Electroacupuncture Research Articles

Acupuncture attenuates myocardial ischemia/reperfusion injury-induced ferroptosis via the Nrf2/HO-1 pathway

AimsTo observe the effect of electro-acupuncture (EA) on cardiomyocytes ferroptosis induced by myocardial ischemia/reperfusion injury (MIRI) in mice and to investigate whether this effect occurs via the nuclear factor-E2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) signalling pathway.Materials and methodsFirstly, Fe2+ in the hearts and serum of mice from both the sham-operated (SO) group and MIRI group was measured to ascertain whether ferroptosis had occurred in the cardiomyocytes of mice in MIRI group. In the second phase, EA was administered, with sham acupuncture (SA) group as the comparator, to investigate the protective effects of EA on ferroptosis in MIRI cardiomyocytes and cardiac function. Additionally, we studied the levels of Nrf2 and HO-1 within the myocardium. In the third phase, Nrf2 inhibitor ML385 and agonist DMF were applied to observe the impact of inhibiting Nrf2 on the therapeutic efficacy of EA.ResultsCompared with SO group, MIRI group showed increased iron deposition, along with a significant decrease in Nrf2 and HO-1 levels. Compared with MIRI group, MIRI + EA group exhibited significantly improved cardiac function and reduced cardiac iron deposition, accompanied by increased Nrf2 and HO-1 levels. Furthermore, the therapeutic effect of MIRI + EA group was superior to that of MIRI + SA group. Administration of ML385 partially blocked the anti-ferroptotic and cardioprotective effects of EA, while EA treatment exhibited similar effects to dimethyl fumarate (DMF) intervention.ConclusionEA alleviates ferroptosis-induced damage in MIRI in mice via the Nrf2/HO-1 pathway, providing modern scientific evidence for the application of acupuncture in the treatment of cardiovascular diseases.

EXPRESS: Electroacupuncture intervention relieves pain by stimulating the STING/IFN-1 pathway in rat models of cancer-induced bone pain.

This study aimed to evaluate the effects of electroacupuncture (EA) on cancer-induced bone pain (CIBP) and investigate its interaction with the STING/IFN-1 pathway. A CIBP model was established in female rats. EA was administered for six consecutive days at bilateral L3-L5 Jia Ji points (EX-B2). EA-induced antinociception was evaluated through mechanical, thermal, and cold sensitivity assessments. EA significantly increased the paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) in rats with CIBP (P<0.01). In the spinal cord of CIBP model rats, western blot analysis demonstrated that the application of EA upregulated the expression of STING, IRF3, and IFNAR (P<0.05). The ELISA results indicated that EA significantly increased the expression of IFN-α (P<0.005) and IFN-β (P<0.01) and reduced the expression of TNF-α and IL-1β (P<0.05). Immunofluorescence analysis revealed that STING was predominantly localized in microglia, with a minimal presence in neuronal cells. Furthermore, intrathecal administration of the STING antagonist C-176 attenuated the analgesic effects of EA in CIBP (P<0.05). In particular, EA reduced pro-inflammatory cytokines and increased anti-inflammatory cytokines, whereas the STING agonist, despite its analgesic effects, aggravated the inflammatory response. Both EA and STING agonists were effective in alleviating pain in rats with CIBP, possibly through the activation of the STING/IFN-1 pathway. In addition, EA exhibited an anti-inflammatory effect, underscoring its therapeutic potential in the management of CIBP.

The mechanism of electroacupuncture-mediated improvement in Parkinson’s disease by inhibiting ferroptosis through activating the Nrf2/GPX4 signal pathway

IntroductionFerroptosis, an iron-dependent regulated cell death pathway, shares several features of Parkinson’s disease (PD) physiopathology, and efficient neuroprotective therapies are required to prevent DAergic neuron death initiated by ferroptosis. Electroacupuncture (EA), a treasure of Traditional Chinese Medicine, exerted therapeutic effects against PD to avoid the side effects of dopamine (DA)-based therapies. However, its underlying mechanisms still need to be fully understood.MethodsMPTP-induced PD mice were treated with EA to evaluate its neuroprotective effects. Behavioral assessments, histopathological analysis of DAergic neurons, and quantification of ferroptosis biomarkers-including malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), iron, glutathione (GSH), and mitochondrial integrity-were performed. Protein expression levels of SLC7A11, GPX4, ferritin heavy chain 1 (FTH1), and nuclear factor erythroid 2-related factor 2 (Nrf2) were analyzed via immunoblotting. To validate pathway specificity, the Nrf2 inhibitor trigonelline (AT) was co-administered with EA.ResultsEA treatment significantly mitigated MPTP-induced DAergic neuron loss and motor deficits. Mechanistically, EA suppressed ferroptosis by reducing lipid peroxidation and iron accumulation while restoring GSH levels. It upregulated ferroptosis-suppressive proteins SLC7A11, GPX4, FTH1, and Nrf2, alongside ameliorating mitochondrial dysfunction. Crucially, AT administration abolished EA’s protective effects, confirming Nrf2 pathway dependency.DiscussionThese findings demonstrate that EA exerts neuroprotection in PD by inhibiting ferroptosis through activation of the Nrf2/SLC7A11/FTH1/GPX4 signaling axis. This study not only elucidates a novel mechanism underlying EA’s efficacy in PD but also highlights ferroptosis modulation as a therapeutic strategy, bridging traditional medicine with molecular pathophysiology. This study has provided new ideas for exploring the mechanism of EA in PD treatment.

