Electro-acupuncture modulated miR-214 expression to prevent chondrocyte apoptosis and reduce pain by targeting BAX and TRPV4 in osteoarthritis rats

Osteoarthritis (OA) is a highly prevalent joint disorder characterized by progressive degeneration of articular cartilage, subchondral bone remodeling, osteophyte formation, synovial inflammation, and meniscal damage. Although the etiology of OA is multifactorial, pro-inflammatory processes appear to play a key role in disease pathogenesis. Previous studies indicate that electroacupuncture (EA) exerts chondroprotective, anti-inflammatory, and analgesic effects in preclinical models of OA, but the mechanisms underlying these potential therapeutic benefits remain incompletely defined. This study aimed to investigate the effects of EA on OA development in a rat model, as well as to explore associated molecular mechanisms modulated by EA treatment. Forty rats were divided into OA, EA, antagomiR-214, and control groups. Following intra-articular injection of monosodium iodoacetate to induce OA, EA and antagomiR-214 groups received daily EA stimulation at acupoints around the knee joint for 21 days. Functional pain behaviors and chondrocyte apoptosis were assessed as outcome measures. The expression of microRNA-214 (miR-214) and its downstream targets involved in apoptosis and nociception, BAX and TRPV4, were examined. Results demonstrated that EA treatment upregulated miR-214 expression in OA knee cartilage. By suppressing pro-apoptotic BAX and pro-nociceptive TRPV4, this EA-induced miR-214 upregulation ameliorated articular pain and prevented chondrocyte apoptosis. These findings suggested that miR-214 plays a key role mediating EA's therapeutic effects in OA pathophysiology, and represents a promising OA treatment target for modulation by acupuncture.

Effects and Mechanisms of Electroacupuncture at Zhongwan （CV12） on Gastric Nociceptive Response Induced by Gastric Acid Stimulation in Rats / 中医杂志

ObjectiveTo observe the effects of electroacupuncture at Zhongwan （CV12） on gastric nociceptive response induced by gastric acid stimulation and explore the underlying mechanisms associated with nuclei of the medullary viscerosensory and visceral motor neurons. MethodsTwenty SD rats were given intragastric administration of 0.5 mol/L diluted hydrochloric acid （0.5 ml/100 g） to induce gastric nociceptive response induction. Eight rats were randomly selected to record the gastric slow wave （GSW） area under the curve， and extracellular discharge frequency of neurons in the nucleus of the solitary tract （NTS） and dorsal motor nucleus of the vagus nerve （DMV） before intragastric administration and at 1， 5， 10， 15， 20， 25， 30， 35， 40， 45， 50， 55， and 60 minutes after intragastric administration. The remaining 12 rats received electroacupuncture intervention at Zhongwan within 5 to 25 minutes after intragastric administration of diluted hydrochloric acid， with a duration of one minute. The GSW area under the curve and extracellular discharge frequency of NTS and DMV neurons were compared between the 1-minute intervals before and after electroacupuncture intervention. ResultsCompared to the baseline before intragastric administration， the area under the curve of GSW significantly increased at 1， 5， 10， 15， 20， and 25 minutes after intragastric administration， and the extracellular discharge frequency of excitatory neurons in the NTS （accounting for 90%， 57/63） significantly increased at 1， 5， 10， 15， 20， 25， 30， 35， and 40 minutes， both reaching peak values at 1 minute after intragastric administration （P＜0.01 or P＜0.05）. The extracellular discharge frequency of inhibitory neurons in the DMV （accounting for 91%， 20/22） showed a non-significant increase at 1 minute after intragastric administration （P＞0.05）， but significantly decreased at other timepoints （P＜0.05）. Compared to the baseline before electroacupuncture intervention， the GSW area under the curve and the extracellular discharge frequency of excitatory neurons in the NTS significantly decreased （P＜0.05）， while the extracellular discharge frequency of inhibitory neurons in the DMV showed no significant difference （P＞0.05）. ConclusionElectroacupuncture at Zhongwan can improve gastric nociceptive response induced by gastric acid stimulation， possibly by reducing the transmission of visceral sensation and decreasing the excitability of NTS neurons in the medulla.

Electroacupuncture Promotes Functional Recovery after Facial Nerve Injury in Rats by Regulating Autophagy via GDNF and PI3K/mTOR Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To explore the mechanism of electroacupuncture (EA) in promoting recovery of the facial function with the involvement of autophagy, glial cell line-derived neurotrophic factor (GDNF), and phosphatidylinositol-3-kinase (PI3K)/mammalian target of rapamycin (mTOR) signaling pathway.@*METHODS@#Seventy-two male Sprague-Dawley rats were randomly allocated into the control, sham-operated, facial nerve injury (FNI), EA, EA+3-methyladenine (3-MA), and EA+GDNF antagonist groups using a random number table, with 12 rats in each group. An FNI rat model was established with facial nerve crushing method. EA intervention was conducted at Dicang (ST 4), Jiache (ST 6), Yifeng (SJ 17), and Hegu (LI 4) acupoints for 2 weeks. The Simone's 10-Point Scale was utilized to monitor the recovery of facial function. The histopathological evaluation of facial nerves was performed using hematoxylin-eosin (HE) staining. The levels of Beclin-1, light chain 3 (LC3), and P62 were detected by immunohistochemistry (IHC), immunofluorescence, and reverse transcription-polymerase chain reaction, respectively. Additionally, IHC was also used to detect the levels of GDNF, Rai, PI3K, and mTOR.@*RESULTS@#The facial functional scores were significantly increased in the EA group than the FNI group (P<0.05 or P<0.01). HE staining showed nerve axons and myelin sheaths, which were destroyed immediately after the injury, were recovered with EA treatment. The expressions of Beclin-1 and LC3 were significantly elevated and the expression of P62 was markedly reduced in FNI rats (P<0.01); however, EA treatment reversed these abnormal changes (P<0.01). Meanwhile, EA stimulation significantly increased the levels of GDNF, Rai, PI3K, and mTOR (P<0.01). After exogenous administration with autophagy inhibitor 3-MA or GDNF antagonist, the repair effect of EA on facial function was attenuated (P<0.05 or P<0.01).@*CONCLUSIONS@#EA could promote the recovery of facial function and repair the facial nerve damages in a rat model of FNI. EA may exert this neuroreparative effect through mediating the release of GDNF, activating the PI3K/mTOR signaling pathway, and further regulating the autophagy of facial nerves.

