Electroacupuncture Alleviates Memory Deficits in APP/PS1 Mice by Targeting Serotonergic Neurons in Dorsal Raphe Nucleus

Objective Alzheimer’s disease(AD)has become a significant global concern,but effective drugs able to slow down AD progression is still lacked.Electroacupuncture(EA)has been demonstrated to ameliorate cognitive impairment in individuals with AD.However,the underlying mechanisms remains poorly understood.This study aimed at examining the neuroprotective properties of EA and its potential mechanism of action against AD.Methods APP/PS1 transgenic mice were employed to evaluate the protective effects of EA on Shenshu(BL 23)and Baihui(GV 20).Chemogenetic manipulation was used to activate or inhibit serotonergic neurons within the dorsal raphe nucleus(DRN).Learning and memory abilities were assessed by the novel object recognition and Morris water maze tests.Golgi staining,western blot,and immunostaining were utilized to determine EA-induced neuroprotection.Results EA at Shenshu(BL 23)and Baihui(GV 20)effectively ameliorated learning and memory impairments in APP/PS1 mice.EA attenuated dendritic spine loss,increased the expression levels of PSD95,synaptophysin,and brain-derived neurotrophic factor in hippocampus.Activation of serotonergic neurons within the DRN can ameliorate cognitive deficits in AD by activating glutamatergic neurons mediated by 5-HT1B.Chemogenetic inhibition of serotonergic neurons in the DRN reversed the effects of EA on synaptic plasticity and memory.Conclusion EA can alleviate cognitive dysfunction in APP/PS1 mice by activating serotonergic neurons in the DRN.Further study is necessary to better understand how the serotonergic neurons-related neural circuits involves in EA-induced memory improvement in AD.

微小核糖核酸mmu-miR-8114和mmu-miR-3473b在细粒棘球蚴致敏小鼠中的调控作用

目的探索微小核糖核酸(miRNAs)在通过细粒棘球蚴感染引起的小鼠过敏反应中的调节作用。方法建立小鼠体内棘球蚴病动物模型,收集脾脏单核细胞进行转录组测序。比较过敏性小鼠和非过敏小鼠之间mRNAs和miRNAs的差异表达。使用Targetscan和miRanda软件预测mRNAs和miRNAs之间的靶向调控关系,并通过蛋白质—蛋白质相互作用(PPI)网络分析,识别核心基因,通过富集分析探讨核心基因的生物学作用。最后利用qRT-PCR和Western blot验证关键结果。结果过敏与非过敏小鼠之间存在1642个差异表达的mRNAs。对18个差异表达的miRNAs进行分析,鉴定出60个差异表达的靶标基因,其中在PPI网络中连通度最大的前10个基因被认定为核心基因。富集分析显示核心基因主要参与NF-kappa B信号通路。基于miRNAs与mRNAs之间的负调控作用以及双荧光素酶报告系统发现mmu-miR-8114与Atf3以及mmu-miR-3473b与Myd88具有靶向调控关系,并与NF-kappa B信号通路有关。通过qRT-PCR验证了mmu-miR-8114和mmu-miR-3473b在过敏小鼠中下调表达,Atf3、Myd88、Socs3在过敏小鼠中上调表达。qRT-PCR和Western blot结果发现NF-kappa B信号通路在过敏小鼠中的活性增加。结论本研究揭示了mmu-miR-8114和mmu-miR-3473b通过NF-kappa B信号通路调控导致小鼠过敏反应的重要作用,为理解细粒棘球蚴感染引起的过敏反应机制提供了新的见解。

NF-kappa B p65和VEGF在原发性肝癌中的表达及意义

[目的]探讨核转录因子(NF-kappa B p65)和血管内皮生成因子(VEGF)在晚期原发性肝癌(HCC)组织和正常肝脏组织中的表达及其意义。[方法]采用免疫组织化学染色SP法检测65例晚期HCC组织切片(其中36例直径≥5cm,16例5cm>直径>2cm,13例直径≤2cm;28例肝内病灶数目大于1个,37例肝内病灶数目为1个)和30例正常肝脏组织(对照组)中NF-kappa B p65和VEGF抗体的表达水平,采用秩和检验方法分析其差异性,并再以直线相关方法分析NF-kappa B p65和VEGF在HCC中表达的相关性。[结果]各肿瘤组随肿瘤直径的增大,NF-kappa B p65和VEGF表达水平均出现递增的趋势。肝内(肿瘤数目>1个)NF-kappa B p65和VEGF表达水平强于肝内无转移组(P<0.05)。NF-kappa B p65和VEGF在HCC中表达呈正相关。[结论]VEGF有促进肝癌生长和侵袭转移作用,而NF-kappaBp65在HCC生长中有促进VEGF表达上调的作用。

