PurposeSemaglutide is effective for type 2 diabetes mellitus (T2DM) but limited by dose-dependent gastrointestinal adverse effects. As electroacupuncture (EA) possesses glucose-lowering effects, this study investigates the therapeutic potential and mechanism of combining EA with semaglutide to facilitate dose reduction while maintaining efficacy.MethodsMale db/db mice were divided into: the model group(T2DM), the high/normal/low-dose semaglutide group (SH, SN, SL), the normal/low-dose semaglutide + EA at ST25 group (ST25N, ST25L), and the EA at ST25 group (ST25). The db/m mice served as the control group (CON). Blood glucose regulation was assessed via random/fasting blood glucose and oral glucose tolerance test (OGTT). To evaluate glycemic control and lipid profiles, HbA1c and insulin levels were quantified via ELISA, and T-CHO, TG, LDL-C, and HDL-C via biochemical kits. GLP-1R expression and β-cell abundance were assessed by immunofluorescence, and pancreatic apoptosis by TUNEL staining. PKA, pPKA, Bcl-2, Bax, and GLP-1R were analyzed by Western blot. Male Sprague–Dawley (SD) rats were separated into 2 groups: the normal-dose semaglutide rats group (RSN) and the normal-dose semaglutide + EA at ST25 rats group (RST25N). LC-MS/MS was employed to measure the concentrations of semaglutide in the plasma and pancreas, thereby evaluating the influence of EA.ResultsEA enhances the effects of semaglutide on improving glycemic control and lipid metabolism, with significant reductions in random/fasting blood glucose, HbA1c, TCHO, TG and LDL-C levels, improvements in OGTT, insulin and HDL-C levels, and longer duration of controlled blood glucose within the target range, Furthermore, EA combined with semaglutide showed a marked increase in pancreatic β-cells, with the increased expression of pancreatic GLP-1R, PKA, pPKA and Bcl-2, the reduced expression of Bax, increased concentration of semaglutide in plasma and pancreas.ConclusionEA combined with normal- or low-dose semaglutide achieved glycemic control comparable to, or even better than, high-dose semaglutide alone, and its mechanism may be related to its enhancement of GLP-1R to inhibit β-cell apoptosis and the increased concentration of semaglutide in plasma and pancreas. EA thus has great potential to reduce the dose of semaglutide to alleviate its side effects while ensuring its clinical efficacy.

Myocardial infarction (MI) triggers maladaptive sympathetic overactivation, which acts as a key driver of adverse cardiac remodeling and progression to heart failure. Electroacupuncture (EA) at the Neiguan (PC6) acupoint exhibits reliable cardioprotective effects, yet its central neural mechanisms remain incompletely clarified. This study investigated whether EA at PC6 improves post-MI outcomes by modulating the hypothalamic paraventricular nucleus-superior cervical ganglion (PVN-SCG) sympathetic axis. MI was surgically induced by permanent ligation of the left anterior descending coronary artery in mice, followed by consecutive EA treatment at PC6. Cardiac function was assessed by echocardiography, while sympathetic activity was evaluated via heart rate variability (HRV) analysis and norepinephrine levels of serum and heart. Myocardial fibrosis and key molecular markers within the PVN-SCG axis were systematically analyzed. In this mice MI model, EA at PC6 significantly improved cardiac function as indicated by ejection fraction and attenuated ventricular dilation. Mechanistically, EA suppressed systemic sympathetic activity, as evidenced by decreased serum and cardiac norepinephrine levels as well as normalized HRV parameters. Notably, EA markedly inhibited neuronal hyperactivity (c-fos expression) in both the PVN and SCG. Peripherally, EA mitigated cardiac sympathetic remodeling by downregulating tyrosine hydroxylase (TH) and synaptophysin (SYN) expression, accompanied by reduced myocardial fibrosis and collagen deposition. These findings demonstrate that EA at PC6 exerts s potent cardioprotection against post-MI injury by specifically inhibiting sympathetic hyperactivation at both central (PVN-SCG axis) and peripheral levels. This study identifies a novel neuromodulatory mechanism underlying EA therapy and supports PC6 EA as a promising non-pharmacological strategy for post-MI cardiac protection.

