Yoshida Koichi, Kawai Motofumi, Utsunomiya Tsugiaki, Ishibazawa Akihiro, Song Young-Seok, Udo Mariana Sayuri B, Tasaki Yoshikazu, Yoshida Akitoshi
Department of Ophthalmology, Asahikawa Medical University, Asahikawa, Hokkaido 078-8510, Japan.
Department of Hospital Pharmacy & Pharmacology, Asahikawa Medical University, Asahikawa, Hokkaido 078-8510, Japan.
Int J Ophthalmol. 2022 Mar 18;15(3):388-393. doi: 10.18240/ijo.2022.03.03. eCollection 2022.
To investigate changes in extracellular matrix (ECM) gene expression in human trabecular meshwork (HTM) cells in response to mechanical fluid flow stimulation.
HTM cells were grown on a glass plate coated with 0.02% type I collagen (COL) and exposed to shear stress (0, 0.2, 1.0 dyne/cm) for 12h. Changes in genes related to the ECM were evaluated by real-time reverse transcriptase-polymerase chain reaction. Phosphorylation of Smad2 protein was investigated by Western blotting.
After mechanical stimulation, COL type 4 alpha 2, COL type 6 alpha 1, and fibronectin-1 mRNA were significantly higher than the static control (<0.05, <0.05, and <0.01, respectively). The metalloproteinase-2 and plasminogen activator inhibitor-1 mRNA were significantly higher than the static control (<0.05 and <0.01, respectively), while the differences in the tissue inhibitors of metalloproteinases-2 mRNA were not significant. The phosphorylation of Smad2 levels was significantly higher compared to the static control cells.
Changes in the expressions of genes associated ECM metabolism result in HTM cells after mechanical stimulation. The mechanical stimulation of the aqueous humor to the trabecular meshwork may promote ECM turnover and contribute to intraocular pressure homeostasis.
研究人小梁网(HTM)细胞在机械性流体流动刺激下细胞外基质(ECM)基因表达的变化。
将HTM细胞培养在涂有0.02% I型胶原(COL)的玻璃板上,并暴露于剪切应力(0、0.2、1.0达因/平方厘米)下12小时。通过实时逆转录聚合酶链反应评估与ECM相关基因的变化。通过蛋白质印迹法研究Smad2蛋白的磷酸化。
机械刺激后,IV型胶原α2、VI型胶原α1和纤连蛋白-1 mRNA显著高于静态对照组(分别为<0.05、<0.05和<0.01)。金属蛋白酶-2和纤溶酶原激活物抑制剂-1 mRNA显著高于静态对照组(分别为<0.05和<0.01),而金属蛋白酶组织抑制剂-2 mRNA的差异不显著。与静态对照细胞相比,Smad2水平的磷酸化显著更高。
机械刺激后HTM细胞中与ECM代谢相关的基因表达发生变化。房水对小梁网的机械刺激可能促进ECM周转并有助于眼压稳态。