The Electroacupuncture Treatment Causes Earlier Recovery of Motor Nerve Dysfunction After Eccentric Exercise

PurposeThe purpose of this study was to investigate the effect of electroacupuncture (EA) on muscle and nerve function after eccentric exercise.MethodsFifteen young subjects were included in this study. All participants performed 100 eccentric contractions (ECCs) on the flexor pollicis brevis muscle of both hands and EA or rest intervention was randomly conducted on each hand. Maximal voluntary contraction torque (MVC), delayed onset muscle soreness (DOMS), range of motion (ROM), muscle stiffness, thickness, echo intensity, and motor nerve conduction velocity (MCV) were measured before, immediately after ECCs (only for MVC), immediately after intervention, day 1, 2, and 5 after ECCs. ResultsThe results showed that MVC, DOMS, ROM, muscle stiffness, muscle thickness, and echo intensity did not differ between the groups. However, MCV in the control group demonstrated a significant decrease immediately after ECCs and continued until 2 days post-exercise compared to the EA group (immediately after; p = 0.009, day 1; p < 0.001, day 2; p = 0.002). In addition, no significant change in MCV was observed in the EA group (before exercise; 59.8 ± 10.1 m/s, immediately after EA; 63.0 ± 11.2 m/s, day 1; 64.0 ± 7.7 m/s, day 2; 64.1 ± 7.2 m/s, and day 5; 63.4 ± 9.8 m/s). ConclusionEA intervention after strenuous exercise may cause earlier recovery from nerve dysfunction, although the detailed mechanisms need to be examined.

Electroacupuncture Inhibits Ferroptosis by Modulating Iron Metabolism and Oxidative Stress to Alleviate Cerebral Ischemia–Reperfusion Injury

Ischemic stroke (IS) is one of the leading causes of mortality and long-term disability worldwide. Electroacupuncture (EA) is commonly used in the treatment of IS, meaning that may reduce cerebral ischemia–reperfusion injury (CIRI). The middle cerebral artery occlusion/reperfusion (MCAO/R) rat models were created by the modified Zea Longa suture method. EA treatment was performed for 7 consecutive days at the acupoints Neiguan (PC6), Shuigou (GV26), and Sanyinjiao (SP6). The neurological function was assessed using the Zausinger six-point neurological deficiency score. The cerebral infarct volume was detected by 2,3,5-triphenyl tetrazolium chloride (TTC) staining. Hematoxylin and eosin (HE) staining was employed to observe the pathological changes in brain tissues. Prussian blue staining was employed to investigate iron deposition within the brain tissues. Transmission electron microscopy (TEM) was utilized to examine the morphological characteristics of mitochondria. Simultaneously, flow cytometry was utilized to detect the fluorescence intensity of reactive oxygen species (ROS). Assay kits were employed to measure the levels of Fe2+ and glutathione (GSH). Additionally, western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR) assays were performed to evaluate the expression levels of proteins associated with ferroptosis. Compared with the MCAO/R group, both the MCAO/R + EA and MCAO/R + DFO groups exhibited significant improvements in neurological function following cerebral ischemia–reperfusion (CIR), attenuated the pathological brain tissue injury, and reduced the cerebral infarct volume and iron deposition in brain tissue. Furthermore, both the MCAO/R + EA and MCAO/R + DFO groups displayed a marked reduction in mitochondrial injury. There was a substantial decrease in Fe2+ and ROS levels, accompanied by a notable increase in GSH level and glutathione peroxidase 4 (GPX4) activity. Compared with the MCAO/R group, the levels of ferroportin1 (FPN1) protein and mRNA expression were significantly increased in the MCAO/R + EA and MCAO/R + DFO groups, and the expression levels of transferrin (TF), transferrin receptor 1 (TFR1), divalent metal transporter 1 (DMT1) protein and mRNA, as well as ferritin (FER) protein, were significantly decreased. EA inhibits ferroptosis by modulating iron metabolism and oxidative stress to alleviate CIRI, exerting neuroprotective effects.Graphical

The safety and efficacy of acupuncture in treating nonalcoholic fatty liver disease: A systematic review and meta-analysis based on randomized controlled trials.