BACKGROUND:Acupuncture at different traditional acupuncture point locations activates different autonomic pathways and modulates systemic inflammation. Electroacupuncture preconditioning (EAP) at PC6 or ST36 has been demonstrated to be cardioprotective during acute myocardial ischemia (MI) injury, but the mechanism remains unclear. As inflammation is crucial for repair post-MI, we aimed to investigate whether the protective effect of EAP at PC6 or ST36 is related to the suppression of inflammation and whether autonomic pathways are involved. METHODS:Mice underwent ligation of the left anterior descending (LAD) coronary artery to generate model MI after 3 days of EAP at PC6 or ST36. Echocardiography and triphenyltetrazolium chloride (TTC) staining were used to evaluate cardiac function and myocardial infarction area. Cardiac troponin (cTn)I and norepinephrine (NE) were measured by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was employed to determine macrophage M1/M2 phenotypic polarization, and the levels of pro- and anti-inflammatory cytokines in cardiac tissue were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). In addition, unilateral cervical vagotomy and chemical sympathetic blockade were employed to clarify specific autonomic pathways. RESULTS:EAP at PC6 or ST36 preserved cardiac function, reduced myocardial infarct size, decreased serum cTnI levels, promoted M2 macrophage polarization and decreased the cardiac pro-inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-18 and nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3). Compared to EAP at ST36, EAP at PC6 yielded a better cardioprotective effect and regulated the anti-inflammatory cytokine transforming growth factor (TGF)-β. Moreover, sympathetic nerves were involved in the protective effect of EAP at PC6, while the vagus nerve was necessary for EAP at ST36. CONCLUSION:Our findings suggest that EAP at PC6 or ST36 activates different autonomic pathways and modulates M2 macrophage polarization, resulting in the suppression of myocardial inflammation.

Ischemic stroke seriously endangers both the health and quality of life of patients. The gut microbiota, which plays a crucial role in modulating communication between the gut and the nervous system, has emerged as a promising target for therapeutic interventions in stroke. Electroacupuncture (EA), which is associated with intestinal immunity, has been proven to exert significant beneficial effects in ischemic stroke, but its exact mechanism remains unclear. In this study, we investigated the regulatory mechanism of EA on the microbiome-gut-brain axis following ischemic stroke. In rat models of ischemic stroke, EA treatment significantly reduced cerebral infarct volume and neuronal damage following cerebral ischemia-reperfusion injury, and also modulated the composition, diversity, and taxonomic distribution of the gut microbiota. Fecal microbiota transplantation from EA-treated donors significantly reduced cerebral infarct volume and neuronal damage in the ischemic hemisphere of recipient mice, and likewise upregulated Treg cell expression to suppress immune-inflammatory responses in the brain. These results indicate that, through modulation of the gut microbiota, which in turn regulates Treg-mediated immune-inflammatory responses, EA ameliorates cerebral ischemic injury to thereby improve the prognosis of ischemic stroke patients. This study provides new perspectives on the efficacy of EA in the treatment of ischemic stroke.

Ethnopharmacological relevance: Zuo Gui Wan (ZGW) is a well-known traditional Chinese medicine decoction used for approximately 400 years to treat age-related degenerative conditions, including cognitive impairment in older adults, osteoporosis, and general aging. However, the mechanism of action for ZGW remains unclear. Aims of the study: This study aims to investigate the efficacy of ZGW in improving cognitive function in Alzheimer's disease (AD) animal models and to explore the underlying mechanisms, presenting a novel perspective in the field. Materials and methods: Six-month-old male APP/PS1 mice were divided into three groups that received either metformin (200 mg/kg daily) or ZGW (6 and 12 g/kg daily). High-performance liquid chromatography was conducted for ZGW's quality control. Cognitive function was assessed using the Morris water maze test. Neuronal loss, synaptic plasticity, and beta-amyloid (A beta) deposition were evaluated through Western blot or immunofluorescence staining. The underlying molecular mechanisms were investigated using ELISA, Western blot, qRT-PCR, co-immunoprecipitation assay, ATP assay, and cytochrome c oxidase assay. Results: ZGW, administered in both low and high doses, significantly enhanced cognitive performance, notably decreased neuronal loss and A beta deposition, and reduced levels of A beta 1-40/42. It also inhibited excessive mitochondrial division primarily by suppressing phosphorylated dynamin-related protein 1 (Drp1), especially at high doses of ZGW. Co-immunoprecipitation experiments further confirmed that ZGW inhibited the interaction between A beta and p-Drp1. Furthermore, similar to the effects of the AMP-activated Protein Kinase (AMPK) activator metformin, ZGW led to a marked increase in the mitochondrial DNA copy number and upregulated the AMPK/ PGC-1 alpha/NRF1/TFAM pathway. Improvements in mitochondrial function were evident from the increased ATP production, elevated expression of superoxide dismutase 2, and upregulated cytochrome c oxidase activity. Additionally, the excess byproduct of reactive oxygen species, 4-hydroxy-2-nonenal, decreased in the group treated with ZGW. Conclusion: This study provides compelling evidence that ZGW improves cognitive impairment in APP/PS1 mice by activating AMPK/PGC-1 alpha-regulated mitochondrial bioenergetics and inhibiting A beta-induced mitochondrial fragmentation, highlighting its potential as an effective therapeutic strategy for AD.

