Purpose: To characterize prejunctional 5-HT heteroreceptors which modulate nacetylcholine (3 H-ACh release) in isolated rabbit and human iris-ciliary bodies (ICBS) . Methods: ICB tissue segments were incubated with H-...Purpose: To characterize prejunctional 5-HT heteroreceptors which modulate nacetylcholine (3 H-ACh release) in isolated rabbit and human iris-ciliary bodies (ICBS) . Methods: ICB tissue segments were incubated with H-choline, superfused and electrically stimulated four times (S1,S2,S3,S4) at 3 - 10 Hz for 1 min to elicit 3H-ACh. secretion. Test agents (5-HT agonists and antagonists) were added before S2,S3 and S4 and their effects determined by the stimulation ratio (Sx/S1) of evoked 3H-ACh. release. 3H-ACh in super-fusate fractions was fractionated and quantified by ion exchange chromatography. Results: In rabbit ICBs, evoked 3H-ACh. release was enhanced in a concentration-dependent manner by 5-HT (10- 9 - 10-5 M, EC50 = 5.8× 10-8 M). The maximum effect of 5-HT (10-6M) corresponded to a 45. 14 ± 7.40%) (n = 6) increase in 3H-ACh release. Higher concentrations of 5-HT ( > 10- M) induced desensitization. The response to 5-HT ( 10-6 M) in the presence of the 5-HT3/4 antagonist tropisetron (10-9 M) ,展开更多
BACKGROUND Melanocortin 3 and 5 receptors(i.e.,MC3R and MC5R)belong to the melanocortin family.However,data regarding their role in inflammatory bowel diseases(IBD)are currently unavailable.AIM This study aims to asce...BACKGROUND Melanocortin 3 and 5 receptors(i.e.,MC3R and MC5R)belong to the melanocortin family.However,data regarding their role in inflammatory bowel diseases(IBD)are currently unavailable.AIM This study aims to ascertain their expression profiles in the colonic mucosa of Crohn’s disease(CD)and ulcerative colitis(UC),aligning them with IBD disease endoscopic and histologic activity.METHODS Colonic mucosal biopsies from CD/UC patients were sampled,and immunohisto-chemical analyses were conducted to evaluate the expression of MC3R and MC5R.Colonic sampling was performed on both traits with endoscopic scores(Mayo endoscopic score and CD endoscopic index of severity)consistent with inflamed mucosa and not consistent with disease activity(i.e.,normal appearing mucosa).RESULTS In both CD and UC inflamed mucosa,MC3R(CD:+7.7 fold vs normal mucosa,P<0.01;UC:+12 fold vs normal mucosa,P<0.01)and MC5R(CD:+5.5 fold vs normal mucosa,P<0.01;UC:+8.1 fold vs normal mucosa,P<0.01)were significantly more expressed compared to normal mucosa.CONCLUSION MC3R and MC5R are expressed in the colon of IBD patients.Furthermore,expression may differ according to disease endoscopic activity,with a higher degree of expression in the traits affected by disease activity in both CD and UC,suggesting a potential use of these receptors in IBD pharmacology.展开更多
目的:5-HT1A基因C-1019G多态性与多种神经精神疾病如抑郁症、精神分裂症、焦虑症等的发生风险、症状严重程度及治疗效果存在关联。本研究旨在建立一种直接巢式PCR结合测序检测该位点的方法。方法:以口腔上皮细胞粗处理物为材料,通过直...目的:5-HT1A基因C-1019G多态性与多种神经精神疾病如抑郁症、精神分裂症、焦虑症等的发生风险、症状严重程度及治疗效果存在关联。本研究旨在建立一种直接巢式PCR结合测序检测该位点的方法。方法:以口腔上皮细胞粗处理物为材料,通过直接巢式PCR扩增包含5-HT1A基因C-1019G位点的靶片段,PCR产物经桑格测序鉴定基因型。结果:所检样本均能扩增出预期大小的PCR产物,测序峰图清晰。结论:成功建立了一种直接巢式PCR结合测序鉴定5-HT1A基因型的方法,有良好的应用前景。Objective: The C-1019G polymorphism of the 5-HT1A gene has been associated with the risk of occurrence, symptom severity, and treatment outcome of a variety of neuropsychiatric disorders, such as depression, schizophrenia, and anxiety disorders. The aim of this study was to establish a direct nested PCR combined with sequencing to detect this locus. Methods: The target fragment containing the C-1019G locus of the 5-HT1A gene was amplified by direct nested PCR using crude processed oral epithelial cells, and the PCR product was genotyped by Sanger sequencing. Results: PCR products of expected size were amplified from all the samples tested, and the sequencing peaks were clear. Conclusion: A direct nested PCR combined with sequencing method was successfully established to identify the genotypes of 5-HT1A gene, which has good prospects for application.展开更多
