Low-frequency Electroacupuncture Research Articles

An attempt to use low-frequency electroacupuncture to treat a patulous Eustachian tube

An attempt to use low-frequency electroacupuncture to treat a patulous Eustachian tube

Ovarian Innervation Coupling With Vascularity: The Role of Electro-Acupuncture in Follicular Maturation in a Rat Model of Polycystic Ovary Syndrome

Low-frequency electro-acupuncture (EA) has been shown to restore ovulation in patients with polycystic ovary syndrome (PCOS), and previous animal experiments showed that EA improves ovarian blood flow and angiogenesis. We performed EA for 4 weeks in dihydrotestosterone (DHT)-induced PCOS-like rats and investigated the three-dimensional (3D) ovarian innervation to determine the role of innervation in folliculogenesis and vascularity. Ovarian tissues were made transparent following the CUBIC 3D tissue-clearing protocol and were immunostained using antibodies against platelet endothelial cell adhesion molecule-1 and tyrosine hydroxylase to visualize the ovarian vasculature and innervation, respectively. This was followed by 3D imaging using lightsheet microscopy and analysis using the Imaris software. In control rats, ovarian innervation increased with age, and the neuronal branching started from the ovarian hilum and reached the individual follicles at different follicle stages. At the individual follicle level, each follicle was mainly innervated by one neuronal fiber. Compared with control rats, ovaries from DHT-treated PCOS-like rats had more antral follicles and fewer preovulatory follicles and corpora lutea. Furthermore, PCOS ovaries showed decreased innervation of blood vessels near the hilum and the surrounding individual antral follicles. EA in PCOS-like rats led to increased numbers of preovulatory follicles and corpora lutea together with increased innervation of blood vessels near the hilum. To determine the role of ovarian innervation, we further performed unilateral sectioning of the superior ovarian nerve (SON) in PCOS + EA rats and found that the left sectioned ovary had fewer preovulatory follicles and corpora lutea compared with those in the right non-sectioned ovary. In conclusion, ovarian innervation likely played an important role in folliculogenesis, and EA might restore PCOS pathophysiology by regulating ovarian innervation, at least partially mediated through the SON.

Effects of electroacupuncture of different frequencies on electromyography, NOS and ICC of colon in rats with slow transit constipation

To observe the effects of electroacupuncture (EA) of different frequencies on transmission function, electromyography, nitric oxide synthase (NOS) content and interstitial cells of Cajal (ICC) expression of colon in rat models with slow transit constipation (STC). Of the 50 healthy male Wistar rats, 10 were randomly selected as a normal group and fed with ordinary diet, and the remaining 40 rats were fed with the diet added with the compound diphenoxylate at a dose of 8 mg/(kg·bw) each day for continuous 120 d. The 40 successfully established STC rat models were randomly divided into a model group, a low-frequency EA group (2 Hz), a high-frequency EA group (100 Hz), and a variable-frequency EA group (2 Hz/100 Hz), with 10 rats in each group. Rats in the normal and the model groups were not given any treatment; the low-frequency EA and the high-frequency EA groups were given EA at Tianshu (ST 25), Zusanli (ST 36) and Zhigou (TE 6) with continuous wave at the designated frequency, and the variable-frequency EA group received sparse-dense wave (2 Hz/100 Hz) EA at the same acupoints, once a day for a total of 15 d. After treatment, the colonic transmission function, electromyography, NOS content and ICC expression (calculated by the difference in the area of the C-kit positive cells) of the rats in each group were measured. For the colonic transmission function, compared with the normal group, the first black stool excretion durations of rats in the other groups were significantly prolonged (all P<0.05); compared with the model group, the first black stool excretion durations of rats in the three EA groups were significantly shortened (all P<0.05), which was significantly shorter in the variable-frequency EA group than in the low-frequency EA and high-frequency EA groups (both P<0.05). For the colonic electromyography, compared with the normal group, the amplitude was significantly increased and the frequency was accelerated in rats of the other groups (all P<0.05); compared with the model group, the amplitude was significantly reduced and the frequency was slowed down in the three EA groups (both P<0.05); compared with the low-frequency EA and the high-frequency EA groups, the amplitude was reduced and the frequency was significantly reduced in rats of the variable-frequency EA group (both P<0.05). Compared with the normal group, the colonic NOS contents were significantly increased in the other groups (all P<0.05); compared with the model group, the NOS contents were significantly reduced in the three EA groups (all P<0.05); compared with the low-frequency EA and the high-frequency EA groups, the content was significantly reduced in the variable-frequency EA group (all P<0.05). For the area of rat colonic C-kit-positive cells, compared with the normal group, the areas were significantly reduced in rats of other groups (all P<0.05); compared with the model group, the areas were increased significantly in the three EA groups (all P<0.05); compared with the low-frequency EA group, the area was increased significantly in the variable-frequency EA group (P<0.05). EA, especially EA at the 2 Hz/100 Hz variable-frequency, has a positive treatment effect on the STC model rats. It may improve rats’ colonic function by regulating the electromyography, NOS content and ICC expression of colon.