ABSTRACT Background Astragaloside IV (AS‐IV) exhibits therapeutic potential in central nervous system (CNS) disorders. However, its brain bioavailability is restricted by P‐glycoprotein (P‐gp)‐mediated efflux at the blood–brain barrier (BBB). This study aims to investigate whether electroacupuncture (EA) can enhance the cerebral delivery of AS‐IV by inhibiting nuclear factor‐kappa B (NF‐κB) nuclear translocation to downregulate P‐gp expression. Methods The permeability of the BBB was assessed using Evans blue (EB) staining and transmission electron microscopy. The intracerebral concentration of AS‐IV was quantified by liquid chromatography–tandem mass spectrometry (LC–MS/MS). The therapeutic efficacy of AS‐IV in combination with EA was evaluated using triphenyltetrazolium chloride (TTC) staining and neurological function assessments. mRNA and protein expression levels of relevant factors were analyzed through real‐time quantitative polymerase chain reaction (RT‐qPCR), western blot, and immunofluorescence. Results In normal mice, EA effectively increases the levels of EB and AS‐IV in brain tissue. The combination of EA and AS‐IV significantly activates the Wnt/β‐catenin signaling pathway and promotes the expression of zonula occludens protein (ZO)‐1 and Claudin‐5. Furthermore, EA inhibits NF‐κB nuclear translocation and mitigates AS‐IV‐induced upregulation of P‐gp expression. Notably, EA significantly elevates the AS‐IV content in ischemic brain tissue. The combination therapy demonstrates enhanced therapeutic effects in ischemic rats, including reductions in neurological function scores, infarct size, and pro‐inflammatory factor levels, along with increased expression of anti‐inflammatory factors. Additionally, this combination significantly inhibits NF‐κB nuclear translocation and reduces P‐gp expression in the brain tissue of ischemic stroke rats. Conclusion These findings indicate that EA enhances the brain uptake and neuroprotective effects of AS‐IV, irrespective of BBB integrity. The underlying mechanism involves P‐gp‐mediated transcellular transport rather than the paracellular pathway.

Electroacupuncture (EA) has been shown to ameliorate brain injury and protect against intestinal injury after ischemic stroke. These protective effects are closely associated with the enhancement of regulatory T (Treg) cell numbers and function in the intestine, as well as the inhibition of intestinal γδ T cell production and their migration to the brain. This study aimed to elucidate the potential mechanism by which EA regulates intestinal Treg cell differentiation after stroke. Sprague–Dawley rats were divided into three groups: the sham group, the middle cerebral artery occlusion (MCAO) group, and the MCAO plus EA (MEA) group. The MCAO model was generated by occluding the middle cerebral artery. EA was applied to Baihui (GV20) acupoint once daily. Samples were collected 3 days after reperfusion. Our results showed that EA reduced the inflammatory response in the brain and intestine after ischemic stroke. EA treatment increased the percentage of Treg cells in the small intestine of rats. EA increased the levels of SCFAs, while also inhibiting histone deacetylase activity (HDAC). Additionally, acetylated Foxp3 protein in the small intestine was increased after EA treatment. These results suggest that EA at GV20 alleviates brain and intestinal inflammatory injury in stroke rats, potentially through the enhancement of SCFA-mediated Foxp3 acetylation in Treg cells.

