Matsuda A, Tagawa Y, Matsuda H, Nishihira J
Department of Ophthalmology, Hokkaido University School of Medicine, Sapporo, Japan.
Invest Ophthalmol Vis Sci. 1997 Jul;38(8):1555-62.
The study was conducted to evaluate the expression of macrophage migration inhibitory factor (MIF) during penetrating corneal injury.
A penetrating linear incision (2 mm) was made in the center of the right cornea with a razor blade. The expression of MIF in the lacerated eye and in the contralateral eye was examined by immunohistochemistry at 3, 6, 24, 48, and 72 hours after injury. Concentrations of MIF in the aqueous humor of the injured and contralateral eyes were also measured by enzyme-linked immunosorbent assay. The expression of MIF messenger RNA (mRNA) in the injured cornea was quantified by reverse transcription-polymerase chain reaction and subsequent Southern blot analysis.
Positive migration inhibitory factor staining was observed in the basal cells of epithelial and endothelial cells of the normal rat cornea. The positive staining of the central corneal epithelium diminished at 3 hours after injury. At 6 hours after injury, positive MIF staining reappeared on the basal cells of the injured area, whereas the staining of the contralateral eye remained unchanged. Enzyme-linked immunosorbent assay of the aqueous humor revealed that the MIF concentration was elevated in both the injured and the contralateral eyes. The maximum concentration of aqueous MIF was observed at 6 hours after injury in both eyes. Reverse transcription-polymerase chain reaction and Southern blot analysis revealed that MIF-mRNA expression in the injured cornea increased from 6 to 48 hours after injury.
The results of immunohistochemistry suggest the possibility that MIF is released from the corneal epithelial cells of the injured eye within 3 hours. Conversely, the MIF-mRNA level of the injured cornea is increased from 6 to 48 hours after injury and then diminished. In addition, unilateral corneal injury induces bilateral upregulation of MIF in the aqueous humor.
本研究旨在评估穿透性角膜损伤过程中巨噬细胞移动抑制因子（MIF）的表达情况。
用剃须刀片在右眼角膜中央制作一个2毫米的穿透性线性切口。在损伤后3、6、24、48和72小时，通过免疫组织化学检查受伤眼和对侧眼中MIF的表达。还通过酶联免疫吸附测定法测量受伤眼和对侧眼房水中MIF的浓度。通过逆转录-聚合酶链反应和随后的Southern印迹分析对受伤角膜中MIF信使核糖核酸（mRNA）的表达进行定量。
在正常大鼠角膜的上皮和内皮细胞的基底细胞中观察到阳性移动抑制因子染色。损伤后3小时，角膜中央上皮的阳性染色减弱。损伤后6小时，受伤区域的基底细胞上再次出现MIF阳性染色，而对侧眼的染色保持不变。房水的酶联免疫吸附测定显示，受伤眼和对侧眼的MIF浓度均升高。双眼在损伤后6小时观察到房水MIF的最大浓度。逆转录-聚合酶链反应和Southern印迹分析显示，受伤角膜中MIF-mRNA的表达在损伤后6至48小时增加。
免疫组织化学结果提示，MIF可能在3小时内从受伤眼的角膜上皮细胞中释放出来。相反，受伤角膜的MIF-mRNA水平在损伤后6至48小时升高，然后下降。此外，单侧角膜损伤可导致房水中MIF的双侧上调。
