Effect of electroacupuncture on myocardial electrical remodeling in rats with acute myocardial infarction. / ç”µé’ˆå¯¹æ€¥æ€§å¿ƒè‚Œæ¢—æ­»å¤§é¼ å¿ƒè‚Œç”µé‡æž„çš„å½±å“åŠå ¶æœºåˆ¶ç ”ç©¶.

OBJECTIVES: To investigate the mechanism of electroacupuncture (EA) at "Neiguan" (PC6) in impro-ving myocardial electrical remodeling in rats with acute myocardial infarction (AMI) by enhancing transient outward potassium current. METHODS: A total of 30 male SD rats were randomly divided into control, model and EA groups, with 10 rats in each group. The AMI model was established by subcutaneous injection with isoprenaline (ISO, 85 mg/kg). EA was applied to left PC6 for 20 min, once daily for 5 days. Electrocardiogram (ECG) was recorded after treatment. TTC staining was used to observe myocardial necrosis. HE staining was used to observe the pathological morphology of myocardial tissue and measure the cross-sectional area of myocardium. Potassium ion-related genes in myocardial tissue were detected by RNA sequencing. The mRNA and protein expressions of Kchip2 and Kv4.2 in myocardial tissue were detected by real-time fluorescence quantitative PCR and Western blot, respectively. RESULTS: Compared with the control group, cardiomyocyte cross-sectional area in the model group was significantly increased (P<0.01), the ST segment was significantly elevated (P<0.01), and QT, QTc, QTd and QTcd were all significantly increased (P<0.05, P<0.01). After EA treatment, cardiomyocyte cross-sectional area was significantly decreased (P<0.01), the ST segment was significantly reduced (P<0.01), and the QT, QTc, QTcd and QTd were significantly decreased (P<0.01, P<0.05). RNA sequencing results showed that a total of 20 potassium ion-related genes co-expressed by the 3 groups were identified. Among them, Kchip2 expression was up-regulated most notablely in the EA group. Compared with the control group, the mRNA and protein expressions of Kchip2 and Kv4.2 in the myocardial tissue of the model group were significantly decreased (P<0.01, P<0.05), while those were increased in the EA group (P<0.01, P<0.05). CONCLUSIONS: EA may improve myocardial electrical remodeling in rats with myocardial infarction, which may be related to its functions in up-regulating the expressions of Kchip2 and Kv4.2.

Effects of electroacupuncture on rats with cognitive impairment: An iTRAQ-based proteomics analysis.

OBJECTIVE: The study explores the effects of electroacupuncture (EA) at the governing vessel (GV) on proteomic changes in the hippocampus of rats with cognitive impairment. METHODS: Healthy male rats were randomly divided into 3 groups: sham, model and EA. Cognitive impairment was induced by left middle cerebral artery occlusion in the model and EA groups. Rats in the EA group were treated with EA at Shenting (GV24) and Baihui (GV20) for 7 d. Neurological deficit was scored using the Longa scale, the learning and memory ability was detected using the Morris water maze (MWM) test, and the proteomic profiling in the hippocampus was analyzed using protein-labeling technology based on the isobaric tag for relative and absolute quantitation (iTRAQ). The Western blot (WB) analysis was used to detect the proteins and validate the results of iTRAQ. RESULTS: Compared with the model group, the neurological deficit score was significantly reduced, and the escape latency in the MWM test was significantly shortened, while the number of platform crossings increased in the EA group. A total of 2872 proteins were identified by iTRAQ. Differentially expressed proteins (DEPs) were identified between different groups: 92 proteins were upregulated and 103 were downregulated in the model group compared with the sham group, while 142 proteins were upregulated and 126 were downregulated in the EA group compared with the model group. Most of the DEPs were involved in oxidative phosphorylation, glycolipid metabolism and synaptic transmission. Furthermore, we also verified 4 DEPs using WB technology. Although the WB results were not exactly the same as the iTRAQ results, the expression trends of the DEPs were consistent. The upregulation of heat-shock protein ß1 (Hspb1) was the highest in the EA group compared to the model group. CONCLUSION: EA can effect proteomic changes in the hippocampus of rats with cognitive impairment. Hspb1 may be involved in the molecular mechanism by which acupuncture improves cognitive impairment.

[Involvement of hippocampal miR-664-3p in electroacupuncture induced improvement of learning-memory ability of cerebral ischemia-reperfusion rats].

