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Serotonin receptor 2B induces visceral hyperalgesia in rat model and patients with diarrhea-predominant irritable bowel syndrome

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World Journal of Gastroenterology 2024年第10期30卷 1431-1449页

作者： Zheng-Yang Li Yu-Qing Mao Qian Hua Yong-Hong Sun Hai-Yan Wang Xuan-Guang Ye Jing-Xian Hu Ya-Jie Wang Miao Jiang Department of Gastroenterology Jinshan Hospital of Fudan UniversityShanghai 201508China Department of Gastroenterology Shanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China Department of Gastroenterology Dalian Friendship HospitalDalian 116001Liaoning ProvinceChina Department of Pathology Jinshan Hospital of Fudan UniversityShanghai 201508China

BACKGROUND Serotonin receptor 2B(5-HT2B receptor)plays a critical role in many chronic pain conditions.The possible involvement of the 5-HT2B receptor in the altered gut sensation of irritable bowel syndrome with diarrhea(IBS-D)was investigated in the present study.AIM To investigate the possible involvement of 5-HT2B receptor in the altered gut sensation in rat model and patients with IBS-D.METHODS Rectosigmoid biopsies were collected from 18 patients with IBS-D and 10 patients with irritable bowel syndrome with constipation who fulfilled the Rome IV criteria and 15 healthy controls.The expression level of the 5-HT2B receptor in colon tissue was measured using an enzyme-linked immunosorbent assay and correlated with abdominal pain scores.The IBS-D rat model was induced by intracolonic instillation of acetic acid and wrap restraint.Alterations in visceral sensitivity and 5-HT2B receptor and transient receptor potential vanilloid type 1(TRPV1)expression were examined following 5-HT2B receptor antagonist adminis-tration.Changes in visceral sensitivity after administration of the TRPV1 antago-INTRODUCTION Irritable bowel syndrome(IBS)is a chronic functional bowel disorder characterized by recurrent abdominal pain with altered bowel habits that affects approximately 15%of the population worldwide[1].IBS significantly impacts the quality of life of patients.Although the pathogenesis of IBS is not completely understood,the role of abnormal visceral sensitivity in IBS has recently emerged[2,3].5-Hydroxytryptamine(5-HT)is known to play a key role in the physiological states of the gastrointestinal tract.Plasma 5-HT levels in IBS with diarrhea(IBS-D)patients were greater than those in healthy controls[4],suggesting a possible role of 5-HT in the pathogenesis of IBS-D.The serotonin receptor 2(5-HT2 receptor)family comprises three subtypes:5-HT2A,5-HT2B,and 5-HT2c.All 5-HT2 receptors exhibit 46%-50%overall sequence identity,and all of these receptors preferentially bind to Gq/11 to increase inositol phosphates and intracell

关键词： Diarrhea-predominant irritable bowel syndrome Serotonin receptor 2B Transient receptor potential vanilloid type-1 visceral hypersensitivity Abdominal pain

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Electroacupuncture diminishes P2X_2 and P2X_3 purinergic receptor expression in dorsal root ganglia of rats with visceral hypersensitivity

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Neural Regeneration Research 2013年第9期8卷 802-808页

作者： Zhijun Weng Luyi Wu Yuan Lu Lidong Wang Linying Tan Ming Dong Yuhu Xin Shanghai Academy of Traditional Chinese Medicine Fudan University Shanghai Cancer Center

Electroacupuncture at Shangjuxu （ST37） and Tianshu （ST25） can improve visceral hypersensitivity in rats. Colorectal distension was used to establish a rat model of chronic visceral hypersensitivity. Immunohistochemistry was used to detect P2X2 and P2X3 receptor expression in dorsal root ganglia from rats with chronic visceral hypersensitivity. Results demonstrated that abdominal withdrawal reflex scores obviously increased following establishment of the model, indicating visceral hypersensitivity. Simultaneously, P2X2 and P2X3 receptor expression increased in dorsal root ganglia. After bilateral electroacupuncture at Shangjuxu and Tianshu, abdominal withdrawal reflex scores and P2X2 and P2X3 receptor expression decreased in rats with visceral hypersensitivity. These results indicated that electroacupuncture treatment improved visceral hypersensitivity in rats with irritable bowel syndrome by reducing P2X2 and P2X3 receptor expression in dorsal root ganglia.

