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Endogenous orphanin FQ: evidence for a role in the modulation of electroacupuncture analgesia and the development of tolerance to analgesia produced by morphine and electroacupuncture.

1. Our previous work has demonstrated that exogenously administered orphanin FQ (OFQ) antagonizes morphine analgesia and electroacupuncture analgesia (EAA) in the brain and potentiates morphine analgesia and EAA in the spinal cord of the rat. In the present study we evaluated the role of endogenously released OFQ in the development of tolerance to morphine and electroacupuncture (EA) and the analgesia produced by electroacupuncture, by use of the IgG fraction of an anti-OFQ antibody (OFQ-Ab) microinjected into the rat central nervous system (CNS). 2. EAA was produced by stimulating rats at a frequency of 100 Hz. Rats were classified as either high responders (HR) or low responders (LR) based on the analgesic effects of EA. LRs could be converted into HRs by the intracerebroventricular (i.c.v.) microinjection of OFQ-Ab at both 1:1 and 1:10 dilutions but not 1:100. HRs could be changed into LRs by the intrathecal (i.t.) injection of OFQ-Ab at both 1:1 and 1:10 dilutions, but not 1:100. 3. Acute morphine tolerance was induced in rats by repeated subcutaneous (s.c.) injections of morphine (5 mg kg, every 2 h) for 16 h. When injected i.c.v. the OFQ-Ab (1:1 dilution) had no effect on the development of acute morphine tolerance. 4. Chronic morphine tolerance was produced in rats by repeated injection of morphine (5-60 mg kg, s.c., 3 x a day) for 6 days. I.c.v. injection of OFQ-Ab (1:1 dilution) reversed this type of morphine tolerance in rats by 50% (P < 0.01). 5. Acute tolerance to the analgesia produced by EA developed after 6 h of continuous (100 Hz, 3 mA) stimulation. This tolerance was almost completely reversed by the i.c.v. injection of OFQ-Ab (1:1 dilution) (P < 0.05). 6. Chronic tolerance to the analgesic effect of EA was produced by repeatedly administering increasing current (1, 2 and 3 mA, each lasting for 10 min, for a total of 30 min) at a frequency of 100 Hz once a day for 6 days. I.c.v. injection of OFQ-Ab (1:1 dilution) reversed this kind of tolerance by 50% (P < 0.01). 7. Together these results suggest that 100 Hz EA may enhance the release of endogenous OFQ in the CNS of the rat, which in turn may act to antagonize EA-produced analgesia in the brain but potentiate EA produced analgesia in the spinal cord. Therefore, OFQ appears to play an important role in the development of tolerance to the analgesic effects produced by EA. 8. The mechanisms underlying the development of acute morphine tolerance and chronic morphine tolerance appear to be different. Central OFQ may play an important role in the development of tolerance after chronic morphine administration.

Adaptation, Physiological↗

Increased release of immunoreactive CCK-8 by electroacupuncture and enhancement of electroacupuncture analgesia by CCK-B antagonist in rat spinal cord.

Cholecystokinin octapeptide (CCK-8) in CNS has been shown to function as a neuropeptide with potent anti-opioid activity. It hinders opioid analgesia and facilitates opioid tolerance. The present study showed that electroacupuncture (EA) stimulation produced a marked increase of the CCK-8 immunoreactivity (ir) in the perfusate of the rat spinal cord. The increase of CCK-8-ir was most marked in response to EA of 100 Hz and 15 Hz, and less marked in response to EA of 2 Hz. Since CCK-8 has been shown to possess potent anti-opioid activity at the spinal level, blockade of the spinal CCK effect would be expected to potentiate EA-induced analgesia which is known to be opioid-mediated. Intrathecal (i.t.) administration of CCK-B antagonist L-365,260 per se did not affect tail flick latency (TFL) to any significant extent, yet it potentiated EA induced analgesia in a dose- and frequency-dependent manner. The potentiation was most marked at a dose range of 2.5-5.0 ng (i.t.) and at a frequency rank order of 100 Hz > 15 Hz > 2 Hz. The results suggest that an increased release of CCK-8 following EA may limit the effect of opioid peptides, and that the CCK-B receptor mediates the anti-opioid effect of CCK-8 in rat spinal cord.

Acupuncture Analgesia↗

Electroacupuncture up-regulates natural killer cell activity Identification of genes altering their expressions in electroacupuncture induced up-regulation of natural killer cell activity.