Low frequency electroacupuncture alleviates neuropathic pain by activation of spinal microglial IL-10/β-endorphin pathway

Electroacupuncture produces analgesia in chronic pain patients and animal models of pain hypersensitivity. The current study aims to illustrate the mechanisms underlying electroacupuncture-attenuated neuropathic pain. Neuropathic rats, induced by tight ligation of L5/L6 spinal nerves, markedly reduced mechanical thresholds in the ipsilateral hindpaws relative to the contralateral hindpaws. Low frequency (2 Hz) electroacupuncture stimulation for a period of 20 min alleviated neuropathic pain in the ipsilateral hindpaws of neuropathic rats in a time-dependent manner. The same electroacupuncture treatment also stimulated spinal gene and protein expression of IL-10 and β-endorphin but not dynorphin A, measured by real-time quantitative PCR and ELISA kits. Intrathecal injection of the specific IL-10 antibody in neuropathic rats completely blocked electroacupuncture-increased spinal expression of β-endorphin, but the β-endorphin antibody failed to alter electroacupuncture-stimulated spinal IL-10 expression. Using a double fluorescence immunostaining technique, we observed that electroacupuncture stimulated spinal IL-10 and β-endorphin expression in microglia but not in neurons or astrocytes in the spinal dorsal horn of neuropathic rats. Pretreatment with intrathecal injection of the microglial inhibitor minocycline, specific IL-10 antibody and β-endorphin antiserum (but not the dynorphin A antibody), or selective μ-opioid receptor antagonist CTAP (but not κ- or δ-opioid receptor antagonist) completely blocked electroacupuncture-induced attenuation of neuropathic pain. These results suggest that low frequency electroacupuncture alleviates neuropathic pain through stimulation of the spinal microglial expression of IL-10 and subsequent expression of β-endorphin.

Low-frequency electroacupuncture at acupoints guanyuan (CV4) and zhongji (CV3) lengthen ejaculatory latency and improves sexual behavior in male rats.

Acupuncture is a key component of Traditional Chinese Medicine in which needles are inserted into specific areas in the body to stimulate certain physiological reactions of the body. Clinical research shows that acupuncture is beneficial in treating a variety of illnesses, such as erectile dysfunction. Therefore, we investigated the effect of electroacupuncture (EA) stimulation of both low- and high frequencies at Guanyuan (CV4) and Zhongji (CV3) acupoints on male copulatory behavior in sexually experienced male rats. The animals were randomly divided into four groups: control, sham EA, EA, and only acupuncture. The administered low and high EA were 2 Hz and 80 Hz, respectively, for which the respective intensities were 1.5 mA (30 min/day for 5 days) and 1.5 mA (30 min once). The only acupuncture-administered group received acupunctures at CV4 and CV3 without electrical stimulation. Following the acupuncture treatments, copulatory behavior test was conducted. Eventually, animals were sacrificed and blood samples were collected for testing the serum hormonal profile including luteinizing hormone (LH), testosterone (T), and serotonin (5-HT). Results showed that low-frequency EA-treated rats exhibited increasing intromissions and ejaculation latency compared to control, sham EA, and acupuncture groups, while high-frequency EA-treated rats displayed lower intromissions and ejaculation frequency when compared with those in controls. Furthermore, serum levels of 5-HT and LH in low-frequency EA-treated rats were higher than all the other groups. These results indicate that compared to high-frequency EA, the low-frequency EA might be efficacious in the treatment of premature ejaculation, thereby improving sexual behavior.

Lower frequency electroacupuncture is better in promoting recovery of limb locomotion in rats with sciatic nerve injury by reducing local inflammatory reaction

