CAJ | 학술논문

目的探讨组织蛋白酶S(Cathepsin S,Cat S)在高血压性心脏病心室重构发生发展中的表达及其活性动态变化与心功能的关系.方法 7周龄雄性Dahl盐敏感(DS)大鼠分别给予高负荷食盐(8%)或正常负荷食盐(0.3%)为12周和19周建立心肌肥厚(H-LvF)模型,心力衰竭模型(H-HF)及同龄对照组(12W-C和19W-C).另设人体心肌生检标本作研究对象(高血压代偿性心肌肥厚患者7例,心力衰竭患者18例和正常对照6例).RT-PCR和免疫组化分别检测Cat S mRNA及蛋白表达,酶谱法测定大鼠心肌组织中的Cat S活性,免疫印迹杂交检测心肌组织中的白介素1β蛋白表达,细胞荧光染色法检测培养心肌细胞Cat S的表达,心脏石蜡切片苦味酸-酸性品红染色法和van Gieson's染色法分析胶原蛋白和弹性蛋白含量,心脏超声和血流动力学测定心功能和心室重构.结果 H-HF组大鼠和患者心肌中的Cat S mRNAs的表达显著高于19W-C组和H-LVF组大鼠和人(P＜0.01).免疫组化法和in situ法分析显示增高的Cat SmRNA和蛋白主要表达于心肌细胞.H-HF组大鼠心肌的弹性蛋白分解能也显著高于19W-C(4.1倍).H-LVF大鼠心肌间质弹性蛋白含量明显增加,而在H-HF大鼠则下降低于对照组水平.H-HF组心肌白介素IL1β蛋白含量显著高于19W-C.IL1β刺激心肌细胞的Cat S蛋白表达.心肌中的弹性蛋白酶表达与左心室射血分数呈显著负相关(r=-0.88,P＜0.05).结论 Cat S参与心室病理改变,是导致间质改变、心功能异常的主要因素之一,提示心肌Cat S在H-HF时心室重构发展中起重要作用.
목적 탐토조직단백매S(Cathepsin S,Cat S)재고혈압성심장병심실중구발생발전중적표체급기활성동태변화여심공능적관계.방법 7주령웅성Dahl염민감(DS)대서분별급여고부하식염(8%)혹정상부하식염(0.3%)위12주화19주건립심기비후(H-LvF)모형,심력쇠갈모형(H-HF)급동령대조조(12W-C화19W-C).령설인체심기생검표본작연구대상(고혈압대상성심기비후환자7례,심력쇠갈환자18례화정상대조6례).RT-PCR화면역조화분별검측Cat S mRNA급단백표체,매보법측정대서심기조직중적Cat S활성,면역인적잡교검측심기조직중적백개소1β단백표체,세포형광염색법검측배양심기세포Cat S적표체,심장석사절편고미산-산성품홍염색법화van Gieson's염색법분석효원단백화탄성단백함량,심장초성화혈류동역학측정심공능화심실중구.결과 H-HF조대서화환자심기중적Cat S mRNAs적표체현저고우19W-C조화H-LVF조대서화인(P＜0.01).면역조화법화in situ법분석현시증고적Cat SmRNA화단백주요표체우심기세포.H-HF조대서심기적탄성단백분해능야현저고우19W-C(4.1배).H-LVF대서심기간질탄성단백함량명현증가,이재H-HF대서칙하강저우대조조수평.H-HF조심기백개소IL1β단백함량현저고우19W-C.IL1β자격심기세포적Cat S단백표체.심기중적탄성단백매표체여좌심실사혈분수정현저부상관(r=-0.88,P＜0.05).결론 Cat S삼여심실병리개변,시도치간질개변、심공능이상적주요인소지일,제시심기Cat S재H-HF시심실중구발전중기중요작용.
Objective To observe myocardial cathepsin (Cat) S expression and activity in hypertensive heart failure rats.Methods The expression and activity of Cat S were determined in the left ventricular (LV) myocardium (LVM) of Dahl salt-sensitive rats fed either a high-salt(HS,8%)or low-salt(LS,0,3%,controls) diet starting at age 7 weeks for 12 weeks(hypertrophy model,H-LVH) or 19 weeks(heart failure model,H-HF).Age-matched rats served as controls and human normal,hypertensive and heart failure myocardial specimen were also examined for changes on the expression and activity of Cat S.Resuits Reverse transcription and real-time polymerase chain reaction analysis revealed significantly upregulated Cat S mRNA in rats with H-HF than in rats with H-LVH or in control rats and Cat S mRNA expression is negetively correlated with LVEF(r=-0.88.P＜0.05).In situ and immunohistochemistry examinations showed that Cat S was localized predominantly in cardiac myocytes(CMCs)and coronary vascular smooth muscle cells(SMC).Elastic lamina fragmentations and Cat S-dependent elastolvtic activity were significantly increased in H-HF-rats.The expression of interleukin-1β was also increased in the LVM of H-HF rats,and this cytokine was found to increase the Cat S protein expression in cultlife neonatal CMCs.Similar results were revealed in human myocardial specimens.Conclusion Elastolytic Cat S might play an important role in the pathogenesis of myocardial remodeling and heart failure and Cat S might serve as a novel therapheutic target in preventing or reversing hypertension induced LV remodeling and heart failure.