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采用光谱荧光法研究血管钠肽(vasonatrin peptide,VNP)对心肌细胞内钙瞬变的作用及其机制,观察钠尿肽鸟苷酸环化酶(guanylate cyclase,GC)受体的特异性阻断剂(HS-142-1)、8-溴-环磷酸鸟苷(8-Br-cGMP)和镁蓝(methylene blue,MB)对心肌细胞内钙瞬变的影响。结果显示,异丙肾上腺素(isoproterenol,Iso)(10~(-10)～10~(-6)mol/L)可剂量依赖性地引起心肌细胞内钙瞬变增强,相对于对照组分别增强(13±8)%(P>0.05)、(26±13)%(P<0.05)、(66±10)%(P<0.01)、(150±10)%(P<0.01)和(300±25)%(P<0.01)。此效应可被β肾上腺素受体阻断剂普萘洛尔(10~(-6)mol/L)所阻断。VNP(10~(-10)～10~(-6)mol/L)可剂量依赖性地抑制Iso(10~(-8)mol/L)引起的心肌细胞内钙瞬变幅值的升高,相对于Iso(10~(-8)mol/L)分别减弱(99±3)%(P>0.05)、(96±2)%(P<0.05)、(84±6)%(P<0.01)、(66±3)%(P<0.01)和(62±3)%(P<0.01)。8-Br-cGMP(10~(-7)～10~(-3)mol/L)也可剂量依赖性地抑制Iso(10~(-8)mol/L)引起心肌细胞内钙瞬变的增强。HS-142-1(2×10~(-5)mol/L)使VNP的作用几乎完全消失。MB是GC的抑制剂,10~(-5)mol/L MB不但使VNP的作用完全消失,而且增强Iso对心肌细胞内钙瞬变的效应。VNP和HS-142-1本身对心肌细胞内钙瞬变无显著影响。而MB使心?
The effect and mechanism of vasonatrin peptide (VNP) on intracellular calcium transients were studied by spectrofluorometry. The specific blockade of the receptor of guanylate cyclase (GC) was observed (HS-142-1), 8-Br-cGMP and methylene blue (MB) on intracellular calcium transients were investigated. The results showed that isoproterenol (Iso) (10 ~ (-10) ~ 10 ~ (-6) mol / L) increased the intracellular calcium transients in a dose-dependent manner, which were increased compared with the control group (13 ± 8)% (P> 0.05), (26 ± 13)% (P <0.05), (66 ± 10)% ± 25)% (P <0.01). This effect was blocked by propranolol, a beta adrenergic receptor blocker (10 -6 mol / L). VNP (10 ~ (-10) ~ 10 ~ (-6) mol / L) inhibited the increase of intracellular calcium transients induced by Iso (10 ~ (-8) mol / L) (99 ± 3)% (P> 0.05), (96 ± 2)% (P <0.05), (84 ± 6)% respectively compared with Iso (10 -8 mol / L) 0.01), (66 ± 3)% (P <0.01) and (62 ± 3)% respectively (P <0.01). 8-Br-cGMP (10 -7 -10 -3 mol / L) also inhibited the intracellular calcium transients induced by Iso (10 -8 mol / L) in a dose-dependent manner Enhanced. HS-142-1 (2 × 10 ~ (-5) mol / L) almost completely disappeared the role of VNP. MB is an inhibitor of GC, 10-5 mol / L MB not only completely disappeared the role of VNP, and enhance the effect of Iso on intracellular calcium transients. VNP and HS-142-1 alone have no significant effect on intracellular calcium transients. MB makes heart?