As an important cellular component of the innate immune system, NK cells constitute a first line of defense against various infections and malignancies. Previous studies have reported electroacupuncture (EA) modulation of natural killer cell (NK cell) activities. Our study confirmed that EA treatment increases NK cell activity using (51)Cr release assay. Furthermore, in order to better understand the activation mechanism of NK cell by EA, we employed a cDNA microarray technique to elucidate how EA alters gene expressions in the spleen of rats. We screened EA responsive genes using a high-throughput screening and identified 154 genes. Among those genes we selected 4 genes that are known to play a crucial role in NK cell activation and examined their mRNA expressions after EA treatment using RT-PCR. Our data shows that EA treatment increased CD94, PTK and VCAM-1 expressions while decreased PTP and SHP-1. These results imply that EA treatment increase PTK expression, which increases NK cell activity, through induction of CD94 while decreases SHP-1, which inhibits NK cell activity, simultaneously so that it activates NK cell with high efficacy. It seems that increased VCAM-1 expression is due to INF-gamma produced by activated NK cell. Increased production of VCAM-1 is expected to play an important role in binding of NK cell to the target cell. The result of our study may provide key insights in understanding the mechanisms of activation of NK cell induced by EA.

Acupuncture Points↗

Nocistatin potentiates electroacupuncture antinociceptive effects and reverses chronic tolerance to electroacupuncture in mice.

Nocistatin (NST) and nociception/orphanin FQ (OFQ) are peptides derived from the same precursor that play opposing roles in pain modulation. OFQ antagonizes morphine analgesia and electroacupuncture (EA)-induced antinociceptive effect. The present study investigates whether NST potentiates EA-induced antinociceptive effect and reverses chronic tolerance to EA in mice. Injection of NST (0.5, 5.0 and 50.0 ng) intracerebroventricularly had no effect on basal thermal latency, but produced a dose-dependent potentiation of EA-induced antinociceptive effect in mice with the maximum response at 5.0 ng. NST (5.0 ng) partly reversed chronic tolerance to EA. These results suggest that NST in the brain might play roles in EA-induced antinociceptive effect and the development of chronic tolerance to EA in mice.

Analgesics↗

[A comparison between the pain relief effect of electroacupuncture, regional never block and electroacupuncture plus regional never block in frozen shoulder].

BACKGROUND: Frozen shoulder is a kind of spontaneous, progressive peri-arthritis over the shoulder joint. The etiology is not yet clear. Traditional treatments for frozen shoulder included conservative medical therapy, physical therapy, nerve block and acupuncture and so on. The purpose of our study is to determine the pain relief effect of electroacupuncture (EAP), regional nerve block (RNB) and the combination of EAP + RNB for frozen shoulder. MATERIALS & METHODS: One hundred and fifty patients with fresh frozen shoulder were, randomly divided into 3 groups. Group I patients (n = 50) had RNB with stellate ganglion block and suprascapular nerve block by 1% xylocaine 10 ml. Group II patients (n = 50) had EAP with local acupoint--Chien-Yu, Chien-Ching, Chien-Nei-Ling, Ah-Shih Hsueh treatment and Group III (n = 50) patients had RNB+EAP performed with acupuncture first, then followed by the regional nerve block. Six vectors of movements were checked in all methods. Four graded Bromage score was used for pain assessment, Grade 1 means complete painless; Grade 2 means slight pain (i.e. pain on motion); Grade 3 means moderate pain (i.e. pain on silence); Grade 4 means severe pain (i.e. need analgesics). The range of shoulder joint was also recorded. Patients were requested for second treatment if pain recurred. The onset (time from injection to maximal pain relief), duration (time from injection to grade 3) Bromage score and side effects were recorded. RESULTS: The results showed that the combined EAP and RNB method had significant high pain control quality, longer duration, and better range of movement of the shoulder joint than that of EAP or RNB performed alone.

Analgesia↗

Electroacupuncture suppression of a nociceptive reflex is potentiated by two repeated electroacupuncture treatments: the first opioid effect potentiates a second non-opioid effect.

Electroacupuncture (EA) produces suppression of the nociceptive tail flick reflex in rats. Two 10 min treatments given 90 min apart, cause a potentiation of the EA effect produced by the second treatment. This potentiation can be prevented by giving naltrexone before the first and second treatments (either intravenously or intrathecally administered). However, this potentiation cannot be reversed by naltrexone give only once, before the second EA treatment. These results suggest that the first EA effect which is opioid in nature, sets up a cascade effect, potentiating a second non-opioid EA effect.

