Hepatocellular carcinoma (HCC) is the most prevalent form of primary liver cancer and ranks as the third leading cause of cancer-related fatalities worldwide. Recently, prochlorperazine (PCZ), a synthetic antipsychotic medication, has gained attention for its potential anticancer properties. However, the specific mechanisms underlying its anticancer effects remain unclear. In this study, we have identified that PCZ effectively inhibits the proliferation of HCC cells and diminishes the number and size of HCC foci in the mouse model. Subsequent studies have revealed that PCZ can directly bind to β-catenin and enhance the phosphorylation level of β-catenin. This, in turn, inhibits the Wnt signaling pathway, thereby exerting its anticancer effects. This novel finding sheds light on the potential molecular mechanisms of PCZ as a therapeutic strategy for HCC. In summary, our study provides fresh insights into the utility of PCZ as a therapeutic option for HCC and elucidates its molecular mechanisms.

Feline panleukopenia is an acute and highly contagious disease caused by feline panleukopenia virus (FPV). Conventional therapeutic approaches often yield suboptimal outcomes in managing leukopenia, which consequently contributes to its high mortality rate. Mesenchymal stem cells (MSCs) are a type of multipotent stem cell characterized by their multidirectional differentiation potential and immunomodulatory capabilities. Studies have shown that MSCs possess the potential to treat inflammatory and immune-mediated diseases, support and promote hematopoiesis, and facilitate tissue repair. Therefore, this study was to investigate the clinical efficacy of combining MSCs with feline panleukopenia monoclonal antibody to propose a potential treatment for FPV. MSCs were extracted from a newborn kitten’s umbilical cord tissue. Fifteen healthy unimmunized kittens aged 2 to 4 months were divided evenly into three groups: control group, conventional treatment group, and MSCs group (conventional treatment combined with MSCs therapy, involving daily intravenous administration of 1 × 107 MSCs for 3 consecutive days). Treatments were commenced once evident clinical symptoms manifested and white blood cell counts declined below the normal range. The subject cats were assessed for clinical signs, complete blood count (CBC), blood biochemistry, serum amyloid A (SAA), pathology and viral load in major organs. The results revealed that in the control group and conventional treatment group, cats exhibited a rapid decline in white blood cell count following disease onset, ultimately resulting in mortality. Conversely, in the MSCs group, four cats demonstrated an increase in white blood cell count post-treatment, subsequently returning to normal levels. However, one cat did not exhibit a significant increase and died on the third day of treatment. These findings highlight the therapeutic potential of MSCs in elevating leukocyte counts, improving clinical symptoms, and ultimately leading to a significant enhancement of survival rates among FPV-affected cats.

Quality of life is markedly influenced by inflammatory pain, with current therapeutic options frequently necessitating extended treatment periods and resulting in many undesirable side effects. In the present study, a Complete Freund's Adjuvant (CFA)- induced model was employed to examine the therapeutic potential of electroacupuncture (EA) in conjunction with subhypnotic doses of dexmedetomidine (DEX) administered at acupoints for managing inflammatory pain. The experimental design involved 36 healthy male C57BL/6 mice equally divided into six experimental treatments: saline (SAL), CFA, diclofenac sodium standard treatment (ST), DEX, EA and combined DEX-EA, with six mice in each group. Treatments commenced 48h following CFA administration, consisting of four treatment sessions at two-day intervals. Following each treatment session, nociceptive thresholds were evaluated. After the last session, paw tissue samples collected from affected paws were subjected to histopathological and immunohistochemical examination, and RNA seq was performed on spinal cord specimens. Results revealed that the DEX-EA therapy produced significantly higher nociceptive thresholds compared to the CFA, DEX monotherapy, and EA monotherapy (P<0.05). Additionally, inflammatory indices showed significant improvements with the DEX-EA, with decreased levels of CD45, MPO, TNF-alpha, IL-6, and IL-1 beta compared to the CFA, DEX monotherapy, and EA monotherapy (P<0.05). Compared to the CFA, non-significant variations were found in nociceptive thresholds and IL-6 level in the DEX and EA monotherapies; however, a significant decrease was found in the level of CD45, MPO, and IL-1 beta (P<0.05). Transcriptomic data revealed 65 differentially expressed genes (DEGs), demonstrating enhanced expression of genes associated with GABAergic neurotransmission, degradative pathways, and autophagic mechanisms. The findings demonstrate that combining EA with DEX produces synergistic anti-nociceptive and anti-inflammatory effects mediated through spinal cord mechanisms.

