Roles of innate immunity in genesis of heart failure

本研究的目的是探讨先天免疫在心肌功能不全发生或发展中的作用。特别是，我们试图阐明Toll样受体（TLRs）在心肌细胞中的表达及其功能意义.鸡TLR 2的分子克隆和表达分析：我们在培养的10日龄鸡胚（CEVMs）心室肌细胞中克隆了与哺乳动物TLR 2同源的cDNA。该cDNA已证明对应于鸡TLR 2 1型（福井等）。采用北方印迹法检测TLR 2在10日龄雏鸡心脏和CEVMs中的表达.大鼠血管紧张素II（A-IO）输注模型：Sprague-Dawley大鼠皮下连续注射A-LI，建立心室肥厚/衰竭模型。在A-Il灌注的大鼠中观察到胎儿基因重编程和Ca^<2+>的改变.大鼠野百合碱（MCT）输注模型：雄性大鼠也皮下连续给予MCT，以建立肺动脉高压和右心室衰竭。胎儿基因重编程发生在右心室和左心室，尽管观察到一些差异，提示心室相互作用的作用.心肌细胞内Ca^2+的测定：心肌细胞内Ca^2+的瞬时变化用肌浆网特异性表达的flou-3或cameleon记录。

The objectives of this study were to investigate roles of innate immunity in the genesis or progression of myocardial dysfunction. Particularly, we tried to elucidate expression and functional significance of toll-like receptors (TLRs) in cardiac myocytes.1. Molecular cloning and expression analysis of the chick TLR2: we have cloned a cDNA that is homologous to mammalian TLR2s in cultured ventricular myocytes from 10-day-old chick embryos (CEVMs). This cDNA has turned out to correspond to the chick TLR2 type 1 (Fukui, et al.). Total TLR2 expression was detected by Northern blotting in the 10-day-old chick hearts and CEVMs.2. Rat angiotenisn II (A-IO) infusion model: Sprague-Dawley rats were injected subcutaneously and continuously with A-LI to create a ventricular hypertrophy/failure model. Fetal gene reprogramming and alteration of Ca^<2+> were observed in the A-Il-infused rats.3. Rat monocrotaline (MCT) infusion model: MCT was also given subcutaneously and continuously to male rats to create pulmonary hypertension and right ventricular failure. Fetal gene reprogramming occurred in both the right and left ventricles, although some differences were observed, suggesting a role of ventricular interaction.4. Measurements of Ca^<2+> in cardiac myocytes: Ca^<2+> transient in cardiac myocytes was recorded with flou-3 or cameleon, which can be expressed specifically in sarcoplasmic reticulum.