Nonalcoholic fatty liver disease (NAFLD), the most common chronic liver ailment globally, remains a significant concern. Acupuncture has been increasingly utilized for the treatment of NAFLD in recent years. However, current evidence is insufficient to support its efficacy and safety. The aim of this study was to perform a recent and thorough meta-analysis concerning the impacts of acupuncture on NAFLD. Randomized controlled trials (RCTs) investigating acupuncture for NAFLD were retrieved from 6 databases. The search encompassed the period from the inception of each database until February 28, 2024. We conducted literature screening based on predefined inclusion and exclusion criteria, resulting in the selection of 30 articles. Analysis of the data was carried out utilizing Review Manager 5.4 and Stata 15.1. The meta-analysis revealed a marked enhancement in the overall clinical effectiveness rate within the acupuncture group in comparison to the control group (OR = 3.36; 95% CI:2.62 to 4.31; P < .00001, I2 = 0%). Acupuncture exhibited positive impact on liver function recovery, blood lipid reduction, glucose regulation, improvement in insulin levels, antiliver fibrosis treatment, and imaging outcomes. Comprehensive analysis revealed that acupoint embedding (OR = 3.14; 95% CI: 2.113 to 4.62; P < .00001, I2 = 0%) demonstrated the most effective and stable therapeutic effect, followed by manual acupuncture (MA) (OR = 3.27; 95% CI: 2.19 to 4.90; P < .00001, I2 = 0%) and electroacupuncture (EA) (OR = 3.32; 95% CI: 1.69 to 6.52; P < .0005, I2 = 0%). In contrast, acupoint injection (AI) (OR = 5.74; 95% CI: 2.23 to 14.883; P < .0003, I2 = 0%) exhibited relatively modest effects, particularly in lipid reduction. No significant adverse reactions were observed with acupuncture treatments. Acupuncture has demonstrated safety and efficacy in the treatment of NAFLD, significantly improving hepatic function, lowering glucose and lipid levels, and mitigating liver fibrosis. Nevertheless, these findings necessitate validation through large-scale, rigorously designed randomized controlled trials.

The effect of acupuncture combined with hyperbaric oxygenation compared with hyperbaric oxygenation alone for patients with traumatic brain injury: a systematic review and meta-analysis

ObjectiveThis study aimed to evaluate the effects of acupuncture combined with hyperbaric oxygenation (HBO) compared with HBO alone in improving the disturbance of consciousness (DOC) of people with traumatic brain injury (TBI).MethodsThe Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed in this study. In accordance with the preestablished search strategy, all the literature was obtained from eight online databases. Following the stringent application of inclusion and exclusion criteria, two researchers conducted an independent extraction of valid data from eligible randomized controlled trials (RCTs). The risk of bias in each study was assessed using the Cochrane Risk of Bias 2.0 tool. The meta-analysis was conducted utilizing the RevMan software. Adverse events were determined based on data from each study assessing the safety of acupuncture treatment.ResultsA total of 11 RCTs with 896 participants were included in the analysis. Overall, the methodological quality of the RCTs encompassed within this meta-analysis was below standard. The pooled data demonstrated that acupuncture treatment combined with HBO was significantly superior to HBO alone, based on the Glasgow Coma Scale (GCS) scores [mean difference (MD) = 2.13, 95% confidence interval (CI): 1.64–2.62, p &amp;lt; 0.00001]. We also found that electroacupuncture (EA) combined with HBO improved GCS scores more than HBO alone in TBI patients (MD = 2.15, 95% CI: 1.95–2.36). The early intervention (MD = 3.09, 95% CI, 2.66–3.52) demonstrated significantly more significant improvement in GCS scores following combination therapy compared with the late intervention group (MD = 1.86, 95% CI, 1.47–2.25). Furthermore, compared with HBO alone, acupuncture combined with HBO significantly improved patients’ consciousness rate (CR) (RR = 2.26, 95% CI: 1.48–3.46). Statistical analysis also revealed that acupuncture combined with HBO improved the effective rate (ER) (RR = 1.47, 95% CI: 1.27–2.69). Additionally, no studies reported any significant adverse events.ConclusionCompared with HBO alone, acupuncture combined with HBO has a more substantial positive effect on GCS scores, AR, and ER in patients with TBI. However, given the limited availability of high-quality evidence and the dearth of RCTs in this area, the conclusions drawn herein warrant validation through additional research endeavours.Clinical trial registrationhttps://www.crd.york.ac.uk/PROSPERO/view/CRD42024576067, CRD42024576067.