BackgroundImbalances between Bcl-2 and caspase-3 are significant evidence of apoptosis, which is considered an influential factor in rapidly occurring neuronal cell death and the decline of neurological function after stroke. Studies have shown that acupuncture can reduce poststroke brain cell damage via either an increase in Bcl-2 or a reduction in caspase-3 exposure. The current study aimed to investigate whether acupuncture could modulate Bcl-2 and caspase-3 expression through histone acetylation modifications, which could potentially serve as a neuroprotective mechanism.MethodsThis study used TTC staining, Nissl staining, Clark neurological system score, and Evans Blue (EB) extravasation to evaluate neurological damage following stroke. The expression of Bcl-2/caspase-3 mRNA was detected by real-time fluorescence quantification of PCR (real-time PCR), whereas the protein expression levels of Bcl-2, Bax, caspase-3, and cleaved caspase-3 were assessed using western blotting. TUNEL staining of the ischemic cortical neurons determined apoptosis in the ischemic cortex. Histone acetyltransferase (HAT) and histone deacetylase (HDAC) activities, along with the protein performance of AceH3, H3K9ace, and H3K27ace, were detected to evaluate the degree of histone acetylation. The acetylation enrichment levels of H3K9 and K3K27 in the Bcl-2/caspase-3 gene were assessed using Chromatin Immunoprecipitation (ChIP) assay.ResultsOur data demonstrated that electroacupuncture (EA) exerts a significant neuroprotective effect in middle cerebral artery occlusion (MCAO) rats, as evidenced by a reduction in infarct volume, neuronal damage, Blood-Brain Barrier (BBB) disruption, and decreased apoptosis of ischemic cortical neurons. EA treatment can promote the mRNA and protein expression of the Bcl-2 gene in the ischemic brain while reducing the mRNA and protein expression levels of caspase-3 and effectively decreasing the protein expression levels of Bax and cleaved caspase-3. More importantly, EA treatment enhanced the level of histone acetylation, including Ace-H3, H3K9ace, and H3K27ace, significantly enhanced the occupancy of H3K9ace/H3K27ace at the Bcl-2 promoter, and reduced the enrichment of H3K9ace and H3K27ace at the caspase-3 promoter. However, the Histone Acetyltransferase inhibitor (HATi) treatment reversed these effects.ConclusionsOur data demonstrated that EA mediated the expression levels of Bcl-2 and caspase-3 in MCAO rats by regulating the occupancy of acetylated H3K9/H3K27 at the promoters of these two genes, thus exerting a cerebral protective effect in ischemic reperfusion (I/R) injury.

Electroacupuncture (EA) at the Neiguan acupoint (PC6) has shown significant cardioprotective effects. Sympathetic nerves play an important role in maintaining cardiac function after myocardial infarction (MI). Previous studies have found that EA treatment may improve cardiac function by modulating sympathetic remodeling after MI. However, the mechanism in how EA affects sympathetic remodeling and improves cardiac function remains unclear. The aim of this study is to investigate the cardioprotective mechanism of EA after myocardial ischemic injury by improving sympathetic remodeling and promoting macrophage M2 polarization. We established a mouse model of MI by occluding coronary arteries in male C57/BL6 mice. EA treatment was performed at the PC6 with current intensity (1 mA) and frequency (2/15 Hz). Cardiac function was evaluated using echocardiography. Heart rate variability in mice was assessed via standard electrocardiography. Myocardial fibrosis was evaluated by Sirius red staining. Levels of inflammatory factors were assessed using RT‐qPCR. Sympathetic nerve remodeling was assessed through ELISA, western blotting, immunohistochemistry, and immunofluorescence staining. Macrophage polarization was evaluated using flow cytometry. Our results indicated that cardiac systolic function improved significantly after EA treatment, with an increase in fractional shortening and ejection fraction. Myocardial fibrosis was significantly mitigated in the EA group. The sympathetic nerve marker tyrosine hydroxylase and the nerve sprouting marker growth‐associated Protein 43 were significantly reduced in the EA group, indicating that sympathetic remodeling was significantly reduced. EA treatment also promoted macrophage M2 polarization, reduced levels of inflammatory factors TNF‐α, IL‐1β, and IL‐6, and decreased macrophage‐associated nerve growth factor in myocardial tissue. To sum up, our results suggest that EA at PC6 attenuates sympathetic remodeling after MI to promote macrophage M2 polarization and improve cardiac function.

