Can electroacupuncture affect the sympathetic activity, estimated by skin temperature measurement? A functional MRI study on the effect of needling at GB 34 and GB 39 on patients with pain in the lower extremity.

The Purpose of this study was to investigate the possible needling effect on sympathetic activity by using functional MRI (fMRI). Twelve patients with left lower extremity pain were enrolled in our study. Each was given deep needling at left GB34 (yanglingquan) and GB39 (xuanzhong) points simultaneously. All patients got the strong DeQi sensation by manipulating the needles, and then electroacupucture (EA) was given and lasted for thirty minutes before fMRI scan. Then the fMRI scan was performed by scanning the whole brain with five blocks lasting 2 minutes each. The patients' palm skin temperatures were tested every five minutes as indication of the sympathetic activity from the beginning of EA to the end of our fMRI scan. Functional images were processed by using FEAT software at different levels including the simple single subject analysis, multi-subjects group mean analysis, and multi-subjects unpaired two difference analysis. Finally, 9 of 12 patients' palm temperatures increased gradually but the other three decreased. In comparison of two different palm skin temperature change groups, more significant activation over bilateral caudate head, right lentiform and periaqueductal gray (PAG) were found in the temperature-increased group, but palm temperature-decreased patients revealed more significant activation over bilateral anterior cingulated cortex (ACC), insula, primary somatosensory gyrus (SI), orbitofrontal cortex, occipital cortex, hippocampus and amygdala formation. Our study suggested that needling at analgesic points may modulate the sympathetic activity and such evident difference on brain responses may correlate with the clinical analgesia effects.

Sympathetic nervous system mediates surgical trauma stress-induced splenocyte apoptosis in rats.

Surgical trauma stress has been reported to induce immunosuppression. The mechanisms involved are still unclear. The present study was designed to assess the role of the sympathetic nervous system in regulating splenocyte apoptosis induced by surgical trauma stress. Our results showed that the rats that underwent surgical trauma stress exhibited a significant reduction in splenic cellularity, the loss of splenocytes was likely mediated by apoptosis, for a substantial increase in apoptosis was observed by using DNA gel electrophoresis and TUNEL assay. At the same time, an increase in Fas(CD95/Apo-1) protein expression in splenocytes was also observed. These effects were significantly abolished by either chemical sympathectomy or beta-adrenergic receptor antagonist propranolol. The data clearly revealed that the sympathetic nervous system especially beta-adrenergic receptors was involved in surgical trauma-induced immune alterations via a mechanism of apoptotic cell death.

Electroacupuncture combined with clomipramine enhances antidepressant effect in rodents.

The present study was designed to evaluate the antidepressant effect of electroacupuncture (EA) and the potential additive or synergistic effects of EA and clomipramine (CLO, a tricyclic antidepressant) in the mouse forced swimming test (FST) and chronic mild stress (CMS) induced depression-model rats. The FST is an antidepressant screening procedure performed initially to observe the immediate effects of EA and/or CLO on the immobility time. CLO (2.5, 5, 10, 20 and 60mg/kg intraperitoneally) were administered at 23, 6 and 1h respectively prior to each test. EA was given at the 'Bai-Hui' (Du 20) and unilateral 'An-Mian' (EX 17) acupoints 1h before each test. Immobility time was significantly reduced by EA and CLO at 2.5, 5, 10, 20 or 60mg/kg, respectively. EA combined with 2.5mg/kg CLO exhibited additive effects on the immobility time. In addition, rats were exposed chronically (1st-11th week) to a variety of mild unpredictable stressors. Depressed mood and anhedonia were recognized as a decrease in sucrose intake in the CMS rats. CLO at 2.5, 5mg/kg and EA at the same acupoints and parameters were administrated on the CMS rats once every other day for 6 weeks (5th-11th week). The intake of 1% sucrose solution was reduced by CMS, which was restored to normal level after 6 weeks treatment with 5mg/kg CLO or EA combined with 2.5mg/kg CLO. However, neither the sucrose intake nor the sucrose preference in the depressive rats was significantly changed by the treatment with EA or 2.5mg/kg CLO alone. These results demonstrated that EA combined with CLO at low doses has an additive or synergistic antidepressant action, and this combination may provide an effective strategy for depression management.