非小细胞肺癌中NF-kappa B的表达与凋亡的关系

目的:研究核因子kappa B(nuclear factor kappa B,NF-kappa B)在非小细胞肺癌中的表达,探讨不同NF-kappa B表达的非小细胞的凋亡率,明确NF-kappa B在非小细胞肺癌中的临床意义。方法:应用Western blot检测了45例从2005年10月到2005年12月手术切除的非小细胞肺癌的新鲜标本,并用Tunel法测其凋亡率。结果:在45例患者中,NF-Kappa B的相对量是0.6047±0.3572,而且分化差的非小细胞肺癌中的NF-kappa B的表达比分化好的非小细胞肺癌高(P<0.05);在NF-kappa B高表达中肺癌细胞的凋亡率为56.4%,而在低表达中则为76.7%(P<0.05)。结论:NF-kappa B的表达与非小细胞肺癌的分化相关,在非小细胞肺癌中,NF-kappa B抑制凋亡,并在非小细胞肺癌的形成和发展中起重要作用,或许可以作为基因治疗的靶点为非小细胞肺癌的诊治提供新的思路。

离子硅对成骨细胞NF-kappa B核内转录因子活性的影响

目的研究离子硅对体外培养的成骨细胞NF-kappa B核转录因子活性的影响。方法分别用终浓度为1 mmol/L和2 mmol/L离子硅(SiO32)-处理MC3T3-E1成骨细胞,处理时间分别为6、12、24和48 h,设置对照组(不加处理因素);采用流式细胞术检测细胞周期,计算细胞的增殖指数;Westernblotting方法检测NF-kappa B信号通路的相关蛋白表达量及其变化。结果流式细胞术结果显示,与对照组相比,1 mmol/L浓度的离子硅处理24 h组和48 h组,MC3T3-E1细胞增殖明显;Western blot结果显示,1 mmol/L浓度的离子硅促进成骨细胞增殖与p-NF-kappa B表达上升密切相关。结论骨材料中释放的微量的硅不会引起成骨细胞损伤,相反,微量的硅酸盐可能通过激活NF-kappa B诱导成骨细胞增殖。

Effects of ginkgolide B on neuronal discharges in paraventricular nucleus of rat hypothalamic slices

Objective To study the central role of ginkgolide B （BN52021） in regulating cardiovascular function of nerve center by examining the effects of ginkgolide B on the electrical activity of rat paraventricular nucleus （PVN） neurons in hypothalamic slice preparation and to elucidate the mechanism involved. Methods Extracellular single-unit discharge recording technique. Results （1） In response to the application of ginkgolide t3 （0.1, 1, 10 μmol/L; n = 27） into the perfusate for 2 rain, the spontaneous discharge rates （SDR） of 26 （26/27, 96.30%） neurons were significantly decreased in a dose-dependent manner. （2） Pretreatment with L-glutamate （L-Glu, 0.2 mmol/L） led to a marked increase in the SDR of all 8 （100%） neurons in an epileptiform pattern. The increased discharges were suppressed significantly after ginkgolide B （1 μmol/L） was applied into the perfusate for 2 min. （3） In 8 neurons, perfusion of the selective L-type calcium channel agonist, Bay K 8644 （0.1 μmol/L）, induced a significant increase in the discharge rates of 8 （8/8, 100%） neurons, while ginkgolide B （1μmol/L） applied into the perfusate, could inhibit the discharges of 8 （100%） neurons. （4） In 8 neurons, the broad potassium channels blocker, tetraethylammonium （TEA, 1 mmol/L） completely blocked the inhibitory effect of ginkgolide B （1 μmol/L）. Conclusion These results suggest that ginkgolide B can inhibit the electrical activity of paraventricular neurons. The inhibitory effect may be related to the blockade of L-type voltage-activated calcium channel and potentially concerned with delayed rectifier potassium channel （KDR）.