5-HT1A基因编码5-羟色胺(血清素)的G蛋白偶联受体(属于5-羟色胺受体亚家族)。C-1019G多态位点是5-HT1A基因启动子区一个重要的功能性多态位点,人群中存在3种基因型即CC、CG、GG,它与个体的恋爱关系及抑郁症、焦虑症等精神疾病密切相关...5-HT1A基因编码5-羟色胺(血清素)的G蛋白偶联受体(属于5-羟色胺受体亚家族)。C-1019G多态位点是5-HT1A基因启动子区一个重要的功能性多态位点,人群中存在3种基因型即CC、CG、GG,它与个体的恋爱关系及抑郁症、焦虑症等精神疾病密切相关。本文对5-HT1A基因C-1019G多态位点的相关机制、检测方法、研究进展等进行综述。The 5-HT1A gene encodes a G protein-coupled receptor for serotonin, which belongs to the 5-hydroxytryptamine receptor subfamily. C-1019G polymorphism is an important functional poly-morphism in the promoter region of 5-HT1A gene. There are three genotypes (CC, CG and GG) in the population, which are closely related to individual’s romantic relationship and mental disorders such as depression and anxiety. In this paper, the mechanism, detection methods and research progress of C-1019G polymorphism of 5-HT1A gene were reviewed.展开更多
BACKGROUND N6-methyladenosine(m6A)modification represents the predominant alteration found in eukaryotic messenger RNA and plays a crucial role in the progression of various tumors.However,despite its significance,the...BACKGROUND N6-methyladenosine(m6A)modification represents the predominant alteration found in eukaryotic messenger RNA and plays a crucial role in the progression of various tumors.However,despite its significance,the comprehensive investigation of METTL5,a key m6A methyltransferase,in colorectal cancer(CRC)remains limited.AIM To investigate the role of METTL5 in CRC.METHODS We assessed METTL5 expression levels in clinical samples obtained from CRC patients as well as in CRC cell lines.To elucidate the downstream targets of METTL5,we performed RNA-sequencing analysis coupled with correlation analysis,leading us to identify Toll-like receptor 8(TLR8)as a potential downstream target.In vitro functional assessments of METTL5 and TLR8 were conducted using CCK-8 assays,scratch assays,as well as assays measuring cell migration and invasion.RESULTS Our findings reveal a pronounced upregulation of METTL5 expression in both CRC cells and tissues,which correlated significantly with an unfavorable prognosis.In vitro experiments unequivocally demonstrated the oncogenic role of METTL5,as evidenced by its promotion of CRC cell proliferation,invasion,and migration.Notably,we identified TLR8 as a downstream target of METTL5,and subsequent down-regulation of TLR8 led to a significant inhibition of CRC cell proliferation,invasion,and tumor growth.CONCLUSION The heightened expression of METTL5 in CRC is strongly associated with clinicopathological features and a poor prognosis,thereby underscoring its potential utility as a critical marker for facilitating early diagnosis and prognostication in CRC.展开更多
Objective:To investigate the potential of N-acetylcysteine(NAC)and zinc sulphate(ZnSO_(4))in mitigating reproductive dysfunction caused by di-2-ethylhexyl phthalate(DEHP)in rats and to understand the underlying mechan...Objective:To investigate the potential of N-acetylcysteine(NAC)and zinc sulphate(ZnSO_(4))in mitigating reproductive dysfunction caused by di-2-ethylhexyl phthalate(DEHP)in rats and to understand the underlying mechanisms,specifically oxidative stress and sex hormone receptor activity.Methods:Thirty-five male Wistar rats were randomly divided into five equal groups(n=7 per group).Group 1 was administered 0.5 mL of distilled water and served as the control group.Group 2 was given only DEHP(750 mg/kg/day),while group 3,4 and 5 were given DEHP(750 mg/kg/day)plus NAC(100 mg/kg/day),DEHP(750 mg/kg/day)plus ZnSO_(4)(0.5 