To observe the effect of different frequencies (2 Hz, 100 Hz) of electroacupuncture (EA) on limb locomotion and the expression of inflammatory factors IL-1β, IL-6, TNF-α in sciatic nerve, and nuclear factor kappa B (NF-κB) in lumber(L)4-L5of spinal cord in rats with sciatic nerve injury (SNI), so as to reveal its mechanisms underlying improvement of SNI. A total of 48 SD rats (half male and half female) were equally divided into blank control, model, low frequency (2 Hz) EA and high frequency (100 Hz) EA groups. The SNI model was established by clamping the spinal nerve. EA intervention (2 Hz, 100 Hz, 1 mA), starting on the 8th day after modeling, was applied to "Huantiao" (GB30) on the injured side for 15 min, once daily for 14 consecutive days. The sciatic function index (SFI) was calculated to assess the injured hindlimb recovery with reference to BAIN's and colleagues' methods. Histopathological changes of the sciatic nerve were displayed by H.E. staining. The expressions of IL-1β, IL-6 and TNF-α in the sciatic nerve tissue were detected by immunohistochemistry, and the expression of NF-κB in the spinal cord was detected by using Western blot. After modeling, the SFI level on day 8 was significantly decreased in the model group (P<0.01), and no significant differences were found among the model, low frequency EA and high frequency EA groups before the EA intervention (P>0.05). Following the treatment (at the 22nd day), the SFI values of both low frequency EA and high frequency EA groups were significantly increased (P<0.01), suggesting an improvement of the limb motor function, and the SFI of the low frequency EA group was notably higher than that of the high frequency EA group (P<0.01). In comparison with the blank control group, the expression levels of IL-1β, IL-6, TNF-α in the sciatic nerve and NF-κB protein in the spinal cord were significantly up-regulated (P<0.05). Following EA intervention, the increased expression levels of IL-1β, IL-6, TNF-α and NF-κB proteins were significantly down-regulated in both low frequency EA and high frequency EA groups (P<0.05), and the therapeutic effect of low frequency EA was markedly superior to that of high frequency EA in down-regulating the expression levels of IL-1β, IL-6, TNF-α and NF-κB protein (P<0.05). H.E. staining showed increase of Schwann cells in number, cellular swelling, and disintegration of the axons and myelin sheath, and appearance of vacuolar degeneration in the model group, which was relatively milder in both low frequency EA and high frequency EA groups, particularly in the low frequency EA group. EA of GB30 at 2 Hz and 100 Hz can promote the recovery of hindlimb motor function in SNI rats, which is probably related to its function in inhibiting the inflammatory response, and facilitating the repair of the damaged sciatic nerve. 2 Hz EA is better than 100 Hz EA in the therapeutic effect.

Acupuncture for polycystic ovarian syndrome

Background Polycystic ovarian syndrome (PCOS) is characterised by the clinical signs of oligo-amenorrhoea, infertility and hirsutism. Conventional treatment of PCOS includes a range of oral pharmacological agents, lifestyle changes and surgical modalities. Beta-endorphin presents in the follicular fluid of both normal and polycystic ovaries. It was demonstrated that the beta-endorphin levels in ovarian follicular fluid of otherwise healthy women who were undergoing ovulation were much higher than the levels measured in plasma. Given that acupuncture has an impact on beta-endorphin production, which may affect gonadotropin-releasing hormone (GnRH) secretion, it is postulated that acupuncture may have a role in ovulation induction and fertility. Objectives To assess the effectiveness and safety of acupuncture treatment of oligo/anovulatory women with polycystic ovarian syndrome (PCOS). Search methods We identified relevant studies from databases including the Cochrane Central Register of Controlled Trials (CENTRAL), Ovid MEDLINE, EMBASE, PsycINFO, CNKI and trial registries. The data are current to 19 October 2015. Selection criteria We included randomised controlled trials (RCTs) that studied the efficacy of acupuncture treatment for oligo/anovulatory women with PCOS. We excluded quasi- or pseudo-RCTs. Primary outcomes were live birth and ovulation (primary outcomes), and secondary outcomes were clinical pregnancy, restoration of menstruation, multiple pregnancy, miscarriage and adverse events. We assessed the quality of the evidence using GRADE methods. Data collection and analysis Two review authors independently selected the studies, extracted data and assessed risk of bias. We calculated Mantel-Haenszel odds ratios (ORs) and mean difference (MD) and 95% confidence intervals (CIs). Main results We included five RCTs with 413 women. They compared true acupuncture versus sham acupuncture (two RCTs), true acupuncture versus relaxation (one RCT), true acupuncture versus clomiphene (one RCT) and electroacupuncture versus physical exercise (one RCT). Four of the studies were at high risk of bias in at least one domain. No study reported live birth rate. Two studies reported clinical pregnancy and found no evidence of a difference between true acupuncture and sham acupuncture (OR 2.72, 95% CI 0.69 to 10.77, two RCTs, 191 women, very low quality evidence). Three studies reported ovulation. One RCT reported number of women who had three ovulations during three months of treatment but not ovulation rate. One RCT found no evidence of a difference in mean ovulation rate between true and sham acupuncture (MD -0.03, 95% CI -0.14 to 0.08, one RCT, 84 women, very low quality evidence). However, one other RCT reported very low quality evidence to suggest that true acupuncture might be associated with higher ovulation frequency than relaxation (MD 0.35, 95% CI 0.14 to 0.56, one RCT, 28 women). Two studies reported menstrual frequency. One RCT reported true acupuncture reduced days between menstruation more than sham acupuncture (MD 220.35, 95% CI 252.85 to 187.85, 146 women). One RCT reported electroacupuncture increased menstrual frequency more than no intervention (0.37, 95% CI 0.21 to 0.53, 31 women). There was no evidence of a difference between the groups in adverse events. Evidence was very low quality with very wide CIs and very low event rates. Overall evidence was low or very low quality. The main limitations were failure to report important clinical outcomes and very serious imprecision. Authors' conclusions Thus far, only a limited number of RCTs have been reported. At present, there is insufficient evidence to support the use of acupuncture for treatment of ovulation disorders in women with PCOS.