Acupuncture Therapy↗

Cholecystokinin octapeptide (CCK-8): antagonism to electroacupuncture analgesia and a possible role in electroacupuncture tolerance.

The analgesic effect produced by electroacupuncture (EA) stimulation in the rat was dose-dependently antagonized by cholecystokinin octapeptide (CCK-8) administered intracerebroventricularly (i.c.v.) or intrathecally (i.th) at a dose range of 0.25-4 ng. This effect had an immediate onset and lasted for at least 4 h. CCK-8 per se, however, did not affect baseline tail flick latency. Rats subjected to prolonged EA stimulation developed EA tolerance as well as cross-tolerance to morphine. These tolerances could be postponed or reversed by i.c.v. or i.th injection of antiserum against CCK-8. While CCK-8 antagonized opioid analgesia, it did not affect analgesia induced by 5-hydroxytryptamine (5-HT) or norepinephrine (NE). Moreover, CCK-8 antiserum did not alter the basic level of nociception, nor did it potentiate EA analgesia in naive rats. It is concluded that prolonged EA stimulation results in a profound release of opioids which may trigger the release of CCK-8 in the central nervous system to counteract the opioid component of EA analgesia. This mechanism may account, at least in part, for the development of EA tolerance.

Acupuncture Therapy↗

The difference between electroacupuncture only and electroacupuncture with manipulation on analgesia in rats.

Plain acupuncture uses manipulation (rotation or varying the depth of insertion of the needle) to increase its effect. However, in commonly used electroacupunture (EA), variable manipulations have not been used. This study was performed to investigate the possibility of an increase in analgesic effect by adding manipulation to EA. The pain index used was the Tail-Flick latency (TFL) of the rat, which was lightly anesthetized with thiopental sodium (intraperitoneally). Four types of manipulation were used. Rotation and varying the depth of the needle (RN and VN) was employed using two different types of manipulation during each 20 min stimulation of EA. Each manipulation persisted for 1 min out of every 5 min (long - duration and long - interval: LDLI) or 12 s every 1 min (short - duration and short interval: SDSI). EA produced an increase in TFL; peak value was 49.7+/-12.2% of the pre - EA and occurred immediately after cessation of 20 min of EA stimulation. Performing RN or VN combined with EA also increased TFL more than just EA and a greater peak increase in TFL was observed with a SDSI - RN and SDSI - VN as compared to a LDLI - RN and LDLI - VN (77.5+/-13.8, 79.2+/-19.8 and 67.3+/-14.0%, 65.6+/-23.7% of the pre - EA, respectively). These results indicate that manipulation combined with EA produces a more potent antinociception than when only EA is applied.

Acupuncture Analgesia↗

[Clinical observation on electroacupuncture combined with medicine for treatment of acute gouty arthritis].

OBJECTIVE: To observe clinical therapeutic effect of electroacupuncture combined with medicine on acute gouty arthritis and effect on blood uric acid content. METHODS: One hundred cases of acute gouty arthritis were randomly divided into 3 groups, an electroacupuncture group treated by acupuncture at Zusanli (ST 36), Fenglong (ST 40) and Ashi points, a Western medicine group by oral administration of indometacin and benzbromarone, and a combined electroacupuncture and medicine group by electroacupuncture combined with the Western medicine. The changes of their blood uric acid levels before and after treatment were detected and their therapeutic effects were compared. RESULTS: There was a significant difference among the three groups in clinical therapeutic effect (P < 0.05), and the effective rate was respectively 97.5%, 90.0% and 86.7% in the combined electroacupuncture and medicine group, the electroacupuncture group and the Western medicine group, the combined electroacupuncture and medicine group being significantly higher than the electroacupuncture group and the Western medicine group; after treatment, the blood uric acid level in all the 3 groups had significant change (P < 0.05), the blood uric acid in the combined electroacupuncture and medicine group had more obvious decrease as compared with those in the electroacupuncture group or the Western medicine group. CONCLUSION: Electroacupuncture combined with the Western medicine has a definite therapeutic effect on acute gouty arthritis.

Acupuncture Therapy↗

[Effect of electroacupuncture on metabolism of lipids in rats of obesity induced by sodium glutamate].