Neuropathic pain severely affects patients' quality of life. Limited treatments offer relief but often involve long-term use and side effects. This study aimed to explore the electroacupuncture (EA) and subanesthetic alfaxalone (ALF) combination as a novel therapeutic substitute for the treatment of neuropathic pain in a spared nerve injury (SNI) mouse model. Sixty healthy C57BL/6 male mice were randomized into 5 equal-sized groups: sham (n = 12) group, spared nerve injury (SNI; n = 12) group, SNI-ALF (n = 12), SNI-EA (n = 12), and SNI-EA-ALF (n = 12). The SNI mice received the treatment regimens at 8 days postoperatively once every 2 days for 7 treatments in total. The mechanical and thermal pain thresholds were tested after each treatment. Spinal cord samples were collected after the fourth and seventh treatments for detection of diazepam-binding inhibitor (DBI) and γ-aminobutyrate A1 isoform receptor (GABAA1). Mice in the SNI-EA-ALF group showed a significant increase in mechanical and thermal pain thresholds as compared to those in the SNI-ALF and SNI-EA groups during the treatment period (P < 0.05). A significant increase in GABAA1 expression was observed after the fourth and seventh treatments in the SNI-EA-ALF group compared to the SNI-ALF and SNI-EA groups (P < 0.05). In addition, the DBI expression level was significantly lower in the SNI-ALF-EA group than the SNI, SNI-ALF, and SNI-EA groups (P < 0.05). Our results support the use of EA, combined with ALF, to synergistically relieve pain in preclinical models of NP.

Ketosis is a prevalent metabolic disorder in periparturient dairy cows and severely impairs production performance. It has been established that ketotic cows manifest hepatic oxidative damage; however, the precise molecular mechanisms underlying this pathology remain elusive. The present results demonstrate that ketotic dairy cows exhibit severe hepatic oxidative damage and ferroptosis. This pathological state is characterized by aberrant hepatic iron accumulation, driven by an imbalance between increased transferrin receptor 1 (TFR1)-mediated iron uptake and decreased ferroportin (FPN)-mediated iron export. Furthermore, we observed a substantial accumulation of lipid peroxides due to both an impaired nuclear factor erythroid 2-related factor 2 (NFE2L2)/glutathione peroxidase 4 (GPX4) antioxidant pathway and enhanced acyl-CoA synthetase long-chain family member 4 (ACSL4)-mediated lipid peroxidation. In vitro, β-hydroxybutyrate (BHB) induced oxidative injury and ferroptosis in bovine hepatocytes, and the molecular patterns were consistent with those in vivo. Mechanistically, BHB disrupted TFR1/FPN-mediated iron metabolism, impaired the NFE2L2-mediated GSH synthesis pathway, and promoted ACSL4-mediated lipid peroxidation. Notably, inhibition of ferroptosis can significantly alleviate BHB-caused oxidative damage in bovine hepatocytes. In summary, these data demonstrate that BHB-induced ferroptosis is mechanistically associated with hepatic oxidative injury in ketotic dairy cows. These findings not only contribute to mechanistic insights into hepatic oxidative injury of ketotic cows but also highlight the potential of targeting ferroptosis for therapeutic intervention in ketosis.

Aiming to enhance disease modeling and drug screening for fatty liver disease, a bovine liver organoid model was developed using R-spondin-1 conditioned medium. Adult stem cells from calf liver were cultured and differentiated into organoids, exhibiting specific liver markers and functions. These organoids, treated by a mixture of oleic acid and palmitic acid, demonstrated significant accumulation of total cholesterol and triglycerides, effectively mimicking the fatty liver condition observed in high-yielding dairy cows during the periparturient period. Treatment with 5 natural compounds (silymarin, gastrodin, cordycepin, curcumin, and polydatin) and a positive control (atorvastatin) showed significantly reduced inflammation and lipid accumulation in the bovine liver organoid. Specifically, anti-inflammatory factors were upregulated, pro-inflammatory factors downregulated, and genes related to lipid synthesis were downregulated. Collectively, the application of organoid technology exhibits its potential in simulating disease mechanisms and evaluating therapeutic interventions, notably reducing the need for live animal experiments. Our study establishes a bovine liver organoid model and validates its effectiveness as a tool for advancing disease research and pharmaceutical development.