OBJECTIVE: To explore whether the regulation of matrix metalloproteinase 9(MMP-9)/tissue inhibitors of MMPs(TIMPs) gene expression through histone acetylation is a possible mechanism by which electroacupuncture(EA)protects blood-brain barrier(BBB) integrity in a middle cerebral artery occlusion(MCAO) rat model.METHODS: Male Sprague-Dawley rats were divided into four groups: the sham group, the MCAO group, the MCAO + EA(MEA) group, and the MCAO + EA + HAT inhibitor(HATi) group. The MCAO model was generated by blocking the middle cerebral artery. EA was applied to Baihui(GV20). Samples were collected 1 or 3 d after reperfusion. Neurological function scores and Evans blue extravasation were employed to evaluate the poststroke injury. The effect of EA on MMP-9/TIMPs gene expression was assessed by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR) and chromatin immunoprecipitation(ChIP). RESULTS: Our results showed that EA treatment prominently improved neurological function and ameliorated BBB disruption. The RT-qPCR assay showed that EA reduced the expression of MMP-9 and promoted TIMP-2 mRNA expression, but HATi reversed these effects of EA. In addition, ChIP results revealed that EA decreased the enrichment of H3K9ace/H3K27ace at MMP-9 promoters and notably stimulated the recruitment of H3K9ace/H3K27ace at TIMP-2 promoter. CONCLUSION: EA treatment at Baihui(GV20) regulates the transcription of MMP-9 and TIMP-2 through histone acetylation modification in the acute stage of stroke, which preserves the structural integrity of the BBB in MCAO rats. These findings suggested that the histone acetylation-mediated transcriptional activity of target genes may be a crucial mechanism of EA treatment in stroke.

目的 观察电针对心肌缺血损伤小鼠心功能及心脏组织中巨噬细胞极化、TLR4、MyD88等表达的影响,探讨电针促进心肌损伤保护的可能机制.方法 27只雄性C57BL/6J小鼠随机分为假手术组、模型组、电针组,每组9只.模型组、电针组通过结扎心脏冠状动脉左前降支建立心肌缺血模型.电针组于造模成功后,电针小鼠双侧"内关"穴治疗:2/15 Hz,疏密波,1 mA,20 min·次-1,每日1次,共治疗7 d.运用超声心动图评价小鼠心脏射血分数(EF)及短轴收缩率(FS),天狼星红染色观察小鼠心脏纤维化情况,流式细胞术检测心肌组织中中性粒细胞、巨噬细胞数量及各表型巨噬细胞占比,qPCR法检测心肌组织中TNF-α、IL-1β的mRNA表达量,Western blot法检测心肌组织中TLR4、MyD88、IL-1β、IL-17A的蛋白表达水平.结果 与假手术组相比,模型组小鼠EF、FS值下降(P<0.0001),胶原容积分数升高(P<0.001),心脏组织中中性粒细胞、巨噬细胞数增多(P<0.0001,P<0.001),同时M1型巨噬细胞占比增高(P<0.05),心肌组织中TNF-α、IL-1β的mRNA表达量上升(P<0.01,P<0.001),TLR4、MyD88、IL-1β、IL-17A蛋白表达水平上升(P<0.05,P<0.01);与模型组相比,电针组小鼠EF、FS值升高(P<0.01,P<0.001),胶原容积分数降低(P<0.001),心脏组织中中性粒细胞、巨噬细胞数减少(P<0.0001,P<0.01),其中M2型巨噬细胞占比增高(P<0.05),心肌组织中TNF-α、IL-1β的mRNA表达量下降(P<0.001),TLR4、MyD88、IL-1β、IL-17A蛋白表达水平降低(P<0.05,P<0.001).结论 电针可能通过降低TLR4、MyD88在心肌组织中的表达,促进心肌缺血损伤后心脏巨噬细胞向M2型极化,减轻局部炎症,抑制心肌纤维化,实现心肌保护效应.

