE-1) and macrophage mar-ker CD68 (CD68) in the myocardial tissue were detected by immunofluorescence assay. The protein expression levels of vascular endothelial growth factor C (VEGF-C) and VEGFR-3 were detected by Western blot., Results: Compared with the sham operation group, the ECG-ST level, the contents of serum LDH and cTnI, and the contents of IL-18 and IL-6 in the myocardial tissue were significantly increased ( P <0.01), the expression of LYVE-1 and the protein expression levels of VEGF-C and VEGFR-3 in the myocardial tissue were significantly decreased ( P <0.01), while the number of CD68 positive cells was significantly increased ( P <0.01) in the model group. Compared with the model, inhibitor and inhibitor+EA groups, the ECG-ST level, the contents of serum LDH and cTnI, and the contents of IL-18 and IL-6 in the myocardial tissue were decreased ( P <0.01), the expression of LYVE-1 and the expression level of VEGFR-3 protein were increased ( P <0.01), while the number of CD68 positive cells was significantly decreased ( P <0.01) in the EA group. Compared with the model and inhibitor groups, the expression level of VEGF-C was increased ( P <0.01) in the EA group. Outcomes of H.E. staining showed that the myocardial fibers were disordered with a large number of inflammatory cell infiltration in the model group, which was milder in the EA group., Conclusion: Acupuncture can improve the inflammatory injury of AMI mice, which may be related to activate VEGF-C/VEGFR-3 pathway to promote lymphangiogenesis, reduce macrophage infiltration and inflammatory factors.

Objective: To observe the effect of electroacupuncture (EA) on the expression of myocardial protein kinase B (Akt) and mammalian target of rapamycin (mTOR) in acute myocardial ischemia (AMI) rats., Methods: Thirty male SD rats were randomly divided into control, model and EA groups ( n =10 in each group). The AMI model was established by occlusion of the descending anterior branch (DAB) of the left coronary artery. EA (2 Hz, 1-2 mA) was applied to bilateral "Shenmen" (HT7) and "Tongli" (HT5) for 20 min, once daily for consecutive 7 days. The electrocardiogram (ECG) of nape-xiphoid lead was recorded for assessing changes of myocardial ischemia. Histopathologic changes of the ischemic myocardial tissue were observed after H.E. staining and ultra-microstructural changes of cardiomyocytes observed by transmission electron microscopy (TEM). The expression levels of Akt, phosphorylated-Akt (p-Akt), mTOR and phosphorylated-mTOR (p-mTOR) in the myocardium were detected by Western blot, followed by calculating the ratios of p-Akt/Akt and p-mTOR/mTOR., Results: Following ligature of DAB, the ECG-ST level was significantly increased in the model group in comparison with the control group ( P <0.01). At 30 min after treatment, the ECG-ST level decreased significantly compared with the model group ( P <0.01). At the end of the 7-day treatment period, the ECG-ST level increased compared with the model group ( P <0.05). The levels of myocardial p-Akt and p-mTOR protein expression, and the ratios of p-Akt/Akt and p-mTOR/mTOR were significantly lower in the model group than those in the control group ( P <0.01), and considerably increased in the EA group than in the model group ( P <0.01). No significant differences were found among the three groups in the expression levels of Akt and mTOR proteins ( P >0.05). Outcomes of H.E. staining and TEM showed damage of mitochondria and occurrence of a large number of autophagosomes in myocardiocytes in the model group, which was milder in the EA group., Conclusion: EA at HT5 and HT7 can improve AMI in AMI rats, which may be related to its effect in facilitating Akt/mTOR signaling.

Objective: To explore the molecular mechanism of locus coeruleus（LC） involved in electroacupuncture (EA) anti myocardial ischemia., Methods: Twenty-four SD rats were randomly divided into sham-operation, model, EA and EA +lesion groups, with 6 rats in each group. The acute myocardial ischemia (AMI) model was established by ligation of the left anterior descending branch of coronary artery. EA (2 Hz/15 Hz, 1 mA) was applied to bilateral "Shenmen" (HT7) -"Tongli" (HT5) and the middle-point between HT7 and HT5 for 30 min, once daily for 3 days. For rats of the EA +lesion group, the virus (300 nL) was injected into bilateral LC before EA treatment. Serum aspartate aminotransferase (AST) was detected by ELISA. The gene expression profiles of rat heart were detected by transcriptome sequencing, the differentially expressed genes were screened, and Gene Ontology (GO) functional classification and Kyoto Encyclopedia of genes and genomes (KEGG) metabolic pathway enrichment analysis were performed., Results: Compared with the sham-operation group, serum AST content was significantly increased in the model group ( P <0.01). Following the intervention, serum AST was significantly reduced in the EA group ( P <0.01), while the serum AST in the EA + lesion group was significantly higher compared with the EA group ( P <0.05). Differential expression analysis showed that 1 138 differentially expressed genes were screened out between the model group and the sham-operation group, 1 330 differentially expressed genes between model and EA group, and 804 differentially expressed genes between EA and EA + lesion group. Among them, 218 differential genes were involved in the regulation of EA anti-myocardial ischemia in LC. GO functional classification analysis showed that these differentially expressed genes mainly involved in cell processes, metabolic processes and biological regulation in biological processes. KEGG pathway analysis showed that these differentially expressed genes were enriched in sulfur relay system, thiamine metabolism, glutathione metabolism, C5 branch dicarboxylic acid metabolism, cell adhesion molecules and Th1 and Th2 cell differentiation., Conclusion: EA intervention has a positive effect in anti-myocardial ischemia, which may be related to the sulfur relay system, thiamine metabolism, glutathione metabolism, C5 branch dicarboxylic acid metabolism, cell adhesion molecules and Th1 and Th2 cell differentiation involved in LC.

