Sakimoto Tohru, Shoji Jun, Kanno Hitoshi, Sawa Mitsuru
Department of Ophthalmology, Nihon University School of Medicine, Tokyo, Japan.
Jpn J Ophthalmol. 2004 Jan-Feb;48(1):17-22. doi: 10.1007/s10384-003-0022-2.
To determine the expression of gelatinase A [matrix metalloproteinase (MMP)-2] and gelatinase B (MMP-9) in ocular surface disorders, we carried out examinations of the eyes of patients with such disorders.
The cases consisted of a normal group comprising three eyes as the control and a patient group comprising six eyes of patients with ocular surface disorders. These groups underwent the following examinations. (1) Gelatin zymography: Tear samples taken by using the Schirmer I test were eluted and analyzed by gelatin zymography. (2) Immunohistochemical study: By using the alkaline phosphate-anti-alkaline phosphatase or avidin-biotin complex method, corneal or conjunctival tissue was examined for the presence of MMP-2, MMP-9, and membrane type 1 (MT1)-MMP, which activates MMP-2. (3) Reverse transcription-polymerase chain reaction (RT-PCR): Total RNA was eluted from the specimens, and the expression patterns of MMP-2, MMP-9, and MT1-MMP mRNA were analyzed.
(1) Gelatin zymography: Only the proforms of MMP-2 and MMP-9 were detected in the normal group. Both active MMP-2 and active MMP-9 were detected in all studied cases in the patient group. (2) Immunohistochemical study: In the normal group, only MMP-2-positive cells were present in conjunctival epithelium. In the patient group, cells positive for MMP-2, MMP-9 and MT1-MMP were present in the epithelium of the conjunctiva or cornea except in one thermal burn case. (3) RT-PCR: In the normal group, only MMP-2 expression was observed. MMP-2, MMP-9, and MT1-MMP mRNA expression was observed in three cases in the patient group.
MMP-2, MMP-9, and MT1-MMP, which activates MMP-2, were expressed in the patients with various ocular surface disorders. In ocular surface disorders, therefore, the expression of gelatinases is upgraded.
为了确定明胶酶A[基质金属蛋白酶（MMP）-2]和明胶酶B（MMP-9）在眼表疾病中的表达情况，我们对患有此类疾病的患者眼睛进行了检查。
病例包括作为对照的由三只眼睛组成的正常组和由患有眼表疾病患者的六只眼睛组成的患者组。这些组进行了以下检查。（1）明胶酶谱法：使用泪液分泌试验采集的泪液样本经洗脱后用明胶酶谱法进行分析。（2）免疫组织化学研究：采用碱性磷酸酶-抗碱性磷酸酶或抗生物素蛋白-生物素复合物方法，检查角膜或结膜组织中MMP-2、MMP-9以及激活MMP-2的膜型1（MT1）-MMP的存在情况。（3）逆转录-聚合酶链反应（RT-PCR）：从标本中洗脱总RNA，并分析MMP-2、MMP-9和MT1-MMP mRNA的表达模式。
（1）明胶酶谱法：正常组仅检测到MMP-2和MMP-9的前体形式。患者组所有研究病例中均检测到活性MMP-2和活性MMP-9。（2）免疫组织化学研究：正常组结膜上皮中仅存在MMP-2阳性细胞。患者组中，除1例热烧伤病例外，结膜或角膜上皮中存在MMP-2、MMP-9和MT1-MMP阳性细胞。（3）RT-PCR：正常组仅观察到MMP-2表达。患者组中有3例观察到MMP-2、MMP-9和MT1-MMP mRNA表达。
激活MMP-2的MMP-2、MMP-9和MT1-MMP在各种眼表疾病患者中均有表达。因此，在眼表疾病中，明胶酶的表达上调。
