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肺动脉高压模型大鼠肺动脉平滑肌RyR结合特性及基因C-末端序列的研究
引用本文:承伟,叶兆伟,王佩琪,赵文明,张壮,李智.肺动脉高压模型大鼠肺动脉平滑肌RyR结合特性及基因C-末端序列的研究[J].中国药理学通报,2008,24(10).
作者姓名:承伟  叶兆伟  王佩琪  赵文明  张壮  李智
作者单位:1. 中国医科大学药学院天然药物研究室,辽宁,沈阳,110001
2. 辽宁医学院药学院,辽宁,锦州,121000
3. 辽宁医学院第三附属医院,辽宁,锦州,121000
摘    要:目的分析肺动脉高压时肺动脉平滑肌Ryanodine受体(ryanodine receptor,RyR)的结合功能改变及基因序列的变化。方法梯度离心法提取肺动脉平滑肌肌浆网膜蛋白组分,放射配基结合方法测定平衡解离常数(Kd)、最大结合容量(Bmax)的变化;体外提取肺动脉平滑肌细胞总RNA,反转录合成RyR2cDNA,以cDNA为模板使用特异性引物对目的片段进行PCR扩增,切胶回收后Sanger双脱氧末端终止法测定序列。结果野百合碱(monocrotaline,MCT)所致肺动脉高压组RyR最大结合容量(Bmax)为(197±21.4)nmol.g-1protein,较对照组(168±13.7)nmol·g-1protein明显增加(P<0.01);平衡解离常数(Kd)为(0.276±0.026)nmol·L-1与对照组(0.282±0.031)nmol·L-1相比差异无显著性(P>0.05);野百合组RyR2C-末端基因序列与对照组无差异。结论MCT所致肺动脉高压大鼠RyR最大结合容量(Bmax)增加,提示受体的数量发生了上调;Kd无明显变化,说明与受体的亲和力无改变。MCT所致肺动脉高压大鼠2型RyR(RyR2)C-末端基因序列无变化。

关 键 词:肺动脉高压  Ryanodine受体  肌浆网  放射配基  平衡解离常数  最大结合容量  基因序列

Studies on binding features and C-terminal gene sequence analysis of ryanodine receptor in pulmonary artery hypertension rats
CHENG Wei,YE Zhao-wei,WANG Pei-qi,ZHAO Wen-ming,ZHANG Zhuang,LI Zhi.Studies on binding features and C-terminal gene sequence analysis of ryanodine receptor in pulmonary artery hypertension rats[J].Chinese Pharmacological Bulletin,2008,24(10).
Authors:CHENG Wei  YE Zhao-wei  WANG Pei-qi  ZHAO Wen-ming  ZHANG Zhuang  LI Zhi
Institution:CHENG Wei1,YE Zhao-wei2,WANG Pei-qi2,ZHAO Wen-ming2,ZHANG Zhuang3,LI Zhi1(1. College of Pharmacology,China Medical University,Shenyang 110001,China,2. College of Pharmacology,Liaoning Medical College,3.the Third Affiliated Hospital,Jinzhou Liaoning 121000,China)
Abstract:Aim To investigate the changes in binding features and C-terminal gene sequence of ryanodine receptors in the pulmonary artery smooth muscle of pulmonary arterial hypertension (PAH) rats induced by monocrotaline (MCT). Methods Sarcoplasmic reticulum (SR) were fractionated with density gradient centrifugation and the characteristic of ryanodine receptor was assayed with a method of radioreceptor binding assay; The total RNA was obtained from the PAMSC of rat. The purity of total RNA was detected by agarose e...
Keywords:pulmonary artery hypertension  ryanodine receptor  sarcoplasmic reticulum  radioreceptor binding  Kd  Bmax  DNA sequence  
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