Electroacupuncture Attenuates Cerebral Ischemia-Reperfusion Injury by Inhibiting Ferroptosis via the p53/SLC7A11 Pathway.

Acupuncture has demonstrated efficacy in treating post-stroke complications. Electroacupuncture (EA) ameliorates neurological outcomes in cerebral ischemia models, yet its mechanisms remain unclear. This study investigated EA's role in reducing cerebral ischemia-reperfusion injury (CIRI) in a rat model, focusing on ferroptosis. A CIRI model was established via the MCAO/R method. Rats were randomly assigned to five experimental groups: Sham, MCAO/R, MCAO/R + COTI-2, MCAO/R + EA and MCAO/R + COTI-2 + EA. We evaluated neurological function with Zausinger scoring. 2,3,5-Triphenyltetrazolium chloride (TTC) staining assessed infarct size, while haematoxylin-eosin (HE) staining examined neuronal damage. Transmission electron microscopy analysed mitochondrial changes associated with ferroptosis, and Perl staining measured iron levels in neurons. The biomarkers associated with ferroptosis, including glutathione (GSH), reactive oxygen species (ROS) and malondialdehyde (MDA), were measured. The expression of p53, SLC7A11 and GPX4 was assessed by qRT-PCR and Western blot. EA enhanced neurological function, reduced the infarct size, alleviated excessive serum iron accumulation, increased antioxidant markers (GSH, GPX4) and decreased lipid peroxidation levels (ROS, MDA), attenuating lipid peroxidation. Additionally, it reversed mitochondrial morphological changes associated with ferroptosis. qRT-PCR and Western blot analyses revealed that EA downregulated p53 expression while upregulating SLC7A11 and GPX4 expression. In summary, ferroptosis was activated after CIRI, and EA ameliorated neurological deficits in cerebral ischemia models by modulating the p53/SLC7A11 axis to counteract oxidative stress-induced ferroptosis, ultimately providing neuroprotective benefits.

Electroacupuncture ameliorates cognitive impairment and white matter injury in vascular dementia rats via activating HIF-1α/VEGF/VEGFR2 pathway.

Electroacupuncture ameliorates cognitive impairment and white matter injury in vascular dementia rats via activating HIF-1α/VEGF/VEGFR2 pathway.

Effect of electrical stimulation in the treatment on patients with foot drop after stroke: a systematic review and network meta-analysis.

Effect of electrical stimulation in the treatment on patients with foot drop after stroke: a systematic review and network meta-analysis.

PTK2B inhibitor PF-431396 inhibits inflammatory response and apoptosis of ovarian granulosa cells by targeting AKT1 phosphorylation in premature ovarian insufficiency.

PTK2B inhibitor PF-431396 inhibits inflammatory response and apoptosis of ovarian granulosa cells by targeting AKT1 phosphorylation in premature ovarian insufficiency.

Effect of electroacupuncture at Zusanli reduced Olanzapine-induced lipid disturbances in mice via potential liver-brain interaction.

Olanzapine-induced weight gain and disruptions in lipid metabolism represent multifactorial processes that compromise treatment adherence in schizophrenia and contribute to substantial clinical challenges. While acupuncture has been reported to promote weight loss, the neural substrates mediating its metabolic effects in olanzapine-induced models remain largely undefined. In this study, we evaluated the efficacy of electroacupuncture (EA) at the Zusanli (ST36) acupoint in ameliorating olanzapine-induced obesity and dyslipidemia, and investigated the underlying liver-brain regulatory mechanisms. We further assessed the effects of EA through Reverse Transcription Polymerase Chain Reaction (RT-PCR), ELISA, and lipidomics analysis. Comprehensive evaluations were conducted using three-dimensional fine behavioral analysis and metabolic cages. Additionally, ultrasound functional imaging (fUS), retrograde tracing with pseudorabies virus 152 (PRV152), and c-Fos mapping techniques were employed to investigate liver-brain interactions. EA at ST36 effectively reduced body weight, reversed Olanzapine-induced adipocyte hypertrophy and hepatocyte damage, and restored disturbances in free fatty acids (FFA) and liver lipid metabolism. These effects also involved the restoration of olanzapine-induced disruptions in respiratory exchange rate (RER) during the dark cycle. We further explored the potential regulatory mechanisms, and we successfully demonstrated that EA stimulation significantly deactivated Agouti-related protein (AGRP)-expressing neurons in the lateral septal nucleus (LS), nucleus accumbens (NAc), and pallidum, suggesting that AGRP+ neurons in these brain regions are involved in the liver-brain interaction during EA treatment. These findings highlight the critical role of metabolism-regulating neurons in the acupuncture-mediated liver-brain axis and provide insights into the mechanisms of information integration from the brain to the liver.