Electroacupuncture at Sensitized Acupoints Relieves Somatic Referred Pain in Colitis Rats by Inhibiting Sympathetic-Sensory Coupling to Interfere with 5-HT Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate whether electroacupuncture (EA) at sensitized acupoints could reduce sympathetic-sensory coupling (SSC) and neurogenic inflammatory response by interfering with 5-hydroxytryptamine (5-HT)ergic neural pathways to relieve colitis and somatic referred pain, and explore the underlying mechanisms.@*METHODS@#Rats were treated with 5% dextran sodium sulfate (DSS) solution for 7 days to establish a colitis model. Twelve rats were randomly divided into the control and model groups according to a random number table (n=6). According to the "Research on Rat Acupoint Atlas", sensitized acupoints and non-sensitized acupoints were determined. Rats were randomly divided into the control, model, Zusanli-EA (ST 36), Dachangshu-EA (BL 25), and Xinshu (BL 15) groups (n=6), as well as the control, model, EA, and EA + GR113808 (a 5-HT inhibitor) groups (n=6). The rats in the control group received no treatment. Acupuncture was administered on 2 days after modeling using the stimulation pavameters: 1 mA, 2 Hz, for 30 min, with sparse and dense waves, for 14 consecutive days. GR113808 was injected into the tail vein at 5 mg/kg before EA for 10 min for 7 consecutive days. Mechanical sensitivity was assessed with von Frey filaments. Body weight and disease activity index (DAI) scores of rats were determined. Hematoxylin and eosin staining was performed to observe colon histopathology. SSC was analyzed by immunofluorescence staining. Immunohistochemical staining was performed to detect 5-HT and substance P (SP) expressions. The calcitonin gene-related peptide (CGRP) in skin tissue and tyrosine hydroxylase (TH) protein levels in DRG were detected by Western blot. The levels of hyaluronic acid (HA), bradykinin (BK), prostaglandin I2 (PGI2) in skin tissue, 5-HT, tryptophan hydroxylase 1 (TPH1), serotonin transporters (SERT), 5-HT 3 receptor (5-HT3R), and 5-HT 4 receptor (5-HT4R) in colon tissue were measured by enzyme-linked immunosorbent assay (ELISA).@*RESULTS@#BL 25 and ST 36 acupoints were determined as sensitized acupoints, and BL 15 acupoint was used as a non-sensitized acupoint. EA at sensitized acupoints improved the DAI score, increased mechanical withdrawal thresholds, and alleviated colonic pathological damage of rats. EA at sensitized acupoints reduced SSC structures and decreased TH and CGRP expression levels (P<0.05). Furthermore, EA at sensitized acupoints reduced BK, PGI2, 5-HT, 5-HT3R and TPH1 levels, and increased HA, 5-HT4R and SERT levels in colitis rats (P<0.05). GR113808 treatment diminished the protective effect of EA at sensitized acupoints in colitis rats (P<0.05).@*CONCLUSION@#EA at sensitized acupoints alleviated DSS-induced somatic referred pain in colitis rats by interfering with 5-HTergic neural pathway, and reducing SSC inflammatory response.

Research progress on the modulation mechanism of electroacupuncture for learning and memory impairment after ischemic stroke / 中国针灸

Electroacupuncture may play a role in treatment of learning and memory impairment after ischemic stroke by regulating phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) signaling pathway, cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA)/cAMP response element binding protein (CREB) signaling pathway, nerve growth factor (NGF)/tyrosine kinase-A (TrkA) signaling pathway, Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway, Notch signaling pathway, erythropoietin-producing hepatocyte (Eph)/ephrin signaling pathway. The interactions among these pathways should be further explored in treatment of learning and memory impairment after ischemic stroke.

GAO Wei-bin's clinical experience in treatment of neurogenic bladder with acupuncture / 中国针灸

The paper introduces GAO Wei-bin's clinical experience in acupuncture treatment for neurogenic bladder. In association with the etiology, the location and types of neurogenic bladder and in accordance with nerve anatomy and meridian differentiation, the acupoints are selected accurately in treatment. Four acupoint prescriptions are allocated. For frequent urination and urinary incontinence, the foot-motor-sensory area of scalp acupuncture, Shenshu (BL 23) and Huiyang (BL 35) are used. For all kinds of urine retention, especially the patients who are not suitable for acupuncture at the lumbar region, Zhongji (CV 3), Qugu (CV 2), Henggu (KI 11) and Dahe (KI 12) are selected. For all kinds of urine retention, Zhongliao (BL 33) and Ciliao (BL 32) are applicable. For the patients with both dysuria and urinary incontinence, Zhongliao (BL 33), Ciliao (BL 32) and Huiyang (BL 35) are chosen. In treatment of neurogenic bladder, both biao (root causes) and ben (primary symptoms) are considered, as well as the accompanying symptoms; and electroacupuncture is combined accordingly. During the delivery of acupuncture, the sites where the acupoints located are detected and palpated so as to rationally control the depth of needle insertion and the operation of reinforcing and reducing needling techniques.