关键词： neural regeneration acupuncture and moxibustion P2X2 P2X3 visceral hypersensitivity irritablebowel syndrome electroacupuncture P2 purinergic receptors abdominal withdrawal reflex scoresacupuncture and moxibustion peripheral nerve injury grants-supported paper photographscontaining paper neuroregeneration

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Fructo-oligosaccharide intensifies visceral hypersensitivity and intestinal inflammation in a stress-induced irritable bowel syndrome mouse model

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World Journal of Gastroenterology 2017年第47期23卷 8321-8333页

作者： Bin-Rui Chen Li-Jun Du Hui-Qin He John J Kim Yan Zhao Ya-Wen Zhang Liang Luo Ning Dai Department of Gastroenterology Sir Run Run Shaw Hospital School of Medicine Zhejiang University Division of Gastroenterology Loma Linda University Medical Center

AIM To determine whether fructo-oligosaccharide(FOS) affects visceral sensitivity, inflammation, and production of intestinal short-chain fatty acids(SCFA) in an irritable bowel syndrome(IBS) mouse model.METHODS Mice were randomly assigned to daily oral gavage of saline solution with or without FOS(8 g/kg body weight) for 14 d. Mice were further assigned to receive either daily one-hour water avoidance stress(WAS) or sham-WAS for the first 10 d. After 2 wk, visceral sensitivity was measured by abdominal withdrawal reflex in response to colorectal distension and mucosal inflammation was evaluated. Gas chromatography, real-time reverse transcription PCR, and immunohistochemistry assays were used to quantify cecal concentrations of SCFA, intestinal cytokine expression, and number of intestinal mast cells per high-power field(HPF), respectively.RESULTS Mice subjected to WAS exhibited visceral hypersensitivity and low-grade inflammation. Among mice subjected to WAS, FOS increased visceral hypersensitivity and led to higher cecal concentrations of acetic acid(2.49 ± 0.63 mmol/L vs 1.49 ± 0.72 mmol/L, P visceral sensitivity, intestinal inflammation, or cecal SCFA levels was detected with or without FOS administration in mice subjected to sham-WAS.CONCLUSION FOS administration intensifies visceral hypersensitivity and gut inflammation in stress-induced IBS mice, but not in t

关键词： Fructo-oligosaccharide Stress Irritable bowel syndrome visceral hypersensitivity Intestinal inflammation Short chain fatty acids FODMAP

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Effect of NMDA receptors NR2B subunit on chronic visceral hypersensitivity

Effect of NMDA receptors NR2B subunit on chronic visceral hy...

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2008北京国际生理科学学术大会

作者： LIN Chun~1,LIN Guo-Wei~2,ZHENG Wei~3,CHEN Ai-Qin~1,YU Chang-Xi~4 1 Laboratory for Pain Research,Department of Physiology and Pathophysiology,Research Center of Neuroscience 2 Department of Physiology,Fujian Vocational and Technical College of Health 3 Department of Pathology 4 College of Pharmacology,Fujian Medical University,China

Aim:To explore effects of peripheral and spinal NMDA receptor NR2B subunit on chronic visceral hypersensitivity.Methods:Model rats received daily noxious colorectal distensions(CRD) on postnatal day 8-15. visceral hypersensitivity of rats were evaluated by abdominal withdrawal reflex"AWR"when they were 8 weeks. Expression of NMDA receptor NR2B subunit in dorsal root ganglions and spinal visceral related neurons was compared between control and model rats by immunohistochemistry method.Responses of primary afferents to CRD were compared before and after systemic administration of AP-7,an NMDA antagonist,in control and model rats.Results:(1) AWR scores significantly increased in adult model rats.(2) Image analysis showed that average optical density which represents the expression of NR2B subunit in dorsal root gangalia was significantly increased in model rats than that in control rats.(3) Image analysis shows that average optical which represents the expression of NR2B subunit in spinal visceral related neurons was significantly higher in model rats than those in control rats.(4) The percent inhibition of the responses of lunbosacral dorsal root to CRD caused by AP-7 was significantly higher in model rats than those in control rats.Conclusion:NMDA-NR2B may play a role in the peripheral and spinal mechanism of chronic visceral pain.