mg/kg/day),and DEHP(750 mg/kg/day)plus NAC(100 mg/kg/day)as well as ZnSO_(4)(0.5 mg/kg/day),respectively.All treatments lasted for 21 days.Samples were obtained after the rats were sacrificed,and hormones levels in the serum and markers of oxidative stress in the testicles were analyzed using the enzyme-linked immunosorbent assay.The amount of androgen receptors in the testicles was determined by immunohistochemistry,and the susceptibility of testosterone and DEHP to bind to androgen receptor and 5α-reductase was determined by molecular docking studies.Results:DEHP decreased reproductive hormones,testicular antioxidant enzymes,increased malondialdehyde levels,and negatively impacted histology of the pituitary and testes.NAC or ZnSO_(4) treatment showed a marked improvement in testicular antioxidant status and hormone levels,as well as a positive effect on the histology of the pituitary and testes.The combination of both treatments appeared to be more effective.The affinity of DEHP to bind to androgen receptors may lead to disruption of androgen receptor signaling,which can further result in dysfunction of hormones related to androgen.However,NAC is more likely to form stronger binding interactions with follicle stimulating hormone and luteinizing hormone receptors,as well as gonadotropin-releasing hormone receptors,when compared to DEHP.Conclusions:The possibility that NAC and ZnSO_(4) could downregulate DEHP-induced sex hormone changes is suggested by their potential to reduce toxicity.展开更多
It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and ...It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and different 5-HT receptors on pentobarbital-induced sleep by using the loss-of-righting reflex method. The results showed that augmentative effect of tetrandrine on pentobarbital hypnosis in mice were potentiated by the p-MPPI （5-HT1A receptor antagonist） （1 mg/kg, i.p.） and ketanserin （5-HT2A/2C receptor antagonist） （1.5 mg/kg, i.p.）, respectively. Pretreatment with either 8-OH-DPAT （5-HT1A receptor agonist） （0.1 mg/kg, s.c.） or DOI （5-HT2A/2C receptor agonist） （0.2 mg/kg, i.p.） significantly decreased pentobarbital-induced sleep time, and tetrandrine （60 mg/kg, i.g.） significantly reversed this effect. These results suggest that both the 5-HTLA and 5-HT2A/2C subfamily may be involved in the potentiating mechanism of tetrandrine＇s effects on pantobarbital hypnosis.展开更多
基金This study was supported by NIH grant EYO5246,the Kentucky Lions Eye Foundation and an unrestricted grant from Research to Prevent Blindness.Inc
文摘Purpose: To characterize prejunctional 5-HT heteroreceptors which modulate nacetylcholine (3 H-ACh release) in isolated rabbit and human iris-ciliary bodies (ICBS) . Methods: ICB tissue segments were incubated with H-choline, superfused and electrically stimulated four times (S1,S2,S3,S4) at 3 - 10 Hz for 1 min to elicit 3H-ACh. secretion. Test agents (5-HT agonists and antagonists) were added before S2,S3 and S4 and their effects determined by the stimulation ratio (Sx/S1) of evoked 3H-ACh. release. 3H-ACh in super-fusate fractions was fractionated and quantified by ion exchange chromatography. Results: In rabbit ICBs, evoked 3H-ACh. release was enhanced in a concentration-dependent manner by 5-HT (10- 9 - 10-5 M, EC50 = 5.8× 10-8 M). The maximum effect of 5-HT (10-6M) corresponded to a 45. 14 ± 7.40%) (n = 6) increase in 3H-ACh release. Higher concentrations of 5-HT ( > 10- M) induced desensitization. The response to 5-HT ( 10-6 M) in the presence of the 5-HT3/4 antagonist tropisetron (10-9 M) ,
基金The study was conducted in accordance with the Declaration of Helsinki and approved by the Ethics Committee of University of Campania Luigi Vanvitelli(Protocol code 795 on December 23,2019).