Electroacupuncture Accelerates the Delayed Intestinal Transit in POI by Suppressing M1 Like Muscularis Macrophages and IL6 Secretion

Background: Electroacupuncture (EA) at ST-36 could accelerate the delayed gastrointestinal (GI) motility in many GI motility dysfunction models, but the definite effect and mechanisms are unclear. In this study, We intended to investigate the effects of EA on intestinal manipulation (IM) mice model and involved mechanisms. Methods: Male C57BL/6 mice were randomized into five groups: normal control, intestinal manipulation (IM), IM with sham EA (SEA), IM with high-frequency EA (HEA) and IM with low-frequency EA (LEA). The GI transit was evaluated. The infiltration of muscularis macrophages (MMφ) and its phenotype were analysed with flow cytometry. Magnetic-activated cell sorting was applied to isolate MMφ, and the relationship between the MMφ and interstitial cells of Cajal (ICCs) was further investigated. Results: (1) Compared with the IM group, HEA and LEA attenuated the delayed intestinal transit. (2) Both the HEA and LEA obviously reduced the MMφ and supressed the M1 activation of the MMφ in the ileum. (3) EA restored the disrupted ICC networks through inhibiting the release of IL6 by the MMφ. Conclusion: (1)EA at acupoint ST-36 could accelerate the delayed intestinal transit in the IM murine model by restoring the ICC networks. (2)EA protected the ICCs through reducing the MMφ, inhibiting its M1 polarization and its IL6 secretion. Funding Statement: The work was supported by grants from the National Natural Science Foundation of China (nos. 81570488; nos. 81770536). Declaration of Interests: The authors declare they have no conflict interests. Ethics Approval Statement: This study was carried out in accordance with the principles of the Guide for the Care and Use of Laboratory Animals of Tongji Medical College, the Committee of Experimental Animals of Tongji Medical College of Huazhong University of Science and Technology.

Electroacupuncture in rats normalizes the diabetes-induced alterations in the septo-hippocampal cholinergic system.

Diabetes induces early sufferance in the cholinergic septo-hippocampal system, characterized by deficits in learning and memory, reduced hippocampal plasticity and abnormal pro-nerve growth factor (proNGF) release from hippocampal cells, all linked to dysfunctions in the muscarinic cholinergic modulation of hippocampal physiology. These alterations are associated with dysregulation of several cholinergic markers, such as the NGF receptor system and the acetylcholine biosynthetic enzyme choline-acetyl transferase (ChAT), in the medial septum and its target, the hippocampus. Controlled and repeated sensory stimulation by electroacupuncture has been proven effective in counteracting the consequences of diabetes on cholinergic system physiology in the brain. Here, we used a well-established Type 1 diabetes model, obtained by injecting young adult male rats with streptozotocin, to induce sufferance in the septo-hippocampal system. We then evaluated the effects of a 3-week treatment with low-frequency electroacupuncture on: (a) the expression and protein distribution of proNGF in the hippocampus, (b) the tissue distribution and content of NGF receptors in the medial septum, (c) the neuronal cholinergic and glial phenotype in the septo-hippocampal circuitry. Twice-a-week treatment with low-frequency electroacupuncture normalized, in both hippocampus and medial septum, the ratio between the neurotrophic NGF and its neurotoxic counterpart, the precursor proNGF. Electroacupuncture regulated the balance between the two major proNGF variants (proNGF-A and proNGF-B) at both gene expression and protein synthesis levels. In addition, electroacupuncture recovered to basal level the pro-neurotrophic NGF receptor tropomyosin receptor kinase-A content, down-regulated in medial septum cholinergic neurons by diabetes. Electroacupuncture also regulated ChAT content in medial septum neurons and its anterograde transport toward the hippocampus. Our data indicate that repeated sensory stimulation can positively affect brain circuits involved in learning and memory, reverting early impairment induced by diabetes development. Electroacupuncture could exert its effects on the septo-hippocampal cholinergic neurotransmission in diabetic rats, not only by rescuing the hippocampal muscarinic responsivity, as previously described, but also normalizing acetylcholine biosynthesis and NGF metabolism in the hippocampus.

