AIM: To assess corticotropin-releasing factor receptor 2 (CRF 2 ) expression in the colon of healthy subjects and patients with ulcerative colitis (UC). METHODS: We examined CRF2 gene and protein expression in the dis...AIM: To assess corticotropin-releasing factor receptor 2 (CRF 2 ) expression in the colon of healthy subjects and patients with ulcerative colitis (UC). METHODS: We examined CRF2 gene and protein expression in the distal/sigmoid colonic mucosal biopsies from healthy subjects and patients with UC (active or disease in remission), human immunodeficiency virus (HIV) and functional bowel disease (FBD) by reverse transcriptionpolymerase chain reaction and immunofluorescence. RESULTS: Gene expression of CRF2 was demonstrated in the normal human colonic biopsies, but not in the human colorectal adenocarcinoma cell line Caco2. Receptor protein localization showed immunoreactive CRF 2 receptors in the lamina propria and in the epithelial cells of the distal/sigmoid biopsy samples. Interestingly, CRF 2 immunoreactivity was no longer observed in epithelial cells of patients with mild-moderately active UC and disease in remission, while receptor protein expression did not change in the lamina propria. No differences in CRF 2 expression profile were observed in distal/sigmoid intestinal biopsies from HIV infection and FBD patients, showing no signs of inflammation. CONCLUSION: The down-regulation of the CRF2 receptor in the distal/sigmoid biopsies of UC patients is indicative of change in CRF 2 signalling associated with the process of inflammation.展开更多
目的:从结肠、海马及下丘脑中促肾上腺皮质激素释放因子(corticotropin releasing factor,CRF)和CRF受体1(CRF receptor 1,CRFR1)角度探讨健脾化湿颗粒改善腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome,D-IBS)模...目的:从结肠、海马及下丘脑中促肾上腺皮质激素释放因子(corticotropin releasing factor,CRF)和CRF受体1(CRF receptor 1,CRFR1)角度探讨健脾化湿颗粒改善腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome,D-IBS)模型大鼠结肠运动和内脏敏感性的作用机制.方法:采用番泻叶灌胃结合束缚应激法建立D-I B S大鼠模型,应用健脾化湿颗粒进行干预,采用酶联免疫法(ELISA)检测大鼠结肠中CRF含量,采用免疫组织化学法检测结肠中CRFR1及海马、下丘脑中CRF,CRFR1阳性表达,采用RT-PCR法检测结肠、海马中CRF m RNA和CRFR1 m RNA的表达水平.结果:与正常组相比,模型组结肠中CRF含量(67.1±3.8 vs 36.0±3.0),海马、下丘脑中CRF阳性表达(0.23±0.02 vs 0.09±0.01,0.17±0.02 v s 0.09±0.01)明显升高(P<0.01);结肠、海马、下丘脑中C R F R1阳性表达(0.17±0.01 vs 0.03±0.01,0.20±0.02 vs 0.09±0.01,0.19±0.02 vs 0.07±0.01)明显升高(P<0.01);结肠、海马中C R F m RNA和CRFR1 m RNA的表达(结肠:0.89±0.04 vs 0.09±0.01,1.09±0.09 vs 0.21±0.04;海马:0.56±0.01 vs 0.15±0.05,1.26±0.14 vs 0.23±0.06)显著升高(P<0.01).与模型组相比,各治疗组结肠、海马中C R F(51.0±3.4,54.6±4.1,45.1±4.7,43.3±3.9 vs 67.1±3.8;0.18±0.02,0.19±0.02,0.15±0.02,0.11±0.01 vs 0.23±0.02)显著下降(P<0.01),阳性对照组、中、高剂量组下丘脑中CRF(0.15±0.02,0.13±0.01,0.12±0.01 vs 0.17±0.02)下降显著(P<0.05,P<0.0 1);阳性对照组、中、高剂量组结肠、海马、下丘脑中C R F R1表达(结肠:0.10±0.01,0.08±0.01,0.05±0.01 vs 0.17±0.01;海马:0.16±0.01,0.14±0.02,0.13±0.01 vs 0.20±0.02;下丘脑:0.15±0.02,0.13±0.01,0.11±0.01 vs 0.19±0.02)下降显著(P<0.05,P<0.01);结肠中CRF m RNA表达(0.63±0.04,0.76±0.06,0.32±0.06,0.13±0.03 v s 0.89±0.04)及中、高剂量组海马中CRF m RNA表达(0.76±0.11,0.67±0.10 v s 1.09±0.09)显著降低(P<0.01);阳性对照组、中、高剂量组结肠中C R F R1m RNA表达(0.47±0.03,0.40±0.06,0.24±0.06 vs 0.56±0.01)及中、高剂量组海马中CRFR1 m RNA表达(0.62±0.06,0.60±0.07vs 1.26±0.14)显著降低(P<0.05,P<0.01).结论:健脾化湿颗粒可能通过下调结肠、海马及下丘脑中CRF、CRFR1表达来改善D-IBS模型大鼠结肠运动和内脏敏感性.展开更多
