||The effect of H9c2 cell-mediated HGF transgene on the improvement of cardiac relaxation in a rat model of myocardial infarction
||Department of Physiology
hepatocyte growth factor
本論文研究的目的，在探討大鼠心肌梗塞(myocardial infarction)動物模式下，注射感染肝生長因子(hepatocyte growth factor)的胚胎心肌細胞(H9c2)，對心臟功能及動脈形態的影響。
以縫線將大鼠心臟冠狀動脈左下降枝(left antirior descending coronary artery)血流給予阻斷，造成急性冠狀心肌梗塞(acute myocardial infarciton)。心肌梗塞14天之後，老鼠隨機分成五組，分別給予細胞培養基質(medium)(50μl)、腺病毒(adenovirus carrying LacZ)(5x105 Pfu)、腺病毒攜帶肝細胞因子(adenovirus carrying HGF)(1x109 )、胚胎心肌細胞(5x105 )以及胚胎心肌細胞感染肝生長因子(H9c2 cell carrying HGF)(5x105 )。實驗結果發現：1) 與細胞培養基質組比較，在胚胎心肌細胞感染肝生長因子和腺病毒攜帶肝細胞因子兩組中，心臟纖維組織(scar)有明顯的減少；2) 與細胞培養基質組相比，在胚胎心肌細胞感染肝生長因子和腺病毒攜帶肝細胞因子兩組中，心肌等容舒張能力(diastolic dp/dt; isovolumetric relaxation)有顯著的增加；3) 與細胞培養基質組相比，胚胎心肌細胞感染肝生長因子組中，直徑大於100μm的動脈數目沒有變化; 4) 和細胞培養基質組相比，腺病毒攜帶肝細胞因子組中，動脈外膜纖維組織(adventitia)層與平滑肌層的面積比有明顯增加。本論文証實了注射感染肝生長因子(hepatocyte growth factor)的胚胎心肌細胞(H9c2)，能減少大鼠心肌梗塞所造成的心臟纖維組織的形成，並增加心肌舒張功能。
Backgroud- The purpose of this study was to evaluate the effects of embryonic cardiomyocytes mediated-HGF gene therapy on ventricular function of infarcted hearts, and arterial development.
Methods and Resutls-In rats, left anterior descending coronary artery ligation was operated to create MI. Two weeks after ligation, rats were randomly divided into five groups: (1) the medium group for control (injection of 50μm culture medium, n=3), (2) the Ad-LacZ group (injection of 5*10^5 pfu Ad.LacZ, n=3), (3) the HGF group (injection of 1*10^9 pfu Ad.HGF, n=3), (4) the cell group (1*10^5 H9c2 cells, n=3), and (5) the C-HGF group (1*10^5 H9c2 cells infected with Ad.HGF, n=3). Scar formation was reduced after either the HGF group or the C-HGF group compared to the medium group. Cardiac functions determinated by Millar demonstrated that end-diastolic pressure was significantly decreased in two treatment groups when compared with the medium group. Further, the minimal dp/dt was significantly increased in the C-HGF group compared to the medium group. In vascular system, the arterial density was no changed among groups. The ratio of adventitia area to smooth muscle cell area was increased in the HGF group compared to the medium group.
Conclusions-The present study proves that the application of H9c2 cell-mediated HGF gene therapy affected cardiac function by reducing scar expansion and improving cardiac relaxation. The intervention of this combine therapy may contribute to the prevention of heart failure from a chronic MI.
MATERIAL AND METHODS 9
AIMⅠ: THE MORPHOLOGICAL CHARACTERIZATION OF INFARCTED HEARTS IN RATS 15
Objective 1: to investigate the effect of coronary occlusion on myocardial infarction by Masson’s trichrome staining 15
AIMⅡ: THE THERAPEUTIC EFFECT OF CELL-MEDIATED HGF GENE THERAPY ON VENTRICULAR REMODELING 16
Objective 1: to insure the infection efficiency of Ad.HGF in H9C2 cells by cell staining 16
Objective 2 to confirm the distribution of H9c2 cell-carried LacZ transgene in the ligated hearts by X-gal staining 16
Objective3: to evaluate the effect of cell-mediated HGF gene therapy on ventricular geometry by Massion’s Trichrome staining. 16
Objective 4: to measure the effect of cell-mediated HGF gene therapy on cardiac functions by Millar Instrument. 17
AIMⅢ: THE THERAPEUTIC EFFECT OF CELL-MEDIATED HGF GENE THERAPY ON VASCULAR REMODELING 18
Objective 1: to study the effect of acute myocardial infarction on arterial morphology by SMCα-actin staining 18
Objective 2: to evaluate the effect of cell-mediated HGF gene therapy on the arterial density by SMCα-actin staining. 18
Objective 3: to evaluate the effect of cell-mediated HGF gene therapy on arterial remodeling by Massion’s Trochrome staining 18
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