Efficacy of electroacupuncture in regulating the imbalance of AMH and FSH to improve follicle development and hyperandrogenism in PCOS rats

Polycystic ovary syndrome (PCOS) is characterized by hyperandrogenism and follicular arrest. These two characteristics may result from an imbalance between anti-Müllerian hormone and follicle stimulating hormone. Electroacupuncture is effective in improving hyperandrogenism and follicular arrest in PCOS; however, the mechanism is not sufficiently clear. This study aimed to elucidate whether electroacupuncture in PCOS is exerted by regulating an imbalance of anti-Müllerian hormone and follicle stimulating hormone. In this study, a rat model of polycystic ovary syndrome was treated with low-frequency electroacupuncture at acupoints (CV-3 and CV-4). To observe the mechanism of electroacupuncture in PCOS, we first observed the estrous cycle. We then observed ovarian morphology by hematoxylin-eosin staining and evaluated levels of testosterone, estradiol, P450arom, follicle stimulating hormone and its receptor, and anti-Müllerian hormone and its receptor by enzyme-linked immunosorbent assay, western blotting, double immunofluorescence assay and real-time PCR. Our results showed that in 80% of rats in the electroacupuncture acupoints group, their estrous cycle recovered, ovarian morphology significantly improved, testosterone level significantly decreased, and levels of estradiol and P450arom significantly increased in peripheral serum after 14 consecutive days of treatment (P < 0.01). The expression of anti-Müllerian hormone and anti-Müllerian hormone type II receptor decreased (P < 0.05), whereas the expression of follicle stimulating hormone receptor increased (P < 0.05). These results indicated that electroacupuncture improved hyperandrogenism and follicular arrest by decreasing the excessive expression of AMH to regulate FSH and AMH imbalance in granulosa cells in PCOS.

Paired Associative Electroacupuncture and Transcranial Magnetic Stimulation in Humans.

Pairing transcutaneous electric nerve stimulation (TENS) and transcranial magnetic stimulation (TMS) with specific stimulus-intervals induces associative motor plasticity at the primary motor cortex (M1). Electroacupuncture (EA) is an established medical technique in the eastern countries. This study investigates whether EA paired with TMS induces distinct M1 motor plasticity. Fifteen healthy, right-handed subjects (aged 23.6 ± 2.0 years, eight women) were studied. Two-hundred and twenty-five pairs of TMS of the left M1 preceded by right EA at acupoint “Neiguan” [Pericardium 6 (PC6), located 2 decimeters proximal from the wrist wrinkle] were respectively applied with the interstimulus interval (ISI) of individual somatosensory evoked potential (SSEP) N20 latency plus 2 ms (N20+2) and minus 5 ms (N20-5) with at least 1-week interval. The paired stimulation was delivered at a rate of 0.25 Hz. Sham TMS with a sham coil was adopted to examine the low-frequency EA influence on M1 in eleven subjects. M1 excitability was assessed by motor-evoked potential (MEP) recruitment curve with five TMS intensity levels, short-interval intracortical inhibition (SICI), intracortical facilitation (ICF) and cerebellar inhibition (CBI) at the abductor pollicis brevis (APB) muscle of the right hand before and after the EA-M1 paired associative stimulation (PAS). In addition, median nerve SSEPs and H-reflex were respectively measured to monitor somatosensory and spinal excitability. The MEP showed significantly facilitated after the sham EA-M1 PAS while tested with 80% of the TMS intensity producing on average 1 mV amplitude (i.e., MEP1 mV) in the resting APB muscle. It was also facilitated while tested with 90% MEP1 mV irrespective of the stimulation conditions. The SSEP showed a higher amplitude from the real EA-M1 PAS compared to that from the sham EA-M1 PAS. No significant change was found on SICI, ICF, CBI and H-reflex. Findings suggest that repetitive low frequency EA paired with real TMS did not induce spike-timing dependent motor plasticity but EA paired with sham TMS induced specific M1 excitability change. Complex sensory afferents with dispersed time locked to the sensorimotor cortical area could hamper instead of enhancing the induction of the spike-timing dependent plasticity (STDP) in M1.

Electroacupuncture promotes the gastrointestinal motility of diabetic mice by CNP/NPR-B-cGMP and PDE3A-cGMP signaling.

Electroacupuncture (EA) can promote gastrointestinal (GI) motility of diabetic mice, but the mechanism is not clearly elucidated. Natriuretic peptides (NPs) were related to the diabetes-induced gut dysfunction of mice, which may be associated with ICC (interstitial cells of cajal). Besides, EA could increase the ICC of diabetic mice. Our aim was to explore whether EA can promote the gut motility by CNP/NPR-B-cGMP and PDE3A-cGMP signaling in diabetic mice, and the relationship between NPs and ICC. Wild C57BL/6 male mice were divided into five groups: control group, diabetic mellitus (DM group), diabetic mellitus plus sham EA group (SEA), diabetic mellitus plus low-frequency EA group (LEA), and diabetic mellitus plus high-frequency group (HEA). Gastrointestinal motility was assessed by gastric emptying and GI transit test. Immunofluorescence staining was applied to assess the expression level of CNP, NPR-B, and c-Kit. Western blot, PCR, and ELISA were used to detect the level of CNP, NPR-B, PDE2A, PDE3A, c-Kit, mSCF, and cGMP content. The correlativity between NPR-B and mSCF was evaluated by Pearson's correlation and linear regression analyses. (a) EA could improve the GI dysfunction of diabetic mice. (b) CNP, NPR-B, and cGMP contents were decreased, but the level of PDE3A, c-Kit, and mSCF was increased in the EA groups. (c) There was a negative correlation between NPR-B and mSCF among the groups. Electroacupuncture promotes the GI function by CNP/NPR-B-cGMP and PDE3A-cGMP signaling in diabetic mice; up-regulated mSCF/c-Kit signaling by EA may be mediated partially via down-regulation of CNP/NPR-B signaling.

