CAJ | 학술논문

β-肾上腺素能受体%β3-AR激动剂%心肌成纤维细胞%磷脂酰肌醇(-3)激酶-蛋白激酶B信号传导途径%细胞增殖%Ⅰ,Ⅲ型胶原纤维%心脏重构%心力衰竭 β-腎上腺素能受體%β3-AR激動劑%心肌成纖維細胞%燐脂酰肌醇(-3)激酶-蛋白激酶B信號傳導途徑%細胞增殖%Ⅰ,Ⅲ型膠原纖維%心髒重構%心力衰竭
β-신상선소능수체%β3-AR격동제%심기성섬유세포%린지선기순(-3)격매-단백격매B신호전도도경%세포증식%Ⅰ,Ⅲ형효원섬유%심장중구%심력쇠갈
β3-adrenergic receptor%β3-adrenergic receptor agonist%Cardiac fibroblast%Phosphatidylinositol 3 kinase-protein kinase B signal transduction pathway%Cell proliferation%Type Ⅰ and Ⅱ collagen fiber%Cardiac remodeling%Heart failure

目的探讨β3-肾上腺素能受体(β3-adrenergic receptor,β3-AR)激动剂BRL37344是否通过细胞内磷脂酰肌醇(-3)激酶-蛋白激酶B(phosphatidylinositol 3 kinase-protein kinase B,PI3K-Akt)信号传导途径促进大鼠心肌成纤维细胞(cardiac fibroblasts,CFbs)增殖及胶原纤维增生.方法哈尔滨医科大学附属第一医院中心实验室无菌条件下剪取新生1～3 d的Wistar大鼠心脏,采用酶消化法、差速贴壁法获取CFbs.随机数字法分为5组:①空白组:不给予任何药物干预.②BRL组:给予BRL37344孵育.③LY组:给予LY294002(PI3K抑制剂)预孵育1 h后联合BRL37344共同孵育.④Akt-Ⅰ组:给予Akt-Ⅰ(Akt抑制剂)预孵育1 h后联合BRL37344共同孵育.⑤L-A组:LY294002和Akt-Ⅰ预孵育1 h后联合BRL37344共同孵育.通过MTT比色法观察细胞的增殖情况,RT-PCR法观察Ⅰ、Ⅲ型胶原表达情况.组间进行单因素方差分析,组间比较采用Tukey检验.以P＜0.05为差异具有统计学意义.结果 ①CFbs中存在β3-AR.②与BRL组比较,LY组和Akt-Ⅰ组增殖减低(P＜0.01);L-A组较LY组和Akt-Ⅰ组增殖减低更明显(P＜0.01).③与空白组比较,BRL组Ⅰ、Ⅲ型胶原mRNA表达均明显增加,给药48 h增加最显著.选取48 h点为作用时间点,与BRL组比较,LY组、Akt-Ⅰ组Ⅰ、Ⅲ型胶原mRNA表达减低,L-A组较LY组和Akt-Ⅰ组表达减低更显著(P＜0.01).结论 BRL37344通过PI3K-Akt信号传导途径促进心肌成纤维细胞增殖和Ⅰ,Ⅲ型胶原表达增加.
목적 탐토β3-신상선소능수체(β3-adrenergic receptor,β3-AR)격동제BRL37344시부통과세포내린지선기순(-3)격매-단백격매B(phosphatidylinositol 3 kinase-protein kinase B,PI3K-Akt)신호전도도경촉진대서심기성섬유세포(cardiac fibroblasts,CFbs)증식급효원섬유증생.방법 합이빈의과대학부속제일의원중심실험실무균조건하전취신생1～3 d적Wistar대서심장,채용매소화법、차속첩벽법획취CFbs.수궤수자법분위5조:①공백조:불급여임하약물간예.②BRL조:급여BRL37344부육.③LY조:급여LY294002(PI3K억제제)예부육1 h후연합BRL37344공동부육.④Akt-Ⅰ조:급여Akt-Ⅰ(Akt억제제)예부육1 h후연합BRL37344공동부육.⑤L-A조:LY294002화Akt-Ⅰ예부육1 h후연합BRL37344공동부육.통과MTT비색법관찰세포적증식정황,RT-PCR법관찰Ⅰ、Ⅲ형효원표체정황.조간진행단인소방차분석,조간비교채용Tukey검험.이P＜0.05위차이구유통계학의의.결과 ①CFbs중존재β3-AR.②여BRL조비교,LY조화Akt-Ⅰ조증식감저(P＜0.01);L-A조교LY조화Akt-Ⅰ조증식감저경명현(P＜0.01).③여공백조비교,BRL조Ⅰ、Ⅲ형효원mRNA표체균명현증가,급약48 h증가최현저.선취48 h점위작용시간점,여BRL조비교,LY조、Akt-Ⅰ조Ⅰ、Ⅲ형효원mRNA표체감저,L-A조교LY조화Akt-Ⅰ조표체감저경현저(P＜0.01).결론 BRL37344통과PI3K-Akt신호전도도경촉진심기성섬유세포증식화Ⅰ,Ⅲ형효원표체증가.
Objective To explore the effects of the β3 adrenergic receptor (β3-AR) agonist BRL37344 on cardiac fibroblast proliferation and collagen fiber hyperplasia in Wistar rats by promoting the phosphaticlylinositol 3 kinase-protein kinase B(PI3K-Akt) signal transduction pathway. Method Cardiac fibroblasts(CFbs) were isolated from 1 - 3 day-old Wistar rats under the sterile environment in the laboratory of the First Affiliated Hospital of Hasbin Medical University, by using enzyme digestion and an modified technique of differential anchoring velocity.The cultured myocardial cells were randomly (random number) divided into five groups. ① In blank group, rats were not treated with drug; ②in BRL group, rats were treated with BRL37344; ③in LY group, rats were treated with LY294002(PI3K antagonist) for one hour before treated with BRL37344;④in Akt-Ⅰ group,rats were treated with Akt-Ⅰ (Akt antagonist) for one hour before treated with BRL37344;⑤in L-A group, rats were treated with LY294002 and Akt-Ⅰ for one hour before treated with BRL37344. MTT colorimetric method and RT-PCR were used to observe the role of β3-AR agonist with or without PI3K antagonist and/or Akt antagonist in cardiac fibroblast proliferation (CFP) and collagen fiber hyperplasia (CFH). Comparisons between groups were made by one-way ANOVA and Tukey test. Results ①β3-AR was present in the CFbs. ②Compared with BRL group, the CFP and CFH in LY and Akt-Ⅰ groups were lower (P ＜0.01) and those in L-A group were lowest (P ＜ 0.01). ③Compared with blank group,the expressions of type Ⅰ and type Ⅲ fiber mRNA obliviously increased in BRL group (P ＜ 0.01),and at 48 hours,the expressionrs reached peak. At 48 hours,compared with BRL group,the expressions in LY and Akt-Ⅰ groups were lower, and were lowest in L-A group ( P ＜0.01). Conclusions BRL37344 promotes cardiac fibroblast proliferation and expressions of type Ⅰ and Ⅲ collagen fiber mRNA by activating the PI3K-Akt signal transduction pathway.