NF-kappa B抑制剂PDTC抑制IL-1β诱导软骨细胞生成NO的实验研究

为探讨PDTC对IL 1β诱导的大鼠软骨细胞生成NO的影响 ,寻找抑制NO生成的新途径。分离并培养大鼠软骨细胞 ,加入不同度的PDTC ,以IL 1β刺激软骨细胞 ,RT PCR检测iNOS的mRNA表达 ,NO检测试剂盒检测NO的含量。结果显示PDTC能显著抑制IL 1β诱导的iNOSmRNA的表达及NO的生成且呈剂量依赖性。表明PDTC能抑制IL 1β诱导的NO生成 。

Expression of NF-kappaB in rotavirus-induced damage to the liver and biliary tract in neonatal mice

BACKGROUND: Biliary atresia, the etiology of which still remains unclear, occurs exclusively in newborns and most are infected with rotavirus. In this study, we aimed to investigate the histopathological patterns of different kinds of rotavirus in the liver and biliary tract of neonatal mice and the expression of NF-kappa B in the liver and biliary tract of infected mice. METHODS: Twenty-three adult mice (8 were male and 15 female) were divided into 8 breeding pairs, and each pair (I male and 2 females) was housed in a cage in a laminar flow hood. Newborn mice, 24-48 hours old were randomly divided into A, B and C groups. The A and B groups were respectively inoculated with MMU18006 and SA11 rotavirus through the intraperitoneal route, while group C as blank control was only inoculated with culture medium. The liver was dissected after 5, 10, 15, 21 and 28 days; the weight of each mouse and the histopathological patterns in the liver were recorded. The expression of NF-kappa B in the liver and intrahepatic bile ducts was detected by immunohistochemical staining and the expression intensity was analyzed with a GT-2 imaging instrument. RESULTS: The average increase in weight of infected mice was significantly slower than that of the normal control, while the growth rate of group A (injected with MMU18006 rotavirus) was slower than that of group B (SA11 rotavirus). In infected mice, the acute and chronic inflammation of liver and intra- and extra-hepatic bile ducts was more significant in group A. Stenosis was found in most intrahepatic bile ducts, and sporadically in extrahepatic bile ducts. The expression of NF-kappa B in infected mice was dramatically higher than that of the normal control, while the expression in group A was higher than in group B. CONCLUSIONS: Significant damage to the liver and biliary tract of neonatal mice can be induced by inoculating MMU18006 rotavirus through the intraperitoneal route, which is very similar to the pathology of biliary atresia in the newborn human. Similar inoculation with SA11 rotavirus can only result in moderate impairment that disappears quickly. The difference of pathogenicity between the two rotaviruses may depend on their differing capacities to increase the expression of NF-kappa B in the liver and biliary tract.

Activation of TLR-4 and liver injury via NF-kappa B in rat with acute cholangitis

BACKGROUND: Toll-like receptors (TLRs) are a family of type 1 transmembrane receptors, which can recognize different pathogen-associated molecular patterns. Among them, TLR-4 is specific to lipopolysaccharide. It transfers the infection signal into the cell and promotes the translocation of nuclear factor kappa B (NF-kappa B) to the nucleus and the subsequent transcriptional activation of genes encoding pro- and anti-inflammatory cytokines and chemokines. Acute cholangitis (AC) is a common biliary tract infection in oriental countries, and often leads to liver injury. The activation of TLR-4 and its significance in liver injury in rats with AC remain unclear. METHODS: Rat models of AC (biliary tract obstruction+E. coli injection, n=36) and control models (biliary tract obstruction+saline, n=18) were made. Liver tissue injury was investigated by pathological examination. The levels of serum TNF-alpha and IL-10 were measured by enzyme-linked immunosorbent assay, and the expressions of TLR-4, NF-kappa B mRNAs and proteins in the liver were detected by RT-PCR, immunohistochemical staining and Western blotting, respectively. RESULTS: Severe liver tissue injury in rats with AC was evident as shown by pathological examination. TLR-4 and NF-kappa B were strongly expressed in the cytoplasm of hepatocytes in the AC group. They were negative or slightly positive in the control group. TLR-4 mRNA and protein in the liver of rats with AC increased 1 hour after biliary tract ligation and E. coli injection, and peaked at 6 hours after surgery. Twenty-four hours later, they began to decrease. The expression of TLR-4 was paralleled by that of NF-kappa B in the liver and TNF-alpha in serum. CONCLUSION: The higher expression of TLR-4 in the liver of rats with AC may be involved in liver injury through the activation of NF-kappa B and release of cytokines such as TNF-alpha.