OBJECTIVE: To explore mechanisms of acupuncture for reducing of body weight. METHODS: The rat obesity model was developed by subcutaneous injection of sodium glutamate solution, and the experimental rats were divided into a blank control group, a model group, an electroacupuncture group and a sibutramine group; blood TC, TG, HDL-C, LDL-C contents and lipoprotein lipase (LPL) activity, and serum leptin and insulin levels were determined. RESULTS: The body weight and Lee's index, and TG, TC and LDL-C contents in the electroacupuncture group decreased significantly as compared with those in the model group (all P<0.01), the actions in the electroacupuncture group being superior to those in the sibutramine group; the HDL-C contents in both the electroacupuncture and the sibutramine groups were higher than that of the model group (P<0.01) with no significant difference between the electroacupuncture group and the sibutramine group; LPL activity in the electroacupuncture group increased significantly as compared with that in the model group (P<0.01); serum leptin and insulin levels in both the electroacupuncture group and the sibutramine group were lower than that in the model group (P<0.05, P<0.01), with no significant difference between the electroacupuncture group and the sibutramine group in the effect on insulin content, and the decrease of leptin level in the electroacupuncture group was more as compared with the sibutramine group (P<0.01). CONCLUSION: Electroacupuncture can improve the status of high blood lipids, increase lipoprotein lipase activity, and regulate serum leptin and insulin levels in the obestic rat.

Acupuncture Points↗

Analgesia and c-Fos expression in the periaqueductal gray induced by electroacupuncture at the Zusanli point in rats.

The need to use anaesthetised or restrained animals in acupuncture research in laboratory animals may represent a confounding variable, since both anaesthesia and stress alter the pain threshold and the activity of pain-related brain areas. In the current study we assessed the participation of the periaqueductal gray (PAG) in electroacupuncture's (EA) analgesic effects applied to the Zusanli point (36S) under carefully controlled stress conditions. Repeated immobilisation protocols (6 days, 1 h/day and 13 days, 2 h/day) were used to diminish the influence of acute immobilisation stress on c-Fos expression and analgesia (tail-flick test) induced by electroacupuncture on the 36S point (EA36S). Animals submitted to immobilisation alone (IMMO) or to electroacupuncture (100 Hz, 2-4 V, faradic wave) on a non-point region (EANP) were compared with animals submitted to electroacupuncture on the 36S point. In animals not previously submitted to repeated immobilisation, electroacupuncture on the 36S point induced analgesia and c-Fos expression in the PAG was not different from that induced by electroacupuncture at a non-acupuncture point. In animals submitted to repeated immobilisation (repeated immobilisation for 6 days or repeated immobilisation for 13 days), however, electroacupuncture on point 36S led to higher levels of analgesia and c-Fos expression, specifically in the ventrolateral PAG (vlPAG), as compared with animal groups subjected only to immobilisation or to electroacupuncture on a non-point. Our findings endorse previous results, and point to a specific part of the PAG involved in the effects of electroacupuncture at the Zusanli point.

Acupuncture Analgesia↗

[Analgesic effects of electroacupuncture of different pulse waveforms in the rat of adjuvant arthritis].

OBJECTIVE: To explore the best pulse waveform of electroacupuncture for treatment of adjuvant arthritis (AA). METHODS: Forty Wistar rats were randomly divided into 5 groups. The electroacupuncture groups of AA were treated with different pulse waveforms of electroacupuncture, respectively. Pain threshold, swelling degrees of joint, and contents of 5-HT, beta-EP and LEK in local tissues of inflammation were observed. RESULTS: After the treatment, pain thresholds of the electroacupuncture groups with different pulse waveforms were higher than that of the model group (P < 0.05, P < 0.01); the local swelling degrees of inflammation in the electroacupuncture group were higher than that in the normal group (P < 0. 01), but was lower than that in the model group (P < 0.05, P < 0. 01); the contents of 5-HT in local tissues of inflammation in the electroacupuncture groups were lower than that in the model group (P < 0.05, P < 0.01); in comparison with the model group, the electroacupuncture of sound-electric wave increased the contents of beta-EP and LEK (P < 0.01, P < 0.05) and the electroacupuncture of intermittent wave increased the content of beta-EP in local tissues of inflammation (P < 0.05). CONCLUSION: Sound-electric wave is the best pulse waveform of electroacupuncture for treatment of adjuvant arthritis.

Acupuncture Analgesia↗

[Clinical study on electroacupuncture combined with manual acupuncture for treatment of simple obesity].