文摘BACKGROUND Melanocortin 3 and 5 receptors(i.e.,MC3R and MC5R)belong to the melanocortin family.However,data regarding their role in inflammatory bowel diseases(IBD)are currently unavailable.AIM This study aims to ascertain their expression profiles in the colonic mucosa of Crohn’s disease(CD)and ulcerative colitis(UC),aligning them with IBD disease endoscopic and histologic activity.METHODS Colonic mucosal biopsies from CD/UC patients were sampled,and immunohisto-chemical analyses were conducted to evaluate the expression of MC3R and MC5R.Colonic sampling was performed on both traits with endoscopic scores(Mayo endoscopic score and CD endoscopic index of severity)consistent with inflamed mucosa and not consistent with disease activity(i.e.,normal appearing mucosa).RESULTS In both CD and UC inflamed mucosa,MC3R(CD:+7.7 fold vs normal mucosa,P<0.01;UC:+12 fold vs normal mucosa,P<0.01)and MC5R(CD:+5.5 fold vs normal mucosa,P<0.01;UC:+8.1 fold vs normal mucosa,P<0.01)were significantly more expressed compared to normal mucosa.CONCLUSION MC3R and MC5R are expressed in the colon of IBD patients.Furthermore,expression may differ according to disease endoscopic activity,with a higher degree of expression in the traits affected by disease activity in both CD and UC,suggesting a potential use of these receptors in IBD pharmacology.
文摘目的:5-HT1A基因C-1019G多态性与多种神经精神疾病如抑郁症、精神分裂症、焦虑症等的发生风险、症状严重程度及治疗效果存在关联。本研究旨在建立一种直接巢式PCR结合测序检测该位点的方法。方法:以口腔上皮细胞粗处理物为材料,通过直接巢式PCR扩增包含5-HT1A基因C-1019G位点的靶片段,PCR产物经桑格测序鉴定基因型。结果:所检样本均能扩增出预期大小的PCR产物,测序峰图清晰。结论:成功建立了一种直接巢式PCR结合测序鉴定5-HT1A基因型的方法,有良好的应用前景。Objective: The C-1019G polymorphism of the 5-HT1A gene has been associated with the risk of occurrence, symptom severity, and treatment outcome of a variety of neuropsychiatric disorders, such as depression, schizophrenia, and anxiety disorders. The aim of this study was to establish a direct nested PCR combined with sequencing to detect this locus. Methods: The target fragment containing the C-1019G locus of the 5-HT1A gene was amplified by direct nested PCR using crude processed oral epithelial cells, and the PCR product was genotyped by Sanger sequencing. Results: PCR products of expected size were amplified from all the samples tested, and the sequencing peaks were clear. Conclusion: A direct nested PCR combined with sequencing method was successfully established to identify the genotypes of 5-HT1A gene, which has good prospects for application.
文摘5-HT1A基因编码5-羟色胺(血清素)的G蛋白偶联受体(属于5-羟色胺受体亚家族)。C-1019G多态位点是5-HT1A基因启动子区一个重要的功能性多态位点,人群中存在3种基因型即CC、CG、GG,它与个体的恋爱关系及抑郁症、焦虑症等精神疾病密切相关。本文对5-HT1A基因C-1019G多态位点的相关机制、检测方法、研究进展等进行综述。The 5-HT1A gene encodes a G protein-coupled receptor for serotonin, which belongs to the 5-hydroxytryptamine receptor subfamily. C-1019G polymorphism is an important functional poly-morphism in the promoter region of 5-HT1A gene. There are three genotypes (CC, CG and GG) in the population, which are closely related to individual’s romantic relationship and mental disorders such as depression and anxiety. In this paper, the mechanism, detection methods and research progress of C-1019G polymorphism of 5-HT1A gene were reviewed.
基金Supported by Natural Science Foundation in Anhui Province of China,No.2008085MH279Key Project of Anhui Translational Medicine Research Institute,No.2022zhyx-B08.
