目的探讨Apelin受体与G蛋白之间的相互作用。方法在HEK293T细胞中建立Apelin受体与各亚型G蛋白的过表达体系,利用荧光共振能量转移技术(fluorescence resonance energy transfer technology,FRET)实时、动态、连续的监测Apelin受体与G...目的探讨Apelin受体与G蛋白之间的相互作用。方法在HEK293T细胞中建立Apelin受体与各亚型G蛋白的过表达体系,利用荧光共振能量转移技术(fluorescence resonance energy transfer technology,FRET)实时、动态、连续的监测Apelin受体与G蛋白偶联的机制。结果加入激动剂Apelin-13刺激后,Apelin受体与Gαi1亚基的FRET值没有发生明显的变化(P>0.05),Apelin受体与Gαi2、Gαi3亚基的FRET信号显著增加(P<0.05),而Apelin受体与Gαo、Gαq亚基的FRET信号相对于刺激前显著降低(P<0.05)。结论 Apelin受体在其激动剂Apelin-13作用下,能够激活G蛋白的Gαi2、Gαi3、Gαo、Gαq亚基,并引起Gαi2、Gαi3亚基构象重排进而与Gγ亚基相互靠近,Gαo、Gαq亚基远离Gγ2亚基,而不与Gαi1亚基发生相互作用。展开更多
Objective To investigate the effect of nitric oxide （NO） on the expression of apelin receptor mRNA, as well as their correlation, in the caudate nucleus of rat. Methods L-Arginine （L-Arg）, N^G-nitro-L-arginine met...Objective To investigate the effect of nitric oxide （NO） on the expression of apelin receptor mRNA, as well as their correlation, in the caudate nucleus of rat. Methods L-Arginine （L-Arg）, N^G-nitro-L-arginine methyl ester （L-NAME） and normal saline （NS） was separately microinjected into rat caudate nucleus. Expressions of neuronal NO synthase （nNOS） mRNA and apelin receptor mRNA were detected by RT-PCR at 4, 8, 12, 24 and 48 h after microinjection, and their correlation was determined. Results The expressions of nNOS mRNA and apelin receptor mRNA were both significantly increased after microinjection of L-Arg, but significantly decreased after microinjection of L-NAME compared with the NS control group. The nNOS mRNA had a positive correlation with the expression of apelin receptor mRNA after microinjection of L-Arg and L-NAME. Conclusion The activity of NOS in the central nervous system, especially in the caudate nucleus, is one of the key factors for NO to exert many kinds of biological actions, such as modulation of central pain, as a neurotransmitter. The neurobiological action of NO in rat caudate nucleus may be associated with apelin receptors.展开更多
文摘目的探讨Apelin受体与G蛋白之间的相互作用。方法在HEK293T细胞中建立Apelin受体与各亚型G蛋白的过表达体系,利用荧光共振能量转移技术(fluorescence resonance energy transfer technology,FRET)实时、动态、连续的监测Apelin受体与G蛋白偶联的机制。结果加入激动剂Apelin-13刺激后,Apelin受体与Gαi1亚基的FRET值没有发生明显的变化(P>0.05),Apelin受体与Gαi2、Gαi3亚基的FRET信号显著增加(P<0.05),而Apelin受体与Gαo、Gαq亚基的FRET信号相对于刺激前显著降低(P<0.05)。结论 Apelin受体在其激动剂Apelin-13作用下,能够激活G蛋白的Gαi2、Gαi3、Gαo、Gαq亚基,并引起Gαi2、Gαi3亚基构象重排进而与Gγ亚基相互靠近,Gαo、Gαq亚基远离Gγ2亚基,而不与Gαi1亚基发生相互作用。
文摘Objective To investigate the effect of nitric oxide （NO） on the expression of apelin receptor mRNA, as well as their correlation, in the caudate nucleus of rat. Methods L-Arginine （L-Arg）, N^G-nitro-L-arginine methyl ester （L-NAME） and normal saline （NS） was separately microinjected into rat caudate nucleus. Expressions of neuronal NO synthase （nNOS） mRNA and apelin receptor mRNA were detected by RT-PCR at 4, 8, 12, 24 and 48 h after microinjection, and their correlation was determined. Results The expressions of nNOS mRNA and apelin receptor mRNA were both significantly increased after microinjection of L-Arg, but significantly decreased after microinjection of L-NAME compared with the NS control group. The nNOS mRNA had a positive correlation with the expression of apelin receptor mRNA after microinjection of L-Arg and L-NAME. Conclusion The activity of NOS in the central nervous system, especially in the caudate nucleus, is one of the key factors for NO to exert many kinds of biological actions, such as modulation of central pain, as a neurotransmitter. The neurobiological action of NO in rat caudate nucleus may be associated with apelin receptors.