OBJECTIVE: To explore the effect of electroacupuncture (EA) on learning-memory ability and its correlation with miR-664-3p levels in the hippocampus in cerebral ischemia-reperfusion injury (CI/RI) rats. METHODS: A total of 27 male SD rats were randomly divided into sham operation, CI/RI model and EA groups (n＝9 in each group). The CI/RI model was established by occlusion of the middle cerebral artery (MACO) according to the modified Longa methods. EA was applied to "Baihui" (GV20) and "Shenting" (GV24) for 20 min, once a day for 7 days. The Morris water maze tests (place navigation tasks and spatial probe trials) were used to evaluate the rats' learning-memory ability. After EA intervention, the hippocampal tissue was collected to measure the mRNA and miRNA expression profiles by using high-throughput next generation sequencing. Bioinformatics analyses including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to screen the most abundant differentially-expressed miRNAs and mRNAs, particularly the target genes of miR-664-3p. Their biological functional categories including the key biochemical metabolic pathways and signaling pathways involved in CI and EA effect in the left hippocampus tissue were analyzed. RESULTS: The average escape latency of place navigation tests was significantly longer in the CI/RI model group than in the sham operation group (P<0.01), and obviously shorter in the EA group than in the model group (P<0.01); and the times of escape-platform quadrant crossing of the spatial probe trials were considerably fewer in the CI/RI model group than in the sham operation group (P<0.05), and markedly more in the CI/RI EA group than in the model group (P<0.05). Combined analysis of miRNA and mRNA showed that the differentially-expressed target genes of miR-664-3p were significantly up-regulated after CI/RI relevant to the sham operation rats (P<0.05) and notably down-regulated in the EA group relevant to the CI/RI model group (P<0.05). The GO analysis displayed that the target genes of miR-664-3p were significantly enriched in the category of cell transportation activity, ion binding，and transmembrane transporter activity, the production of precursor metabolites and energy, transportation, signal transduction，localization, etc. KEGG analysis showed that the target genes of miR-664-3p in this study were mainly enriched in the thyroid hormone pathway, Hippo signaling pathway, MAPK signal transduction pathway，cAMP signaling pathway, neurotrophic factor signal transduction pathway, etc. CONCLUSION: EA of GV20 and GV24 can improve the learning-memory ability of CI/RI rats, which is closely related to its effect in down-regulating the levels of hippocampal target genes of miR-664-3p involving cellular transmembrane transportation，cell to environment interaction，cell-cell communication, etc.

[Changes of Skin Temperature of Acupoint Regions of the Governor Meridian after Moxibustion Stimulation].

OBJECTIVE: To observe changes of skin temperature along the Governor Vessel (GV) after moxibustion stimulation. METHODS: A total of 36 healthy volunteer subjects were recruited in the present study. The skin temperature of Mingmen (GV 4), Dazhui (GV 14), Shenzhu (GV 12), Zhiyang (GV 9), Zhongshu (GV 7), Yaoyangguan (GV 3) regions and their bilateral control sites (1.5 cun lateral to GV 4) was detected by using a Thermo Tracer TH 9100. RESULTS: Following moxibustion stimulation of GV 4 region, a marked infrared radiant track along the central part of the back covering the GV was observed in the healthy volunteer subjects, and the skin temperature levels of both GV 4 and its bilateral control points were significantly increased (P < 0.001). The skin temperature of GV 4 was remarkably higher than those of both left and right control points (P < 0.001). After moxibustion stimulation, the average skin temperature levels of GV 9, GV 7, GV 4 and GV 3 were all up-regulated in comparison with pre-moxibustion (P < 0.01). CONCLUSION: Moxibustion stimulation of Mingmen (GV 4) can induce an infrared radiant track along the central part of the back covering the GV and raise skin temperature of acupoint areas of the GV.

