Rozek R J, Kuo T H, Giacomelli F, Wiener J
J Mol Cell Cardiol. 1983 Mar;15(3):173-87. doi: 10.1016/0022-2828(83)90297-3.
Two-kidney, one clip Goldblatt hypertension of 2, 4 and 8 weeks duration was induced in 100-g male Wistar-Kyoto rats. Nucleic acid content was determined in the isolated cardiac muscle cells from the left ventricle. The profile for several major proteolytic activities in either isolated cardiac muscle cells or left ventricle preparations was also studied, using [3H]acetyl-casein as substrate. From the soluble fraction of the tissue or cell preparation, a pH 6 proteolytic activity, two forms of calcium-activated protease as well as cathepsin D were identifiable by inhibitor assay or DEAE-cellulose chromatography. From the myofibrillar fraction of the same preparation, two kinds of proteolytic activity were detected at alkaline pH: a phenylmethylsulfonyl fluoride (PMSF) inhibitable activity that was serine protease-like and the other a N-ethylmaleimide (NEM) inhibitable activity that resembled Ca2+-activated protease. At 2 weeks of hypertension, there was a significant increase in the pH 6 proteolytic activity as well as the calcium-activated protease I and the NEM-inhibitable alkaline protease activities, while the other identifiable proteolytic activities remained unchanged. Lysosomal cathepsin D showed a rise in activity only after 8 weeks of hypertension. These results may be related to the development of myocyte necrosis and lysis that occur in this model of hypertensive cardiomyopathy.
在100克雄性Wistar-Kyoto大鼠中诱发持续2周、4周和8周的双肾单夹型戈德布拉特高血压。测定左心室分离心肌细胞中的核酸含量。还使用[3H]乙酰酪蛋白作为底物研究了分离的心肌细胞或左心室制剂中几种主要蛋白水解活性的概况。通过抑制剂测定或DEAE-纤维素色谱法，可从组织或细胞制剂的可溶部分中鉴定出一种pH 6蛋白水解活性、两种形式的钙激活蛋白酶以及组织蛋白酶D。从同一制剂的肌原纤维部分中，在碱性pH下检测到两种蛋白水解活性：一种是苯甲基磺酰氟（PMSF）抑制的类似丝氨酸蛋白酶的活性，另一种是N-乙基马来酰亚胺（NEM）抑制的类似Ca2+激活蛋白酶的活性。在高血压2周时，pH 6蛋白水解活性以及钙激活蛋白酶I和NEM抑制的碱性蛋白酶活性显著增加，而其他可鉴定的蛋白水解活性保持不变。溶酶体组织蛋白酶D仅在高血压8周后活性升高。这些结果可能与该高血压性心肌病模型中发生的心肌细胞坏死和溶解的发展有关。