An analysis of electroacupuncture as adjunctive treatment for insomnia: a new perspective targeting GABA-mediated microbiome-gut-brain axis

IntroductionInsomnia is a prevalent psychiatric disorder that significantly impacts mental and physical health. Emerging evidence suggests that gut microbiota, particularly γ-aminobutyric acid (GABA) producing bacteria, plays a critical role in the microbiome-gut-brain axis (MGBA), influencing sleep and mental states. Electroacupuncture (EA) has been shown to have favorable anti-insomnia effects. This research protocol outlines a randomized controlled trial (RCT) designed to investigate the efficacy of EA in modulating GABA levels via the MGBA to alleviate insomnia.Methods and analysisThe study will use glutamate (Glu) concentrations of p-chlorophenylalanine (PCPA) -induced insomnia rat model to examine whether EA at specific acupoints can increase GABA levels in the brain and plasma by enhancing the abundance of GABA-producing gut bacteria. A second experiment will transplant stool from insomnia rats into germ-free mice to evaluate the causal role of microbiota in insomnia pathology. Primary outcome measures include changes in GABA and Glu levels, data about the open field test, and gut microbiota composition.DiscussionThis study will provide insights into the therapeutic mechanisms of EA targeting the MGBA in the treatment of insomnia and contribute to the development of novel therapeutic strategies.Ethics and disseminationThis study has been approved by the Laboratory Animal Center of Xiamen University (No. XMULAC20220100). The study findings will be submitted for publication in peer-reviewed academic journals.

BNSTGABA Neurons Regulate Autophagy to Alleviate Depression with Gastric Dysfunction Symptoms.

BNSTGABA Neurons Regulate Autophagy to Alleviate Depression with Gastric Dysfunction Symptoms.

Effect of electroacupuncture on the primary input level of dorsal root ganglion neurons under different stimuli observed by in vivo calcium imaging

To observe the effect of electroacupuncture (EA) on the neuronal activity of primary afferent dorsal root ganglion (DRG) caused by different mechanical stimuli such as brush, clamp [clamping hind paw or "Zusanli" (ST36) acupoint] or stimulation at different temperatures (43 ℃, 51 ℃, 4 ℃), so as to explore the interaction between EA signals and different primary afferent signals at the DRG level. The strength threshold of activated A fiber was determined by electrophysiological technique. DRG neurons were labeled by intrathecal injection of AAV9 virus, and the response of DRG neurons to different primary afferent stimuli was observed and recorded by calcium imaging. EA (0.5 mA, 0.2 ms, 2 Hz) was applied to "Zusanli" (ST36) on the right side for 10 min. The changes of the number, percentage of neurons of different sizes and fluorescence intensity of DRG neurons activated under different stimulation before and after EA were compared. Clamping stimulation, brush stimulation, or different temperature stimulation could all activate DRG neurons of large, medium, and small classes. EA at ST36 could significantly decrease the percentage of activated small neurons and large neurons in DRG induced by clamping the hind paw (P<0.05), and reduce the fluorescence intensity of neurons with different diameters (P<0.000 1), suggesting that EA could suppress the neuronal activity induced by noxious mechanical stimulation of the hind paw. However, EA at ST36 had no significant effect on the percentage of activated DRG neurons and fluorescence intensity caused by clamping the anterior tibial muscle. Meanwhile, EA at ST36 could significantly reduce the percentage of activated neuron in DRG induced by brush stimulation (P<0.05), and reduce the fluorescence intensity of neurons with different diameters (P<0.000 1). In temperature stimulation, EA could significantly inhibit the fluorescence intensity of DRG neurons under 4 °C cold stimulation (P<0.01), but had no significant effect on the percentage and fluorescence intensity of DRG neurons activated by 43 °C and 51 °C. EA at ST36 can inhibit the nociceptive mechanical input of the hind paw and the input of 4 °C cold stimulation.