Neuronal specificity of needling acupoints at same meridian: a control functional magnetic resonance imaging study with electroacupuncture.

The purpose of this study was to investigate the neuronal specificity of needling acupoints at same meridian by functional Magnetic Resonance Imaging (fMRI). The selected acupoints GB34 (Yanglinquan) and GB39 (Xuanzhong) were at the same gallbladder meridian based on traditional Chinese medicine. In our study we devise three distinct EA (electroacupuncture) manipulations: real EA (deep needling at acupoints), sham EA (deep needling at no-meridian points) and shallow EA (subcutaneous needling at acupoints). Twelve healthy volunteers with right-handiness were enrolled and received three different EA manipulations in counter-balanced orders. DeQi scores were used to evaluate the degree of needling sensation. We found real EA can induce significant stronger needling sensation than sham EA and shallow EA. Multisubjects group mean analysis showed that pain-related cortex including primary and secondary somatosensory cortex (SI and S II), anterior cingulated cortex (ACC), insula were involved in three EA stimulation. Bilateral activation of prefrontal gyrus and occipital cortex were exclusively found in real EA. Deactivation over the rostral segment of ACC was also shown in real and shallow EA. Further paired two difference analysis indicated that real EA induced higher activation than sham EA over bilateral prefrontal gyrus, right-side occipital gyrus and deactivation over the rostral segment of ACC. In the comparing with real EA versus shallow EA, there was right-side activation over the SI, S II, motor cortex, ACC, insula, thalamus, hippocampus, occipital cortex, and cerebellum; also activation over bilateral prefrontal gyrus, caudate and pons. Although no significant activation was found over periaqueductal gray (PAG), further analysis showed the mean and maximal signal changes were different under three EA manipulations. We concluded that EA at analgesic acupoints of same meridian maybe involved the pain-related neuromatrix especially the hypothalamus-limbic system; deep EA at meridian points could elicit stronger needling sensation and modulate the pain-related neuromatrix more effectively than EA at nonmeridian points or shallow EA at meridian points.

Sucrose preference is restored by electro-acupuncture combined with chlorimipramine in the depression-model rats.

The present study was to investigate the effect of electroacupuncture (EA) combined with chlorimipramine on the sucrose preference of depressive rats induced by chronic mild stress (CMS). Rats were exposed chronically (1st - 4th week) to a variety of mild unpredictable stresses. The tricyclic antidepressant chlorimipramine and EA were administrated on these depressive rats for 6 weeks (5th - 11th week). EA was applied at points of "Bai-Hui" (Du 20) and "An-Mian" (EX 17) (right side), by EA apparatus (Model 6805-2, Shanghai). Dense (60Hz/5sec)-sparse (4Hz/2.5sec) frequency of the wave was selected and the current intensity (< or = 1mA) was adjusted to provoke slight twitches of the rat's ear. The preference for 1% sucrose solution and the immobility time in the forced swimming test were measured as the symptoms of anhedonia and depressed mood, which were central features of major depression. The preference for 1% sucrose solution was reduced by CMS, but could be restored to normal level after 6 weeks treatment with chlorimipramine at 5mg/kg or EA combined with chlorimipramine at 2.5mg/kg. In the forced swimming test, the immobility time of depressive rats was decreased in both groups. However, the preference for sucrose and the immobility time in the depressive rats were not significantly changed by the treatment with only EA or chlorimipramine at 2.5 mg/kg. The results suggested that EA could potentiate the antidepressant effect of chlorimipramine in low dose, and EA combined with antidepressant might be a better method in treating depression.

Electroacupuncture suppresses surgical trauma stress-induced lymphocyte apoptosis in rats.