Effects of Dan-shao-hua-xian on expression of PPAR-gamma and NF-kappa B in rat liver fibrosis

BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPAR-gamma) and nuclear factor kappa B (NF-kappa B) play important roles in liver fibrosis. This study aimed to investigate the effects of Dan-shao-hua-xian, a preparation of traditional Chinese medicine, on the expression of PPAR-gamma and NF-kappa B in the fibrotic livers of rats. METHODS: Seventy Wistar rats were randomly divided into 4 groups: treatment (model, 8 weeks+treatment, 8 weeks; group A), natural recovery (model, 8 weeks+ saline, 8 weeks; group B), model (model only, 8 weeks; group Q, and control (normal, untreated, 16 weeks; group D). Each group consisted of 20 rats (except for group D, which had 10). Fibrotic liver models were induced in rats by subcutaneous injection of CCI4, oral administration of alcohol and a high-lipid/low-protein diet for 8 weeks. After the models were established, the rats in group A were orally given Dan-shao-hua-xian capsules daily for another 8 weeks. Then, the liver indices serum hyaluronic acid (HA), tumor necrosis factor-alpha (TNF-alpha) and alanine aminotransferase (ALT) were measured. The degree of hepatic fibrosis was evaluated by optical microscopy. Hydroxyproline (Hyp) in the liver tissue was determined. The expression of PPAR-gamma was detected by immunohistochemical techniques. The protein levels of PPAR-gamma and NF-kappa B were determined by Western blotting. RESULTS: The concentrations of serum HA, TNF-alpha and Hyp in group C increased compared with group D (P<0.05), and they decreased in group A compared with group C (P<0.05). The expression of PPAR-gamma in group C decreased compared with group D (P<0.05), and it increased in group A compared with groups B and C (P<0.05). Similarly, Western blotting showed that the expression of PPAR-gamma in group C decreased compared with group D, and it increased in group A compared with group C. The expression of NF-kappa B increased in group C compared with group D (P<0.05), and it decreased in group A compared with group C (P<0.05). CONCLUSION: Dan-shao-hua-xian capsules enhance the expression of PPAR-gamma but decrease that of TNF-alpha and NF-kappa B in the liver tissues of CCI4-induced hepatic fibrotic rats. These effects may play a role in its activity in treating hepatic fibrosis.

Relationship between the Expression of NF-kappa B and Apoptosis in Non-Small Cell Lung Cancer

OBJECTIVE To study the expression of the nuclear factor kappa B (NF-kappa B) in non-small cell lung cancer (NSCLC),to explore the apoptotic ratio in NSCLC related to different NF-kappa Bs,and to understand the clinical significance of NF-kappa B in NSCLC apoptosis. METHODS NF-kappa B expression in 45 new samples of NSCLC,collected during a period from October to December,2005,was assayed using Western blots,and the apoptotic ratio of NSCLC was determined by the Tunel method. RESULTS Of the 45 patients,the average relative expression of NF-kappa B was 0.6047±0.3572.The expression of NF-kappa B was higher in the poorly differentiated lung cancer cells than in the well-differentiated tumors(P<0.05).The apoptotic ratio was 56.4% in the lung cancer cells with higher NF-kappa B expression,and was 76.7% in those with lower NF-kappa B expression(P<0.05). CONCLUSION The expression of NF-kappa B is correlated with the differentiation of NSCLC.NF-kappa B inhibits apoptosis in NSCLC.As a transcription factor,NF-kappa B plays a very important role both in formation and in development of NSCLC. NF-kappa B might serve as a target for NSCLC gene therapy.

Effects of ivabradine on Notch and NF-kappa B signaling pathway in myocardial cells of rats with myocardial infarction

