超氧阴离子对培养的牛动脉平滑肌细胞内钙及收缩性的影响

The effects of superoxide anion on intracellular Ca^(2+) concentration and contractility in cultured bovine aortic smooth muscle cells

目的研究超氧阴离子对牛主动脉平滑肌细胞内钙及收缩性的影响。方法采用Ｆｕｒａ－２测定牛肺动脉平滑肌细胞内钙及采用胶原凝胶实验方法分析平滑肌细胞的收缩性。结果超氧阴离子作用于平滑肌细胞后使ＡＴＰ（１０μｍｏｌ·Ｌ－１）诱导的细胞内钙浓度的增加从（２０６± １０） ｎｍｏｌ·Ｌ－１降到（１４７±１５） ｎｍｏｌ·Ｌ－１。 ５ ｍｉｎ内细胞内钙浓度增加的积分（∫ △［Ｃａ２＋］ｉ·ｄｔ）从（１２．２± ０．５）μｍｏｌ· Ｌ－１·ｓ－１降至（９．８± ０．８）μｍｏｌ·Ｌ－１·ｓ－１。ｔｈａｐｓｉｇａｒｇｉｎ在无钙溶液中诱导的细胞内钙浓度的增加不受超氧阴离子的影响，而在含钙的Ｋｒｅｂａ溶液中，超氧阴离子能使随后的钙释放激活的钙进入（△［Ｃａ２＋］ｉＣＲＡＣ）从（２７．３ ±１．０） ｎｍｏｌ·Ｌ－１降到（１３．５ ±１．０） ｎｍｏｌ·Ｌ－１。ＡＴＰ诱导的凝胶面积减少的百分比从２４％±５％降到 ７．４％ ±０．２％；在无钙 Ｋｒｅｂｓ溶液中，ｔｈａｐｓｉｇａｒｇｉｎ诱导的凝胶面积减少的百分比从 ３．５％± ０．６％降到－０．５％±０．７％。结论超氧阴离子通过影响平滑肌细胞的内钙动员而降低它的收缩性。

AIM To study the effects of superox- ide anion (O ) on Ca2+ homeostasi and contractility in cultured bovine aortic smooth muscle cell. METHODS Using Fura-2 fluorescence technique to determine Ca2+ level and collagen gel contraction system to analyze muscle contractility. RESULTS ATP (10 μmol· L-1 )-induced Ca2+ transient was smaller in xanthine oxidase treated cells(X/XO) than control. The mean peak increment of [Ca2+ ]i (△[Ca2+ ], peak) and the time integral of the elevated [Ca2+ ], (∫ △[Ca2+ ]i dt) for 5 min were decreased from (206. 1 ± 10.2) to (147.4 ± 14.7) nmol· L-1, and from (12.2 ±0.5) to (9.8 ± 0.8) μmol·L-1 .s-1. △ [Ca2+ ], peak induced by thapsigargin(1 μmol. L- 1 ) in Ca2+ -free solution was not affected by X/XO, but was decreased from (27. 3 ± 1 .0) nmol· L－l to (13 .5 ± 1 .0 ) nmol· L- １ in Ca2+ -containing solution be cause of the activation of CRAC(△［Ｃa2+ ], CRAC). X/XO accelerated the velocity of thapsigargin-induced Ca2+ leak from (78.7 ± 3.4) s to (64.8 ± 4.40) s. Gel contraction area in X/XO-treated cells induced by ATP or thapsigargin (in Ca2+ free solution and in Krebs solution) was decreased from 23.6% ± 4.6% to 7.4% ±0.2%, from 3.5% ±0.6% to － l.0% ± 0.5%, and from 7.9% ± l.4% to － 0.5% ± 0.7%, respectively. CONCLUTION O2- attenuats smooth muscle contraction by impairing some of Ca2+ mobilization pathways.