Electroacupuncture at Baliao points for erectile dysfunction after stroke: a randomized controlled trial / 中国针灸

OBJECTIVE@#To observe the clinical effect of electroacupuncture at Baliao points in patients with erectile dysfunction after stroke.@*METHODS@#A total of 58 patients with erectile dysfunction after stroke were randomly divided into an observation group (29 cases, 1 case dropped off, 1 case discontinued) and a control group (29 cases, 1 case dropped off). Both groups were given basic treatment, including routine medical treatment, routine acupuncture treatment, rehabilitation training and pelvic floor biofeedback electrical stimulation treatment. The observation group was treated with electroacupuncture at Baliao points, and the control group was treated with shallow acupuncture combined with electroacupuncture at the control points (8 points at 20 mm horizontally beside Baliao points), continuous wave, frequency in 50 Hz, current intensity in 1-5 mA, 5 times a week for 4 weeks. The 5-item version of the international index of erectile function (IIEF-5) score, erectile dysfunction effect on quality of life (ED-EQoL) score and pelvic floor muscle contraction amplitude were compared between the two groups before and after treatment.@*RESULTS@#After treatment, the IIEF-5 scores and the contraction amplitude of fast muscle fiber, comprehensive muscle fiber and slow muscle fiber in the two groups were higher than those before treatment (P<0.05), the ED-EQoL scores were lower than those before treatment (P<0.05), and the changes of above indexes in the observation group were larger than the control group (P<0.05).@*CONCLUSION@#Electroacupuncture at Baliao points can improve the erectile function of patients with erectile dysfunction after stroke, increase the contraction amplitude of pelvic floor muscles, and promote the quality of life of patients.

Transcutaneous electrical acupoint stimulation combined with electroacupuncture for rapid recovery after abdominal surgery: a randomized controlled trial / 中国针灸

OBJECTIVE@#To observe the effect of transcutaneous electrical acupoint stimulation (TEAS) combined with electroacupuncture (EA) on rehabilitation after abdominal surgery.@*METHODS@#A total of 320 patients undergoing abdominal surgery were randomly divided into a combination group (80 cases), a TEAS group (80 cases, 1 case discontinued), an EA group (80 cases, 1 case discontinued) and a control group (80 cases, 1 case discontinued). The patients in the control group received enhance recovery after surgery (ERAS) standardized perioperative management. On the basis of the treatment in the control group, the TEAS group was treated with TEAS at Liangmen (ST 21) and Daheng (SP 15); the EA group was treated with EA at Neiguan (PC 6), Hegu (LI 4), Zusanli (ST 36), Shangjuxu (ST 37) and Xiajuxu (ST 39); the combination group was treated with TEAS combined with EA, with continuous wave, 2-5 Hz in frequency, and the intensity was tolerable to the patients, 30 min each time, once a day, from the first day after surgery, until the anus resumed spontaneous defecation and the oral intake of solid food was tolerated. The gastrointestinal-2 (GI-2) time, first exhaust time, first defecation time, first tolerance of oral intake of solid food time, time of first get out of bed and hospital stay were observed in all the groups; the pain visual analogue scale (VAS) score and incidence rates of nausea and vomiting 1, 2, 3 days after surgery were compared in all the groups; after treatment, the acceptability of each treatment was evaluated by patients in each group.@*RESULTS@#Compared with the control group, the GI-2 time, first exhaust time, first defecation time, first tolerance of oral intake of solid food time were shortened (P<0.05), the VAS scores 2, 3 days after surgery were decreased (P<0.05) in the combination group, the TEAS group and the EA group; those in the combination group were shorter and lower than the TEAS group and the EA group (P<0.05). Compared with the control group, the time of hospital stay in the combination group, the TEAS group and the EA group were shortened (P<0.05), and that in the combination group was shorter than the TEAS group (P<0.05).@*CONCLUSION@#TEAS combined with EA can accelerate the recovery of gastrointestinal function in patients after abdominal surgery, relieve postoperative pain, and shorten hospital stay.

Effect of electroacupuncture at different frequencies on brain insulin signaling transduction pathway in Alzheimer's disease mice / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) at different frequencies on learning and memory functions, as well as the relevant proteins of brain insulin signal transduction pathway in Alzheimer's disease (AD) mice and explore the effect mechanism of EA in treatment of AD.@*METHODS@#Seventy-two SPF Kunming male mice were randomized into a blank group, a sham-operation group, a model group, a 2 Hz EA group, a 15 Hz EA group and a 30 Hz EA group, 12 mice in each one. In the model group and each EA group, AD model were established by the injection with streptozotocin (ST2) solution (8 mg/kg) into the left lateral ventricles. In the sham-operation group, 0.9% sodium chloride solution of the same volume was injected into the left lateral ventricles. After successful modeling, in each EA group, EA was applied at "Baihui" (GV 20), "Dazhui" (GV 14) and "Shenshu" (BL 23) with corresponding frequencies, once daily. One course of EA intervention consisted of 7 treatments and 2 courses were given totally at interval of 1 day. After modeling and intervention, Morris water maze test was conducted for the mice of each group. Using immunohistochemistry and Western blot method, the protein expression of insulin receptor (IR), insulin receptor substrate-1 (IRS-1) and phosphatidylinositol 3-kinase (PI3K) was detected in the hippocampal of the mice after intervention.@*RESULTS@#Compared with the blank group, in the model group, the 2 Hz, 15 Hz and 30 Hz EA groups, the escape latency and the first time of crossing the platform were all extended (P<0.01), and the number of crossing the platform was reduced (P<0.01) after modeling. When compared with the blank group, the escape latency and the first time of crossing the platform were all extended (P<0.01), and the number of crossing the platform was reduced (P<0.01) in the model group after intervention. In the 2 Hz, 15 Hz and 30 Hz EA groups, the escape latency and the first time of crossing the platform were all shortened (P<0.01), and the number of crossing the platform was increased (P<0.05, P<0.01) after intervention when compared with the model group. The escape latency and the first time of crossing the platform were all shortened (P<0.01, P<0.05), and the number of crossing the platform was increased (P<0.05) in the 15 Hz and 30 Hz EA groups in comparison with the 2 Hz EA group. The protein expression levels of IR, IRS-1 and PI3K were reduced in the model group when compared with those of the blank group (P<0.01, P<0.05); and these protein expression levels were increased in the 15 Hz and 30 Hz EA groups compared with the model group (P<0.05, P<0.01). Compared with the 2 Hz EA group, the protein expression levels of IR, IRS-1 and PI3K were all elevated in the 15 Hz and 30 Hz EA groups (P<0.05).@*CONCLUSION@#The learning and memory function of AD mice may be improved through regulating brain insulin signaling transconduction pathway with electroacupuncture, and electroacupuncture at 15 Hz and 30 Hz obtains the overall better effect compared with the intervention at 2 Hz.