Bone marrow-derived interstitial cells of cajal are increased by electroacupuncture in the colon of diabetic mice.

Insufficiency of interstitial cells of cajal (ICC) in the muscular plexus of colon has been proved in diabetic mice. Electroacupuncture (EA) at ST36 can accelerate ICC supplement in the colon of diabetic mice, but the source of increased ICC has not been clearly explored. Bone marrow cells possess the potential of migration and differentiation into ICC of the intestine. Our aim is to explore the effects of EA on variations of bone marrow-derived ICC in the colon of diabetic mice as well as its mechanism. Wild C57BL/6 mice were divided into six groups with random assignment method: control group, diabetic mellitus (DM) group, bone marrow transplantation (BMT)+DM group, BMT+DM+sham EA group, BMT+DM+low-frequency EA group, and BMT+DM+high-frequency EA group. Flow cytometric method was adopted to identify the chimera model. The specific location and expression levels of c-Kit+ green fluorescent protein (GFP+) cells in colon were detected by immunofluorescence. Western blot and quantitative polymerase chain reaction were used to assess the expression level of c-Kit, GFP, membrane-band stem cell factor (mSCF), p-ERK, p-c-Jun, ETV1, stromal cell-derived factor 1 (SDF-1), CXCR4, transforming growth factor β1 (TGF-β1), and smad3. c-Kit+ GFP+ cells in the muscular plexus of colon were apparently increased in the EA groups; the protein and mRNA expression level of CXCR4, SDF-1, TGF-β1, smad3, c-kit, mSCF, p-ERK, p-c-Jun, and ETV1 were elevated in the EA groups. Electroacupuncture can effectively increase bone marrow-derived ICC in the colon by SDF-1/CXCR4, TGF-β1/Smad3, and mSCF/Kit-p-ERK/p-c-Jun-ETV1 signal pathway.

Observations on the Therapeutic Effect of Different Frequency Electroacupuncture on Poststroke Hemidysesthesia

Objective to observethe clinical effect of electroacupuncture with different frequency in the treatment of hemidysesthesia after stroke. Methods Sixty patients with hemidysesthesia after stroke were randomly divided into treatment group and control groups, with 30 cases in each group. Both groups were treated with electroacupuncture with the same acupoint selection. The electroacupuncture frequency was 100 Hz in the treatment group and 1 Hz in the control group. The clinical effficacy of the two groups was observed after 30 days of treatment. Results The total effective rate and the recovery rate of the treatment group were 93.3% and 80.0%, respectively, and the control group were 86.0% and 43.3%, respectively The difference in the total effective rate between the two groups was statistically significant (p < 0.05). And the difference in the recovery rate between the two groups was statistically significant (p < 0.05). There were was a statistically significant differences in the scores of limb dysesthesia score between the two groups after treatment and before treatment (p < 0.01). There was a statistically significant difference in the limb dysesthesia score after treatment between the treatment group and control group (p < 0.05). Conclusions Electroacupuncture is an effective method to treat poststroke hemidysesthesia. High frequency electroacupuncture is more obvious than low frequency electroacupuncture.

Low-frequency electroacupncture improves locomotor function after sciatic crushed nerve injury in rats.

Sciatic crushed nerve injury (SCI) causes pain-related gait and swelling in the affected limb. Electroacupuncture (EA) is a modified acupuncture technique, and analgesic effect of EA on different types of pain has been documented. Scientific functional index (SFI) is a mathematical formula to represent parameters of normal and experimental footprints. We investigated the effect of low-frequency EA on functional recovery following SCI in rats. For this study, immunohistochemistry for c-Fos in the ventral lateral periaqueductal gray (vlPAG) and paraventricular nucleus (PVN) and western blot for neurofilament (NF) and brain-derived neurotrophic factor (BDNF) in the sciatic nerve were conducted. To induce crush injury on the sciatic nerve, sciatic nerve was crushed for 30 sec using a surgical clip. The rats in the acupuncture groups received acupuncture bilaterally at respective site, once a day for 14 days. The rats in the EA group received 100-Hz electrical stimulation for 10 min once a day during 14 days. SCI decreased SFI value, in contrast, EA increased SFI value. c-Fos expression in the vlPAG and PVN was increased following SCI, in contrast, EA suppressed c-Fos expression. NF expression in the sciatic nerve was decreased by SCI, in contrast, EA increased NF expression. BDNF expression in the sciatic nerve was increased by SCI, in contrast, EA suppressed BDNF expression. In the present study, EA showed effectiveness on functional recovery from SCI.