文摘BACKGROUND N6-methyladenosine(m6A)modification represents the predominant alteration found in eukaryotic messenger RNA and plays a crucial role in the progression of various tumors.However,despite its significance,the comprehensive investigation of METTL5,a key m6A methyltransferase,in colorectal cancer(CRC)remains limited.AIM To investigate the role of METTL5 in CRC.METHODS We assessed METTL5 expression levels in clinical samples obtained from CRC patients as well as in CRC cell lines.To elucidate the downstream targets of METTL5,we performed RNA-sequencing analysis coupled with correlation analysis,leading us to identify Toll-like receptor 8(TLR8)as a potential downstream target.In vitro functional assessments of METTL5 and TLR8 were conducted using CCK-8 assays,scratch assays,as well as assays measuring cell migration and invasion.RESULTS Our findings reveal a pronounced upregulation of METTL5 expression in both CRC cells and tissues,which correlated significantly with an unfavorable prognosis.In vitro experiments unequivocally demonstrated the oncogenic role of METTL5,as evidenced by its promotion of CRC cell proliferation,invasion,and migration.Notably,we identified TLR8 as a downstream target of METTL5,and subsequent down-regulation of TLR8 led to a significant inhibition of CRC cell proliferation,invasion,and tumor growth.CONCLUSION The heightened expression of METTL5 in CRC is strongly associated with clinicopathological features and a poor prognosis,thereby underscoring its potential utility as a critical marker for facilitating early diagnosis and prognostication in CRC.
文摘Objective:To investigate the potential of N-acetylcysteine(NAC)and zinc sulphate(ZnSO_(4))in mitigating reproductive dysfunction caused by di-2-ethylhexyl phthalate(DEHP)in rats and to understand the underlying mechanisms,specifically oxidative stress and sex hormone receptor activity.Methods:Thirty-five male Wistar rats were randomly divided into five equal groups(n=7 per group).Group 1 was administered 0.5 mL of distilled water and served as the control group.Group 2 was given only DEHP(750 mg/kg/day),while group 3,4 and 5 were given DEHP(750 mg/kg/day)plus NAC(100 mg/kg/day),DEHP(750 mg/kg/day)plus ZnSO_(4)(0.5 mg/kg/day),and DEHP(750 mg/kg/day)plus NAC(100 mg/kg/day)as well as ZnSO_(4)(0.5 mg/kg/day),respectively.All treatments lasted for 21 days.Samples were obtained after the rats were sacrificed,and hormones levels in the serum and markers of oxidative stress in the testicles were analyzed using the enzyme-linked immunosorbent assay.The amount of androgen receptors in the testicles was determined by immunohistochemistry,and the susceptibility of testosterone and DEHP to bind to androgen receptor and 5α-reductase was determined by molecular docking studies.Results:DEHP decreased reproductive hormones,testicular antioxidant enzymes,increased malondialdehyde levels,and negatively impacted histology of the pituitary and testes.NAC or ZnSO_(4) treatment showed a marked improvement in testicular antioxidant status and hormone levels,as well as a positive effect on the histology of the pituitary and testes.The combination of both treatments appeared to be more effective.The affinity of DEHP to bind to androgen receptors may lead to disruption of androgen receptor signaling,which can further result in dysfunction of hormones related to androgen.However,NAC is more likely to form stronger binding interactions with follicle stimulating hormone and luteinizing hormone receptors,as well as gonadotropin-releasing hormone receptors,when compared to DEHP.Conclusions:The possibility that NAC and ZnSO_(4) could downregulate DEHP-induced sex hormone changes is suggested by their potential to reduce toxicity.
基金National Natural Science Foundation of China(Grant No.30772556 and 30640070)Research Fund of Janssen Research Council and the‘985'Project in Peking University.
文摘It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and different 5-HT receptors on pentobarbital-induced sleep by using the loss-of-righting reflex method. The results showed that augmentative effect of tetrandrine on pentobarbital hypnosis in mice were potentiated by the p-MPPI （5-HT1A receptor antagonist） （1 mg/kg, i.p.） and ketanserin （5-HT2A/2C receptor antagonist） （1.5 mg/kg, i.p.）, respectively. Pretreatment with either 8-OH-DPAT （5-HT1A receptor agonist） （0.1 mg/kg, s.c.） or DOI （5-HT2A/2C receptor agonist） （0.2 mg/kg, i.p.） significantly decreased pentobarbital-induced sleep time, and tetrandrine （60 mg/kg, i.g.） significantly reversed this effect. These results suggest that both the 5-HTLA and 5-HT2A/2C subfamily may be involved in the potentiating mechanism of tetrandrine＇s effects on pantobarbital hypnosis.