Hypoglycemic effect of electroacupuncture at "Tianshu" (ST 25) combined with metformin on rats with type 2 diabetes mellitus based on AMPK / 中国针灸

OBJECTIVE@#To observe the hypoglycemic effect of electroacupuncture (EA) at "Tianshu" (ST 25) combined with metformin on rats with type 2 diabetes mellitus (T2DM) as well as its effect on expression of adenosine monophosphate activated protein kinase (AMPK) in liver and pancreas.@*METHODS@#Thirty-six male SD rats were randomly divided into a blank group (6 rats) and a model establishing group (30 rats). The rats in the model establishing group were fed with high-fat diet and treated with intraperitoneal injection of low-dose streptozotocin (STZ) to establish T2DM model. The rats with successful model establishment were randomly divided into a model group, a control group, a metformin group, an EA group and a combination group, 6 rats in each group. The rats in the EA group were treated with EA at "Tianshu" (ST 25), dense-disperse wave, 2 Hz/15 Hz in frequency and 2 mA in current intensity, 20 min each time. The rats in the metformin group were treated with intragastric administration of metformin (190 mg/kg) dissolved in 0.9% sodium chloride solution (2 mL/kg). The rats in the combination group were treated with EA at "Tianshu" (ST 25) and intragastric administration of metformin. The rats in the control group were treated with intragastric administration of 0.9% sodium chloride solution with the same dose. All the treatments were given once a day for 5 weeks. After the intervention, the body mass and random blood glucose were detected; the serum insulin level was detected by ELISA; the expression of AMPK and phosphorylated adenosine monophosphate activated protein kinase (p-AMPK) in liver and pancreas was detected by Western blot method; the expression of protein gene product 9.5 (PGP9.5) was detected by immunofluorescence.@*RESULTS@#①Compared with the blank group, the body mass in the model group was decreased (P<0.05); compared with the model group, the body mass in the EA group and the combination group was decreased (P<0.05); the body mass in the EA group and the combination group was lower than the metformin group (P<0.05). Compared with the blank group, the random blood glucose in the model group was increased (P<0.01); compared with the model group, the random blood glucose in the metformin group, the EA group and the combination group was decreased (P<0.01). The random blood glucose in the combination group was lower than the metformin group and the EA group (P<0.05). ②Compared with the blank group, the insulin level in the model group was decreased (P<0.05); compared with the model group, the insulin level in the metformin group, the EA group and the combination group was all increased (P<0.05). The insulin level in the combination group was higher than the metformin group and the EA group (P<0.05). ③Compared with the blank group, the protein expression of AMPK and p-AMPK in liver tissue was decreased (P<0.05), and the protein expression of AMPK and p-AMPK in pancreatic tissue was increased (P<0.05) in the model group. Compared with the model group, the protein expression of AMPK and p-AMPK in liver tissue in the metformin group, the EA group and the combination group was increased (P<0.05, P<0.01); the protein expression of AMPK in pancreatic tissue in the metformin group was increased (P<0.05); the protein expression of AMPK in pancreatic tissue in the EA group and the combination group was decreased (P<0.05); the protein expression of p-AMPK in pancreatic tissue in the metformin group, the EA group and the combination group was decreased (P<0.05). The protein expression of AMPK and p-AMPK in liver tissue in the combination group was higher than that in the metformin group and the EA group (P<0.05); the protein expression of AMPK in pancreatic tissue in the EA group and the combination group was less than that in the metformin group (P<0.05), and the expression of p-AMPK protein in pancreatic tissue in the combination group was less than that in the metformin group and the EA group (P<0.05). ④Compared with the blank group, the expression of PGP9.5 in pancreatic tissue in the model group was increased (P<0.01); compared with the model group, the expression of PGP9.5 in pancreatic tissue in the metformin group, the EA group and the combination group was decreased (P<0.05, P<0.01). The expression of PGP9.5 in pancreatic tissue in the EA group was lower than the metformin group and the combination group (P<0.05).@*CONCLUSION@#Electroacupuncture at "Tianshu" (ST 25) could promote the effect of metformin on activating AMPK in liver tissue of T2DM rats, improve the negative effect of metformin on AMPK in pancreatic tissue, and enhance the hypoglycemic effect of metformin. The mechanism may be related to the inhibition of pancreatic intrinsic nervous system.

