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目的 探讨地塞米松对培养的人眼小梁细胞生长的影响及抑制小梁细胞表达表皮生长因子 (epidermalgrowthfactorEGF)mRNA的情况。方法 取人眼小梁组织进行小梁细胞体外培养 ,对传3代的小梁细胞进行地塞米松处理实验。实验组在传代后的培养液中按 30 0 μg/ml加入地塞米松 ,另一组作为对照组进行常规培养 ,观察生长 5d后的细胞情况 ,取培养 7d的两组小梁细胞分别提取RNA ,用EGFcDNA探针 ,α 32 P同位素标记进行斑点杂交 ,放射自显影。对显影片用计算机激光密度扫描 ,测定吸光度A值相对值进行组间比较。结果 加入地塞米松 30 0 μg/ml实验组 ,小梁细胞生长明显受到抑制 ,5d时对照组细胞已经融合 ,地塞米松组的细胞仍呈集落状态。从对照组小梁细胞提取RNA 2 2 5 μg ,地塞米松组提取RNA 14μg ,取 14μg两组等量RNA ,用EGFcDNA探针进行斑点杂交 ,结果阳性。激光密度扫描值地塞米松组明显低于对照组。结论 地塞米松对培养的人眼小梁细胞有明显的生长抑制作用 ,通过抑制总RNA转录及EGFmRNA表达而抑制小梁细胞生长。提示糖皮质激素性青光眼是因抑制了小梁细胞的多种代谢和生理功能所致。
Objective To investigate the effects of dexamethasone on cultured human trabecular meshwork cells and the inhibition of trabecular cells expression of epidermal growth factor (EGF) mRNA. Methods Trabecular cells were cultured in vitro and trabecular cells were treated with dexamethasone for 3 passages. In the experimental group, dexamethasone was added at 30 0 μg / ml in the culture medium after the passage, and the other group was used as a control group to conduct routine culture. After 5 days of growth, the cells in the culture were observed. Two trabecular cells cultured for 7 days were used to extract RNA , Dot blot hybridized with EGF cDNA probe and α 32 P isotope, and autoradiographed. On the film using computer laser scanning density, absorbance A value relative value was compared between groups. Results The dexamethasone 30 0 μg / ml experimental group, trabecular cell growth was significantly inhibited, 5 d the control group cells have been fused, the dexamethasone group cells still showed a colony. RNA was extracted from the trabecular cells of control group, and the RNA was extracted from dexamethasone group. The amount of RNA was 14μg, and 14μg of the same amount of RNA was obtained from the two groups. The result of dot blot hybridization with EGF cDNA probe showed positive results. Laser densitometry scan dexamethasone group was significantly lower than the control group. Conclusion Dexamethasone can significantly inhibit the growth of human trabecular meshwork cells and inhibit the growth of trabecular cells by inhibiting the transcription of total RNA and the expression of EGFmRNA. Suggest that glucocorticoid-induced glaucoma is due to inhibition of trabecular cells in a variety of metabolic and physiological functions.