Electroacupuncture improves synaptic plasticity and cognitive dysfunction via down-regulating HDAC3 in mice of Alzheimer's disease

To observe the effect of electroacupuncture(EA) on histone deacetylase 3 (HDAC3), synaptic plasticity and N-methyl-D-aspartate (NMDA) receptors in the hippocampus of mice with Alzheimer's disease(AD), so as to explore the underlying mechanism of EA in treatment of AD. 5XFAD mice were randomly divided into EA group, model group and sham-acupuncture group, with 13 mice in both the EA group and the model group, and 7 mice in the sham-acupuncture group. Thirteen wild-type mice from the same litter were taken as the normal control group. The mice in the EA group received EA at "Baihui" (GV20)and "Dazhui" (GV14) for 15 min once daily, 6 times a week for 4 weeks. The mice in the sham-acupuncture group received sham EA, i.e., the needle was inserted into the rubber clay which was placed on the surface of the corresponding acupoints. The novel object recognition(NOR), Y-maze and Morris water maze(MWM) tests were used to observe the cognitive functions of mice. Electrophysiological technique was used to detect long-term potentiation (LTP) of the hippocampal neurons and Western blot was used to detect the relative expressions of HDAC3 and NMDAR-related receptors (NMDAR1, NMDAR2A, NMDAR2B) in the hippocampus. Compared with the normal control group, 5XFAD mice in the model group showed decreased(P<0.01, P<0.05) preference index for new object recognition, alternative arm ratio (AAR), number of times crossing the original platform, percentage of time and distance traveled in the target quadrant, NMDAR2B, NMDAR2A and NMDAR1 protein expression levels, with prolonged(P<0.01) escape latency, and increased (P<0.05) protein relative expression of HDAC3. At the same time, with high-frequency stimulation, the slope of fEPSP was decreased(P<0.01, P<0.05)in the 5XFAD mice. After EA intervention, comparison between the EA and the model groups revealed that, the preference index for new object recognition, AAR were increased (P<0.01, P<0.05) in the EA group, the escape latency was shortened (P<0.05), and the number of times crossing the platform, percentage of time and distance traveled in the target quadrant, the slope of fEPSP, and the protein relative expressions of NMDAR2B, NMDAR2A and NMDAR1 in the hippocampus were increased (P<0.01, P<0.05), while the protein relative expression of HDAC3 decreased (P<0.01). Compared with sham-acupuncture group, the above indexes improved to different degree in the EA group (P<0.01, P<0.05). EA of GV20 and GV14 can restore the impaired LTP and improve the cognitive impairment, which may be related to increasing the expressions of NMDA-related receptor proteins and down-regulating the expression of HDAC3 in the hippocampus of 5XFAD mice.

Effects of electroacupuncture on cortactin and cytoskeletal stability in lung tissue of mice with chronic obstructive pulmonary disease

To observe the effects of electroacupuncture (EA) at "Zusanli" (ST36) and "Feishu" (BL13) points on Cortactin and cytoskeletal stability of pulmonary endothelial cells in mice with chronic obstructive pulmonary disease (COPD), and to explore the mechanism by which EA alleviates inflammatory damage in COPD. The C57BL/6 mice were randomly divided into 5 groups:normal group, model group, EA group, Cortactin small interfering RNA (si-Cortactin) group, and si-Cortactin + EA group, with 10 mice in each group. A COPD model was established through 12 weeks of cigarette exposure. Nasal instillation of Cortactin siRNA was performed 24 h before EA treatment. EA at bilateral ST36 and BL13 with each EA session lasted for 30 min, conducted once every other day for a continuous period of 2 weeks. Pulmonary function was assessed using a small animal lung function analyzer and HE staining was used to observe lung histopathology. ELISA was employed to measure the contents of tumor necrosis factor-α (TNF-α) and Caspase-3 in bronchoalveolar lavage fluid (BALF) and serum. TUNEL assay was used to detect apoptosis of pulmonary endothelial cells, and double immunofluorescence staining was performed to assess cytoskeletal stability in pulmonary endothelial cells. The expression levels of Caspase-3 and Cortactin mRNA in lung tissue were evaluated using qPCR, while Western blot was used to measure the protein expression levels of Caspase-3 and Cortactin in lung tissues. Compared to the normal group, the model group exhibited decreases in forced vital capacity (FVC), forced expiratory volume in 0.05 s (FEV0.05), forced expiratory volume in 0.1 s (FEV0.1), FEV0.05/FVC, and FEV0.1/FVC (P<0.001). Lung tissue showed severe inflammatory infiltration, accompanied by increased contents of TNF-α and Caspase-3 in serum and BALF (P<0.001). The expression levels of Caspase-3 mRNA and protein were elevated, while Cortactin mRNA and protein levels were reduced in lung tissue (P<0.001). There was significant apoptosis of pulmonary endothelial cells and disruption of the cytoskeletal structure (P<0.001). After treatment, the EA group demonstrated marked improvement in these parameters compared to the model group (P<0.001, P<0.01, P<0.05). Following intervention with si-Cortactin, the indicators in the si-Cortactin group worsened further (P<0.01, P<0.001, P<0.05). The si-Cortactin + EA group showed improvements in these parameters relative to the si-Cortactin group (P<0.001, P<0.05, P<0.01). EA can alleviate inflammatory pathological damage in the lung tissues of mice with COPD, potentially through up-regulating Cortactin, stabilizing the cytoskeleton of pulmonary endothelial cells, and inhibiting apoptosis.

