四氢生物蝶呤通过蛋白激酶Cε途径对醋酸脱氧皮质酮-盐型高血压小鼠左心室舒张功能的影响  

Impact of tetrahydrobiopter on left ventricular diastolic function by upregulating protein kinase Cεsignaling pathway in desoxycorticosterone acetate-salt hypertensive mice

作　　者：许涵[1] 王琼英[1] 杨米娜[1] 马瑞新[1] 李宁荫[1] 李秀丽[1] 林欣[1] 张小卫[1] 余静[1] 

机构地区：[1]兰州大学第二附属医院高血压病科,甘肃兰州730030

出　　处：《中华高血压杂志》2015年第3期245-251,共7页Chinese Journal of Hypertension

基　　金：国家自然科学基金资助课题(81270332);甘肃省科技支撑项目(1104FKCA150)

摘　　要：目的探讨四氢生物蝶呤(BH4)对醋酸脱氧皮质酮(DOCA)-盐型高血压小鼠左心室舒张功能及蛋白激酶Cε(PKCε)表达的影响。方法雄性C57BL/6小鼠分为手术组(n=50)与假手术组(n=40),手术组小鼠切除左侧肾脏后在颈部埋入DOCA药片,术后以普通饲料及1.05%NaCl溶液饲养,至术后14d,排除死亡小鼠6只后随机分为DOCA组(n=22)与DOCA+BH4组(n=22,2.5%的BH4溶液0.1mL/10g灌胃7d),假手术组小鼠仅游离左侧肾脏而不切除,并在颈部埋入安慰剂药片,术后以普通饲料及常规不含盐净化水饲养,然后随机分为假手术组(n=20)及假手术+BH4组(n=20)。动物在术后22d时处死,采用酶联免疫吸附试验、Western-blot以及高效液相色谱法(HPLC)测定心肌组织中环磷酸鸟苷(cGMP)、丙二醛、BH4和PKCε等指标的变化。处死前行鼠尾动脉压、心脏超声及血流动力学检查。结果与假手术组相比,DOCA组血压升高(P<0.05),DOCA+BH4组与DOCA组相比,血压差异无统计学意义(P>0.05)。与假手术组相比,DOCA组二尖瓣舒张早期血流速度与二尖瓣环舒张早期运动速度比值(E/E’)、舒张末期压力-容积关系(EDPVR)及Tau指数增高[分别为(38.49±3.91)比(25.77±5.21),(0.22±0.05)比(0.15±0.02)mm Hg/μL,(14.27±0.79)比(10.60±0.52)ms,均P<0.05]。与DOCA组相比,DOCA+BH4组EDPVR及Tau指数减小[(0.16±0.04)比(0.22±0.05)mm Hg/μL,(12.05±1.35)比(14.27±0.79)ms,均P<0.05]。与假手术组相比,DOCA组超氧化物歧化酶(SOD)及一氧化氮减少,给予BH4干预后,SOD及一氧化氮增多。Western-blot结果显示,与假手术组相比,DOCA组PKCε的表达减少(P<0.05),与DOCA组相比,DOCA+BH4组PKCε的表达增加(P<0.05)。结论 BH4无明显降压作用,但可改善左心室舒张功能不全,这可能与其通过PKCε途径降低体内氧化应激水平、提高一氧化氮表达等有关。Objective To identify the impact of tetrahydrobiopterin （BH4） on left ventricular diastolic function and the expression of protein kinase Cε （PKCε） in desoxycorticosterone acetate （DOCA）-salt hypertensive mice. Methods Male C57BL/6 mice were divided into the operation group （n=50） or the sham operation group （n=40）. In the operation group, left kidney was resected and DOCA pill was implanted in cervical part; then mice were fed with normal fodder and normal saline （NS） and were randomly divided into the DOCA group （n= 22） and the DOCA＋ BH4 group （n=22, 2.5% BH_4, 0. 1 mL/10 g intragastrical administration for 7 d） without death （n=6） until the 14th day. Left kidney in mice of the sham operation group was only separated and placebo pill was implanted in the cervical part; then mice were fed with normal fodder and normal water without salt and all of them were randomly separated into the sham operation group （Sham group, n=20） or the Sham＋BH4 group （n=20）. Arterial pressure,echocardigraphy and hemodynamic method were used to test whether DOCA model was successfully established, and cardiac structure and function were assessed on the 21st day after operation. Cyclic guanosine monophosphate （cGMP）, malonaldehyde, BH4 and PKCε were detected by enzyme linked immunosorbent assay, Western-blot or high-performance liquid chromatography in cardiac tissues of all enthanized mice the next day. Results Compared to the Sham group, systolic and diastolic blood pressure （SBP and DBP） in DOCA group were significant ly increased （P〈0.05）, but no significance was revealed between the DOCAq-BH4 group and the DOCA group （P〉0.05）. The ratio of left-ventrieular early diastolic filling velocity to early diastolic mitral annular velocity （E/E＇） （ 38.49±3.91 vs 25.77 ±5.21 ）, end-diastolic pressure-volume relation （ EDPVR ） [ （ 0.22 ± 0.05 ） vs（0. 15±0.02）mm Hg/μL] and Tau index[- （14. 27±0.79） vs （10. 60±0. 52）ms] were significantl

关 键 词：四氢生物蝶呤 醋酸脱氧皮质酮-盐型高血压小鼠 高血压 左心室舒张功能不全 蛋白激酶CΕ 

分 类 号：R544.1[医药卫生—心血管疾病]