关键词： chronic visceral hypersensitivity AP-7 NMDA-NR2B

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Bladder-colon chronic cross-sensitization involves neuro-glial pathways in male mice

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World Journal of Gastroenterology 2022年第48期28卷 6935-6949页

作者： Karim Atmani Fabien Wuestenberghs Maximilien Baron Illona Bouleté Charlène Guérin Wafa Bahlouli David Vaudry Jean Claude do Rego Jean-Nicolas Cornu Anne-Marie Leroi Moïse Coëffier Mathieu Meleine Guillaume Gourcerol Nutrition Gut and Brain Unit(Inserm U1073)Institute for Research and Innovation in BiomedicineUniversitéde Rouen NormandieRouen 76000France Department of Gastroenterology and Hepatology UniversitéCatholique de LouvainCHU UCL NamurYvoir 5530Belgium Department of Physiology CHU RouenUniversitéde Rouen NormandieRouen 76031France Department of Urology CHU RouenUniversitéde Rouen NormandieRouen 76000France Inserm UMR 1245Team Epigenetics and Pathophysiology of Neurodevelopmental DisordersUniversitéde Rouen NormandieRouen 76000France Behavioural Analysis Platform(SCAC) HeRacLeS Inserm US51-CNRS UAR2026Institute for Research and Innovation in BiomedicineUniversitéde Rouen NormandieRouen 76000France Department of Nutrition CHU RouenUniversitéde Rouen NormandieRouen 76000France Inserm U1107 NeuroDolClermont Auvergne UniversityClermont-Ferrand 63000France

BACKGROUND Irritable bowel syndrome and bladder pain syndrome often overlap and are both characterized by visceral hypersensitivity.Since pelvic organs share common sensory pathways,it is likely that those syndromes involve a cross-sensitization of the bladder and the colon.The precise pathophysiology remains poorly understood.AIM To develop a model of chronic bladder-colon cross-sensitization and to investigate the mechanisms involved.METHODS Chronic cross-organ visceral sensitization was obtained in C57BL/6 mice using ultrasound-guided intravesical injections of acetic acid under brief isoflurane anesthesia.Colorectal sensitivity was assessed in conscious mice by measuring intracolonic pressure during isobaric colorectal distensions.Myeloperoxidase,used as a marker of colorectal inflammation,was measured in the colon,and colorectal permeability was measured using chambers.c-Fos protein expression,used as a marker of neuronal activation,was assessed in the spinal cord(L6-S1 level)using immunohistochemistry.Green fluorescent protein on the fractalkine receptor-positive mice were used to identify and count microglia cells in the L6-S1 dorsal horn of the spinal cord.The expression of NK1 receptors and MAPK-p38 were quantified in the spinal cord using western blot.RESULTS visceral hypersensitivity to colorectal distension was observed after the intravesical injection of acetic acid vs saline(Pvisceral hypersensitivity was associated with the increased expression of c-Fos protein in the spinal cord(P<0.0001).In green fluorescent protein on the fractalkine receptor-positive mice,intravesical acetic acid injection resulted in an increased number of microglia cells in the L6-S1 dorsal horn of the spinal cord(P<0.0001).NK1 receptor and MAPK-p38 levels were increased in the spinal cord up to 7 d after injection(P=0.007 and 0.023 respectively).Color

关键词： Cross-organ sensitization MAPK-p38 Microglia NK1 receptor Pain visceral hypersensitivity

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P2X3 in primary afferent neurons mediates the relief of visceral hypersensitivity by electroacupuncture in an irritable bowel syndrome rat model

P2X3 in primary afferent neurons mediates the relief of visc...

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2019中国生理学会学术年会暨张锡钧基金第十五届全国青...