[Effect of Electroacupuncture Stimulation of Acupoints at the Distal Limbs on Heart Function of Volunteers with Acute Hypoxia].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation of different acupoints at the distal ends of the limbs on cardiac function in volunteers with acute hypoxia, so as to determine if its actions are realized by way of segmental innervations or meridians. METHODS: Twenty healthy volunteers were divided into control, Quze (PC 3), Shousanli (LI 10), Guangming (GB 37) and Zusanli (ST 36) groups (both PC 3 and LI 10 are innervated by spinal C3-C6, and both ST 36 and GB 37 innervated by L5-S1). Acute hyoxia (simulating the conditions of about 5,000 m height above the sea level) was induced by asking the volunteers to inhale low-oxygen gas mixture (10.8% O2 + 89.2% N2) for 30 min, when, the participants' cardiac output (CO), heart rate (HR), left cardiac work (LOW), left ventricular ejection time (LVET)were measured using a ICG Monitor and EA stimulation (10 Hz/20 Hz, 1-2 V) was also conducted for 20 min following inhaling low-oxygen for 10 min. RESULTS: Before low-oxygen inhale, the levels of CO, HR, LCW and LVET ratios (test value/basic value) of the control, PC 3, LI 10, ST 36 and GB 37 groups were comparable (P > 0.05). After inhaling low-oxygen gas mixture for 10 min, the levels of CO, HR, and LCW ratios were significantly increased, and the LVET ratios were notably decreased in the five groups (P < 0.05). Compared with the 10 min-low-oxygen inhale of the same one group, CO and HR ratios at both EA 10 min and 20 min in the PC 3 and ST 36 groups, LCW ratios at EA 10 min in both PC 3 and ST 36 groups were notably down-regulated (P < 0.05), while the LVET ratios of both PC 3 and ST 36 groups was significantly prolonged (P < 0.05). No significant changes of CO, HR, LCW and LVET ratios were found in the LI 10 and GB 37 groups after EA for 10 min and 20 min (P > 0.05). CONCLUSION: EA stimulation of Quze (PC 3) and Zusanli (ST 36), but not Shousanli (LI 10) and Guangming (GB 37) can lower CO, HR and LCW levels and increase LVET in volunteer subjects undergoing acute hypoxia, suggesting that the therapeutic effect of EA maybe not rely on the segmental innervations, but rather, depend on the meridians to which the acupoints belong.

[Relationship between regional mast cell activity and peripheral nerve discharges during manual acupuncture stimulation of "Zusanli" (ST 36)].

OBJECTIVE: To observe changes of discharges of the sciatic nerve branch and mast cell activities and collagen fibers in the acupoint area during manual acupuncture stimulation of "Zusanli"(ST 36), so as to reveal the relationship between peripheral nerve and mast cell activities. METHODS: A total of 30 male SD rats were divided into normal, acupuncture control (an acupuncture neidle was inserted into ST 36 without manipulation), manual acupuncture (MA), disodium cromoglycate (DSCG, suppressing mast cell activity) plus acupuncture (MA + DSCG) and col lagenase (dissolving the collagen fibers) plus acupuncture (MA+ collagenase) groups (6 rats/group). After dissection of a branch of the sciatic nerve innervating ST 36 region in the left hind-limb under anesthesia, the ipsilateral ST 36 was stimulated by manipulating the acupuncture needle for 20 min. Discharges of the sciatic nerve branch were recorded by using a pair of metal electrodes and data acquisition system (Power Lab). Skin and muscle tissues of ST 36 area were sampled, sliced and stained with Toluidine Blue for detecting the number of degranulated mast cells. RESULTS: Compared with the control group, the mean power spectrum of d ischarges of the sciatic nerve and the mean rates of the degranulated mast cells in "Zusanli" (ST 36) area in the MA group were significantly increased (P<0.01). Whereas the mean power spectrum of discharges of the sciatic nerve and the mean degranulation rates of mast cells were considerably lower in the MA + DSCG group and MA+ collagenase group than in the MA group (P<0.01). No significant differences were found between the normal and control groups, and between the MA+NDSCG and MA+collagenase groups in the mean power density and degranulation rates of mast cells (P>0.05). CONCLUSION: Manual acupuncture stimulation of Zuai"ST 36 can significantly potentiate the discharge activity of the sciatic nerve and induce degranulation of mast cells at the same time, suggesting an involvement of mast cells in initiating acupuncture signals by peripheral sensory nerve.

[Development of studies on bioeffects of ultrasound-acupuncture therapy and its underlying mechanism].

The so-called ultrasound acupuncture is a therapeutic approach for clinical problems and health care by applying the ultrasound energy to the acupoints of the human body directly or indirectly. It has been applied in clinic for about 30 years since 1980s. In the present paper, the authors review the development of both experimental and clinical researches in the past 30 years. Its clinical application includes allergic rhinitis, local pain, mastitis, angina pectoris of coronary heart disease, stroke, etc. Regarding the researches on the underlying mechanism of ultrasound and ultrasound acupuncture, the authors make a summary from 1) bioeffects (thermal and nonthermal effects) of ultrasound intervention; 2) cell lysis and nonlysis effects of ultrasound intervention; and 3) effects of ultrasound acupuncture on the degranulation of mast cells. Based on the idea that "inflammatory reaction caused by mast cell degranulation is one of the initial factors of acupuncture for inducing therapeutic effects", bioeffects including cellular changes, especially mast cell degranulation caused by ultrasound stimulation, are thought to be the main possible mechanisms underlying the favorable efficacy of ultrasound acupuncture intervention. However, the ultrasound metrology and the specific superiority of ultrasound acupuncture remain unknown up to now.

