Objective To investigate the role of sphingosine-l-phosphate （S1P） and its receptors in cardiomyocyte autophagy, cardiomyocyte hypertrophy and cardiac function. Methods Cardiomyocytes were isolated from neonatal Vis...Objective To investigate the role of sphingosine-l-phosphate （S1P） and its receptors in cardiomyocyte autophagy, cardiomyocyte hypertrophy and cardiac function. Methods Cardiomyocytes were isolated from neonatal Vista rats. Autophagy and hypertrophy of car- diomyocytes were induced via starvation culture and phenylephrine （PE）, respectively, and S 1 P was used to treat the cardiomyocytes. The effect of S1P on cardiomyocyte autophagy was evaluated by the number of autophagosomes, the expression of autophagy-related proteins and autophagic marker genes in cardiomyocytes. The effect of S1P on cardiomyocyte hypertrophy was evaluated by examining the surface area of cardiomyoeytes and the expression of hypertrophic genes. Subsequently, different small interfering RNAs （siRNAs） were used to knockdown the expression of the three types of S 1P receptors on cardiomyocytes and to analyze the type of receptor that mediates S 1P sig- naling in cardiomyocytes. Finally, sphingosine 1 phosphate receptor-1 （S1PR1） was knockout in the mouse cardiomyocytes using the Cas9 technique. The effect of S 1PR1 on cardiac autophagy and cardiac hypertrophy was examined by assessing cardiomyocyte autophagy, car- diomyocyte hypertrophy and cardiac function. Results Starvation-induced cardiomyocyte autophagy and PE -induced cardiomyocyte hy- pertrophy were significantly attenuated by SIP. The results showed that the formation of autophagosomes was decreased, the auto- phagy-associated protein LC3 II/I and the expression of autophagic marker genes Atg5, Atgl2, Beclinl and LC3B decreased after SIP treatment. The surface area of the cardiomyocytes was decreased, and the expression of hypertrophic genes, including atrial natriuretic factor （ANF）, skeletal muscle and cardiac actin （SKA）, myosin heavy chain （β-MHC） and brain natriuretic peptide （BNP） were all decreased after S 1 P treatment. The autophagy and hypertrophy of cardiomyocytes in the S 1PR 1 knocked-down group were significantly increased compared to those in the control group, the SIPR2 and the S1PR3 knocked-down groups. In vivo, the knockout of S1PR1 in cardiomyocytes exacer- bated stress-induced cardiac autophagy, cardiac hypertrophy and the impairment of cardiac function. Conclusion SIP could inhibit car- diomyocyte autophagy, thereby inhibiting cardiomyocyte hypertrophy and protecting cardiac function by activating S1PR1 in pres- sure-overloaded cardiomyocytes in mice.展开更多
目的:观察经外周转染S1PR1基因慢病毒(LV-S1PR1)对实验性自身免疫性脑脊髓炎(EAE)小鼠调节性T细胞比例及IL-17、IFN-γ水平的影响。方法:将30只健康雌性C57BL/6小鼠随机分为空白对照组、EAE模型组、LV-S1PR1转染组,后两组建立EAE模型,LV...目的:观察经外周转染S1PR1基因慢病毒(LV-S1PR1)对实验性自身免疫性脑脊髓炎(EAE)小鼠调节性T细胞比例及IL-17、IFN-γ水平的影响。方法:将30只健康雌性C57BL/6小鼠随机分为空白对照组、EAE模型组、LV-S1PR1转染组,后两组建立EAE模型,LV-S1PR1转染组经尾静脉注射LV-S1PR1。每日进行神经功能缺损评分;免疫印迹法(Western blot)检测外周血S1PR1表达水平;ELISA法检测外周血1-磷酸鞘氨醇(S1P)水平以及脊髓组织IL-17、IFN-γ水平;流式细胞术检测脾脏中调节性T细胞的比例。结果:与EAE模型组相比较,LV-S1PR1转染组神经功能缺损症状改善;外周血S1PR1表达增加( P <0.05);外周血S1P水平、脾脏中调节性T细胞比例及脊髓组织中IL-17和IFN-γ水平降低( P <0.05)。结论:经外周转染LV-S1PR1可以改善EAE小鼠的发病,其防治作用机制可能与上调外周S1PR1的表达,降低外周S1P水平,使T细胞迁移受阻有关。展开更多
The binding of Sphingosine-1-phosphate(S1P)with the S1PR1-5 plays a fundamental physiological role in a number of processes including vascular development and stabilization,lymphocyte migration and distribution.S1P-S1...The binding of Sphingosine-1-phosphate(S1P)with the S1PR1-5 plays a fundamental physiological role in a number of processes including vascular development and stabilization,lymphocyte migration and distribution.S1P-S1PR1 signal axis established roles in immune cell trafficking thus playing a therapeutic role in multiple sclerosis and inflammatory bowel disease.In this study,a series of oxadiazole derivatives were designed and synthesized as S1PR1 agonists based on rational drug design.Among them,compound 9i was identified as a potent and selective S1PR1 agonist with activities onβ-arrestin recruitment(EC50=0.36 nmol/L)and receptor internalization(EC50=8.09 nmol/L).Meanwhile,compound 9i displayed an oral bioavailability up to 93.6%.Based on its excellent activity to S1PR1 and pharmacokinetic properties,compound 9i effectively alleviated dextran sulfate sodium(DSS)-induced ulcerative colitis in mice at a dose of 0.1 mg/kg.展开更多