OBJECTIVE: To compare therapeutic effects of electroacupuncture combined with manual acupuncture and simple electroacupuncture on simple obesity. METHODS: One hundred and twenty cases were randomly divided into an electroacupuncture plus manual acupuncture group (observation group, n = 63) and a simple electroacupuncture group (electroacupuncture group, n = 57). Their clinical therapeutic effects and the time starting the therapeutic effect were compared. RESULTS: In the observation group 57 cases (90.5%) were cured and the total effective rate was 100.0%, and in the electroacupuncture group 42 cases (73.7%) were cured and the total effective rate was 96.5%, with significant difference in the cured rate between the two groups (P < 0.05); and the time starting the therapeutic effect in the observation group was superior to the electroacupuncture group. CONCLUSION: Electroacupuncture combined with manual acupuncture is superior to the simple electroacupuncture in the cured rate and the time starting the effect in treatment of simple obesity.

Acupuncture Therapy↗

[Effects of electroacupuncture at "Zusanli" (ST 36) on ERK1/2 phosphorylation in the dorsal horn of spinal cord of the rat].

OBJECTIVE: To observe the effect of electroacupuncture at Zusanli (ST 36) on phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) in the dorsal horn of spinal cord induced by plantar inflammation in the rat. METHODS: All the rats were randomly divided into 5 groups: normal control group, simple electroacupuncture group, formalin group, formalin plus ipsilateral electroacupuncture group and formalin plus contralateral electroacupuncture group. The acute inflammation animal model was made by injection of 100 microL of 4% formalin into the right posterior foot pad. Electroacupuncture was given at "Zusanli" (ST 36) for 30 min, with sparse-dense waves, frequency 2-15 Hz, and intensity 2-3 mA. One and a half hours latter, the rats were killed under anesthesia, and pERK1/2 expression in the lumbar dorsal horn were investigated with immunohistochemical method. RESULTS: The positive cells were rarely seen (6.45 +/- 1.05) in the superficial spinal cord in the control group; a few cells (14.07 +/- 3.19) in ipsilateral superficial spinal cord were found in the electroacupuncture group. The number of pERK1/2-positive neurons (26.57 +/- 4.93) in lamina I - II0 of the ipsilateral dorsal horn in the formalin group increased significantly. After electroacupuncture at ipsilateral Zusanli (ST 36), the number of positive cells (20.79 +/- 5.21) had a tendency to decrease, but with no statistically significant difference. However, after electroacupuncture at contralateral Zusanli (ST 36), the number of positive cells (14.75 +/- 3.03) significantly decreased as compared with the non-acupuncture group (P < 0.05). CONCLUSION: The inhibition of ERK1/2 phosphorylation in the spinal cord dorsal horn by electroacupuncture is possibly involved in acupuncture analgesic effect.

Acupuncture Analgesia↗

Transcriptomic Changes Associated with Electroacupuncture in a DMCAO Model of Delayed Cognitive Impairment.

INTRODUCTION: Delayed Cognitive Impairment (DCIS) occurs in approximately 31% to 77% of individuals following stroke. Clinical findings have indicated that electroacupuncture may alleviate post-stroke DCIS. However, insights derived from animal models remain limited. The present study utilized a Distal Middle Cerebral Artery Occlusion (DMCAO) mouse model to investigate the potential mechanisms of electroacupuncture through hippocampal transcriptomic analysis. MATERIALS AND METHODS: Adult male BALB/c mice were subjected to DMCAO and received electroacupuncture treatment. High-throughput RNA sequencing of hippocampal tissue was performed to identify Differentially Expressed Genes (DEGs). Enrichment analyses, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, hierarchical clustering, and Protein-Protein Interaction (PPI) network analysis, were performed to elucidate potential biological mechanisms. RESULTS: The DMCAO model exhibited features consistent with DCIS. Electroacupuncture treatment was associated with improved cognitive performance and enhanced hippocampal neuroplasticity. A total of 116 DEGs were identified in the DMCAO group compared with the sham group, while 69 DEGs were identified in the DMCAO + electroacupuncture group compared with the untreated DMCAO group. DISCUSSION: GO enrichment analysis indicated that electroacupuncture modulated biological processes related to nerve fibers, axonal development, neuronal regulation, cellular processes, and cardiovascular protection. KEGG pathway analysis indicated involvement in pathways associated with neuronal recovery and axonal function. The PPI network comprised 28 nodes and 33 interactions, with hub genes such as Gna13, Hipk2, and Stambp playing key roles. Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) results were consistent with RNA sequencing findings. CONCLUSION: Electroacupuncture improved DCIS in the DMCAO mouse model. Transcriptomic analysis of the hippocampus provided preliminary evidence of the potential mechanisms underlying the therapeutic effects of electroacupuncture treatment following ischemic stroke.

Animals↗