Effect of electroacupuncture at Tinghui (GB 2) and Benshen (GB 13) on abnormal auditory response in children with autism spectrum disorder / 中国针灸

OBJECTIVE@#To compare the effect of electroacupuncture at Tinghui (GB 2) and Benshen (GB 13) combined with routine rehabilitation training and simple routine rehabilitation training on abnormal auditory response in children with autism spectrum disorder (ASD).@*METHODS@#A total of 70 children with ASD were randomly divided into an observation group and a control group, 35 cases in each group. The control group was given routine rehabilitation training. On the basis of the treatment in the control group, the observation group was given electroacupuncture at Tinghui (GB 2) and Benshen (GB 13), disperse-dense wave, frequency in 2 Hz/10 Hz, 20 min each time, and 1 day off after 6 days of treatment. Both groups were treated for 12 weeks. The incidence of abnormal auditory response, the scores of childhood autism rating scale (CARS) and autism behavior checklist (ABC) were compared between the two groups before and after treatment.@*RESULTS@#After treatment, the total incidences of abnormal auditory response in the two groups and the incidences of dull and hypersensitive auditory response in the observation group were lower than those before treatment (P<0.05). The total incidence of abnormal auditory response and the incidence of hypersensitive auditory response in the observation group were lower than those in the control group (P<0.05). After treatment, CARS and ABC scores of both groups were lower than those before treatment (P<0.05), and those in the observation group were lower than the control group (P<0.05).@*CONCLUSION@#Electroacupuncture at Tinghui (GB 2) and Benshen (GB 13) combined with routine rehabilitation training can reduce the incidence of abnormal auditory response and improve the core symptoms in children with ASD, and the clinical effect is better than the simple routine rehabilitation training.

Electro-acupuncture promotes gut motility and alleviates functional constipation by regulating gut microbiota and increasing butyric acid generation in mice / 中西医结合学报

OBJECTIVE@#Abnormalities in the gut microbiota and intestinal short-chain fatty acid (SCFA) levels are implicated in the pathogenesis of functional constipation (FC). Electro-acupuncture (EA) has been shown to improve constipation-related symptoms and rebalance the gut microbiota. However, it is currently unknown whether the gut microbiota is a key mechanistic target for EA or how EA promotes gut motility by regulating the gut microbiota and SCFAs. Therefore, we assessed the effects of EA in FC mice and pseudo-germfree (PGF) mice to address these questions.@*METHODS@#Forty female Kunming mice were randomly separated into a normal control group (n = 8), an FC group (n = 8), an FC + EA group (n = 8), a PGF group (n = 8) and a PGF + EA group (n = 8). The FC group and FC + EA group were treated with diphenoxylate to establish the FC model; the PGF group and PGF + EA group were given an antibiotic cocktail to initiate the PGF model. After maintaining the model for 14 d, mice in the FC + EA and PGF + EA groups received EA stimulation at the ST25 and ST37 acupoints, once a day, 5 times per week, for 2 weeks. Fecal parameters and intestinal transit rate were calculated to assess the efficacy of EA on constipation and gastrointestinal motility. Colonic contents were used to quantify gut microbial diversity using 16S rRNA sequencing, and measure SCFA concentrations using gas chromatography-mass spectrometry.@*RESULTS@#EA significantly shortened the first black stool defecation time (P < 0.05) and increased the intestinal transit rate (P < 0.01), and fecal pellet number (P < 0.05), wet weight (P < 0.05) and water content (P < 0.01) over 8 h, compared with the FC group, showing that EA promoted gut motility and alleviated constipation. However, EA treatment did not reverse slow-transit colonic motility in PGF mice (P > 0.05), demonstrating that the gut microbiota may play a mechanistic role in the EA treatment of constipation. In addition, EA treatment restored the Firmicutes to Bacteroidetes ratio and significantly increased butyric acid generation in FC mice (P < 0.05), most likely due to the upregulation of Staphylococcaceae microorganisms (P < 0.01).@*CONCLUSION@#EA-mediated resolution of constipation occurs through rebalancing the gut microbiota and promoting butyric acid generation. Please cite this article as: Xu MM, Guo Y, Chen Y, Zhang W, Wang L, Li Y. Electro-acupuncture promotes gut motility and alleviates functional constipation by regulating gut microbiota and increasing butyric acid generation in mice. J Integr Med. 2023; Epub ahead of print.

The effect of electroacupuncture stimulation at acupoint Zusanli on the model of Staphylococcus aureus osteomyelitis / 中华创伤骨科杂志

Objective:To explore whether the electroacupunture stimulation (ES) at acupoint Zusanli (ST36) can inhibit the bone loss caused by Staphylococcus aureus (SA) infection and its mechanism in a model of SA osteomyelitis.Methods:Twelve male C57 BL/6 mice aged 10 to 12 weeks were randomly divided into 2 even groups ( n=6) for SA infection + ES or SA infection only. After ES at ST36 was conducted for 4 weeks in the model of SA osteomyelitis, samples were harvested from the femora and tibiae. Micro-CT reconstruction was performed to detect trabecular bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), connectivity density (Conn.Dn) to analyze changes in bone mass. Leptin receptor (LEPR) staining was performed to detect osteoblasts. Tartrate resistant acid phosphatase (TRAP) staining was used to detect the changes in osteoclasts. The changes in plasma inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA). Results:Micro-CT results showed that the BV/TV, Tb.N, Tb.Th, and Conn.Dn in the cancellous bone in the target areas in the SA + ES group were all higher than those in the SA group, LEPR immunofluorescence results indicated that the number of osteogenic precursor cells in the ES group was larger than that in the SA group, and serum ELISA indicated a decrease in inflammatory factors in the blood in the SA+ES group compared with the SA group. There were significant differences in the comparisons above ( P<0.05). There was no significant difference in the number of osteoclasts on the surface of trabecular bone between the 2 groups in TRAP staining. Conclusion:ES may slow down infectious bone destruction by inhibiting the inflammatory response induced by SA infection and by inducing aggregation and differentiation of mesenchymal stem cells into trabecular bone.