Electroacupuncture intervention promotes browning of white adipose tissue by regulating LKB1/AMPK pathway in obese mice

To observe the effect of electroacupuncture (EA) on hypothalamic liver kinase B1 (LKB1)/AMP-activated protein kinase (AMPK) signaling pathway, sympathetic nerve activity and browning of white adipose tissue (WAT) in obese mice, so as to explore the possible central mechanisms of EA in improving obesity. C57BL/6J male mice were divided into normal group (n=8) and high fat diet group (n=30) randomly, and fed by normal diet and high fat diet for 8 weeks, respectively. In the high fat diet group, mice with body weight over 20% heavier than that of the normal group were considered as obese mice. The obese mice were divided into model and EA groups (n=8 in each group). Mice in the EA group were treated with EA (2 Hz/15 Hz, 1 mA, 3 mm needling depth) at "Zusanli" (ST36) and "Tianshu" (ST25) for 30 min, 5 times a week for 4 weeks. The mice's body weight and length were measured for calculating the Lee's index. The contents of serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and noradrenaline (NE) were detected by ELISA. The weight of abdominal white adipose tissue was weighed, and the morphological changes of abdominal adipose tissue were observed by HE staining. The mRNA and protein expression levels of LKB1 and AMPK in hypothalamus, peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), uncoupling protein-1 (UCP1), peroxisome proliferator-activated receptor γ (PPAR-γ) and β3-adrenergic receptor (β3-AR) in abdominal white adipose tissue were detected by real-time fluorescence quantitative PCR and Western blot, respectively. Compared with the normal group, the body weight, Lee's index, serum contents of TC, TG and LDL-C, abdominal WAT mass, hypothalamus LKB1, AMPK mRNA and protein expression were increased (P<0.01), while serum HDL-C and NE contents, mRNA and protein expression of β3-AR, PGC-1α, UCP1 and PPAR-γ in adipose tissue were decreased (P<0.01) in the model group. The diameter of adipocytes in abdominal WAT was significantly increased, and the number of cells per unit area was decreased in the model group. Following EA intervention, the body weight, Lee's index, serum contents of TC, TG and LDL-C, abdominal WAT mass, hypothalamus LKB1, AMPK mRNA and protein expression were decreased (P<0.01), while the serum HDL-C and NE contents, mRNA and protein expression of β3-AR, PGC-1α, UCP1 and PPAR-γ were increased (P<0.01). The diameter of adipocytes in abdominal WAT was decreased and the number of cells per unit area was increased in the EA group. EA can reduce the body weight and Lee's index in obese mice, regulate blood lipids, and reduce the weight of abdominal WAT, which may be closely related to its effect in regulating the hypothalamic LKB1/AMPK signaling pathway, activating the adipose tissue sympathetic nerve activity, and promoting the browning of WAT.

Effect of electroacupuncture of "Jiaji"(EX-B2) on autophagy and apoptosis of nucleus pulposus cells in degenerative intervertebral disc rabbits based on PI3K/AKT/mTOR pathway

