Oza N B, Schwartz J H, Goud H D, Levinsky N G
Evans Memorial Department of Clinical Research, Boston University Medical Center, Massachusetts 02118.
J Clin Invest. 1990 Feb;85(2):597-600. doi: 10.1172/JCI114479.
We have studied rat vascular smooth muscle (VSM) cells in culture for the presence of key elements of the glandular kallikrein-kinin system. Direct radioimmunoassay (RIA) using antiserum against rat urinary kallikrein detected a glandular kallikrein-like enzyme (GKLE) in VSM cells and in media. VSM homogenates and culture media had kininogenase activity, generating kinins from dog kininogen. About half of the GKLE was enzymatically inactive which could be activated with trypsin. Kininogenase activity was inhibited completely by aprotinin but only 20% by soybean trypsin inhibitor (SBTI). Trypsin liberated kinins from homogenates and media, demonstrating that VSM cells contain kininogen. Homogenates and media rapidly degrade bradykinin. GKLE, kininogen, and bradykininase activity were all present in VSM cells grown in defined media that contain no serum, thus eliminating any contamination or artefacts from fetal calf serum in standard culture media. Blood vessels of the rat have been reported to contain GKLE. Our observations indicate that GKLE is synthesized by VSM cells, not deposited from plasma. Furthermore, VSM cells synthesize kininogen and bradykininase(s), the other key elements of the glandular kallikrein-kinin system. Thus it is possible that the system functions as an autocoid mechanism that regulates local vascular tone.
我们研究了培养的大鼠血管平滑肌(VSM)细胞中是否存在腺体激肽释放酶 - 激肽系统的关键成分。使用抗大鼠尿激肽释放酶抗血清的直接放射免疫测定(RIA)在VSM细胞和培养基中检测到一种腺体激肽释放酶样酶(GKLE)。VSM匀浆和培养基具有激肽原酶活性,可从犬激肽原产生激肽。大约一半的GKLE无酶活性,可被胰蛋白酶激活。激肽原酶活性被抑肽酶完全抑制,但被大豆胰蛋白酶抑制剂(SBTI)仅抑制20%。胰蛋白酶从匀浆和培养基中释放出激肽,表明VSM细胞含有激肽原。匀浆和培养基能快速降解缓激肽。GKLE、激肽原和缓激肽酶活性都存在于不含血清的限定培养基中生长的VSM细胞中,从而消除了标准培养基中胎牛血清的任何污染或假象。据报道大鼠血管含有GKLE。我们的观察表明,GKLE是由VSM细胞合成的,而非从血浆中沉积而来。此外,VSM细胞合成激肽原和缓激肽酶,它们是腺体激肽释放酶 - 激肽系统的其他关键成分。因此,该系统有可能作为一种自身调节机制来调节局部血管张力。