Cumulative evidences suggested that electroacupuncture (EA) could modulate immune function, but the mechanism needs further study. In the present study, the effect of EA on surgical trauma stress-induced lymphocyte apoptosis was investigated by using DNA gel electrophoresis, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay, and Western blot for Fas protein expression. The results showed that rats with surgical trauma stress exhibited a significant reduction in splenic cellularity. Increase in apoptotic cell death and Fas (CD95/Apo-1) expression in splenic lymphocytes was also observed. EA could suppress the increase of apoptosis and Fas protein expression in splenic lymphocytes induced by the surgical trauma stress. These results implied that EA could decrease splenic lymphocytes apoptosis via inhibiting Fas protein expression; consequently prevent deleterious immunological changes in the post-operative state.

Melatonin enhances lymphocyte proliferation and decreases the release of pituitary pro-opiomelanocortin-derived peptides in surgically traumatized rats.

The present study was undertaken to evaluate the effects of melatonin (MT) on spleen lymphocyte proliferation and release of pituitary pro-opiomelanocortin (POMC)-derived peptides in surgically traumatized rats. MT prevented the depression of lymphocyte proliferation induced by trauma in vivo and in vitro, and prevented the decrease of beta-endorphin and adrenocorticotropin in the pituitary in vivo, but dose-dependently inhibited the release of POMC-derived peptides from the pituitary in vitro. The culture media of the pituitaries derived from the traumatized rats inhibited lymphocyte proliferation of normal rats. These results suggest that MT can improve the trauma-induced depression of lymphocyte proliferation and inhibit the release of POMC-derived peptides. The neuroimmunomodulatory role of MT and POMC-derived peptides deserves further study.

Inhibitory effects of intrathecally administered interleukin-1beta on carrageenan-induced hyperalgesia and spinal c-Fos expression in rats.

The present study investigated the effects of interleukin-1beta (IL-1beta) injected intrathecally (i.t.) on carrageenan-induced thermal hyperalgesia and spinal c-Fos expression. The paw withdrawal latency (PWL) by the thermal stimulus was taken as an index of the thermal hyperalgesia of rats. I.t. injection of 10 ng IL-1beta significantly increased the PWL of the carrageenan-injected paw. Expression of c-Fos induced by intraplantar (i.pl.) injection of carrageenan was examined in the spinal cord with immunohistochemical methods. Three hours after i.pl. injection of carrageenan, the number of c-Fos-like immunoreactive (c-Fos-LI) neurons was significantly increased in laminae I-II, III-IV and V-VI of the ipsilateral spinal cord at L4-5 with the higher density in laminae I-II and V-VI. I.t. pre-injected IL-1beta significantly decreased the number of carrageenan-induced c-Fos-LI neurons in laminae I-II in the ipsilateral spinal cord and also inhibited the hyperalgesia induced by i.pl. carrageenan. These results suggested that i.t. injection of IL-1beta suppressed the central nociceptive input into laminae I-II and produced an antinociceptive effect.

Effect of electroacupuncture on the expression of interlukin-1beta mRNA after transient focal cerebral ischemia.

It has been reported that interleukin-1beta (IL-1beta ) play a key role in the pathogenesis of cerebral ischemia. Acupuncture is an effective traditional medical therapy in China. The aim of present study was to evaluate the effect of electroacupuncture (EA) on IL-1beta mRNA expression after middle cerebral artery occlusion (MCAO) in rats. Using in situ hybridization technique, it was found that in the MCAO group the expression of IL-1beta mRNA was significantly increased at 2h, 6h, 12h after reperfusion in cerebral ischemic cortex compared with normal group. In EA+ MCAO group the expression of IL-1beta mRNA was significantly decreased at 2h, 6h and 12h in ischemic cortex compared with MCAO group. The results indicated that EA might decrease the IL-1beta protein expression by reducing the IL-beta mRNA expression in ischemic cortex.

Neuroimmunal regulation of electroacupuncture (EA) on the traumatic rats.