作者：张方 Zhe Ma Zhijun Weng Min Zhao Huangan Wu HANDan Zheng Luyi Wu Yuan Lu Chunhui Bao Yanan Liu Huirong Liu Shanghai Research Institute of Acupuncture and Meridian Shanghai University of Traditional Chinese Medicine Shanghai University of Traditional Chinese Medicine

Objective: To reveal the neurobiological mechanism by which the P2 X3 receptor of colonic primary sensory neurons in the dorsal root ganglia（DRG） of the lumbosacral segment（L6-S2）is involved in the alleviation of visceral hypersensitivity by electroacupuncture（EA） at the HeMu points in an irritable bowel syndrome（IBS） rat model.Methods: The IBS chronic visceral pain model was established in clean-grade male Sprague-Dawley（SD） rats according to the method of AL-chaer et al.At the 6 th week, bilateral EA at the He-Mu points（2/100 Hz, 1 mA,30 min） was conducted for intervention.The acupoints included the abdominal Tianshu point（ST25） and the Shangjuxu point（ST37）.We conducted behavioral studies, histopathology of the colon, electrophysiology, immunofluorescence histochemistry, and real-time polymerase chain reaction（PCR） assays to observe the role of the P2 X3 receptor in the colon and related DRG in relieving visceral hypersensitivity by EA.Results: 1) EA at the He-Mu points significantly reduced the visceral pain behavior scores of the IBS rats under different levels（20,40, 60, 80 mmHg） of colorectal distention（CRD） stimulation.2) EA downregulated the expression levels of the P2 X3 protein and its messenger RNA（mRNA） in the colon and related DRG of the IBS rats.3) EA regulated the electrical properties of the membranes [the resting membrane potential（RMP）, rheobase and action potential（AP）] of colon-associated DRG neurons in the IBS rats with visceral hypersensitivity.Conclusion: EA can regulate the excitatory properties of DRG neurons by regulating the abnormally elevated P2 X3 receptor protein and mRNA expression levels in the colon and related DRG of IBS rats with visceral pain.The regulation of P2 X3 may be the important neurobiological mechanism by which EA at the He-Mu points relieves visceral hypersensitivity in IBS rats.

关键词： irritable bowel syndrome visceral hypersensitivity peripheral sensitization P2X3 receptor electroacupuncture

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TRPV1 in basolateral amygdala mediates visceral hypersensitivity in adult rats with neonatal maternal deprivation

TRPV1 in basolateral amygdala mediates visceral hypersensiti...

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中国生理学会消化与营养专业委员会成立大会暨第一届学术会...

作者： Ying Xiao Ping-An Zhang Qiya Xu Hang Zheng Youlang Zhou 徐广银 Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases Laboratory of Translational Pain Medicine Institute of Neuroscience Soochow University 苏州大学神经科学研究所

Chronic stress exacerbates or causes relapse of symptoms such as abdominal pain and cramping in patients with irritable bowel syndrome（IBS）.Neonatal maternal deprivation（NMD）as an animal model used to study the mechanism of IBS may cause visceral hypersensitivity in adult rats.A growing body of evidence demonstrates that central amygdala（CeA）plays an important role in the modulation of visceral pain.However as one of the input pathways to CeA the role of basolateral amygdala（BLA）in the modulation of pain remains elusive.There arc highest densities of TRPV1 labeled neurons within BLA and TRPV1 was found to take part in peripheral pain modulation.This study is to determine whether TRPV1 in BLA mediates visceral hypersensitivity induced in adult rats with neonatal maternal deprivation.visceral hypersensitivity was induced by neonatal maternal deprivation（NMD）and was quantified by abdominal withdrawal reflex.Expression of TRPV1 was determined by Western blot.The synaptic transmission of neurons in BLA was recorded by patch clamping.It was found that the expression of TRPV1 in BLA was significantly upregulated in NMD rats;glutamatergic synaptic activities in BLA were increased in NMD rats;application of capsazepine（TRPV1 antagonist）decreased glutamatergic synaptic activities of BLA neurons in NMD slices through a presynaptic mechanism;application of capsaicin（TRPV1 agonist）increased glutamatergic synaptic activities of BLA neurons in control slices through presynaptic mechanism without affecting GABAergic synaptic activities;microinjecting capsazepine into BLA significantly increased colonic distension threshold both in control and NMD rats.Our data suggested that upregulation of TRPV1 in BLA contributes to visceral hypersensitivity of NMD rats through enhancing excitaion of BLA thus identifying a potential target for treatment of chronic visceral pain.

