Maki K, Hayashi S, Nishioka T, Kimura M, Noguch T
Department of Pediatric Dentistry, Kyushu Dental College, 2-6-1 Manazuru, Kokura-kita, Kitakyushu 803-8580, Japan.
Connect Tissue Res. 2000;41(2):109-15. doi: 10.3109/03008200009067663.
Using 4-month-old fetal bovine tissue, the properties of the tibia epiphyseal cartilage matrix vesicles, a type of endochondral ossification tissue, were compared with those from tracheal cartilage. The matrix vesicle fractions, obtained by collagenase digestion and differential centrifugation, were subjected to sucrose-density-gradient centrifugation. Alkaline phosphatase activity, protease activity, and lacatate dehydrogenase activity were assayed for the marker enzyme of the matrix vesicles. Matrix vesicles containing alkaline phosphatase, metalloprotease, and lacatate dehydrogenase were found in the tibia epiphyseal cartilage at a density of 1.11 g/ml. In surprising contrast, we also found matrix vesicle-like vesicles with a high density of 1.24 g/ml in the tracheal cartilage. These also contained alkaline phosphatase and lactate dehydrogenase, but not metalloprotease. The electrophoretic profiles of the lactate dehydrogenase isoenzymes from the matrix vesicle and matrix vesicle-like vesicles were identical with those of chondrocyte cytosolic lactate dehydrogenase. Aldolase, aspartate: 2-oxoglutarate aminotransferase, alanine: 2-oxoglutarate aminotransferase, glucose-6-phosphatase, glutamate dehydrogenase, catalase, and cytosolic enzymes except for lactate dehydrogenase were not detected in these vesicles. These results suggest the presence of a mechanism for specific uptake of cytosolic lactate dehydrogenase in both vesicles. In this study, a new type of matrix vesicles without protease was found in the tracheal cartilage, a kind of permanent cartilage, but not in the tibia epiphyseal cartilage, which is replaced by bone tissue.
利用4月龄胎牛组织,将长骨骨骺软骨基质小泡(一种软骨内成骨组织)的特性与气管软骨的特性进行了比较。通过胶原酶消化和差速离心获得基质小泡组分,再进行蔗糖密度梯度离心。检测碱性磷酸酶活性、蛋白酶活性和乳酸脱氢酶活性,作为基质小泡的标记酶。在长骨骨骺软骨中发现含有碱性磷酸酶、金属蛋白酶和乳酸脱氢酶的基质小泡,其密度为1.11 g/ml。令人惊讶的是,我们在气管软骨中还发现了密度为1.24 g/ml的基质小泡样小泡。这些小泡也含有碱性磷酸酶和乳酸脱氢酶,但不含金属蛋白酶。基质小泡和基质小泡样小泡中乳酸脱氢酶同工酶的电泳图谱与软骨细胞胞质乳酸脱氢酶的电泳图谱相同。在这些小泡中未检测到醛缩酶、天冬氨酸:2-酮戊二酸转氨酶、丙氨酸:2-酮戊二酸转氨酶、葡萄糖-6-磷酸酶、谷氨酸脱氢酶、过氧化氢酶以及除乳酸脱氢酶外的胞质酶。这些结果表明,两种小泡中都存在特异性摄取胞质乳酸脱氢酶的机制。在本研究中,在气管软骨(一种永久性软骨)中发现了一种新型的不含蛋白酶的基质小泡,但在被骨组织替代的长骨骨骺软骨中未发现。