During the periparturient period, reduced feed intake often causes negative energy balance in dairy cows, leading to fat mobilization, hepatic lipid accumulation, and fatty liver disease (FLD), ultimately compromising health and milk production. This study investigated the association between FLD and gut microbiota dysbiosis, with a particular focus on the role of Clostridium perfringens within the gut-liver axis. Metagenomic sequencing of ileal contents revealed a marked decrease in microbial diversity in cows with FLD, along with increased abundances of potential pathogens such as C. perfringens, Enterobacter cloacae, and Vibrio alginolyticus. Functional annotation indicated elevated expression of virulence factors (e.g., Hsp60, flagella, mu-toxin), antibiotic resistance genes (e.g., otrA, lsaC), and pathways related to lipopolysaccharide (LPS) biosynthesis and mitogen-activated protein kinase (MAPK) signaling pathways, suggesting enhanced pro-inflammatory potential. qPCR analysis of ileal tissue demonstrated reduced expression of tight junction proteins (zona occludens 1 (ZO-1), Claudin-1, and Occludin) and increased levels of pro-inflammatory cytokines (Interleukin-1 beta (IL-1β), Interleukin-6 (IL-6), Tumour necrosis factor-alpha (TNF-α)), alongside a decrease in the anti-inflammatory cytokine interleukin-10 (IL-10), indicating compromised intestinal barrier function and local inflammation. Given the significant enrichment of C. perfringens in the ileum of FLD cows, we hypothesized its involvement in disease pathogenesis. To test this, C. perfringens was isolated and orally administered to antibiotic-pretreated mice fed a high-fat diet. These mice developed exacerbated hepatic steatosis, metabolic disturbances, and heightened inflammatory responses. Moreover, Western blot analysis revealed reduced expression of intestinal tight junction proteins (ZO-1, Claudin-1, Occludin), indicating increased intestinal permeability. Quantitative PCR confirmed upregulation of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) and downregulation of IL-10 in both intestinal and hepatic tissues. These findings indicate that C. perfringens may promote FLD by impairing gut barrier integrity and enhancing inflammatory responses. In conclusion, our findings suggest that C. perfringens may contribute to the development of FLD in dairy cows by impairing intestinal barrier integrity and promoting systemic inflammation.

Combining decellularized biological scaffolds with PRP can prevent the rapid inactivation of growth factors and achieve their controlled and sustained release during tissue regeneration. Therefore, the purpose of this study was to evaluate the combined effect of decellularized bovine pericardium (dBP) and leukocyte-rich platelet-rich plasma (LR-PRP) on the bone repair in a rabbit femoral defect model. Bovine pericardium was decellularized using the Trypsin-Triton X-SDS protocol and histologically assessed. Unicortical bone defects were surgically created in the femur of rabbits (n = 6) and randomly assigned to three treatment allocations: (1) untreated control, (2) LR-PRP, and (3) LR-PRP + dBP-treated defects. Bone defect healing was evaluated using quantitative computed tomography (CT) and histopathological analyses. The dBP achieved 99.2

ObjectiveTo investigate the role of quercetin-added pancreatic prescription food in regulating metabolic homeostasis in dogs.MethodsThe experimental dogs were divided into a control diet group and a prescription diet group. The control group was fed regular food, while the prescription group was fed pancreatic prescription food (3.9 g of quercetin was added in per 1 kg of food) for 8 weeks. Canine physical examination, complete blood count, and serum biochemical tests were conducted at 0 w, 4 w, and 8 w. Non-targeted metabolomics tests were performed using plasma samples at 0 w and 8 w.ResultsDogs that received a quercetin-added pancreatic diet supplemented with quercetin showed no changes in the body weight, fasting blood glucose, body condition score, the indexes of whole blood program of red blood cells, white blood cells and platelets, and most blood biochemical indexes, but increased lipase levels in plasma at 8 w. Quercetin significant improved in metabolic homeostasis, especially in fatty acid, amino acid, and bile acid metabolism. Untargeted metabolomics analysis revealed that quercetin activates ABC transport and arginine/proline pathways, suggesting potential benefits for pancreatitis in large animals, while maintaining comparable safety parameters.ConclusionsQuercetin-added prescription food enhances fatty acid and amino acid metabolism, demonstrating its potential to promote pancreatic function and sustain metabolic homeostasis.