关键词： Neonatal maternal deprivation visceral hypersensitivity TRPV1 Basolateral amygdala Presynaptic mechanism

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Sensitization of sodium channels contributes to colonic visceral hypersensitivity in diabetic rats and antinociceptive effects of α-lipoic acid

Sensitization of sodium channels contributes to colonic visc...

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中国生理学会第24届全国会员代表大会暨生理学学术大会

作者：宋珍源 Hong-Hong ZHANG Shu-Fen HU Ji HU Guang-Yin XU Division of Endocrinology the Second Affiliated Hospital of Soochow University Institute of Neuroscience Suzhou Key Laboratory of Pain Research and TherapyDepartment of NeurobiologySoochow University

Background:Intestinal dysfunction is a common chronic complication of diabetes mellitus,the underlying mechanisms of colonic visceral hypersensitivity in diabetic patients remain elusive.Recently,several lines of evidence suggest that the change of ion channels in dorsal root ganglion（DRG）neurons play a significant role in diabetic neuropathic pain.Objective:Our study was designed to investigate the changes in function and expression of voltage-gated sodium channels（VGSCs）of DRG neurons in visceral hypersensitivity of STZ-induced diabetic rats.Moreover,we aimed to explore the effect ofα-lipoic acid on VGSCs activities and visceral hypersensitivity of diabetic rats.Methods:Diabetic model was induced by a single intraperitoneal injection of streptozotocin（STZ;65mg/kg）in adult female rats.Behavioral response to colorectal distention（CRD）was used to examine visceral hypersensitivity by visual observation of abdominal withdrawal reflex（AWR）.Colon-specific DRG neurons were labeled with Di I and acutely dissociated for measuring excitability and sodium channel current under whole-cell patch clamp configurations.Western blot analysis was employed to measure the expression of Na V1.7 and Na V1.8 in colon-specific DRGs.Results:As evidenced by AWR scores responding to CRD of rats,we showed that visceral hypersensitivity was induced 2 weeks after STZ injection and lasted for another 6 weeks.STZ injection enhanced neuronal excitability and sodium current density of colon DRG neurons in diabetic rats.STZ injection also led to a marked upregulation expression of Na V1.7 and Na V1.8 compared with age-matched control rats.In addition,intraperitoneal injection withα-lipoic acid for 1 week,remarkably reduced the expression of Na V1.7 and Na V1.8 in STZ-induced diabetic rats.Furthermore,it also suppressed neuronal excitability,sodium channel current density,and attenuated visceral hypersensitivity in diabetic rats as well.Conclusion:Our findings suggest that STZ injection increases function and expression of VGSCs of colon-specific

关键词： Diabetes visceral hypersensitivity dorsal root ganglion voltage-gated sodium channels α-lipoic acid

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Neonatal reprogramming of miRNA-19 negatively regulates GRK6 expression in adult rats with visceral hypersensitivity

Neonatal reprogramming of miRNA-19 negatively regulates GRK6...

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中国神经科学学会第十二届全国学术会议

作者： Yuan Zhou Xuelian Liu Rui Wu Guang-Yin Xu Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-PsychoDiseases Institute of Neuroscience Soochow University Department of Anesthesiology Affiliated Hospital of Nantong University