EA has a wide range of function, many of them is mediated by the release of the endogenous opioid peptides. Using surgical traumatic stress model, it was observed that EA could improve the depression of cell mediated immune response. Based on the above results, we focused our work on the elucidation of the mechanism of EA in the central nervous system. The results showed that trauma amplified the activity of peritoneal macrophage, but inhibited Orphanin FQ and its receptor NP4 transcripts in the central nervous system, in the mean time, IL-1beta transcripts in the central nervous system was also augmented. EA stimulation of"Zusanli" (St. 36) and "Lanwei" (Extra. 37) points could inhibit all the above responses, but it had no influence on the normal rat. The results suggested that EA could modulate immune response via the interaction between Orphanin FQ and IL-1beta.

Effect of melatonin and electroacupuncture (EA) on NK cell activity, interleukin-2 production and POMC-derived peptides in traumatic rats.

The present study was to evaluate the effect of melatonin (MT) and EA on the cytotoxic activity of natural killer (NK) cells, the dynamic changes of the induction of interleukin-2(IL-2) and the content of POMC-derived peptides, beta-endorphins (betaE) and ACTH in spleen lymphocytes and in plasma of traumatic rats. The results showed that intraperitioneal (i.p.) injection of MT was able to recover the lower levels of NK cell activity and the induction of IL-2 production; MT could also decrease the higher betaE and ACTH levels induced by trauma in spleen lymphocytes and plasma. EA needling of Zusanli (St.36) and Lanwei (Extra.37) points obviously improved the immunosuppression produced by trauma and antagonized the elevation of betaE and ACTH contents induced by trauma stress in spleen lymphocytes and plasma. MT + EA could further modulate the depressed immune function, and there was a significant difference compared with MT (i.p.) or EA alone. MT + EA group further decreased the betaE and ACTH contents in immune cells and plasma. Yet, the mechanisms of the attenuation of MT and EA on immunosuppression induced by trauma need further study.

Modulation of the gene expression in the protective effects of electroacupuncture against cerebral ischemia: a cDNA microarray study.

In the present study, a cDNA microarray technology was used to analyze the gene expression profile in ischemia and in electroacupuncture (EA)-treated ischemia. A focal cerebral ischemia/reperfusion model on Macaca mulatta was performed with a modified middle cerebral artery occlusion method. Among the nearly 8000 genes, approximately 8% of the total number of genes examined were affected after ischemia/reperfusion injury. Major altered genes were downregulated. In EA-treated monkeys, approximately 10% of the total number of genes examined were affected. Major altered genes were upregulated, including signal transduction-, cell-cycle-, metabolism-, stress response-, DNA repair-related genes. One of the representative upregulated genes encodes insulin-like growth factor-1 (IGF-1) was confirmed using in situ hybridization. Results showed that after ischemia/reperfusion injury, IGF-1 mRNA expression decreased in ipsilateral striatum, whereas increased in ipsilateral hippocampus. No expression changes were observed in cortex. EA treatment could obviously upregulate the IGF-1 mRNA expression in striatum, and further enhance its expression in hippocampus. Therefore, the data presented suggest a possible mediator underlying the mechanisms of anti-ischemic effect of acupuncture. In conclusion, the protective mechanisms of EA against stroke include several related pathways and gene expressions. Microarray analysis may provide a framework for understanding these complicated mechanisms and yield valuable, clinically relevant insights and potentially therapeutic targets of stroke.

[History and causes of acupuncture advancing towards the world].

Acupuncture therapy is one of the original innovations in traditional Chinese medicine. The study of acupuncture has gradually advanced towards the world since 1970s. In these years the most important events might be the influence of acupuncture anesthesia, the attention of the World Health Organization, the establishment and development of the World Federation of Acupuncture and Moxibustion Societies, and the consensus development conference on acupuncture held by the National Institutes of Health in America, and so on. The reasons why acupuncture is accepted by the world are both the convinced curative effects and the scientific foundations, due to the great efforts of medical stuff in China guided by the principle of integration of traditional Chinese and western medicine. This article poses several proposals about further extending the influence of acupuncture. In brief, acupuncture research is one of the few fields that authentically impact the western scientific technology. It is of very important responsibility in the new century to impulse the acupuncture study to further progress and expand the international influence of acupuncture.