Neuropathic pain (NP), resulting from nerve damage, is difficult to manage and often requires long-term treatment. However, prolonged use of pain medications can lead to addiction and reduced effectiveness over time. Understanding drug tolerance is essential for developing improved pain management strategies. Dexmedetomidine (DEX) is effective in targeting the α2-adrenergic receptor, providing relief from pain, especially NP. However, its extended use leads to tolerance and hinders its clinical utility. Herein, we investigated tolerance mechanisms and potential applications of this drug in managing NP. Adult C57BL/6 mice (male) were distributed into DEX Dosage Groups (n = 48), DEX Tolerance Model Groups (n = 32), SGK1 Inhibitor GSK650394 Groups (n = 48), and NF-κB Inhibitor PDTC Groups (n = 32) to explore dexmedetomidine's effects on NP and tolerance mechanisms. NP was established via selective ligation of the sciatic nerve branch (SNI), followed by administration of DEX. The results revealed a dose-dependent analgesic effect of DEX, with significant increases in pain thresholds observed compared to the sham group (p < 0.05). Optimal efficacy was found at a dose of 30 μg/kg, indicating its potential as an effective treatment for NP (p < 0.05). However, continuous administration of DEX over 13 days induced analgesic tolerance, evidenced by an initial increase in pain thresholds followed by a gradual decrease (p < 0.05). Despite an initial efficacy in elevating pain thresholds, the analgesic effect of DEX diminished over time, returning to pre-dose levels after 5 days (p < 0.05). Transcriptome sequencing of spinal cord samples from mice receiving multiple DEX injections revealed differential gene expression patterns, notably upregulation of SGK1, NR2A, and NR2B subunits (p < 0.05). Inhibiting SGK1 mitigated DEX-induced tolerance, suggesting its involvement in tolerance development (p < 0.05). Moreover, NF-κB inhibition reversed DEX-induced tolerance and implicated the SGK1-NF-κB pathway in the mediation of analgesic tolerance. To sum up, these findings revealed the molecular mechanism underlying DEX-induced analgesic tolerance in the NP model and offer potential avenues for future therapeutic interventions.

This study evaluated the effects of two novel probiotics on gut microbiota composition, metabolic health, liver function, lipid metabolism, and milk production after calving to evaluate the effects of probiotic supplementation during the early lactation phase over a 60-day period. Thirty cows were randomly assigned to one of three groups: control (no probiotics), Probiotic A (P1; Lactobacillus spp., 1 × 109 CFU/day), or Probiotic B (P2; Bifidobacterium spp., 1 × 109 CFU/day). Fecal, blood, and liver samples were collected at days 0, 15, 30, 45, and 60. Gut microbiota analysis using PCR revealed that from day 15 onwards, P1 and P2 groups exhibited significantly higher abundances of Firmicutes and Bacteroidetes compared to controls (p < 0.05). Metabolic health assessments showed that blood glucose levels were significantly higher (p < 0.05) and non-esterified fatty acids (NEFA) and beta-hydroxybutyrate (BHB) levels were significantly lower (p < 0.05) in probiotic-supplemented groups, particularly in P2. Liver function markers (ALT, AST, and hepatic triacylglycerol content) were significantly improved (p < 0.05) in probiotic-fed cows. Histopathology confirmed reduced hepatocyte swelling and lipid accumulation in the probiotic groups. Milk production significantly increased (p < 0.05) in probiotic-supplemented cows, with P2 achieving the highest yields. Data were analyzed using repeated measures ANOVA followed by Tukey's post hoc test (p < 0.05). These results demonstrate that probiotic supplementation, particularly P 1, effectively modulates gut microbiota, improves metabolic and liver health, and enhances productivity in dairy cows.