BACKGROUND:Accumulating evidence suggests that acupuncture may serve as a potent strategy to mitigate the deleterious effects of ischemic stroke on neural tissue. The present investigation delineated the neuroprotective potential of electroacupuncture (EA) administered pre-and post-stroke, with a focus on determining the commonalities and disparities between these two therapeutic approaches in ameliorating ischemic stroke-induced brain injury. The ultimate objective is to inform optimal timing for acupuncture intervention in the clinical management and prevention of stroke.METHODS:The extent of cerebral infarction was quantified with 2,3,5-triphenyltetrazolium chloride staining. The integrity of the blood-brain barrier was assessed by evaluating the extravasation of Evans blue (EB) dye, while neurological function was appraised using the Longa neurological scoring system. RNA sequencing was employed to examine the transcriptomic landscape of ischemic brain tissue, with subsequent bioinformatics annotation of the sequencing data facilitated by Metascape.RESULTS:(1) A notable decrease in the ischemic infarct volume was observed in both the EA-preconditioned plus middle cerebral artery occlusion (MCAO), EA-preconditioned plus middle cerebral artery occlusion (EAM) and MCAO plus EA-treated (MEA) groups, compared to the MCAO group. Furthermore, the decreased leakage of EB and reduction in neurological function impairment scores were evident in the EAM and MEA groups compared with the MCAO group. (2) Relative to the Sham group, the MCAO group exhibited a total of 4798 differentially expressed genes (DEGs), with 67.84% demonstrating an expression fold change (FC) greater than 1.5, and 34.16% exceeding a FC of 2. The EAM and MEA groups displayed 4020 and 1956 DEGs, respectively, compared to the MCAO group. In both groups, more than 55% of DEGs showed an expression FC surpassing 1.5, whereas only approximately 10% exhibited a change greater than 2-fold. Remarkably, EA preconditioning and EA treatment resulted in the reversal of 18.72% and 28.91% of DEGs, respectively, in the MCAO group. (3) The DEGs upregulated in response to ischemic stroke were predominantly implicated in immune inflammatory processes and cellular apoptosis, whereas the downregulated DEGs were associated with neurogenesis and neuronal signal transduction. The MEA-induced upregulated DEGs were primarily involved in neural transmission and metabolic processes, whereas the downregulated DEGs were linked to excessive inflammatory responses to physical and chemical stimuli, as well as cell matrix adhesion chemotaxis. In the context of EAM, the upregulated DEGs were chiefly related to protein biosynthesis, and energy and metabolic processes, whereas the downregulated genes were connected to gene transcriptional activity, synaptic function, and neuronal architecture.CONCLUSIONS:Both preconditioning and post-event treatment with acupuncture demonstrated efficacy in mitigating pathological damage to brain tissue in a rat model of ischemic stroke, albeit with some divergences in their gene targets. The integration of EA preconditioning and treatment may potentially confer enhanced neuroprotection in the clinical management of stroke patients.

Background: Electro-acupuncture (EA) is an effective and safe treatment for ischemic stroke. It is not only capable of reducing cerebral damage but also alleviating intestinal inflammation. However, its mechanism has not been fully elucidated. Methods: All rats were randomly divided into three experimental groups: the SHAM group, the MCAO group, and the MEA (MCAO+EA) group. Ischemic-reperfusion (I/R) injury was induced by MCAO surgery. Rats in the MEA group were treated with EA stimulation in the "Baihui" acupoint (1 mA, 2/15 Hz, 20 min for each time). The Real-time (RT)-qPCR was used to evaluate the mRNA expression of inflammation factors in the ischemic brain and the small intestine after I/R injury. In addition, our research evaluated the effects of EA on regulatory T cells (Tregs) and gamma delta T cells in the small intestine and brain via Flow cytometry analysis. Finally, we applied CM-Dil and CFSE injection and explored the potential connections of T cells between the ischemic hemisphere and the small intestine. Results: Our results suggested that EA treatment could significantly reduce the inflammation response in the ischemic brain and small intestine 3 days after I/R injury in rats. To be specific, EA increased the percentage of Tregs in the brain and the small intestine and decreased intestinal and cerebral gamma delta T cells. Concomitantly, after EA treatment, the percentage of cerebral CD3+TCR gamma delta+CFSE+ cells dropped from 12.06% to 6.52% compared with the MCAO group. Conclusions: These findings revealed that EA could regulate the Tregs and gamma delta T cells in the ischemic brain and the small intestine, which indicated its effect on inhibiting inflammation. And, EA could inhibit the mobilization of intestinal T cells, which may contribute to the protection of EA after ischemic stroke.