Objective:To investigate the effects of ivabradine on Notch and NF-kappa B signaling pathway in myocardial cells of rats with myocardial infarction.Methods: The model of myocardial infarction was established by ligating the left anterior descending coronary artery. The surviving rats were randomly divided into model group (MI group,n=8) and treatment group (IVA group,n=8). Rats with the same location but without ligation of the left anterior descending coronary artery were used as control group (CON group,n = 8). IVA was administered for 28 d. Hemodynamic and cardiac function indexes of all rats were measured: heart rate (HR), systolic pressure (SBP), diastolic pressure (DBP), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP) and maximum rate of increase and decrease of left ventricular internal pressure (+dp/dt);left ventricular mass index, left ventricular cross-sectional diameter and infarct area;The expression of Notch signaling pathway components mRNA (Notch-1, Dll-4, Hes-1) in rat cardiomyocytes was detected by PT-PCR, and the expression of DICD-1 and P65 protein was detected by western-blot. One-way ANOVA was used for comparison between groups, and SNK was used for comparison between groups.Results: SBP, DBP, MAP, LASP, LVEDP and (+dp/dt) in MI group were lower than those in control group (P<0.05), while IVA was higher than those in MI group (P<0.05). Left ventricular mass index and left ventricular sectional diameter in MI group were significantly higher than those in control group (P<0.05), but lower than those in IVA group (P<0.05). The expression of Notch-1 in MI group was significantly higher than that in control group (P<0.05), but lower than that in IVA group (P<0.05). There was no significant difference in the expression of Dll-4 and Hes-1 mRNA between the three groups (P>0.05). The expression levels of NICD-1 and P 65 in MI group were significantly higher than those in CON group (P<0.05), but lower than those in IVA group (P<0.05). Conclusion: IVA may improve cardiac function and inhibit ventricular remodeling in rats with myocardial infarction through Notch and NF-kappa B signaling pathways.

Effects of gamma-aminobutyric acid receptors on muscarinic receptor-mediated free calcium ion levels in the facial nucleus following facial nerve injury

Muscarinic receptors and nicotine receptors can increase free calcium ion levels in the facial nucleus via different channels following facial nerve injury. In addition, γ-aminobutyric acid A （GABAA） receptors have been shown to negatively regulate free calcium ion levels in the facial nucleus by inhibiting nicotine receptors. The present study investigated the influence of GABAA, γ-aminobutyric acid B （GABAB） and C （GABAc） receptors on muscarinic receptors in rats with facial nerve injury by confocal laser microscopy. GABAA and GABAB receptors exhibited significant dose-dependent inhibitory effects on increased muscarinic receptor-mediated free calcium ion levels following facial nerve injury. Results showed that GABAA and GABAB receptors negatively regulate muscarinic receptor effects and interplay with cholinergic receptors to regulate free calcium ion levels for facial neural regeneration.

Lactate Decreases Bortezomib Sensitivity and Predicts Poor Clinical Outcomes of Multiple Myeloma

Objective:Metabolic disorders are regarded as hallmarks of multiple myeloma(MM)and are responsible for rapid cancer cell proliferation and tumor growth.However,the exact biological roles of metabolites in MM cells have not been fully explored.This study aimed to explore the feasibility and clinical significance of lactate for MM and investigate the molecular mechanism of lactic acid(Lac)in the proliferation of myeloma cells and cell sensitivity to bortezomib(BTZ).Methods:Metabolomic analysis of the serum was carried out to obtain metabolites expression and clinical characteristics in MM patients.The CCK8 assay and flow cytometry were used to detect cell proliferation,apoptosis,and cell cycle changes.Western blotting was used to detect the potential mechanism and apoptosis-and cycle-related protein changes.Results:Lactate was highly expressed in both the peripheral blood and bone marrow of MM patients.It was significantly correlated with Durie-Salmon Staging(DS Staging)and the International Staging System(ISS Staging)and the serum and urinary involved/uninvolved free light chain ratios.Patients with relatively high lactate levels had a poor treatment response.Moreover,in vitro experiments showed that Lac could promote the proliferation of tumor cells and decrease the proportion of G0/G1-phase cells,which was accompanied by an increased proportion of S-phase cells.In addition,Lac could decrease tumor sensitivity to BTZ by disrupting the expression of nuclear factor kappa B subunit 2(NFκB2)and Re1B.Conclusion:Metabolic changes are important in MM cell proliferation and treatment response;lactate could be used as a biomarker in MM and as a therapeutic target to overcome cell resistance to BTZ.

组织芯片研究Notch1与NF-κB在胃癌中的表达和意义

目的:研究Notchl和NF-KB在胃癌中的表达与临床病理特征的关系以及两者间相互作用的关系。方法:通过免疫组化的方法检测了含有168例胃癌和27例正常胃组织的组织芯片中Notchl和NF-κB的表达情况。结果:Notchl在胃癌中表达较正常胃组织明显上调(P<0.01),并与肿瘤大小、分化程度、浸润深度、脉管浸润明显相关;NF-κB在胃癌中表达明显高于正常对照组,并与肿瘤大小、分化程度、浸润深度、远处转移及脉管浸润明显相关。Kaplan-Meier分析显示Notchl和NF-κB阳性组的预后较阴性组差。Cox回归多因素分析显示Notchl是影响胃癌预后的独立因素之一(P<0.05)。结论:NF-κB的过度表达可能参与了Notchl信号通路失调介导的胃癌发生发展机制。Notchl可能成为评价胃癌预后和生存的新的指标,也可能作为胃癌治疗的新靶点。

