ETB and epidermal growth factor receptor stimulation of wound closure in bovine corneal epithelial cells.
ETB and epidermal growth factor receptor stimulation of wound closure in bovine corneal epithelial cells.
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发表时间：
1995-12
影响因子：
4.4
通讯作者：
W. Tao;G. Liou;X. Wu;T. O. Abney;P. Reinach
中科院分区：
文献类型：
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作者：
W. Tao;G. Liou;X. Wu;T. O. Abney;P. Reinach
目的确定内皮素（ET）受体亚型是否存在异质性模式（即，ETA和ETB）基因在牛角膜上皮（BCE）中的表达。确定ET受体亚型刺激是否增加表皮生长因子（EGF）加速牛角膜上皮细胞（BCEC）原代培养物中伤口闭合的有效性。方法采用原位杂交组织化学方法检测BCE中ETA和ETB基因的表达。伤口闭合试验评价了培养4至7天后BCEC的伤口愈合率。[3H]胸苷掺入法和MTT法测定增殖。结果ETA基因在基底细胞的表达明显高于基底上细胞，而ETB基因在基底上细胞的表达则相反。表皮生长因子（5 ng/ml）最大限度地增加了伤口闭合145%以上的控制。与单独使用5 ng/ml EGF相比，10（-9）M ET-1或10（-8）M sarafotoxin-6-c（s-6-c）使伤口闭合增加了39%（P < 0.001）。BQ 123（10（-7）M）不改变ET-1或s-6-c的任何这些作用。表皮生长因子通过选择性增加增殖刺激伤口闭合。ET-1和s-6-c单独使用对增殖和迁移均无影响。结论ETA和ETB基因在BCE中均有表达。然而，仅在BCEC中，ETB刺激增加EGF刺激伤口闭合的有效性。这种反应是由细胞迁移而不是增殖的增加引起的，因为用丝裂霉素C治疗后，ET-1和EGF都不能刺激伤口闭合。
PURPOSE To determine if there is a heterogeneous pattern of endothelin (ET) receptor subtype (i.e., ETA and ETB) gene expression in the bovine corneal epithelium (BCE). To determine if ET receptor subtype stimulation increases the effectiveness of epidermal growth factor (EGF) to accelerate wound closure in a primary culture of bovine corneal epithelial cells (BCEC). METHODS In situ hybridization histochemistry was used to characterize ETA and ETB gene expression in the BCE. A wound closure assay evaluated wound healing rates in BCEC after 4 to 7 days in culture. [3H] thymidine incorporation and MTT assay measured proliferation. RESULTS ETA gene expression was appreciably higher in the basal cells than in the suprabasal cells, whereas the pattern for ETB was reversed. Epidermal growth factor (5 ng/ml) maximally increased wound closure by 145% above the control. With 5 ng/ml EGF, either 10(-9) M ET-1 or 10(-8) M sarafotoxin-6-c (s-6-c) increased wound closure by an additional 39% (P < 0.001) above that measured with 5 ng/ml EGF alone. BQ123 (10(-7) M) did not alter any of these effects of ET-1 or s-6-c. Epidermal growth factor stimulated wound closure through a selective increase in proliferation. Neither ET-1 nor s-6-c alone had any effect on proliferation or migration. CONCLUSIONS Both ETA and ETB genes are expressed in BCE. However, in BCEC only, ETB stimulation increases the effectiveness of EGF to stimulate wound closure. This response was caused by an increase in cell migration rather than proliferation because, after treatment with mitomycin C, neither ET-1 nor EGF stimulated wound closure.