AIM: To explore the role of nesfatin-1 on irritable bowel syndrome (IBS)-like visceral hypersensitivity. METHODS: The animal model of IBS-like visceral hypersensitivity was induced by intracolonic infusion of 0.5% ace...AIM: To explore the role of nesfatin-1 on irritable bowel syndrome (IBS)-like visceral hypersensitivity. METHODS: The animal model of IBS-like visceral hypersensitivity was induced by intracolonic infusion of 0.5% acetic acid (AA) in saline once daily from postnatal days 8-21. Experiments were performed when rats became adults. The visceral sensitivity of rats was evaluated by abdominal withdrawal reflex (AWR) and electromyographic (EMG) activity of the external oblique muscle to graded colorectal distension. The content of nesfatin-1 in serum was determined using enzyme-linked immunosorbent assay. After implantation of an intracerebroventricular (ICV) cannula and two electrodes into the external oblique muscle, model rats were randomly divided into four groups. Animals then received ICV injection of 8 μg of anti-nesfatin-1/ nucleobindin-2 (NUCB2), 50 μg of α-helical cortico-tropin releasing factor (CRF) 9-41 (non-selective CRF receptor antagonist), 50 μg of NBI-27914 (selective CRF1 receptor antagonist) or 5 μL of vehicle. After 1 h of ICV administration, visceral sensitivity of each group was measured again, and comparisons between groups were made. RESULTS: Rats treated with AA showed higher mean AWR scores and EMG activity at all distension pressures compared with controls (P < 0.05). On histopathologic examination, no evidence of inflammation or abnormalities in structure were noted in the colon of either control or AA-treated groups. Myeloperoxidase values were not significantly different between the two groups. The level of nesfatin-1 in serum was significantly higher in the AA-treated group than in the control group (5.34 ± 0.37 ng/mL vs 4.81 ± 0.42 ng/mL, P < 0.01). Compared with rats injected with vehicle, rats which received ICV anti-nesfatin-1/NUCB2, α-helical CRF9-41 or NBI-27914 showed decreased mean AWR scores and EMG activity at all distension pressures (P < 0.05). CONCLUSION: Nesfatin-1 may be associated with IBS-like visceral hypersensitivity, which may be implicated in brain CRF/CRF1 signaling pathways.展开更多
Disordered signalling between the brain and the gut are generally accepted to underlie the functional bowel disorder, irritable bowel syndrome(IBS). However, partly due to the lack of disease-defining biomarkers, unde...Disordered signalling between the brain and the gut are generally accepted to underlie the functional bowel disorder, irritable bowel syndrome(IBS). However, partly due to the lack of disease-defining biomarkers, understanding the aetiology of this complex and multifactorial disease remains elusive. This common gastrointestinal disorder is characterised by alterations in bowel habit such as diarrhoea and/or constipation, bloating and abdominal pain, and symptom exacerbation has been linked with periods of stress, both psychosocial and infection-related. Indeed, a high level of comorbidity exists between IBS and stress-related mood disorders such as anxiety and depression. Moreover, studies have observed alterations in autonomic output and neuro-endocrine signalling in IBS patients. Accumulating evidence indicates that a maladaptive stress response, probably mediated by the stress hormone, corticotropin-releasing factor contributes to the initiation, persistence and severity of symptom flares.Other risk factors for developing IBS include a positive family history, childhood trauma, dietary factors and prior gastrointestinal infection. An emerging role has been attributed to the importance of immune factors in the pathophysiology of IBS with evidence of altered cytokine profiles and increased levels of mucosal immune cells. These factors have also been shown to have direct effects on neural signalling. This review discusses how pathological changes in neural, immune and endocrine pathways, and communication between these systems, contribute to symptom flares in IBS.展开更多
