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
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
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.
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.