文摘Objective To investigate the role of sphingosine-l-phosphate （S1P） and its receptors in cardiomyocyte autophagy, cardiomyocyte hypertrophy and cardiac function. Methods Cardiomyocytes were isolated from neonatal Vista rats. Autophagy and hypertrophy of car- diomyocytes were induced via starvation culture and phenylephrine （PE）, respectively, and S 1 P was used to treat the cardiomyocytes. The effect of S1P on cardiomyocyte autophagy was evaluated by the number of autophagosomes, the expression of autophagy-related proteins and autophagic marker genes in cardiomyocytes. The effect of S1P on cardiomyocyte hypertrophy was evaluated by examining the surface area of cardiomyoeytes and the expression of hypertrophic genes. Subsequently, different small interfering RNAs （siRNAs） were used to knockdown the expression of the three types of S 1P receptors on cardiomyocytes and to analyze the type of receptor that mediates S 1P sig- naling in cardiomyocytes. Finally, sphingosine 1 phosphate receptor-1 （S1PR1） was knockout in the mouse cardiomyocytes using the Cas9 technique. The effect of S 1PR1 on cardiac autophagy and cardiac hypertrophy was examined by assessing cardiomyocyte autophagy, car- diomyocyte hypertrophy and cardiac function. Results Starvation-induced cardiomyocyte autophagy and PE -induced cardiomyocyte hy- pertrophy were significantly attenuated by SIP. The results showed that the formation of autophagosomes was decreased, the auto- phagy-associated protein LC3 II/I and the expression of autophagic marker genes Atg5, Atgl2, Beclinl and LC3B decreased after SIP treatment. The surface area of the cardiomyocytes was decreased, and the expression of hypertrophic genes, including atrial natriuretic factor （ANF）, skeletal muscle and cardiac actin （SKA）, myosin heavy chain （β-MHC） and brain natriuretic peptide （BNP） were all decreased after S 1 P treatment. The autophagy and hypertrophy of cardiomyocytes in the S 1PR 1 knocked-down group were significantly increased compared to those in the control group, the SIPR2 and the S1PR3 knocked-down groups. In vivo, the knockout of S1PR1 in cardiomyocytes exacer- bated stress-induced cardiac autophagy, cardiac hypertrophy and the impairment of cardiac function. Conclusion SIP could inhibit car- diomyocyte autophagy, thereby inhibiting cardiomyocyte hypertrophy and protecting cardiac function by activating S1PR1 in pres- sure-overloaded cardiomyocytes in mice.
文摘目的:观察经外周转染S1PR1基因慢病毒(LV-S1PR1)对实验性自身免疫性脑脊髓炎(EAE)小鼠调节性T细胞比例及IL-17、IFN-γ水平的影响。方法:将30只健康雌性C57BL/6小鼠随机分为空白对照组、EAE模型组、LV-S1PR1转染组,后两组建立EAE模型,LV-S1PR1转染组经尾静脉注射LV-S1PR1。每日进行神经功能缺损评分;免疫印迹法(Western blot)检测外周血S1PR1表达水平;ELISA法检测外周血1-磷酸鞘氨醇(S1P)水平以及脊髓组织IL-17、IFN-γ水平;流式细胞术检测脾脏中调节性T细胞的比例。结果:与EAE模型组相比较,LV-S1PR1转染组神经功能缺损症状改善;外周血S1PR1表达增加( P <0.05);外周血S1P水平、脾脏中调节性T细胞比例及脊髓组织中IL-17和IFN-γ水平降低( P <0.05)。结论:经外周转染LV-S1PR1可以改善EAE小鼠的发病,其防治作用机制可能与上调外周S1PR1的表达,降低外周S1P水平,使T细胞迁移受阻有关。
基金supported in part by the National Natural Science Foundation of China(grants 81825020 and 82150208 to H.L.)the Shanghai Science and Technology Commission Biomedical Science and Technology Support Special Project(grants 21S11907900 and 20S11901000 to Z.Z.)the Fundamental Research Funds for the Central Universities.Honglin Li is also sponsored by National Program for Special Supports of Eminent Professionals and National Program for Support of Top-notch Young Professionals.
文摘The binding of Sphingosine-1-phosphate(S1P)with the S1PR1-5 plays a fundamental physiological role in a number of processes including vascular development and stabilization,lymphocyte migration and distribution.S1P-S1PR1 signal axis established roles in immune cell trafficking thus playing a therapeutic role in multiple sclerosis and inflammatory bowel disease.In this study,a series of oxadiazole derivatives were designed and synthesized as S1PR1 agonists based on rational drug design.Among them,compound 9i was identified as a potent and selective S1PR1 agonist with activities onβ-arrestin recruitment(EC50=0.36 nmol/L)and receptor internalization(EC50=8.09 nmol/L).Meanwhile,compound 9i displayed an oral bioavailability up to 93.6%.Based on its excellent activity to S1PR1 and pharmacokinetic properties,compound 9i effectively alleviated dextran sulfate sodium(DSS)-induced ulcerative colitis in mice at a dose of 0.1 mg/kg.