Hypothalamic Corticotropin-releasing factor (CRF) directly activates the hypothalamic pituitary adrenal axis (HPA axis) during the surgical trauma induced stress response. Electroacupuncture (EA) has been demonstrated...Hypothalamic Corticotropin-releasing factor (CRF) directly activates the hypothalamic pituitary adrenal axis (HPA axis) during the surgical trauma induced stress response. Electroacupuncture (EA) has been demonstrated to have stress relieving effects in breast surgery, colorectal surgery, prostatectomy and craniotomy. This study was aimed to investigate the hypothesis that EA could regulate hypothalamic CRF in surgical trauma rats. In experiment one, Sprague-Dawley (SD) male rats were divided into intact, model (10% partial hepatectomy), sham EA and EA group. Rats from the Sham EA and EA group were stimulated at ST36-Zusanli and SP6-Sanyiniiao acupoints twice, 24 hours before the surgery and immediately after the surgery. Expressions of hypothalamic CRF and CRFR, GABA receptors, glutamate decarboxylase (GAD), serum adrenocorticotropic hormone (ACTH) and Corticosterone (CORT) were observed at 2, 4, 8 and 24 h after the surgery by radioimmunoassay (RIA), western blot, real-time PCR and immunohistochemistry. In the experiment two, SD male rats were divided into the intact, model, model + vehicle, model + L-838,417 EA and EA + L838,417 group. It was found that hypothalamus CRF, serum ACTH and CORT levels were increased in model group compared with the intact group, and those in the EA group decreased in comparison with the model group. Compared with the model group, hypothalamus-aminobutyric acid (GABA) receptor Aα3 mRNA and protein expressions of the EA group raised strikingly. In conclusion, EA alleviated surgical stress response by improving the GABA synthesis in hypothalamus, thus enhancing GABA receptors’ inhibitory regulation of the HPA axis dysfunction in rats with acute surgical trauma.展开更多
基金Supported by Grants-in-Aid for Scientific Research(C)from the Ministry of Education,Culture,Sports,Science,and Tech-nology of Japanthe Foundation for the Promotion of Can-cer Research and Mitsui Life Social Welfare Foundation
文摘AIM: To examine whether commensal bacteria are a contributing cause of stress-related mucosal inflammation.
基金Supported by The National Institute of Diabetes and Digestive and Kidney Diseases R01 grant DK-57238Center Grant DK-41301 (Clinical core)+1 种基金Veteran Administration Research Career Scientist Award (YT)NIH DK-78676 (MM)
文摘AIM: To assess corticotropin-releasing factor receptor 2 (CRF 2 ) expression in the colon of healthy subjects and patients with ulcerative colitis (UC). METHODS: We examined CRF2 gene and protein expression in the distal/sigmoid colonic mucosal biopsies from healthy subjects and patients with UC (active or disease in remission), human immunodeficiency virus (HIV) and functional bowel disease (FBD) by reverse transcriptionpolymerase chain reaction and immunofluorescence. RESULTS: Gene expression of CRF2 was demonstrated in the normal human colonic biopsies, but not in the human colorectal adenocarcinoma cell line Caco2. Receptor protein localization showed immunoreactive CRF 2 receptors in the lamina propria and in the epithelial cells of the distal/sigmoid biopsy samples. Interestingly, CRF 2 immunoreactivity was no longer observed in epithelial cells of patients with mild-moderately active UC and disease in remission, while receptor protein expression did not change in the lamina propria. No differences in CRF 2 expression profile were observed in distal/sigmoid intestinal biopsies from HIV infection and FBD patients, showing no signs of inflammation. CONCLUSION: The down-regulation of the CRF2 receptor in the distal/sigmoid biopsies of UC patients is indicative of change in CRF 2 signalling associated with the process of inflammation.