Effect of early intervention with electroacupuncture on behavior and gut microbiota in mice with post-traumatic stress disorder / 中华行为医学与脑科学杂志

Objective:To investigate the effect of early intervention with electroacupuncture (EA) on the gut microbiota in a mouse model of post-traumatic stress disorder(PTSD).Methods:Totally 32 C57BL/6 mice were randomly assigned to the following 4 groups ( n=8 for each group): Control group, EA group, PTSD group and PTSD+ EA group.After 7 days acclimation, mice in the PTSD group and PTSD+ EA group were subjected to modified single prolonged stress (mSPS). Mice in the EA group and PTSD+ EA group received EA (2/15 Hz, 1 mA, dilatational wave, 30 min/d) on "Baihui" for 7 days. Mice in the Control group and PTSD group received false stimulation (stimulated the same acupiont without electricity) for 7 days. Seven days after the last stimulation, elevated plus maze test and fear conditioning test were conducted to observe the effect of EA on PTSD-like behavior of mice. At the same time, feces of the mice were collected for gut microbiota detection by 16S rRNA sequencing.SPSS 19.0 was used for statistical analysis.One-way ANOVA was used for multiple group comparison and Bonferrani test was done for further pairwise comparision. Results:(1) There were statistically differences in the open arm activity time of the elevated plus maze test and the immobility time in contextual and cued fear conditioning test among the four groups ( F=6.93, 5.26, 14.51, all P<0.01). In the elevated plus maze test, mice in PTSD group ((60.17±15.52) s) showed significant less time in the open arms than mice in Control group((96.37±14.62) s) and PTSD+ EA group ((86.89±15.02) s) (both P<0.05). In the fear conditioning test, mice in PTSD group ((121.99±29.67) s, (130.82±29.11) s) showed significant increased immobility time both in contextual and cued fear conditioning tests than mice in Control group((74.50±26.65) s, (39.50±23.52) s) and PTSD+ EA group ((76.77±22.60) s, (102.17±3.39) s)(both P<0.05). (2) There were no significant differences among the four groups in the alpha diversity of gut microbiota ( F=0.79-2.45, all P>0.05). (3)Correlation analysis showed that 13 gut microbiotas were negatively correlated with the immobility time in contextual fear conditioning test, 2 gut microbiotas were positively correlated with it; 7 gut microbiotas were negatively correlated with the immobility time in cued fear conditioning test, 1 gut microbiota was positively correlated with it; 3 gut microbiotas were positively correlated with time spent in open arms of elevated plus maze test. Conclusion:Early intervention with EA can improve anxiety-fear like behaviors and gut microflora disorder in PTSD model mice.

Effects of electroacupuncture preconditioning of "Neiguan"(PC 6) on myocardial ischemia reperfusion injury via Akt/mTOR pathway / 国际中医中药杂志

Objective:To observe the effects of electroacupuncture preconditioning on the autophagy-related pathway protein kinase B (Akt)/mammalian target of rapamycin (mTOR) in myocardial tissue of rats with myocardial ischemia reperfusion injury (MIRI); To investigate the protective mechanism of "Neiguan"(PC 6) on myocardial injury.Methods:Totally 48 SD rats were randomly divided into blank group, sham-operation group, model group and Neiguan group ( n=12 in each group). The Neiguan group was applied to bilateral "Neiguan"(PC 6) by electroacupuncture for 30 min, once daily for consecutive 7 days before model replication. Except in the blank group, the MIRI model was established by ligation of the descending anterior branch of the left coronary artery in the rest groups after the intervention. The histomorphological changes in the myocardium of the rats were observed by HE staining, and the expression levels of Akt, phosphorylated Akt (p-Akt), mTOR and phosphorylated mTOR (p-mTOR) in the myocardium were measured by protein immunoblotting. The ratio of p-Akt/Akt and p-mTOR/mTOR was calculated. Results:In the blank group, the myocardial fibres were arranged regularly and neatly, and no inflammatory cell infiltration or haemorrhage was seen in the interstitium; in the sham-operation group, the arrangement of myocardial fibers was slightly irregular, no rupture was found, and a small amount of myocardial fiber gap was slightly enlarged; in the model group, the distribution of myocardial fibers was disordered, hypertrophic cardiomyocytes increased, some mitochondria were red and swollen or the outer membrane was ruptured, and inflammatory infiltration and hemorrhage were seen in the interstitium; the extent of myocardial lesions in the Neiguan group was less than that in the model group, with a small amount of interstitial hemorrhage and inflammatory cell infiltration. There was no statistical significance in the levels of Akt and mTOR in the myocardial tissues of the rats in each group ( P>0.05); compared with the sham-operation group, the levels of p-Akt, p-mTOR and p-Akt/Akt, p-mTOR/mTOR in the model group decreased ( P<0.01); compared with the model group, the levels of p-Akt, p-mTOR and p-Akt/Akt, p-mTOR/mTOR in the Neiguan group increased ( P<0.01). Conclusion:Electroacupuncture preconditioning may inhibit excessive autophagy by activating the Akt/mTOR pathway in cardiomyocytes of MIRI rats, thereby exerting a protective effect on the myocardium.

