自分泌酶可能对三维（3D）培养的人眼小梁细胞（HTM）具有与溶血磷脂酸无关的作用。

Autotaxin May Have Lysophosphatidic Acid-Unrelated Effects on Three-Dimension (3D) Cultured Human Trabecular Meshwork (HTM) Cells.

机构信息

Departments of Ophthalmology, School of Medicine, Sapporo Medical University, Sapporo 060-8556, Japan.

Departments of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University, Sapporo 060-8556, Japan.

出版信息

Int J Mol Sci. 2021 Nov 7;22(21):12039. doi: 10.3390/ijms222112039.

PURPOSE

The objective of the current study was to evaluate the effects of the autotaxin (ATX)-lysophosphatidic acid (LPA) signaling axis on the human trabecular meshwork (HTM) in two-dimensional (2D) and three-dimensional (3D) cultures of HTM cells.

METHODS

The effects were characterized by transendothelial electrical resistance (TEER) and FITC-dextran permeability (2D), measurements of size and stiffness (3D), and the expression of several genes, including extracellular matrix (ECM) molecules, their modulators, and endoplasmic reticulum (ER) stress-related factors.

RESULTS

A one-day exposure to 200 nM LPA induced significant down-sizing effects of the 3D HTM spheroids, and these effects were enhanced slightly on longer exposure. The TEER and FITC-dextran permeability data indicate that LPA induced an increase in the barrier function of the 2D HTM monolayers. A one-day exposure to a 2 mg/L solution of ATX also resulted in a significant decrease in the sizes of the 3D HTM spheroids, and an increase in stiffness was also observed. The gene expression of several ECMs, their regulators and ER-stress related factors by the 3D HTM spheroids were altered by both ATX and LPA, but in different manners.

CONCLUSIONS

The findings presented herein suggest that ATX may have additional roles in the human TM, in addition to the ATX-LPA signaling axis.

目的

本研究旨在评估自分泌酶（ATX）-溶血磷脂酸（LPA）信号轴对二维（2D）和三维（3D）培养的人眼小梁细胞（HTM）的影响。

方法

通过跨内皮电阻（TEER）和 FITC-葡聚糖通透性（2D）、大小和硬度测量（3D）以及包括细胞外基质（ECM）分子、其调节剂和内质网（ER）应激相关因子在内的几个基因的表达来表征这些影响。

结果

2D 培养的 HTM 单层中，LPA 诱导的屏障功能增加，1 天暴露于 200 nM LPA 可导致 3D HTM 球体显著缩小，而较长时间暴露则略微增强了这种效果。TEER 和 FITC-葡聚糖通透性数据表明，LPA 诱导 2D HTM 单层的屏障功能增强。1 天暴露于 2mg/L 的 ATX 溶液也导致 3D HTM 球体的大小显著减小，并且观察到硬度增加。3D HTM 球体的几种 ECM、其调节剂和 ER 应激相关因子的基因表达均被 ATX 和 LPA 改变，但方式不同。