Electro-acupuncture (EA) has an anti-inflammatory role in ischemic stroke, but whether the protective effect of EA involves the regulation of the intestine barrier and Treg/ γδ T cells is unclear. Cerebral ischemia-reperfusion (I/R) injury was induced by middle cerebral artery occlusion(MCAO) for 2 h followed by reperfusion for 24 h. The rats have treated with EA at the "Baihui" acupoint(GV20). Triphenyl tetrazolium chloride (TTC) staining and Longa neurologic score were performed to evaluate the outcomes after ischemic stroke. Inflammatory factor expression levels in the serum, ischemic hemisphere brain, and small intestine were detected by ELISA or RT-qPCR. Additionally, the morphology change of the small intestine was evaluated by analyzing villus height and smooth muscle thickness. Meanwhile, the expression of tight-junction proteins, including Zonula Occludens-1 (ZO-1), Occludin, and Claudin-1, were detected to evaluate the impact of EA on mucosal permeability in the small intestine. The percentages of regulatory T cells (Tregs) (CD45+CD4+Foxp3+) and γδ T cells (CD45+CD4-γδ T+) were measured to assess the effect of EA on intestinal T cells. EA decreased the brain infarction volume and intestine barrier injury in ischemic stroke rats. At the same time, it effectively suppressed the post-stroke inflammation in the brain and small intestine. More importantly, EA treatment increased the percentage of Tregs in the small intestine while reducing the rate of γδ T cells, and ultimately increased the ratio of Treg/ γδ T cells. These results demonstrated that EA ameliorated intestinal inflammation damage by regulating the Treg/ γδ T cell polarity shift and improving the intestine barrier integrity in rats with I/R injury. This may be one of the mechanisms underlying the anti-ischemic injury effects of acupuncture on stroke.

Context Yi-Qi Cong-Ming (YQCM) decoction has been widely used to prevent age-related hearing loss (ARHL), the most prevalent neurodegenerative disease in the elderly. Objective To explore the mechanism of YQCM decoction in the treatment of ARHL. Materials and methods The chemical constituents of YQCM were screened from the Traditional Chinese Medicine Systems Pharmacology Database. Potential targets of YQCM against ARHL were predicted by DrugBank, GeneCards, and OMIM database. Protein-protein network and enrichment analysis were used for exploring possible molecular mechanisms. Molecular docking and an in vitro model of ARHL by exposing auditory cells with 100 mu M H2O2 for 3 h were applied. Cell viability and mitochondrial membrane potential (Delta psi M) were detected by CCK-8 and high-content analysis. gamma H2AX and cleaved caspase-3 were detected by Western blot. Results The main compounds have good affinities with hub targets, especially AKT1, PTGS2, and CASP3. GO and KEGG analysis showed that the main biological process and key targets were related to negative regulation of the apoptotic process. H2O2 treatment could reduce the cell viability by 68% and impaired Delta psi M, while 90 mu g/mL YQCM pre-treatment could restore the cell viability by 97.45% and increase Delta psi M (2-fold higher). YQCM pre-treatment also reduced gamma H2AX and cleaved caspase-3 protein levels. Conclusions Our study suggested that YQCM prevents ARHL by modulating the apoptosis process in auditory hair cells. Moreover, this study proved that bioinformatics analysis combined with molecular docking and cell model is a promising method to explore other possible pharmacological interventions of ARHL.

摄食行为是生物从外界获得营养物质维持个体生存、保障身体各器官功能和从事各种活动的能量需要所进行的进食行为〔1〕.因机体摄入过多高热量食物导致体内能量代谢失衡,从而引发肥胖已成为共识;同时,在有关肥胖的基础〔2〕和临床〔3〕研究中也发现体重增加通常伴随着摄食行为的异常,包括摄食总量增加及对高糖高脂食物的摄入偏嗜等,说明摄食行为与肥胖之间具有相互调控的关系,即摄食行为不当为肥胖的重要诱因,肥胖亦可引发机体摄食行为的异常.

