维药异叶青兰总黄酮对血管紧张素Ⅱ诱导的大鼠心肌细胞肥大的影响  

Effect of Dracocephalum heterophyllum Benth Flavonoid on Rat Myocardial Cell Hypertrophy Induced by Angiotensin Ⅱ

作　　者：蒋红[1] 何雯[2] 张晨[1] JIANG Hong;HE Wen;ZHANG Chen(Xinjiang Medical University, Urumqi 830011, China;The Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, China)

机构地区：[1]新疆医科大学,乌鲁木齐830011 [2]新疆医科大学第五附属医院,乌鲁木齐830011

出　　处：《中国药学杂志》2018年第8期594-600,共7页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金项目资助(81560689);新疆医科大学研究生创新创业项目资助(CXCY2017061)

摘　　要：目的观察维药异叶青兰总黄酮(Dracocephalum heterophyllum Benth flavonoid,DHBF)对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导的心肌细胞肥大的作用,并为进一步探讨其作用机制提供依据。方法 SD大鼠,0~3 d龄,体外培养大鼠乳鼠的心肌细胞,实验分为对照组、AngⅡ(1μmol·L^(-1))组、不同浓度的DHBF(10、25、50μmol·L^(-1))+AngⅡ(1μmol·L^(-1))。用AngⅡ1μmol·L^(-1)诱导心肌细胞肥大,DHBF对其进行干预,CCK-8法观察心肌细胞生存率,RT-PCR技术检测心肌肥大基因心房利钠肽(ANP)和脑钠肽(BNP)的mRNA表达水平,其内参因子为甘油醛-3-磷酸脱氢酶(GAPDH);激光共聚焦法检测心肌细胞的表面积和细胞内钙离子浓度[Ca^(2+)]i的浓度;破碎细胞酶促反应测定Ca^(2+)-ATP酶的活性;比色法测定NO的浓度和一氧化氮合酶(NOS)的活性,从而观察上述干预因素对心肌细胞肥大的影响。结果和对照组相比,AngⅡ组中心肌细胞生存率为(85%±5%),当DHBF与AngⅡ同时干预心肌细胞后,细胞生存率显著上升(P<0.05);RT-PCR结果显示,AngⅡ可使肥大标志物基因ANP和BNP的mRNA表达上调,当DHBF与AngⅡ同时干预心肌细胞后,可使ANP和BNP的mRNA表达下调;心肌细胞在AngⅡ的作用下表面积增大,而DHBF与AngⅡ同时作用可逆转这一现象;激光共聚焦法显示,AngⅡ作用于心肌细胞时,心肌细胞内[Ca^(2+)]i浓度增高,而DHBF与AngⅡ同时作用可使心肌细胞内[Ca^(2+)]i浓度明显减小(P<0.05);破碎细胞酶促反应显示AngⅡ作用于心肌细胞时,可使Ca^(2+)-ATP酶的活性下降,而DHBF与AngⅡ同时作用可使Ca^(2+)-ATP酶活性明显增强;比色法显示AngⅡ作用于心肌细胞时,可使NO浓度和NOS活性下降,而DHBF与AngⅡ同时作用可使NO浓度和NOS活性明显增加(P<0.05)。结论 DHBF能提高AngⅡ诱导的心肌细胞肥大的生存率,下调心肌肥大因子ANP和BNP的mRNA表达,减小由AngⅡ诱导的心肌细胞的表面积,且作用机制可能与促进NO的释�OBJECTIVE To observe the effect of Dracocephalum heterophyllum flavonoids（DHBF）of Uygur Medicine on cardiomyocyte hypertrophy,which were induced by angiotensin Ⅱ（AngⅡ）,and it could provide a basis for further study to the mechanism.METHODS SD rats,0-3 d of age,neonatal rat myocardial cells cultured in vitro,the experiment was divided into control group,AngⅡ（1 μmol·L-1）group,different concentrations of DHBF（10,25,50 μmol·L-1）＋AngⅡ（1 μmol·L-1）groups,cardiomyocyte hypertrophy were induced by AngⅡ 1 μmol·L-1 and was intervened using DHBF respectively,CCK-8 method was used to observe the activity of myocardial cells,RT-PCR technique was used to detect the expression of m RNA of cardiac hypertrophy gene atrial natriuretic peptide（ANP）and brain natriuretic peptide（BNP）,the internal factor was glyceraldehyde-3-phosphate dehydrogenase（GAPDH）.Confocal laser scanning was used to detect the surface area of myocardial cell was[Ca2＋]i;the activity of Ca2＋-ATP was measured by the enzymatic reaction of fragmentation cell;the concentration of NO and the activity of NOS were determined by colorimetry.RESULTS Compared with the control group,the activity of myocardial cell was（85%±5%）in the AngⅡgroup,which was increased significantly after it was dealed with DHBF and AngⅡ（P〈0.05）;RT-PCR results showed the expression of mRNA of ANP and BNP were increased by using AngⅡ,which were lower by using DHBF and AngⅡ.The surface area of myocardial cell were increased by using AngⅡ,which could be reversed by using DHBF and AngⅡ.Confocal laser scanning showed the concentration of[Ca2＋]i was increased by using AngⅡ,but which was lower significantly by using DHBF and AngⅡ（P〈0.05）.The enzymatic reaction of fragmentation cell results showed the activity of Ca2＋-ATP was decreased by using AngⅡ,but which was increased significantly by using DHBF and AngⅡ（P〈0.05）.Colorimetry results showed the concentration of NO and the activity of NOS were decreased

关 键 词：心肌细胞 肥大 异叶青兰总黄酮 血管紧张素Ⅱ 

分 类 号：R965[医药卫生—药理学]