Research progress in the treatment of primary insomnia with electroacupuncture / 国际中医中药杂志

Electroacupuncture treatment of primary insomnia is widely used and with confirmed efficacy. The factors influencing the therapeutic effect of electroacupuncture include waveform selection, treatment frequency, stimulation intensity, stimulation time and acupoint selection. The mechanism of electroacupuncture in treating this disease mainly includes regulating the levels of excitatory neurotransmitters, γ-aminobutyric acid, melatonin and interleukin cytokines.

Mechanism underlying reduction of cerebral ischemia-reperfusion injury by electroacupuncture preconditioning in rats: relationship between cannabinoid type 1 receptor and NLRP3 inflammasome / 中华麻醉学杂志

Objective:To evaluate the relationship between cannabinoid receptor 1 (CB1R) and the NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) during the reduction of cerebral ischemia-reperfusion (I/R) injury by electroacupuncture (EA) preconditioning in rats.Methods:Forty SPF healthy male Sprague-Dawley rats, aged 7-9 weeks, weighing 250-280 g, were divided into 5 groups ( n=8 each) according to the random number table method: sham operation group (Sham group), cerebral I/R group (I/R group), EA preconditioning group (EA group), CB1R antagonist AM251+ EA preconditioning group (AM251+ EA group), and CB1R agonist WIN 55, 212-2 group (WIN group). Cerebral I/R was induced by middle cerebral artery occlusion (MCAO) in anesthetized animals. In EA group, EA preconditioning was performed, and the acupoint Baihui (GV20) was stimulated for 30 min with disperse-dense waves, the intensity of 1 mA and frequency of 2/15 Hz once a day for 5 consecutive days, and the model of cerebral I/R injury was developed at 24 h after the last EA. In AM251+ EA group, CB1R antagonist AM251 1 mg/kg was intraperitoneally injected at 30 min before each stimulation, and the remaining operations were the same as those previously described in EA group. CB1R agonist WIN 55, 212-2 1.5 mg/kg was intraperitoneally injected for 5 consecutive days, and the model of cerebral I/R injury was prepared at 24 h after the last injection in WIN group. Neurological behavior was assessed and scored at 3 days of reperfusion. Then the rats were sacrificed, and brains were removed, and the infarct volume was measured by TTC staining, and the tissues in the ischemic penumbra were extracted for determination of the expression of NLRP3, caspase-1 and interleukin-1bata (IL-1β) by Western blot. Results:Compared with Sham group, the percentage of cerebral infarct volume was significantly increased, the neurobehavioral score was decreased, and the expression of NLRP3, caspase-1 and IL-1β was up-regulated in I/R group ( P<0.05). Compared with I/R group, the percentage of cerebral infarct volume was significantly decreased, the neurobehavioral score was increased, and the expression of NLRP3, caspase-1 and IL-1β was down-regulated in EA and WIN groups ( P<0.05). Compared with EA group, the percentage of cerebral infarct volume was significantly increased, the neurobehavioral score was decreased, and the expression of NLRP3, caspase-1 and IL-1β was up-regulated in AM251+ EA group ( P<0.05). Conclusions:EA preconditioning may inhibit the activation of NLRP3 inflammasomes by activating CB1R, thus alleviating cerebral I/R injury in rats.

Electroacupuncture in Treatment of Acute Gastrointestinal Injury in Patients with Severe Traumatic Brain Injury: A Multicenter Randomized Controlled Trial / 中国结合医学杂志

OBJECTIVE@#To evaluate whether electroacupuncture (EA) would improve gastrointestinal function and clinical prognosis in patients with severe traumatic brain injury (TBI) complicocted by acute gastrointestinal injury (AGI).@*METHODS@#This multicenter, single-blind trial included patients with TBI and AGI admitted to 5 Chinese hospitals from September 2018 to December 2019. A total of 500 patients were randomized to the control or acupuncture groups using a random number table, 250 cases in each group. Patients in the control group received conventional treatment, including mannitol, nutritional support, epilepsy and infection prevention, and maintenance of water, electrolytes, and acid-base balance. While patients in the acupuncture group received EA intervention at bilateral Zusanli (ST 36), Shangjuxu (ST 37), Xiajuxu (ST 39), Tianshu (ST 25), and Zhongwan (RN 12) acupoints in addition to the conventional treatment, 30 min per time, twice daily, for 7 d. The primary endpoint was 28-d mortality. The secondary endpoints were serum levels of D-lactic acid (D-lac), diamine oxidase (DAO), lipopolysaccharide (LPS), motilin (MTL) and gastrin (GAS), intra-abdominal pressure (IAP), bowel sounds, abdominal circumference, AGI grade, scores of gastrointestinal failure (GIF), Glasgow Coma Scale (GCS), Acute Physiology and Chronic Health Evaluation (APACHE II), Sequential Organ Failure Assessment (SOFA), and Multiple Organ Dysfunction Syndrome (MODS), mechanical ventilation time, intense care unit (ICU) stay, and the incidence of hospital-acquired pneumonia.@*RESULTS@#The 28-d mortality in the acupuncture group was lower than that in the control group (22.80% vs. 33.20%, P<0.05). Compared with the control group, the acupuncture group at 7 d showed lower GIF, APACHE II, SOFA, MODS scores, D-lac, DAO, LPS, IAP, and abdominal circumference and higher GCS score, MTL, GAS, and bowel sound frequency (all P<0.05). In addition, the above indices showed simillar changes at 7 d compared with days 1 and 3 (all P<0.05) in the EA group.@*CONCLUSION@#Early EA can improve gastrointestinal function and clinical prognosis in patients with severe TBI complicated by AGI. (Registration No. ChiCTR2000032276).