Objective Irritable bowel syndrome（IBS） is a common gastrointestinal disorder characterized by chronic visceral pain in association with altered bowel habits. The pathogenesis of IBS remains elusive and treatment options are very limited. Studies indicate that early-life events constitute a risk factor for the development of IBS in adults. However, its precise mechanism is still poorly understood. MiR-19 a, a vulnerable development-related micro RNA screened out from micro RNA array analysis, is predicted to target 3’UTR of G-protein coupled receptor Kinase 6（GRK6）. The aim of the present study is to explore whether miR19 a is developmentally reprogrammed by early life insults and negatively regulates GRK6 expression in a rat model of adult visceral hypersensitivity induced by neonatal colonic inflammation（NCI）. Methods Distension threshold and abdominal withdrawal reflex（AWR） scores to grade colorectal distention in adult male rats were recorded as a measure of visceral hypersensitivity. Micro RNA microarray was applied to screen out a vulnerable development-related micro RNAs. Fluorescence in situ hybridization（FISH） and Immunofluorescence staining were used to detect the cellular distribution of miR-19 a. Intrathecal injection of miR-19 a antagomir and GRK6-LV was applied to investigate the roles of miR-19 a and GRK6 in NCIinduced visceral hyperalgesia. Methylation specific PCR（MSP） and bisulfite sequencing PCR（BSP） were performed to detect the status of DNA methylation on the Cp G islands of miR-19 a gene promoter. Dual luciferse report gene assay were performed to testify the targeting relationship between miR-19 a and GRK6. Colon-specific DRG neurons were labeled with Dil and acutely dissociated for measuring excitability of GRK6 overexpressed colon-specific neurons with patch-clamp techniques. Results Colonic infusion of acetic acid on postnatal（P） day 10 led to significant visceral hyperalgesia, upregulation of miR-19 a and downregulation of GRK6 in colon associated DRGs of adult rats. In contrast, coloni

关键词： Irritable bowel syndrome visceral hypersensitivity dorsal root ganglion miR-19a GKR6 DNA methyltransferases

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Epigenetic regulations of chronic visceral hypersensitivity in irritable bowel syndrome

Epigenetic regulations of chronic visceral hypersensitivity ...

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中国神经科学学会第十届全国学术会议

作者： Liyan Zhu Ruobing Qu Shufen Hu Ying Xiao Guang-Yin Xu Institute of Neuroscience Soochow University

Irritable bowel syndrome(IBS),characterized by persistent or recurrent abdominal pain with altered bowel movement in the absence of a demonstrable pathology,is one of the most common functional gastrointestinal disorders.The courses of IBS remain unkown and treatment options for chronic visceral pain are limited.Hydrogen sulfide(HS),a third gaseous modulator/mediator,has become recognized as an important endogenous molecular.HS is synthesized from L-cysteine primarily via two endogenous enzymes:cystathionine-β-synthetase(CBS) and cystathionine-γ-lyase,and has been reported to be involved in nociceptive signaling and inflammation.However,the molecular and epigenetic mechanisms of CBS-HS signaling in visceral nociceptive processing are not fully understood.Here we showed that IBS-like chronic visceral hyperalgesia induced by colonic injection of 0.5%acetic acid in 10-day-old rats significantly upregulated expression of CBS at both protein and mRNA levels in rat dorsal root ganglia(DRG).Intraperitoneal administration of CBS antagonist aminooxyacetic acid(AOAA) attenuated visceral hyperalgesia in a dose-dependent manner in rats with visceral hyperalgesia.In addition,application of AOAA reversed the enhanced excitability,reduced the upregulation of voltage-gated sodium channels Na1.7 and Na1.8 in DRGs and decreased total sodium current of DRG neurons in rats with visceral hyperalgesia.We also analyzed cbs gene promoter region by bioinformatics tools and found two CpG sites.Methylation specific PCR demonstrated that promoter region of cbs gene was less methylated in DRG samples from rats with visceral hyperalgesia than that from controls.Neonatal colonic infusion upregulated the expression of thymine DNA glycosylase while expression of DNA methylation transferases was not altered significantly.Our results suggest that epigenetic regulation of CBS expression may contribute to the visceral hyperalgesia,thus identifying a potential therapeutic target for the treatment of chronic visceral pain in patients with irritable bo

关键词： irritable bowel syndrome visceral hypersensitivity dorsal root ganglion hydrogen sulfide epigenetics

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