To observe the effect of electroacupuncture (EA) of bilateral "Jiaji" (EX-B2) on autophagy, apoptosis and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway of nucleus pulposus cells in rabbits with intervertebral disk degeneration (IVDD), so as to explore its possible mechanisms underlying improvement of IVDD. A total of 18 male New Zealand rabbits were randomly and equally divided into sham operation, model and EA groups. The IVDD model was established by axial disc compression, and the modeling effect was confirmed by magnetic resonance imaging (MRI). EA (2 Hz/15 Hz, 1 mA) was applied to bilateral "Jiaji" (EX-B2) of L4-L5 for 20 min, once daily, 6 times a week for 4 weeks. The general condition and body weight change in each group were recorded. The morphology of nucleus pulposus was observed after H.E. staining. The ultrastructure of nucleus pulposus cells was observed by using transmission electron microscope. The apoptosis of nucleus pulposus cells was detected by terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) assay. The expressions of Bax, Bcl-2, Beclin1, light chain 3 (LC3), autophagy-related protein 5 (Atg5), p62 and PI3K/AKT/mTOR signaling pathway proteins were detected by Western blot. Compared with the sham operation group, the general condition of IVDD rabbits was poorer, with decreased MRI signal of intervertebral disc, destruction and necrosis of nucleus pulposus cells, damage of nucleus pulposus cell matrix, and blurred organelles, and the apoptosis rate, expression levels of Bax, Beclin1 and Atg5 proteins and the ratio of LC3- Ⅱ/LC3- Ⅰ were significantly increased (P<0.01, P<0.05), while the body weight, expressions of Bcl-2 and p62, p-PI3K/PI3K, p-AKT/AKT and p-mTOR /mTOR considerably decreased (P<0.01) in the model group. In contrast to the model group, the general condition of the rabbits was improved after EA, with enhanced MRI signal of intervertebral disc, fewer mitochondrial autophagosomes, slightly normal mitochondrial construction, etc., and modeling-induced increase of apoptosis rate, the expression levels of Bax, Beclin1 and Atg5 proteins and the ratio of LC3-Ⅱ/LC3- Ⅰ , and decrease of body weight, the expression levels of Bcl-2, p62, and p-PI3K/PI3K, p-AKT/AKT and p-mTOR /mTOR were all reversed in the EA group (P<0.01, P<0.05). EA stimulation of EX-B2 can delay IVDD by regulating apoptosis and autophagy of nucleus pulposus cells in the intervertebral disc in IVDD rabbits, which may be related to its function in activating PI3K/AKT/mTOR signaling pathway.

Effect of acupotomy on lipid peroxidation of chondrocytes in rabbits with knee osteoarthritis

To observe the effect of acupotomy on expression of long-chain acyl-CoA synthetase 4 (ACSL4), lysophosphatidylcholine acyltransferase 3 (LPCAT3), malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) in knee cartilage of rabbits with knee osteoarthritis (KOA), so as to explore whether acupotomy can treat KOA by inhibiting lipid peroxidation of chondrocytes. Six-month-old New Zealand male rabbits were randomly divided into control, model, acupotomy and electroacupuncture (EA) groups, with 7 rabbits in each group. The KOA model was established by left hindlimb straightening fixation according to Videman method. In the acupotomy group, the high stress points, such as tendons attachment points of the vastus medialis, vastus lateralis, rectus femoris, biceps femoris and pes anserine bursa of the left hind limb knee joint were selected for acupotomy lysis treatment, once a week for 3 weeks. In the EA group, EA (2 Hz/100 Hz, 3 mA) was applied to "Neixiyan" (EX-LE4), "Dubi" (ST35), "Xuehai" (SP10) and "Liangqiu" (ST34) of the left hind limb of rabbits for 20 min, once every other day for 3 weeks. Before and after the treatment, Lequesne MG score and passive range of motion (PROM) of knee joint were detected in each group. After the treatment, safranin O-fast green staining was used to observe the morphological changes of rabbit knee cartilage and Mankin scoring was evaluated. Immunohistochemistry method was used to detect the location and positive expression of ACSL4. Western blot was used to detect the protein expression levels of ACSL4 and LPCAT3, and TBA and ELISA methods were used to detect the expression levels of MDA and 4-HNE in the affected knee cartilage, respectively. Compared to the control group, the Lequesne MG score was significantly increased (P<0.01), while PROM was significantly decreased (P<0.01) in the model group;after the intervention and compared to the control group, the model group exhibited significant increase in Mankin score, enhancement in the positive expression of ACSL4 in knee cartilage (P<0.01), along with significant elevation in ACSL4 and LPCAT3 protein expression levels (P<0.01), as well as elevated contents of MDA and 4-HNE (P<0.01). Compared to the model group, both the acupotomy group and EA group showed significant reductions in Lequesne MG score (P<0.01) and increases in PROM (P<0.01). Additionally, there were marked decreases in Mankin score and the positive expression of ACSL4 in knee cartilage (P<0.01), as well as significant decreases in the protein expression levels of ACSL4 and LPCAT3 (P<0.01). Moreover, the contents of MDA and 4-HNE decreased significantly (P<0.01, P<0.05). Compared to the EA group, the acupotomy group exhibited reduced Lequesne MG score (P<0.01), increased PROM (P<0.01), significant reductions in ACSL4 and LPCAT3 protein expression levels (P<0.01), and lowered content of MDA (P<0.01). Post-intervention, both groups showed significant reductions in Lequesne MG score (P<0.01) and significant increases in PROM (P<0.01) compared to their respective pre-intervention data. Acupotomy intervention can down-regulate the levels of ACSL4, LPCAT3, MDA and 4-HNE in knee cartilage, improve the behavior of affected limbs, indicating that acupotomy therapy may inhibit lipid peroxidation in chondrocytes and slow down cartilage injury to treat KOA.