静压力对人眼筛板细胞Ⅲ型胶原mRNA表达水平的研究

The effect of elevated hydrostatic pressure on expression level of type Ⅲ collagen mRNA by lamina cribrosa cells in vitro

目的观察压力对人眼筛板细胞Ⅲ型胶原ｍＲＮＡ表达水平的影响，探讨筛板细胞外间质的变化在青光眼视神经损害中的作用。方法对筛板细胞进行体外培养并施予一定的静压力，采用ｄｏｔｂｌｏｔ杂交方法和计算机图像分析法测定受压后的筛板细胞在不同时期合成Ⅲ型胶原ｍＲＮＡ表达水平。结果静压力４６７～５３３ｋＰａ（１ｋＰａ＝７．５ｍｍＨｇ）下，加压３天的筛板细胞Ⅲ型胶原ｍＲＮＡ表达量吸光度（Ａ）值０５８２８±００５１７，与同期对照组Ａ值０２８３２±００６１３比较，差异有显著性（Ｐ＜０．０５）；加压６天的筛板细胞Ⅲ型胶原ｍＲＮＡ表达水平Ａ值０４１６２±００５１２，与同期对照组Ａ值０３３６７±００５８９比较，仍有不同程度增高，但差异无显著性（Ｐ＞０．０５）；加压９及１２天的筛板细胞Ⅲ型胶原ｍＲＮＡ表达量分别为Ａ值０３２４６±００９５７及０２１６４±００１７１，均低于同时期对照组Ａ值０４０２９±００３２６、０２６０１±００１３４，差异无显著性（Ｐ＜０．０５）。结论在一定时程的压力下，筛板细胞Ⅲ型胶原ｍＲＮＡ的表达有增强作用。

Objective To investigate the effect of elevated hydrostatic pressure on the expression level of type Ⅲ collagen mRNA by lamina cribrosa cells (LCCs) and approach the action of changes of extracellular stroma on glaucomatous optic nerve damage. Methods LCCs were cultured and submitted to elevated hydrostatic pressure (4.67～5.33 kPa). We observed collagen type Ⅲ mRNA level by dotblot hybridization and computer imaging analysis. Results Under the pressure for 3 days, the amounts of collagen Ⅲ mRNA were 0.582 8±0.051 7 in optical density (OD), compared with the same period controls (0.283 2±0.061 3), the difference was statistically significant (P<0.05). When exposed to pressure up to 6 days, the collagen Ⅲ mRNA level remained somewhat higher than that of the control, but there was no significant difference (P>0.05). After 9 to 12 days of exposure to pressure, the mRNA amounts were decreased respectively to 0.324 6±0.095 7 and 0.216 4±0.017 1 in OD that were lower than that of the control in the respective same peroids, and the differences between the experimental and the control had no significance (P>0.05). Conclusion Elevated hydrostatic pressure may increase expression level of collagen Ⅲ mRNA by LCCs in a certain period.