OBJECTIVE:To compare cumulative dissipated energy (CDE) and postoperative complications between modified prechop and stop-chop techniques in dogs with hard cataracts. ANIMALS STUDIED:Seventy-nine dogs with 123 mature cataract eyes were studied for measuring anteroposterior lens diameters, or 149 dogs with 225 hard cataract eyes were treated with prechop phacoemulsification or stop-chop phacoemulsification surgery. PROCEDURES:A species-specific prechopper (3.5-5.0 mm width) was designed based on canine anteroposterior lens diameter (APLD: 7.40 ± 0.99 mm) and anterior chamber depth (ACD: 2.54 ± 0.75 mm). Among 225 eyes (149 dogs), 110 eyes were randomly selected for prechop phacoemulsification with the specific prechopper, and 115 eyes for stop-chop phacoemulsification. Surgical outcomes, including CDE, postoperative aqueous flare and complications, were evaluated. RESULTS:The prechop group demonstrated less mean CDE and postoperative aqueous flare incidence compared to the stop-chop group (53.93 ± 15.29 vs. 78.32 ± 21.32, p < 0.001; 35.14% vs. 58.59%, respectively). The prechop group had fewer overall complications than the stop-chop group (35.2% vs. 56.7%). Vision retention rates at final follow-up (30-1440 days postoperation) were comparable (prechop: 95.7% vs. stop-chop: 92.9%), but glaucoma incidence was lower in the prechop group than in the stop-chop cohort (5 vs. 3 eyes). CDE correlated strongly with complication severity. CONCLUSIONS:The prechop technique with a novel prechopper offers a safer, more efficient alternative for phacoemulsification of canine cataracts, particularly in eyes requiring high energy nuclear fragmentation.

Contagious ecthyma is a contagious zoonotic disease caused by the Orf virus that can infect farm animals and humans, but no vaccine is available for pregnant mothers. Excessive oxidative stress during pregnancy can suppress the vaccine immune response in pregnant mothers; hence, maternal micronutrient supplementation could effectively improve the immune response, health, and oxidative status during pregnancy. In this study, we employed an 8-week-old pregnant rat model to receive a single intramuscular dose of 200 µg of ORF DNA vaccine with or without vitamin E and selenium supplementation to evaluate their effect on immune responses (specific IgG and IgG isotypes), oxidative stress, liver enzymes, and blood glucose levels in maternal-neonatal serum and milk secretions. Additionally, antioxidant-related gene expressions were analyzed in the maternal placenta and pups’ liver. The results showed that supplementation of vitamin E and selenium with ORF DNA vaccination increased the production of specific antibody and IgG isotypes (IgG1 and IgG2a) and reduced the oxidative stress in neonatal-maternal serum and milk compared to both the control group and those vaccinated without supplementation (p < 0.05). Notably, the ORF DNA vaccine did not cause oxidative stress and hepatic damage. However, combined supplementation of vitamin E and selenium with DNA vaccination significantly decreased serum malondialdehyde (MDA) levels and improved the antioxidant-related enzyme activities of glutathione peroxidase (GPX), superoxide dismutase 1 (SOD1), and selenoprotein P (SELP) in the maternal placenta and liver of pups (p < 0.05). In conclusion, maternal supplementation of vitamin E and selenium enhanced the immune responses of the ORF DNA vaccine by mitigating oxidative stress in pregnant rats and could thus be a promising strategy for better health outcomes for both mothers and neonates.

The prolactin (PRL) hormone is a major regulator of mammary gland development and lactation. However, it remains unclear whether and how PRL contributes to mammary epithelial cell proliferation and secretion. The Boer and Macheng black crossbred goats are superior in reproduction, meat, and milk, and are popular in Hubei province. To elucidate the mechanisms of PRL on mammary growth and lactation, to improve the local goat economic trade, we have performed studies on these crossbred goats during pregnancy and early lactation, and in goat mammary epithelial cells (GMECs). Here, we first found that the amino acid transporters of SNAT1 and SNAT2 expression in vivo and in vitro were closely associated with PRL levels, the proliferation and secretion of GMECs; knockdown and over-expression of SNAT1/2 demonstrated that PRL modulated the proliferation and lactation of GMECs through regulating SNAT1/2 expression. Transcriptome sequencing and qPCR assays demonstrated the effect of PRL on the transcriptional regulation of SNAT1 and SNAT2 in GMECs. Dual-luciferase reporter gene assays further verified that the binding of the potential PRL response element in the SNAT1/2 promoter regions activated SNAT1/2 transcription after PRL stimulation. Additionally, silencing of either PRLR or STAT5 nearly abolished PRL-stimulated SNAT1/2 promoter activity, suggesting PRLR–STAT5 signaling is involved in the regulation of PRL on the transcriptional activation of SNAT1/2. These results illustrated that PRL modulates the proliferation and secretion of GMECs via PRLR–STAT5-mediated regulation of the SNAT1/2 pathway. This study provides new insights into how PRL affects ruminant mammary development and lactation through regulation of amino acid transporters.