Effects of electroacupuncture of different frequencies on SP and VIP expression levels in colon of rats with slow transit constipation

To observe the effects of electroacupuncture (EA) of different frequencies on the expression levels of substance P (SP) and vasoactive intestinal peptide (VIP) in the colon of rats with slow transit constipation (STC). One hundred healthy male Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, a low-frequency EA group, a high-frequency EA group and a variable-frequency EA group, with 20 rats in each group. The rats in the normal group were fed with normal diet. The rats in the other groups were fed with phenethylpiperidine in the diet at a dose of 8 mg/(kg·bw) per day, for 120 d, to establish the STC model. Rats in the normal group and the model group did not receive any treatment; rats in the low-frequency EA group were treated with 2 Hz continuous wave EA, rats in the high-frequency EA group were treated with 100 Hz continuous wave EA, and rats in the variable-frequency EA group were treated with 2 Hz/100 Hz sparse-dense EA. The current intensity of the EA was determined by the slight vibration of the rat limbs without painful screaming. The intervention was performed once a day, 15 min/time for continuous 15 d. After treatment, the intestinal transit function and the expression levels of SP and VIP in the colon of the rats in each group were determined. After treatment, the defecation duration of the first dark stool in the model group was significantly longer than that in the normal group (P<0.05); the defecation durations of the first dark stool in the low-frequency EA group, high-frequency EA group and variable-frequency EA group were significantly shorter than the duration in the model group (all P<0.05); compared with the low-frequency EA group, the first dark stool defecation duration of rats in the variable-frequency EA group was significantly shorter (P<0.05); compared with the normal group, the SP and VIP expression levels in the colon of the model group were significantly decreased (both P<0.01); the SP and VIP expression levels in the colon of the low-frequency EA group, the high-frequency EA group and the variable-frequency EA group were significantly higher than those in the model group (all P<0.05); compared with the high-frequency EA group, the SP expression levels in the colon in the low-frequency EA group and the variable-frequency EA group were significantly increased (both P<0.05); compared with the low-frequency EA group, the VIP expression levels in the colon in the high-frequency EA group and the variable-frequency EA group were significantly increased (both P<0.05). EA improves the intestinal function of STC model rats by regulating the expression levels of SP and VIP in rat colon. The EA stimulation with 100 Hz continuous wave, 2 Hz/100 Hz sparse-dense wave shows a better improvement in the colonic transit function in STC rats, followed by 2 Hz continuous wave.

Low-Frequency Electroacupuncture Alleviates Chronic Constrictive Injury-Induced Mechanical Allodynia by Inhibiting NR2B Upregulation in Ipsilateral Spinal Dorsal Horn in Rats.

To study the effects of electroacupuncture (EA) in chronic constrictive injury (CCI) rat model and the expression of N-methyl-D-aspartate receptor type 2B (NR2B) in ipsilateral spinal dorsal horn in rats to explore the analgesic mechanisms of EA. According to the random number table, totally 180 rats were evenly divided into a sham group, a CCI group, and an EA group. CCI model was conducted with four 4-0 chromic gut ligatures loosely ligated around the left sciatic nerve 1 cm above the trifurcation. Rats in the EA group received 2 Hz EA therapy bilaterally at acupoints of Zusanli (ST 36) and Sanyinjiao (SP 6) once daily (30 min/d) for 30 days after surgery. Paw withdrawal thresholds (PWTs) were measured on 0 (baseline), 1, 3, 7, 15, 30 days after surgery. Rats were sacrificed on 0, 1, 3, 7, 15 and 30 days after surgery, and the L4-5 segments of spinal cord were removed to detect the expression of NR2B by immunohistochemistry and quantitative polymerase chain reaction. PWTs in the CCI group were significantly lower than the sham group at Day 1-30 after surgery, and reached its lowest at Day 1 (P<0.01). After EA treatment, the PWTs recovered rapidly and were significantly higher than those in the CCI group on 3, 7, 15 and 30 days after surgery (P<0.01). The numbers of NR2B-immunoreactive cells of the CCI group significantly increased after CCI surgery compared with the sham group (P<0.01). Compared with the CCI group, stimulation of EA markedly decreased the numbers of NR2B-immunoreactive cells at Day 3, 7, 15 and 30 (P<0.05). In the sham group, NR2B mRNA was expressed at a low level. It increased after CCI surgery, which increased rapidly at Day 7 (P<0.01) and reached its peak value at Day 15 (P<0.01). After EA stimulation, relative quantity of NR2B mRNA expression was less than that in the CCI group at Day 15 and 30 (P<0.05). Low frequency of EA had antinociceptive effect in CCI rat model. The analgesic effects of EA might be through the inhibition of NR2B.

Endogenous Ovarian Angiogenesis in Polycystic Ovary Syndrome-Like Rats Induced by Low-Frequency Electro-Acupuncture: The CLARITY Three-Dimensional Approach.