Hedgehog和NF-κB信号通路在结核分枝杆菌感染Ⅱ型肺泡上皮细胞中的免疫调控作用初探

探讨结核分枝杆菌(Mycobacterium tuberculosis,Mtb)感染Ⅱ型肺泡上皮细胞(Alveolar epithelial typeⅡcells,AECⅡ)的免疫应答过程中,Hedgehog与NF-κB信号通路及炎症细胞因子的表达变化规律。利用结核分枝杆菌BCG(Bacillus Calmette-Guérin,BCG,卡介苗,弱毒株)和H37Rv(国际标准强毒株)株分别感染人AECⅡ细胞株A549,利用q PCR和Western blot检测Hedgehog信号通路以及NF-κB信号通路相关因子和炎症细胞因子的表达变化。结果表明,H37Rv和BCG感染A549细胞后,Hedgehog信号通路中Shh、Ptch和Gli1与NF-κB信号通路中p-NF-κB m RNA的表达水平升高,同时,伴随着炎症细胞因子IL-8和TNF-αm RNA的表达水平也升高。并且,H37Rv感染A549细胞引起信号分子Shh、Ptch、Gli1和NF-κB蛋白的表达水平高于BCG。Mtb感染AECⅡ细胞都能激活Hedgehog信号通路和NF-κB信号通路并促进细胞的炎症因子分泌,强毒株感染对信号通路的活化强于弱毒株。

人参皂苷Rg3对缺氧诱导的Eca-109和786-0细胞血管内皮生长因子和核因子κB表达的影响

目的探讨人参皂苷Rg3对人食管癌细胞株Eca-109和人肾癌细胞株786-0细胞缺氧诱导的血管内皮生长因子(VEGF)和核因子κB(NF-κB)表达的影响。方法在常氧和缺氧环境下,采用噻唑蓝(MTT)法检测Rg3对Eca-109和786-0细胞增殖的影响,实时荧光定量聚合酶链反应(PCR)检测Rg3对VEGF mRNA表达的影响,酶联免疫吸附测定(ELISA)法检测Rg3和NF-κB抑制剂对VEGF蛋白表达的影响,蛋白质印迹(Western)检测Rg3对P65蛋白表达的影响。结果在常氧或缺氧环境中,MTT显示Rg3都能显著抑制Eca-109和786-0细胞增殖,实时荧光定量PCR显示Rg3能显著抑制Eca-109和786-0细胞VEGF mRNA表达,ELISA显示Rg3和NF-κB抑制剂都能显著抑制VEGF蛋白的分泌,Western显示Rg3抑制P65蛋白表达。结论 Rg3可以抑制缺氧和常氧状态下的VEGF表达,其机制与抑制P65蛋白表达密切相关。

Thalamic Nucleus Reuniens Glutamatergic Neurons Mediate Colorectal Visceral Pain in Mice via 5-HT2B Receptors

Irritable bowel syndrome(IBS)is a common functional bowel disorder characterized by abdominal pain and visceral hypersensitivity.Reducing visceral hypersensitivity is the key to effectively relieving abdominal pain in IBS.Increasing evidence has confirmed that the thalamic nucleus reuniens(Re)and 5-hydroxytryptamine(5-HT)neurotransmitter system play an important role in the development of colorectal visceral pain,whereas the exact mechanisms remain largely unclear.In this study,we found that high expression of the 5-HT2B receptors in the Re glutamatergic neurons promoted colorectal visceral pain.Specifically,we found that neonatal maternal deprivation(NMD)mice exhibited visceral hyperalgesia and enhanced spontaneous synaptic transmission in the Re brain region.Colorectal distension(CRD)stimulation induced a large amount of c-Fos expression in the Re brain region of NMD mice,predominantly in glutamatergic neurons.Furthermore,optogenetic manipulation of glutamatergic neuronal activity in the Re altered colorectal visceral pain responses in CON and NMD mice.In addition,we demonstrated that 5-HT2B receptor expression on the Re glutamatergic neurons was upregulated and ultimately promoted colorectal visceral pain in NMD mice.These findings suggest a critical role of the 5HT2B receptors on the Re glutamatergic neurons in the regulation of colorectal visceral pain.