IntroductionEnteritis and dysbiosis are the major causes of high morbidity and mortality of juvenile ostriches. Chicory (CC) has been proven to have excellent antioxidant, anti-inflammatory, and antibacterial activities. However, it’s unclear whether CC could improve the survival rate of juvenile ostriches by relieving enteritis and correcting dysbiosis.Materials and methodsSouth African ostrich hatchlings (Struthio camelus domesticus) were fed with and without a CC-supplemented diet, and the body weight gain and mortality were compared over 4 months of age. Fresh fecal samples of clinically healthy ostriches were collected, and 16S DNAs were analyzed. Moreover, ostrich chicks with LPS-induced enteritis were fed with different dosages (0, 20, 40, and 80 mg/kg) of chicoric acid (CA), a major bioactive component of CC, for five consecutive days. The expression levels of tight junction (TJ)-related proteins and inflammatory mediators in the ilea were detected with western blot and immunofluorescence.ResultsThe ostrich chicks fed on the CC-supplemented diet began to increase in weight at the 1st month of age and became remarkably heavier at the fourth month (p < 0.01) compared with those fed on the non-CC-supplemented diet. Additionally, the mortality percentage was lower in the chicks fed on the CC-supplemented diet than those fed on the non-CC-supplemented diet (19% vs. 36%, respectively). The diet with the CC supplementation significantly increased the abundance of Phascolactobacteria (linear discriminant analysis; LDA >4) and Bacteroidota (26.7% vs. 17.7%, respectively) as well as decreased the enrichment of Clostridium (5.0% vs. 9.1%, respectively) in the ostrich ilea compared to the diet without CC. The supplementation of CA at a dose of 80 mg/kg significantly increased the expression level of ZO-1 and claudin-3 (p < 0.0001) and suppressed the levels of IL-1β, IL-6, and TNF-α (p < 0.0001) in ostriches with LPS-induced ileitis.ConclusionOur results substantiate that CC or CA supplementation in a diet could effectively improve growth performance and reduce mortality in juvenile ostriches via modulating the gut microbiota and attenuating enteritis.

This research aimed to investigate effects of different yeast culture (YC) levels on in vitro fermentation characteristics and bacterial and fungal community under high concentrate diet. A total of 5 groups were included in the experiment: control group without YC (CON), YC1 (0.5% YC proportion of substrate dry matter), YC2 (1%), YC3 (1.5%) and YC4 (2%). After 48 h of fermentation, the incubation fluids and residues were collected to analyze the ruminal fermentation parameters and bacterial and fungal community. Results showed that the ruminal fluid pH of YC2 and YC4 groups was higher (P < 0.05) than that of CON group. Compared with CON group, the microbial protein, propionate and butyrate concentrations and cumulative gas production at 48 h of YC2 group were significantly increased (P < 0.05), whereas an opposite trend of ammonia nitrogen and lactate was observed between two groups. Microbial analysis showed that the Chao1 and Shannon indexes of YC2 group were higher (P < 0.05) than those of CON group. Additionally, YC supplementation significantly decreased (P < 0.05) Succinivibrionaceae_UCG-001, Streptococcus bovis and Neosetophoma relative abundances. An opposite tendency of Aspergillus abundance was found between CON and YC treatments. Compared with CON group, the relative abundances of Prevotella, Succiniclasticum, Butyrivibrio and Megasphaera elsdenii were significantly increased (P < 0.05) in YC2 group, while Apiotrichum and unclassified Clostridiales relative abundances were decreased (P < 0.05). In conclusion, high concentrate substrate supplemented with appropriate YC (1%) can improve ruminal fermentation and regulate bacterial and fungal composition.

Objective:This study evaluates the effect of electro-acupuncture (EA) on visceral hypersensitivity (VH) and the expression of N-methyl-D-aspartate receptor-2B (NMDAR-2B) and glutamate transporter EAAT2 in goats. Methods:Twenty-four goats were divided into four groups: saline, 2, 4, 6-Trinitrobenzenesulfonic acid (TNBS), TNBS + EA, and sham EA. EA was administered at Zusanli (ST36) with 60 Hz and 1-3 mA on specified days. Electromyography (EMG) recorded visceromotor response to colorectal distention (CRD). Spinal cords were collected for immunohistochemistry, western blotting, and RT-PCR. The ileum was examined histologically. Results:The repeated EA administration significantly attenuated VH (P < 0.05) in TNBS-treated goats without similar effects in the sham group. NMDAR-2B expression increased (P < 0.01), and EAAT2 expression decreased (P < 0.01) in the TNBS group compared to saline. EA increased the EAAT2 and decreased the NMDAR-2B expression (P < 0.01) compared to TNBS, with no change in the sham-EA group. Conclusion:EA may alleviate VH by upregulating EAAT2 and downregulating NMDAR-2B in the spinal cord of TNBS-treated goats, indicating its potential for treating chronic visceral pain in gastrointestinal disorders.