Objective: To observe the changes of microvascular structure of acupoints caused by myocardial ischemia, so as to explore the application of photoacoustic imaging technology in the research of acupoint sensitization., Methods: Twelve BALB/c mice were randomly divided into normal, sham operation and acute myocardial ischemia (AMI) model groups, with 4 mice in each group. AMI model was established by ligating the left anterior descending coronary artery. Electrocardiogram (ECG) was recorded by physiological signal acquisition system at 12 h and on the 14 th day after modeling, and serum cardiac troponin T (cTnT) and creatine kinase isoenzyme MB (CK-MB) levels were detected by enzyme-linked immunosorbent assay. The microvascular structure changes of acupoints "Feishu"(BL13), "Jueyinshu"(BL14), "Quze"(PC3) and "Chize"(LU5) were observed by photoacoustic imaging technology, and distance (DM), inflection count metric (ICM), sum of angle metric(SOAM)and microvessel density (MVD) were calculated by microvascular quantification algorithm., Results: Compared with the normal and sham operation groups, the ST segment of ECG was obviously elevated, serum cTnT and CK-MB were significantly increased in AMI model group at 12 h and on the 14 th day after AMI ( P <0.01). The ICM of BL14 in AMI model group was significantly decreased on the 14 th day than that on the 7 th day after AMI. Compared with the normal group, the ICM of BL14 was significantly increased in AMI model group on the 7 th day after AMI( P <0.05). There were no significant changes in DM, ICM, SOAM and MVD at other acupoints on the 7 th and 14 th day ( P >0.05) among the three groups., Conclusion: The change of ICM may be one of the characteristics of acupoint sensitization and photoacoustic imaging technology can be used to study the structure of acupoint microvessels.

Objective: To compare the therapeutic effect of electroacupuncture (EA) of "Shenmen" (HT 7, Yuan point of the Heart Meridian), "Zhizheng" (SI 7, Luo point of the Small Intestine Meridian) and "Xinshu" (BL15, Back- shu point) on the expression of nerve growth factor (NGF) and its receptor, tyrosine kinase A (TrkA) in the cerebral cortex of myocardial ischemia (MI) rats., Methods: A total of 70 male SD rats were randomized into sham operation (sham, n =10), model, HT 7, SI 7, and BL15 groups ( n =15 in each of the latter 4 groups). The MI model was established by ligation of the anterior descending branch of the left coronary artery. EA (1 mA, 2 Hz) was respectively applied to HT 7, SI 7 and BL15 for 15 min, once per day for 1 week. Immunohistochemistry and real-time fluorescent quantitative PCR were used to detect the expression of NGF and TrkA proteins and genes in the cerebral cortex., Results: Compared with the sham group, no significant changes were found in the number of NGF immune-reaction (IR)-positive and TrkA IR-positive cells, and the expression levels of NGF mRNA and TrkA mRNA in the model group ( P >0.05). After EA intervention, the number of NGF and TrkA IR-positive cells and the expression of NGF mRNA and TrkA mRNA in each of the 3 EA groups were significantly increased relevant to the model group ( P <0.01, P <0.05). The effect of EA at BL 15 was significantly superior to that of EA at HT 7 and SI 7 in increasing the number of NGF and TrkA positive cells and up-regulating the expression of their mRNAs ( P <0.01, P <0.05)., Conclusion: EA at HT 7, SI 7 and BL 15 can increase the levels of expression of NGF and TrkA proteins and mRNAs in the cerebral cortex of subacute MI rats and the effects of EA-BL 15 are obviously superior to those of EA-HT 7 and EA-SI 7, suggesting a relative specificity of the effect of EA at different acupoints.

Objective: We have repeatedly demonstrated that electroacupuncture (EA) stimulation of "Shenmen" (HT 7)- "Tongli" (HT 5) segment of the Heart Meridian can improve acute myocardial ischemia (AMI). This study aimed at observing the effect of EA on contents of hippocampal norepinephrine (NE), and interleukin 6 (IL-6), IL-1 β and tumor necrosis factor- α (TNF-α) in AMI rats, so as to explore its underlying mechanism., Methods: SD rats were randomly divided into 3 groups: sham operation (sham), model and EA( n =6 rats in each). The anterior descending branch (ADB) of the left coronary artery was occluded to make an AMI model. For rats of the sham group, a surgical suture was simply threaded beneath the ADB without ligation. EA (2 Hz/15 Hz, 1 mA) was applied to bilateral "Shenmen" (HT 7)- "Tongli" (HT 5) and the middle-point between HT 7 and HT 5 for 30 min, once daily for 3 days. Electrocardiogram (ECG) of the neck-thoracic lead was recorded by using PowerLab 16. The contents of serum creatine kinase (CK), hippocampal IL-6, IL-1 β and TNF-α were assayed by ELISA. The concentration of NE in hippocampal CA 1 area was detected by microdialysis combined with electrochemical detector., Results: Compared with the sham group, the ECG-ST height, serum CK, hippocampal NE, IL-6, IL-1 β and TNF-α contents of the CA 1 region were significantly increased in the model group ( P <0.001). Whereas, after EA intervention, the serum CK, hippocampal NE, IL-6, IL-1 β and TNF-α contents were obviously down-regulated relevant to the model group ( P <0.05, P <0.001), and the IL-6, IL-1 β and TNF-α contents were positively correlated with the NE level ( P <0.001, P <0.01)., Conclusion: EA stimulation of the Heart Meridian may improve ischemic myocardial injury in AMI rats, probably by reducing the proinflammatory factors and hippocampal NE.