Much evidence suggests that gut immunity homeostasis plays a vital role in regulating neuroinflammatory response under ischemic stroke. Acupuncture is an effective treatment for post-stroke rehabilitation. However, little is known about the effect of acupuncture on intestinal injury after stroke. Herein, we observed the intestinal inflammatory response and barrier's impairment of the post-stroke rats to explore the mechanism of electroacupuncture (EA) against cerebral ischemia injury from the brain-gut axis. Rats were randomized to the sham operation group (SHAM), the middle cerebral artery occlusion group (MCAO), and the MCAO plus EA treatment group (MEA). Triphenyl tetrazolium chloride (TTC) staining and Longa neurologic score were performed to evaluate the outcomes after ischemic stroke. Inflammatory factors expression levels in the serum, ischemic hemisphere brain, and small intestine were detected. Additionally, the morphology change of small intestine was evaluated via analyzing villus height and smooth muscle thickness. Meanwhile, the expression of tight-junction proteins, including Zonula Occludens-1(ZO-1), Occludin and Claudin-1, was detected to evaluate the impact of EA on mucosal permeability in the small intestine. The percentage of Treg cells (CD45 + CD4 + Foxp3 + ) and γδ T cells (CD45 + CD4 − γδ T + ) were measured to assess the effect of EA on T cells. Our data showed that EA could significantly decrease the brain infarction volume and intestine barrier injury of the ischemic stroke rats. At the same time, it effectively suppressed the post-stroke inflammation response of the ischemic brain, small intestine, and peripheral serum. More importantly, EA treatment increased the percentage of Treg cells in the small intestine while reducing the rate of γδ T cells, and ultimately increased the ratio of Treg/ γδ T cells. These results suggested that EA could ameliorate intestinal inflammation damage by regulating the Treg/ γδ T cell polarity shift and improving the intestine barrier integrity post-stroke. It may be one of the mechanisms underlying the anti-ischemic injury effects of acupuncture on stroke.

文章通过综述肠道菌群调节T淋巴细胞参与缺血性中风免疫反应的机制,以及针灸对T细胞和肠道微生物稳态的干预现状,进而探讨肠道T细胞分化在针灸抑制脑缺血损伤中的作用,以期从脑肠轴的角度为中风临床治疗和基础研究提供新思考.

Acupuncture promotes the recovery of neurological function by the overall improvement of ischemic brain injury. It is not only regarded as a rehabilitative treatment but also a pretreatment method for stroke. However, its mechanism has not been fully elucidated. In this study, rats were treated with electroacupuncture (EA) at Baihui (GV20) for 30 min/day for 6 days, ahead of conducting cerebral ischemia–reperfusion (I/R) injury. Infarction volume, Evans blue leakage, and neurological deficits were evaluated at 24 h after I/R injury. Then, the ipsilateral ischemic brain was isolated for RNA sequencing (RNA-Seq) to identify molecular consequences. The results showed that EA pretreatment decreased blood–brain barrier (BBB) permeability, reduced brain infarction volume, and improved neurological outcomes. EA pretreatment could upregulate expression of antivirus and immunity activity-associated genes (such as Ifit1 , Ifit3 , Irf7 , and Oasla ) and downregulate expression of matrix disruption-associated genes ( Col24a1 , Col11a1 , Col27a1 , etc.) in healthy rats. In addition, it could partially reverse or ameliorate genome-wide transcription changes of the ipsilateral ischemic brain. For the first time, this study provides insight into genomic network modulation of a healthy rat with EA treatment and a EA-preconditioned rat under subsequent I/R injury, which is helpful in explaining acupuncture precondition-induced ischemic tolerance of stroke. It also provides new strategies and targets for the prevention of ischemic stroke.

Isoflavones are major neuroprotective components of a medicinal herb Astragali Radix, against cerebral ischemia-reperfusion injury but the mechanisms of neuroprotection remain unclear. Calycosin and formononetin are two major AR isoflavones while daidzein is the metabolite of formononetin after absorption. Herein, we aim to investigate the synergistic neuroprotective effects of those isoflavones of Astragali Radix against cerebral ischemia-reperfusion injury. Calycosin, formononetin and daidzein were organized with different combinations whose effects observed in both in vitro and in vivo experimental models. In the in vitro study, primary cultured neurons were subjected to oxygen-glucose deprivation plus reoxygenation (OGD/RO) or l -glutamate treatment. In the in vivo study, rats were subjected to middle cerebral artery occlusion to induce cerebral ischemia and reperfusion. All three isoflavones pre-treatment alone decreased brain infarct volume and improved neurological deficits in rats, and dose-dependently attenuated neural death induced by l -glutamate treatment and OGD/RO in cultured neurons. Interestingly, the combined formulas of those isoflavones revealed synergistically activated estrogen receptor (estrogen receptors)-PI3K-Akt signaling pathway. Using ER antagonist and phosphatidylinositol 3-kinase (PI3K) inhibitor blocked the neuroprotective effects of those isoflavones. In conclusion, isoflavones could synergistically alleviate cerebral ischemia-reperfusion injury via activating ER-PI3K-Akt pathway.