文摘目的:从结肠、海马及下丘脑中促肾上腺皮质激素释放因子(corticotropin releasing factor,CRF)和CRF受体1(CRF receptor 1,CRFR1)角度探讨健脾化湿颗粒改善腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome,D-IBS)模型大鼠结肠运动和内脏敏感性的作用机制.方法:采用番泻叶灌胃结合束缚应激法建立D-I B S大鼠模型,应用健脾化湿颗粒进行干预,采用酶联免疫法(ELISA)检测大鼠结肠中CRF含量,采用免疫组织化学法检测结肠中CRFR1及海马、下丘脑中CRF,CRFR1阳性表达,采用RT-PCR法检测结肠、海马中CRF m RNA和CRFR1 m RNA的表达水平.结果:与正常组相比,模型组结肠中CRF含量(67.1±3.8 vs 36.0±3.0),海马、下丘脑中CRF阳性表达(0.23±0.02 vs 0.09±0.01,0.17±0.02 v s 0.09±0.01)明显升高(P<0.01);结肠、海马、下丘脑中C R F R1阳性表达(0.17±0.01 vs 0.03±0.01,0.20±0.02 vs 0.09±0.01,0.19±0.02 vs 0.07±0.01)明显升高(P<0.01);结肠、海马中C R F m RNA和CRFR1 m RNA的表达(结肠:0.89±0.04 vs 0.09±0.01,1.09±0.09 vs 0.21±0.04;海马:0.56±0.01 vs 0.15±0.05,1.26±0.14 vs 0.23±0.06)显著升高(P<0.01).与模型组相比,各治疗组结肠、海马中C R F(51.0±3.4,54.6±4.1,45.1±4.7,43.3±3.9 vs 67.1±3.8;0.18±0.02,0.19±0.02,0.15±0.02,0.11±0.01 vs 0.23±0.02)显著下降(P<0.01),阳性对照组、中、高剂量组下丘脑中CRF(0.15±0.02,0.13±0.01,0.12±0.01 vs 0.17±0.02)下降显著(P<0.05,P<0.0 1);阳性对照组、中、高剂量组结肠、海马、下丘脑中C R F R1表达(结肠:0.10±0.01,0.08±0.01,0.05±0.01 vs 0.17±0.01;海马:0.16±0.01,0.14±0.02,0.13±0.01 vs 0.20±0.02;下丘脑:0.15±0.02,0.13±0.01,0.11±0.01 vs 0.19±0.02)下降显著(P<0.05,P<0.01);结肠中CRF m RNA表达(0.63±0.04,0.76±0.06,0.32±0.06,0.13±0.03 v s 0.89±0.04)及中、高剂量组海马中CRF m RNA表达(0.76±0.11,0.67±0.10 v s 1.09±0.09)显著降低(P<0.01);阳性对照组、中、高剂量组结肠中C R F R1m RNA表达(0.47±0.03,0.40±0.06,0.24±0.06 vs 0.56±0.01)及中、高剂量组海马中CRFR1 m RNA表达(0.62±0.06,0.60±0.07vs 1.26±0.14)显著降低(P<0.05,P<0.01).结论:健脾化湿颗粒可能通过下调结肠、海马及下丘脑中CRF、CRFR1表达来改善D-IBS模型大鼠结肠运动和内脏敏感性.
基金Supported by Natural Science Foundation of China,No.81070308/H0308
文摘AIM: To explore the role of nesfatin-1 on irritable bowel syndrome (IBS)-like visceral hypersensitivity. METHODS: The animal model of IBS-like visceral hypersensitivity was induced by intracolonic infusion of 0.5% acetic acid (AA) in saline once daily from postnatal days 8-21. Experiments were performed when rats became adults. The visceral sensitivity of rats was evaluated by abdominal withdrawal reflex (AWR) and electromyographic (EMG) activity of the external oblique muscle to graded colorectal distension. The content of nesfatin-1 in serum was determined using enzyme-linked immunosorbent assay. After implantation of an intracerebroventricular (ICV) cannula and two electrodes into the external oblique muscle, model rats were randomly divided into four groups. Animals then received ICV injection of 8 μg of anti-nesfatin-1/ nucleobindin-2 (NUCB2), 50 μg of α-helical cortico-tropin releasing factor (CRF) 9-41 (non-selective CRF receptor antagonist), 50 μg of NBI-27914 (selective CRF1 receptor antagonist) or 5 μL of vehicle. After 1 h of ICV administration, visceral sensitivity of each group was measured again, and comparisons between groups were made. RESULTS: Rats treated with AA showed higher mean AWR scores and EMG activity at all distension pressures compared with controls (P < 0.05). On histopathologic examination, no evidence of inflammation or abnormalities in structure were noted in the colon of either control or AA-treated groups. Myeloperoxidase values were not significantly different between the two groups. The level of nesfatin-1 in serum was significantly higher in the AA-treated group than in the control group (5.34 ± 0.37 ng/mL vs 4.81 ± 0.42 ng/mL, P < 0.01). Compared with rats injected with vehicle, rats which received ICV anti-nesfatin-1/NUCB2, α-helical CRF9-41 or NBI-27914 showed decreased mean AWR scores and EMG activity at all distension pressures (P < 0.05). CONCLUSION: Nesfatin-1 may be associated with IBS-like visceral hypersensitivity, which may be implicated in brain CRF/CRF1 signaling pathways.