We sought to determine the role of ovarian vascularity and neo-angiogenesis in the development of mature follicles in polycystic ovary syndrome (PCOS) and to identify any changes induced by low-frequency electro-acupuncture (EA). Twenty-eight 21-day-old female Wistar rats were randomly divided into four groups—Control, Obesity, PCOS-like, and PCOS-like-EA (n = 7/group). Rats in the Obesity group were fed a high-fat diet throughout the experiment. Rats in the PCOS-like and PCOS-like-EA groups were implanted with a sustained-release tube containing 5α-dihydrotestosterone (DHT) beneath the skin of the neck. Rats in the PCOS-like-EA group received low-frequency EA treatment starting at 70 days for 30 min five times a week for four weeks. At the end of the experiment, all rats were euthanized and perfused with hydrogel. The ovaries were collected for clarification and imaging, and ovarian vascularity and neo-angiogenesis were analyzed. Compared with Control and Obesity rats, the ovaries in DHT-induced PCOS-like rats were smaller in size and had fewer mature follicles and corpora lutea. EA increased angiogenesis in the antral follicles of PCOS-like rats, which in turn promoted follicle maturation, ovulation, and CL formation. Therefore, endogenous ovarian angiogenesis plays a very important role in follicular maturation and might be one of the peripheral and direct mechanisms of EA on PCOS.

Investigation of Interactive Activity of Electro-Acupuncture on Pharmacokinetics of Sildenafil and Their Synergistic Effect on Penile Blood Flow in Rats.

Erectile dysfunction (ED) is a disorder found in males throughout the world, which negatively affects relationships with partners with advancing age. Hence, in this study, we tested a combined novel treatment of electro-acupuncture (EA) and sildenafil citrate against ED. In addition to EA therapy, the sildenafil citrate, a phosphodiesterase 5 inhibitor, is a widely recognized drug that has achieved considerable success in the treatment of ED. However, the combined effect of both the EA and sildenafil has not yet been investigated. Hence, we aimed to examine the effect of EA on the pharmacokinetics and pharmacodynamics of sildenafil in rat plasma. The pharmacokinetic parameters were determined using ultra performance liquid chromatography (UPLC) after EA and sildenafil administration (10 mg/Kg). Following this, the pharmacodynamics was studied via blood flow pattern using developing Doppler images of the lower body and penis. The pharmacokinetic studies demonstrated that sildenafil significantly increases by administration of low-frequency EA. Further, the pharmacodynamic studies using Doppler imaging revealed an elevated blood flow in rat penis compared with lower body during combined treatment of sildenafil and low-frequency EA. These data indicate a synergistic therapeutic effect of EA and sildenafil for the treatment of ED.

F139. Paired associative electroacupuncture and transcranial magnetic stimulation in humans

Introduction Conventional paired associative stimulation (PAS) consists of repeated pairing of a single electric stimulus at median nerve followed by a transcranial magnetic stimulation (TMS) pulse on the contralateral primary motor cortex (M1) with a specific inter-stimulus interval. It induces associative long-term potentiation (LTP) or depression (LTD)-like plasticity at M1. Attention has been found with a positive influence on such plasticity. In this study we adopt electroacupuncture (EA) to replace the transcutaneous electric stimulation, supposedly drawing a strong attention and enhancing the PAS effect. Methods Eleven healthy, right-handed subjects (aged 23.9 ± 2.2 years, 4 men) were studied. Two hundred and twenty-five pairs of TMS of the left M1 preceded by right EA at acupoint ‘Neiguan’ (Pericardium 6 or PC6, located 2 decimeters proximal from the wrist wrinkle) were respectively applied with an interstimulus interval of individual N20 latency plus 2 ms (N20 + 2) and minus 5 ms (N20-5) with at least one week interval. The paired stimulation was delivered at a rate of 0.25 Hz. Sham TMS with a sham coil was performed to exclude any possible effects from the repetitive low-frequency EA stimulation. M1 excitability was assessed by motor-evoked potential (MEP) recruitment curve with five TMS intensity levels (80–120% resting motor threshold), short-interval intracortical inhibition (SICI), intracortical facilitation (ICF) and cerebellar inhibition (CBI) at the abductor pollicis brevis (APB) muscle of the right hand before and after the EA-M1 PAS. In addition, median nerve somatosensory-evoked potentials (SSEP) and H-reflex were measured to monitor somatosensory and spinal excitability, respectively. Results All subjects successfully completed four sessions of the experiment. There was no significant change of the MEP amplitude after the EA-M1 PAS at five TMS intensities. SICI, ICF, CBI, SSEP and H-reflex were also not significantly changed after the EA-M1 PAS intervention (all P > 0.05 by paired t-test). Conclusion Transcutaneous electric stimulation replaced by EA is not feasible to induce long-term plasticity-like effect. Complicated sensory afferents evoked by electroacupuncture might hamper instead of enhancing the spike timing dependent motor plasticity in M1.