硫代磷酸化神经肽Y Y1受体反义寡脱氧核苷酸对清醒大鼠和人体血管的作用。

Effects of phosphorothioated neuropeptide Y Y1-receptor antisense oligodeoxynucleotide in conscious rats and in human vessels.

作者信息

Sun X Y, Zhao X H, Erlinge D, Edvinsson L, Fallgren B, Wahlestedt C, Hedner T

机构信息

Division of Clinical Pharmacology, University of Göteborg, Sweden.

出版信息

Br J Pharmacol. 1996 May;118(1):131-6. doi: 10.1111/j.1476-5381.1996.tb15375.x.

DOI:10.1111/j.1476-5381.1996.tb15375.x

PMID:8733585

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1909502/

Abstract

Metabolically stabilized (phosphorothioate) human and rat NPY Y1 receptor oligodeoxynucleotides (ODNs) complimentary to the rat or human Y1 mRNA were synthesized; [sense (rY1-SODN, 5'-AATTCAACTCTGTTCTCC-3'), antisense (hY1-ASODN, 5'-CCTGGGAAAATAATGTTG-3' and rY1-ASODN, 5'-GGAGAACAGAGTTGAATT-3') and mismatches (hY1-MMODN, 5'-CCTGAGATAA-TAAGGTTG-3' and rY1-MM 5'-GTAGATCAGAGATGAAGT-3')] and used to modulate cardiovascular function in vitro in human vessels as well as in vivo in the rat. 2. The objectives of the experiments were to assess the influence of the NPY Y1 receptor on vasomotor function human resistance arteries in vitro and to investigate the contribution of the NPY receptor system to cardiovascular haemodynamics in vivo. 3. Human subcutaneous resistance arteries removed from patients who underwent surgery for nonvascular diseases were incubated in vitro with the stabilized phosphorothioated hY1-receptor ASODN or MMODN (10(-7) TO 10(-5) M). 4. In human resistance vessels preincubated with hY1-AS (10(-7) to 10(-5) M), the contractile response to NPY was significantly reduced in a dose-dependent fashion. No effects were observed in the hY1-MMODN-incubated vessels at lower concentrations (10(-7) M to 10(-6) M). 5. The haemodynamic effects of the phosphorothioated rY1-ASODN, SODN or MMODN were investigated in conscious rats during 48 h of continuous infusions. The continuous infusion with rY1-ASODN did not change MAP while the rY1-SODN unexpectedly induced an early (10-20) increase in ambulatory MAP and the rY1-MMODN a late (24-44 h) increase. 6. Contractile responses to NPY (2, 4, 8, 16 and 32 micrograms kg-1) were significantly reduced in the rats treated with long-term infusion of rY1-ASODN (2.1 mg kg-1 h-1, i.v. infusion for 48 h) compared with animals treated with rY1-SODN and MMODN, as well as animals treated with saline and glucose. Notably, the group infused with the rY1-SODN showed an exaggerated response to tested doses of NPY. 7. We conclude that the incubation of human subcutaneous arteries with a metabolically stabilized 18 base pair hY1-ASODN and long-term infusion with a corresponding rY1-ASODN attenuate NPY-induced vasoconstriction.

摘要

合成了与大鼠或人Y1 mRNA互补的代谢稳定型（硫代磷酸酯）人及大鼠NPY Y1受体寡脱氧核苷酸（ODN）；[正义链（rY1 - SODN，5'-AATTCAACTCTGTTCTCC - 3'）、反义链（hY1 - ASODN，5'-CCTGGGAAAATAATGTTG - 3'和rY1 - ASODN，5'-GGAGAACAGAGTTGAATT - 3'）以及错配链（hY1 - MMODN，5'-CCTGAGATAA - TAAGGTTG - 3'和rY1 - MM 5'-GTAGATCAGAGATGAAGT - 3'）]，并用于在体外对人血管以及在体内对大鼠调节心血管功能。2. 实验目的是评估NPY Y1受体对体外人阻力动脉血管舒缩功能的影响，并研究NPY受体系统对体内心血管血流动力学的作用。3. 从接受非血管疾病手术患者身上取下的人皮下阻力动脉，在体外与稳定的硫代磷酸化hY1受体反义寡脱氧核苷酸或错配寡脱氧核苷酸（10⁻⁷至10⁻⁵ M）一起孵育。4. 在预先用hY1 - AS（10⁻⁷至10⁻⁵ M）孵育的人阻力血管中，对NPY的收缩反应以剂量依赖性方式显著降低。在较低浓度（10⁻⁷ M至10⁻⁶ M）下，hY1 - MMODN孵育的血管未观察到影响。5. 在清醒大鼠连续输注48小时期间，研究了硫代磷酸化rY1 - ASODN、正义寡脱氧核苷酸或错配寡脱氧核苷酸的血流动力学效应。连续输注rY1 - ASODN未改变平均动脉压（MAP），而rY1 - SODN意外地导致动态MAP早期（10 - 20小时）升高，rY1 - MMODN导致晚期（24 - 44小时）升高。6. 与用rY1 - SODN、错配寡脱氧核苷酸处理的动物以及用生理盐水和葡萄糖处理的动物相比，长期输注rY1 - ASODN（2.1 mg kg⁻¹ h⁻¹，静脉输注48小时）处理的大鼠对NPY（2、4、8、16和32微克 kg⁻¹）的收缩反应显著降低。值得注意的是，输注rY1 - SODN的组对测试剂量的NPY表现出过度反应。7. 我们得出结论，用人皮下动脉与代谢稳定的18个碱基对hY1 - ASODN孵育以及用相应的rY1 - ASODN长期输注可减弱NPY诱导的血管收缩。