Effect of electroacupuncture at "Neiguan" (PC 6) and "Zusanli" (ST 36) on gastrointestinal hormone in the antral tissue of rats with functional dyspepsia / 中国针灸

OBJECTIVES@#To observe the effects of electroacupuncture (EA) at "Neiguan" (PC 6) and "Zusanli"(ST 36) on the gastric emptying rate, the level of serotonin (5-HT) and the protein expression of motilin (MTL), ghrelin, substance P (SP) and vasoactive intestinal peptide (VIP) in the antral tissue of the rats with functional dyspepsia (FD) and explore the effect mechanism of EA in treatment of FD.@*METHODS@#A total of 21 SPF male SD rat pups were randomly divided into a normal group, a model group and an EA group, with 7 rats in each group. In the model group and the EA group, FD model was prepared by the gavage with 0.1% sucrose iodoacetamide solution combined with the modified small platform method. After the successful modeling, EA was applied to "Neiguan" (PC 6) and "Zusanli"(ST 36) in the rats of the EA group, with disperse-dense wave, 20 Hz/100 Hz in frequency, stimulated for 30 min, once daily, for 7 days consecutively. Before and after intervention, the general condition of the rats was observed in each group. After the completion of intervention, the gastric emptying rate was measured, the morphological changes of gastric antral tissue were observed using HE staining, the level of 5-HT was detected with ELISA method, and the protein expression of MTL, ghrelin, SP, and VIP was determined with Western blot method in the antral tissue of rats.@*RESULTS@#In the normal group, the rats were in a good mental state, with lustrous fur, flexible movement and the increase of food intake and body mass. In the model group, the rats were poor in mental state, lack of lustre in fur, preference for the body curled up, reduced activity and response; and a part of rats had loose stool, obviously enlarged gastric body and gastric food retention. In the EA group, the general condition of rats, e.g. the mental state, food intake and activity, were improved, the gastric body got smaller obviously and the gastric food retention was reduced when compared with the model group. The antral structure was intact, the glands were rich and no injury of the gastric mucosa was found, e.g. inflammatory reaction and edema in the rats of each group. Compared with the normal group, the gastric emptying rate was decreased (P<0.01), 5-HT level was increased (P<0.01), the protein expression of MTL and ghrelin was reduced (P<0.01) and that of VIP was elevated (P<0.01) in the rats of the model group. The gastric emptying rate was increased (P<0.01), 5-HT level was decreased (P<0.01), and the protein expression of MTL and ghrelin was elevated (P<0.05, P<0.01) in the rats of the EA group when compared with those in the model group.@*CONCLUSIONS@#Electroacupuncture at "Neiguan" (PC 6) and "Zusanli"(ST 36) may effectively relieve gastric dysfunction, strengthen gastric motility and promote gastric emptying so as to alleviate the symptoms of dyspepsia in FD rats, and its mechanism may be related to the regulation of gastrointestinal hormones in the antral tissue.

Effects of electroacupuncture on the glucose-lipid metabolism and the expression of ZAG and GLUT4 in the femoral quadriceps and adipose tissue in the rats with type 2 diabetes mellitus / 中国针灸

OBJECTIVES@#To observe the effects on the glucose-lipid metabolism and the expression of zinc-α2-glycoprotein (ZAG) and glucose transporter 4 (GLUT4) in the femoral quadriceps and adipose tissue after electroacupuncture (EA) at "Pishu" (BL 20), "Weiwanxiashu" (EX-B 3), "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) in the rats with diabetes mellitus type 2 (T2DM), so as to explore the effect mechanism of EA in treatment of T2DM.@*METHODS@#Twelve ZDF male rats were fed with high-sugar and high-fat fodder, Purina #5008 for 4 weeks to induce T2DM model. After successfully modeled, the rats were randomly divided into a model group and an EA group, with 6 rats in each one. Additionally, 6 ZL male rats of the same months age were collected as the blank group. The rats in the EA group were treated with EA at bilateral "Pishu" (BL 20), "Weiwanxiashu" (EX-B 3), "Zusanli" (ST 36) and "Sanyinjiao" (SP 6), with continuous wave, 15 Hz in frequency, and 2 mA in intensity. The electric stimulation lasted 20 min each time. EA was delivered once daily, 6 times a week for 4 weeks. Separately, the levels of fasting blood glucose (FBG) was measured before modeling, before and after intervention, and the body mass of each rat was weighted before and after intervention. After intervention, the levels of the total cholesterol (TC), triacylglycerol (TG) and free fatty acid (FFA) in serum were detected using enzyme colorimetric method; and the levels of the serum insulin (INS) and ZAG were detected by ELISA. Besides, the insulin sensitivity index (HOMA-ISI) was calculated. With Western blot technique adopted, the protein expressions of ZAG and GLUT4 in the femoral quadriceps and adipose tissue were determined.@*RESULTS@#After intervention, compared with the blank group, the levels of FBG and body mass, and the levels of serum TC, TG, FFA and INS increased (P<0.01), while HOMA-ISI decreased (P<0.01); the level of ZAG in the serum and the protein expressions of ZAG and GLUT4 in the femoral quadriceps and adipose tissue dropped (P<0.01) in the model group. In the EA group, compared with the model group, the levels of FBG and body mass, and the levels of serum TC, TG, FFA and INS were reduced (P<0.01), and HOMA-ISI increased (P<0.01); the level of ZAG in the serum and the protein expressions of ZAG and GLUT4 in the femoral quadriceps and adipose tissue increased (P<0.01, P<0.05).@*CONCLUSIONS@#Electroacupuncture can effectively regulate glucose-lipid metabolism, improve insulin resistance and sensitivity in the rats with T2DM, which is associated with the modulation of ZAG and GLUT4 expression in the skeletal muscle and adipose tissue.