低氧性肺动脉高压大鼠肺动脉平滑肌细胞中Kv2.1基因表达的变化  

Changes of voltage-gated K+ 2.1 gene expression in pulmonary arterial smooth cells in rats with hypoxic pulmonary hypertension

作　　者：林慧庆[1] 黄杰[1] 程邦昌[1] 毛志福[1] 赵华[1] 王志强[1] 

机构地区：[1]武汉大学人民医院胸心外科,湖北省武汉市430060

出　　处：《中国临床康复》2005年第39期92-95,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:探讨肺动脉平滑肌细胞门控钾离子通道Kv2.1基因在缺氧性肺动脉高压发病中的作用。方法:①实验于2003-04/2004-06在武汉大学人民医院胸心血管外科实验室完成。选用健康成年雄性普通级Wistar大鼠48只。随机将大鼠分为6组,分别为对照组、缺氧8h组及缺氧3,7,14和28d组,每组8只。将动物置于常压低氧仓,仓内注入流通氮氧混合气,调节仓内氧浓度保持在(10±5)%,将大鼠造成常压缺氧性肺动脉高压模型。对照组:不造成缺氧性肺动脉高压大鼠模型。缺氧8h组及缺氧3,7,14和28d组:分别将大鼠置于缺氧环境中8h,3,7,14和28d。②记录肺动脉压力波,计算肺动脉平均压。测定血浆中游离钾、钙离子浓度。应用全自动图像心肺血管分析软件测定肺细小动脉中膜平滑肌细胞核密度及肺细小动脉中膜厚度。③对肺动脉平滑肌细胞进行染色及显色后,镜下观察肺动脉平滑肌细胞表面核抗原以及bcl-2抑凋亡因子表达。bcl-2抑凋亡因子表达:0为不表达,+为微弱表达,为中等阳性,为强阳性。④以图像分析系统(IBAS)对杂交点进行密度扫描,以经同样处理的β-actin基因mRNA表达量作为内参照,肺动脉平滑肌细胞Kv2.1基因mRNA表达以两者的积分吸光度(A值)的比值表示。⑤对数据进行方差分析处理,组间比较采用t检验。结果:大鼠48只均进入结果分析。①随着缺氧时间延长,大鼠平均肺动脉压、肺细小动脉中膜平滑肌细胞核密度及肺细小动脉中膜厚度逐渐升高。各缺氧组大鼠平均肺动脉压明显高于对照组(P<0.01),缺氧3,7,14,28d组大鼠肺细小动脉中膜平滑肌细胞核密度及肺细小动脉中膜厚度明显高于对照组(P<0.05～0.01)。②随着缺氧时间延长,大鼠血浆中游离K+浓度逐渐降低、Ca2+浓度逐渐升高。缺氧3,7,14,28d组大鼠血浆中游离K+浓度明显低于对照组(P<0.05～0.01),缺氧7,14,28d组大鼠血浆中游离Ca2+浓度明显高于AIM： To investigate the role of voltage-gated K^＋ （Kv）2.1 channel of pulmonary arterial smooth cells in the attack of hypoxia pulmonary hypertension. METHODS： ① The experiment was carried out in the laboratory of Department of Thoracic ＆ Cardiovascular Surgery, Renmin Hospital of Wuhan　University between April 2003 and June 2006, Forty-eight healthy adult male Wistar rats were randomly divided into 6 groups with 8 rats in each group： control group, hypoxia 3, 7, 14 and 28 days groups, The rats were kept in a normobarie hypoxie chamber, The inspired oxygen fraction was kept at （10±5）%, and the rats, except those in the control group, were made into models of hypoxia pulmonary hypertension for 8 hours and 3, 7, 14 and 28 days, ② The pulmonary artery pressure wave was recorded, average pulmonary artery pressure was calculated, The concentrations of free K^＋ and Ca^2＋ in plasma were detected, The smooth muscle cells （SMCs） nuclear density and media thickness, ③ After staining of pulmonary arterial SMCs, the expressions of surface nuclear antigen and bel-2 inhibiting apoptosis factor were observed under microscope： 0 as no expression, ＋ as weak expression, ＋＋ as moderate positive, as strong positive, ④The image biological analysis system was applied to scan the density of point of intersection, and the same treated β-actin mRNA expression amount was taken as the internal reference, and the pulmonary arterial SMCs Kv2.1 mRNA was expressed with the ratio of the two integrated absorbance （A value）. ⑤ The data were treated with analysis of variance, and the t test was applied in the intergroup comparison, RESULTS： All the 48 rats were involved in the analysis of results. ① With the elongation of hypoxia time, the average pulmonary artery pressure, SMCs nuclear density and media thickness were gradually increased. The average pulmonary artery pressure was obviously higher in the hyporda groups than in the control group （P 〈 0.01）, SMCs nuclear density and media

关 键 词：高血压 肺性 低氧 基因表达 

分 类 号：R563[医药卫生—呼吸系统]