In adjuvant-induced arthritic rats, acupuncture analgesic effects are histamine dependent: potential reasons for acupoint preference in clinical practice.

This study investigated whether immediate acupuncture effects in the acupoint are histamine dependent. Both histamine injection and manual acupuncture stimulation increased the pain threshold (PT) after treatment compared with the model group (P < 0.01), producing an analgesic effect. After pretreatment with clemastine, an H1 receptor antagonist and an antipruritic, the increase in the animals' pain threshold after acupuncture was suppressed compared with the Acu group (P < 0.01); however, there was no interference with the acupuncture-induced degranulation of mast cells. Pretreatment with disodium cromolyn did not suppress the increase in PT induced by the histamine injection at Zusanli (ST-36). We conclude that in adjuvant-induced arthritic rats, acupuncture analgesic effects are histamine dependent, and this histamine dependence determines the acupoint preference of acupoints away from the target site in acupuncture practice.

[Nitric oxide promotes the differentiation of neural stem cells in vitro derived from the subventricular zone of neonatal rats].

OBJECTIVE: To observe the effect of nitric oxide (NO) on the differentiation of neural stem cells (NSCs) derived from subventricular zone (SVZ) of neonatal rats in vitro. METHODS: Conventional method was used to isolate and culture the NSCs from SVZ. Diethylenetriamine/NO(DETA/NO) was used as NO donor and Nitro-L-arginine methylester (L-NAME) was used as inhibitor of nitric oxide synthase (NOS). The immunofluorescence was used to identify the expression of nestin (a marker of NSCs), beta-III-tubulin (Tuj-1, a marker of neurons), glial fibrillary acidic protein (GFAP, a marker of astrocytes) and nNOS. The concentration of NO in medium was measured by Greiss assay. RESULTS: Cultured neurospheres were nestin-, BrdU- and nNOS-positive. After treatment with 40 micromol/L, 50 micromol/L and 60 micromol/L of DETA/NO for 5 days, the concentration of NO released was increased significantly (P < 0.01) as compared with that of the control group. The percentage of both differentiated neurons and astrocytes was increased significantly (P < 0.01 and P < 0.05) as compared with that of the control group. After treatment with 100 micromol/L, 150 micromol/L and 200 micromol/L of L-NAME for 5 days, the concentration of NO released was decreased as compared with that of the control group (P < 0.05). The percentage of both differentiated neurons and astrocytes were decreased as compared with that of the control group (P < 0.05). CONCLUSION: NO could directly promote the differentiation of NSCs derived from rat subventricular zone in vitro.

[Effect of electroacupuncture of different acupoints on electroretinogram and cerebral visual evoked potentials in healthy subjects].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of different acupoints on electroretinogram (ERG) and cerebral visual evoked potentials (VEP) in healthy subjects so as to evaluate the relative specificity of the function of acupoints. METHODS: A total of 33 healthy adult volunteers were observed in the present study. The subject was asked to lie on a testing bed for a while, two disk electrodes were respectively fixed to the suborbital region and the occiput (the crossing point, 5 cm apart from the right median line of the head and 5 cm above the ear), and the reference electrode was fixed to the medial side of the earlobe for recording ERG and cerebral VEP separately. The testing room was asked to keep faint light during the whole recording process. A flash stimulation (frequency 1 Hz) was applied to the subject's eye (with a distance of about 50 cm between the light source and the tested eye, and the other eye was covered by an eyepatch). EA (0.5 Hz, 0.2 ms and 3-4 V) was applied to Guangming (GB 37), Neiguan (PC 6) and Zusanli (ST 36) for 30 min respectively in different testing days (with the interval being 3 days at least). ERG and cerebral VEP were recorded by using Polygraphy (RM-6200). RESULTS: Following EA of Guangming (GB 37), Neiguan (PC 6) and Zusanli (ST 36),the changing values (increase and decrease) of the amplitude of b waves of ERG and P100 waves of cerebral VEP were (19.68 +/- 12.61)% and (12.84 +/- 14.08)%, (6.09 +/- 14.06)% and (5.61 +/- 8.01)%, (2.39 +/- 3.52)% and (5.43 +/- 8.22)%, respectively, while those of b waves of ERG and P100 waves of cerebral VEP of GB 37 were significantly bigger than those of PC 6 and ST 36 (P < 0.01). CONCLUSION: EA of Guangming (GB 37)can significantly change the amplitude of both ERG and cerebral VEP in volunteer subjects, which is markedly bigger than those of EA of Zusanli (ST 36) and Neiguan (PC 6), suggesting a relative specificity of the effect of EA of different acupoints.