Ca^(2+)转运通路对金雀异黄酮舒张大鼠脑血管作用的影响  

Role of Ca^(2+) Transport Pathways in the Effects of Genistein on Serotonin-Activated Cerebrovascular Contraction

作　　者：冯念苹[1] 林静涵[2] 聂雪丹[1] 孙丽娜[2] 张黎明[2] 梁庆成[1] 王正非 郑海洪[4] 

机构地区：[1]哈尔滨医科大学附属第二医院神经内科,黑龙江哈尔滨150086 [2]哈尔滨医科大学附属第一医院神经内科,黑龙江哈尔滨150001 [3]齐齐哈尔市中医医院脑病三科,黑龙江齐齐哈尔161000 [4]哈尔滨医科大学附属第二医院实验动物学部,黑龙江哈尔滨150086

出　　处：《现代生物医学进展》2015年第19期3645-3650,共6页Progress in Modern Biomedicine

基　　金：黑龙江省教育厅科学技术研究项目(12521336)

摘　　要：目的:研究Ca2+转运通路对金雀异黄酮舒张大鼠脑血管作用的影响。方法:75只大鼠被随机分为3组,分别经由二甲亚砜、金雀异黄酮和酪氨酸磷酸化抑制剂A47处理基底动脉及Willis环血管。每组大鼠进一步划分成5个亚组,每个亚组用不同浓度的细胞外Ca2+处理,分为:0、0.6、1.2、1.8和3.6 m M Ca2+组。5-羟色胺诱导血管收缩。测定大鼠基底动脉管壁厚度与官腔周长的比值;荧光成像分析法测定血管平滑肌细胞细胞内Ca2+浓度;免疫印迹分析检测肌球蛋白轻链激酶(MLCK),蛋白质磷酸酶催化亚基1(PP1),肌凝蛋白磷酸酶目标亚基1(MYPT1)的表达来测定血管平滑肌细胞Ca2+敏感性。结果:金雀异黄酮和酪氨酸磷酸化抑制剂A47显著降低大鼠基底动脉管壁厚度与官腔周长的比值(P<0.01),Ca2+内流(P<0.01,P<0.05)及MLCK的表达(P<0.01);增加PP1和MYPT1的表达(P<0.01)。细胞外Ca2+与金雀异黄酮及酪氨酸磷酸化抑制剂A47有协同效应。硝苯地平和毒胡萝卜素可废除该效应。结论:低细胞外Ca2+水平增强了金雀异黄酮和酪氨酸磷酸化抑制剂A47的血管舒张作用。L型电压门控Ca2+通道(L-VGCC)和肌浆网Ca2+库(SR)参与交互效应。Objective： To study the role of Ca2~ transport pathways in the effects of genistein on serotonin [] activated cerebrovascular contraction. Methods： Seventy five rats were randomly divided into 3 groups. Basilar artery （BA） and Willis ring in different groups were incubated with dimethyl sulfoxide, genistein and tyrphostin A47 （Tyr A47）.Each group was further divided into subgroups of 5 rats that were treated with different concentrations of 0, 0.6, 1.2, 1.8 and 3.6 mM Ca2＋. Serotonin induced vasoconstriction. The effects of genistein Tyr A47 on serotonin-activated contraction were examined by measuring the ratio of tube wall thickness and the lumen perimeter of the BA. Intracellular Ca2＋ concentration （[Ca2＋]i） of vascular smooth muscle cells was determined by ratiometric fluorescence imaging analysis. The Ca2＋ sensitivity was determined by measuring the expression of myosin light chain kinase （MLCK）, protein phosphatase catalytic submit 1 （PP1） and myosin phosphatase target submit 1 （MYPT1） by Western blot analysis. Results： Genistein and Tyr A47 significantly reduced the ratio of tube wall thickness and the lumen perimeter of BA （P 〈0.01 ）, [Ca2＋]i （P 〈0.01,P 〈0.05）and the expression of MLCK （P 〈0.01）. Opposite results were observed for PP1 and MYPT1 （P 〈0.01）. Extracellular Ca2＋ had a synergistic effect on genistein and Tyr A47. The effect was abolished by nifedipine and thapsigargin. Conclusions： Low extracellular Ca2＋ enhanced the vasodilatation that was stimulated by genistein and Tyr A47. L-type voltage-gated Ca2＋ channel （L-VGCC） and sarcoplasmic reticulum （SR） Ca2＋ were involved in the interaction.

关 键 词：Ca2+运输 金雀异黄酮 5-羟色胺 脑血管收缩 

分 类 号：Q95-3[生物学—动物学] R743[医药卫生—神经病学与精神病学]