文摘Disordered signalling between the brain and the gut are generally accepted to underlie the functional bowel disorder, irritable bowel syndrome(IBS). However, partly due to the lack of disease-defining biomarkers, understanding the aetiology of this complex and multifactorial disease remains elusive. This common gastrointestinal disorder is characterised by alterations in bowel habit such as diarrhoea and/or constipation, bloating and abdominal pain, and symptom exacerbation has been linked with periods of stress, both psychosocial and infection-related. Indeed, a high level of comorbidity exists between IBS and stress-related mood disorders such as anxiety and depression. Moreover, studies have observed alterations in autonomic output and neuro-endocrine signalling in IBS patients. Accumulating evidence indicates that a maladaptive stress response, probably mediated by the stress hormone, corticotropin-releasing factor contributes to the initiation, persistence and severity of symptom flares.Other risk factors for developing IBS include a positive family history, childhood trauma, dietary factors and prior gastrointestinal infection. An emerging role has been attributed to the importance of immune factors in the pathophysiology of IBS with evidence of altered cytokine profiles and increased levels of mucosal immune cells. These factors have also been shown to have direct effects on neural signalling. This review discusses how pathological changes in neural, immune and endocrine pathways, and communication between these systems, contribute to symptom flares in IBS.
文摘Hypothalamic Corticotropin-releasing factor (CRF) directly activates the hypothalamic pituitary adrenal axis (HPA axis) during the surgical trauma induced stress response. Electroacupuncture (EA) has been demonstrated to have stress relieving effects in breast surgery, colorectal surgery, prostatectomy and craniotomy. This study was aimed to investigate the hypothesis that EA could regulate hypothalamic CRF in surgical trauma rats. In experiment one, Sprague-Dawley (SD) male rats were divided into intact, model (10% partial hepatectomy), sham EA and EA group. Rats from the Sham EA and EA group were stimulated at ST36-Zusanli and SP6-Sanyiniiao acupoints twice, 24 hours before the surgery and immediately after the surgery. Expressions of hypothalamic CRF and CRFR, GABA receptors, glutamate decarboxylase (GAD), serum adrenocorticotropic hormone (ACTH) and Corticosterone (CORT) were observed at 2, 4, 8 and 24 h after the surgery by radioimmunoassay (RIA), western blot, real-time PCR and immunohistochemistry. In the experiment two, SD male rats were divided into the intact, model, model + vehicle, model + L-838,417 EA and EA + L838,417 group. It was found that hypothalamus CRF, serum ACTH and CORT levels were increased in model group compared with the intact group, and those in the EA group decreased in comparison with the model group. Compared with the model group, hypothalamus-aminobutyric acid (GABA) receptor Aα3 mRNA and protein expressions of the EA group raised strikingly. In conclusion, EA alleviated surgical stress response by improving the GABA synthesis in hypothalamus, thus enhancing GABA receptors’ inhibitory regulation of the HPA axis dysfunction in rats with acute surgical trauma.
文摘目的:观察耳电针对糖尿病胃轻瘫大鼠行为学、胃排空率和胃窦组织中酪氨酸激酶受体(c-Kit)和促肾上腺皮质释放激素因子(corticotropin releasing factor,CRF)表达的影响,探讨耳电针改善糖尿病胃轻瘫大鼠胃排空和抑郁样行为的作用机制。方法:将36只SPF级SD大鼠随机分为空白对照组、模型组、耳电针组和假耳针组,每组9只。空白对照组腹腔注射柠檬酸钠缓冲液,其余3组腹腔注射链脲佐菌素(streptozotocin,STZ)溶液,随后继续饲养6周后开始治疗,耳电针组电针双侧耳穴“胃”,每日1次,每周5日,连续2周;假耳针组针刺耳垂,不加电。治疗结束后,进行行为学以及胃排空率检测,qRT-PCR检测胃窦组织CRF、c-Kit m RNA表达,ELISA检测胃窦组织CRF、c-Kit蛋白表达。结果:旷场实验中模型组大鼠进入中央区域次数(CE)、进入中央区域时间百分比(CT%)和进入中央区域距离百分比(CD%)均较空白对照组明显降低(均P<0.05),耳电针组CE、CT%和CD%值均较模型组明显升高(均P<0.05);强迫游泳实验中模型组大鼠漂浮时间百分比(FT%)较空白对照组明显升高(P<0.05),耳电针组FT%值较模型组明显降低(P<0.05);与空白对照组相比,模型组大鼠胃排空率显著降低,胃窦中CRF的mRNA和蛋白表达水平明显升高(P<0.05),c-Kit的mRNA和蛋白表达水平明显降低(P<0.05);与模型组和假耳针组比较,耳针组大鼠胃排空率显著升高,胃窦中CRF的mRNA和蛋白表达水平明显降低(P<0.05),c-Kit的mRNA和蛋白表达水平明显升高(P<0.05)。结论:低频耳电针可以改善糖尿病胃轻瘫大鼠的胃动力与抑郁样行为,其机制可能与调节胃窦组织中相关因子的表达有关。