Beta-adrenoceptor mediated surgery-induced production of pro-inflammatory cytokines in rat microglia cells.

Immunological changes initiated by major operative injury may result in inflammatory responses in both peripheral and central nervous system, which may lead to organ dysfunction. Recent studies indicate that beta-adrenergic receptors (beta-ARs) may mediate production of pro-inflammatory cytokines in the brain. In the present study propranolol (beta-AR antagonist), but not prazosin (alpha1-AR antagonist), antagonized surgical trauma induced pro-inflammatory cytokine production in microglia cells isolated from rats. beta-AR activation in the absence of pro-inflammatory stimuli increased IL-1beta, TNF-alpha and IL-6 mRNA and protein expressions in the primary microglia cell culture. Isoproterenol (beta-AR agonist) treatment induced a time- and concentration-dependent increase of IL-1beta in cells. Both ERK1/2 and P38 MAPK inhibitor, but not PKA and JNK1/2 inhibitor abrogated isoproterenol-induced IL-1beta and IL-6 production in microglia cells. In conclusion, the results suggest that beta-ARs possess pro-inflammatory properties by modulating the functions of microglia cell.

Electroacupuncture downregulates TLR2/4 and pro-inflammatory cytokine expression after surgical trauma stress without adrenal glands involvement.

Cumulative evidences suggest that electroacupuncture (EA) can modulate immune function, but the mechanism needs further study. In the present study, the contribution of EA on toll-like receptors 2 and 4 (TLR2/TLR4) and pro-inflammatory cytokine expression after surgical trauma stress were investigated. The mRNA level of both TLR2/4 and pro-inflammatory cytokine was measured by quantitative real-time PCR. ELISA and Western blot assay were chosen for TLR2/TLR4 protein expression and pro-inflammatory cytokine production, respectively. The results showed that surgical trauma stress increased TLR2 mRNA and TLR2/4 proteins in the spleen and augmented pro-inflammatory cytokines (e.g. IL-1beta) mRNA and protein expression in the spleen and plasma. These effects could be deteriorated by adrenalectomy (ADX). EA at "Zusanli" acupoint significantly inhibited surgical trauma-induced TLR2 mRNA and TLR2/4 protein expression in spleen and pro-inflammatory cytokine expression in the spleen and plasma. ADX, however, could not block the effect of EA. These results suggested that surgical trauma stress primes the innate immune system for enhanced TLR2 expression and pro-inflammatory cytokine production. EA inhibits TLR2/4 and pro-inflammatory cytokines to produce an anti-inflammatory effect in a surgical trauma stress model, without adrenal gland involvement.

Immunomodulatory activity of orphanin FQ/nociceptin on traumatic rats.

AIM: To explore the neuro-immune modulatory effect of orphanin FQ/nociceptin (OFQ) and opioid receptor like 1 (ORL1) receptor on the traumatic rats. METHODS: The quantitative method of immuno-cytochemistry and i n situ hybridization combined with cytokine bioassay were used to detect the expression of endogenous OFQ and ORL1 and the production of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) from peritoneal macrophage. RESULTS: Strong signals for both OFQ immuno-reactive cells and ORL1 mRNA were detected in cerebral cortex, hippocampus, and hypothalamus in normal condition, whereas they were significantly reduced after trauma (P<0.05). However, the production of IL-1 and TNF-alpha from peritoneal macrophage was increased, when expressed as percentage of enhancement, the increment attained to 233 % and 521 % (sample dilution 1:4), 195 % and 566 % (1:8), 233 % and 757 % ( 1:16), 214 % and 622 % (1:32), respectively, after trauma. After icv injection of OFQ at doses of 0.055 nmol, 0.55 nmol, and 2.75 nmol, the units of IL-1 and TNF-alpha were reversed (P<0.05); however, the action of OFQ (0.55 nmol) was blocked by ORL1 selective antagonist [phe1psi(CH2-NH)Gly2]nociceptin-(1-13)-NH2. CONCLUSION: OFQ and ORL1, the new opioid peptide system, are involved in the immune response elicited by traumatic stress.