3,5,3'-三碘-L-甲状腺原氨酸通过αvβ3整合素调节缺氧条件下分化型PC-12细胞中肌动蛋白细胞骨架的动态变化。

3,5,3'-Triiodo-L-Thyronine Regulates Actin Cytoskeleton Dynamic in The Differentiated PC-12 Cells during Hypoxia through An αvβ3 Integrin.

作者信息

Barbakadze Tamar, Kvergelidze Elisabed, Bátor Judit, Szeberényi József, Mikeladze David

机构信息

Faculty of Natural Sciences and Medicine, Ilia State University, Tbilisi, Georgia.

Laboratory of Biochemistry, Ivane Beritashvili Center of Experimental Biomedicine, Tbilisi, Georgia.

出版信息

Cell J. 2023 Apr 1;25(4):247-254. doi: 10.22074/cellj.2022.557501.1059.

DOI:10.22074/cellj.2022.557501.1059

PMID:37210645

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10201356/

Abstract

OBJECTIVE

Thyroid hormones are involved in the pathogenesis of various neurological disorders. Ischemia/hypoxia that induces rigidity of the actin filaments, which initiates neurodegeneration and reduces synaptic plasticity. We hypothesized that thyroid hormones via alpha-v-beta-3 (αvβ3) integrin could regulate the actin filament rearrangement during hypoxia and increase neuronal cell viability.

MATERIALS AND METHODS

In this experimental study, we analysed the dynamics of actin cytoskeleton according to the G/F actin ratio, cofilin-1/p-cofilin-1 ratio, and p-Fyn/Fyn ratio in differentiated PC-12 cells with/without T3 hormone (3,5,3'-triiodo-L-thyronine) treatment and blocking αvβ3-integrin-antibody under hypoxic conditions using electrophoresis and western blotting methods. We assessed NADPH oxidase activity under the hypoxic condition by the luminometric method and Rac1 activity using the ELISA-based (G-LISA) activation assay kit.

RESULTS

The T3 hormone induces the αvβ3 integrin-dependent dephosphorylation of the Fyn kinase (P=0.0010), modulates the G/F actin ratio (P=0.0010) and activates the Rac1/NADPH oxidase/cofilin-1 (P=0.0069, P=0.0010, P=0.0045) pathway. T3 increases PC-12 cell viability (P=0.0050) during hypoxia via αvβ3 integrin-dependent downstream regulation systems.

CONCLUSION

The T3 thyroid hormone may modulate the G/F actin ratio via the Rac1 GTPase/NADPH oxidase/ cofilin1signaling pathway and αvβ3-integrin-dependent suppression of Fyn kinase phosphorylation.

摘要

目的

甲状腺激素参与多种神经疾病的发病机制。缺血/缺氧会导致肌动蛋白丝僵硬，引发神经退行性变并降低突触可塑性。我们推测甲状腺激素通过α - v - β - 3（αvβ3）整合素可在缺氧时调节肌动蛋白丝重排并提高神经元细胞活力。

材料与方法

在本实验研究中，我们使用电泳和蛋白质印迹法，分析了在缺氧条件下，经/未经T3激素（3,5,3'-三碘 - L - 甲状腺原氨酸）处理以及用αvβ3整合素抗体阻断的分化PC - 12细胞中，根据G/F肌动蛋白比率、丝切蛋白 - 1/p - 丝切蛋白 - 1比率和p - Fyn/Fyn比率所反映的肌动蛋白细胞骨架动力学。我们通过发光法评估缺氧条件下的NADPH氧化酶活性，并使用基于酶联免疫吸附测定（G - LISA）的激活检测试剂盒评估Rac1活性。

结果

T3激素诱导Fyn激酶的αvβ3整合素依赖性去磷酸化（P = 0.0010），调节G/F肌动蛋白比率（P = 0.0010）并激活Rac1/NADPH氧化酶/丝切蛋白 - 1（P = 0.0069，P = 0.0010，P = 0.0045）信号通路。T3通过αvβ3整合素依赖性下游调节系统在缺氧期间提高PC - 12细胞活力（P = 0.0050）。