Background Existing remedial approaches for relieving neuropathic pain (NPP) are challenging and open the way for alternative therapeutic measures such as electroacupuncture (EA). The mechanism underlying the antinociceptive effects of repeated EA sessions, particularly concerning the regulation of the Adora3 receptor and its associated enzymes, has remained elusive. Methods This study used a mouse model of spared nerve injury (SNI) to explore the cumulative analgesic effects of repeated EA at ST36 (Zusanli) and its impact on Adora3 regulation in the spinal cord dorsal horn (SCDH). Forty-eight male mice underwent SNI surgery for induction of neuropathic pain and were randomly assigned to the SNI, SNI + 2EA, SNI + 4EA, and SNI + 7EA groups. Spinal cord (L4-L6) was sampled for immunofluorescence, adenosine (ADO) detection and for molecular investigations following repeated EA treatment. Results Following spared nerve injury (SNI), there was a significant decrease in mechanical withdrawal thresholds (PWTs) and thermal nociceptive withdrawal latency (TWL) in the ipsilateral hind paw on the third day post-surgery, while the contralateral hind paw PWTs showed no significant changes. On subsequent EA treatments, the SNI + EA groups led to a significant increase in pain thresholds (p < 0.05). Repeated EA sessions in SNI mice upregulated Adenosine A3 (Adora3) and cluster of differentiation-73 (CD73) expression while downregulating adenosine deaminase (ADA) and enhancing neuronal instigation in the SCDH. Colocalization analysis of Neun-treated cells revealed increased Adora3 expression, particularly in the SNI + 7EA group. Conclusions In conclusion, cumulative electroacupuncture treatment reduced neuropathic pain by regulating Adora3 and CD73 expression, inhibiting ADA and most likely increasing neuronal activation in the SCDH. This study offers a promising therapeutic option for managing neuropathic pain, paving the way for further research.

Neuropathic pain (NPP) is a devastating and unbearable painful condition. As prevailing treatment strategies have failed to mitigate its complications, there remains a demand for effective therapies. Electroacupuncture (EA) has proved a potent remedial strategy in NPP management in humans and mammals. However, past studies have investigated the underlying mechanism of the analgesic effects of EA on NPP, focusing primarily on adenosine receptors in peripheral tissues. Herein, we elucidate the role of the adenosine (Adora-3) signaling pathway in mediating pain relief through EA in the central nervous system, which is obscure in the literature and needs exploration. Specific pathogen-free (SPF) male adult mice (C57BL/6 J) were utilized to investigate the effect of EA on adenosine metabolism (CD73, ADA) and its receptor activation (Adora-3), as potential mechanisms to mitigate NPP in the central nervous system. NPP was induced via spared nerve injury (SNI). EA treatment was administered seven times post-SNI surgery, and lumber (L4–L6) spinal cord was collected to determine the molecular expression of mRNA and protein levels. In the spinal cord of mice, following EA application, the expression results revealed that EA upregulated (p < 0.05) Adora-3 and CD73 by inhibiting ADA expression. In addition, EA triggered the release of adenosine (ADO), which modulated the nociceptive responses and enhanced neuronal activation. Meanwhile, the interplay between ADO levels and EA-induced antinociception, using an Adora-3 agonist and antagonist, showed that the Adora-3 agonist IB-MECA significantly increased (p < 0.05) nociceptive thresholds and expression levels. In contrast, the antagonist MRS1523 exacerbated neuropathic pain. Furthermore, an upregulated effect of EA on Adora-3 expression was inferred when the Adora-3 antagonist was administered, and the EA treatment increased the fluorescent intensity of Adora-3 in the spinal cord. Taken together, EA effectively modulates NPP by regulating the Adora-3 signaling pathway under induced pain conditions. These findings enhance our understanding of NPP management and offer potential avenues for innovative therapeutic interventions.