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Effects of Hippo pathway on differentiation, proliferation, migration of bone marrow mesenchymal stem cells in vitro / 中华危重病急救医学
Chinese Critical Care Medicine ; (12): 750-755, 2019.
Artigo em Chinês | WPRIM | ID: wpr-754049
ABSTRACT
Objective To explore the effects of Hippo pathway on differentiation, proliferation, and migration of bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods BMSCs of C57BL/6 mice were identified using fluorescence-activated cellsorting analysis and the capabilities of osteogenic, chondrogenic and adipogenic differentiation were evaluated. The differentiation of BMSCs to typeⅡalveolar epithelial cells (AECⅡ) was induced by indirect co-culture with mouse lung epithelial cells (MLE-12) and small airway epithelial cell growth medium (SAGM). The Hippo pathway was regulated by 2-deoxy-D-glucose (2-DG) and 9E1, the effects of 2-DG and 9E1 on the expression of BMSCs surface proteins (SPB, SPC and SPD) mRNA and pro-SPC protein were detected by real time quantitative polymerase chain reaction (qRT-PCR) and Western Blot. The effect of Hippo pathway on differentiation of BMSCs to AECⅡ cells was evaluated. The effect of Hippo pathway on the proliferation of BMSCs was evaluated by methyl thiazolyl tetrazolium (MTT) assay (intervention of 0.1, 0.5, 1.0, 5.0 mmol/L 2-DG). The scratch test and Transwell chamber test were used to analyze the effect of Hippo pathway on migration ability of BMSCs to conditioned medium of acute respiratory distress syndrome (ARDS) lung tissue. Results 2-DG could activate Hippo pathway in a dose-dependent manner and promote the differentiation to AECⅡ and proliferation of BMSCs, the maximum effects were observed after 5 mmol/L of 2-DG treatment [SPB mRNA (2-&Delta;&Delta;CT) 2.42±0.28 vs. 1.89±0.11, SPC mRNA (2-&Delta;&Delta;CT) 8.06±0.68 vs. 6.59±0.79, SPD mRNA (2-&Delta;&Delta;CT) 6.45±0.37 vs. 5.27±0.28, pro-SPC/β-actin 5.80±1.86 vs. 4.93±1.18, proliferation rate(145.46±18.18)% vs. (98.91±4.36)%, all P < 0.05], but 9E1 could reverse those effects through inhibition of Hippo pathway [SPB mRNA (2-&Delta;&Delta;CT) 1.32±0.17 vs. 1.89±0.11, SPC mRNA (2-&Delta;&Delta;CT) 3.91±0.34 vs. 6.59±0.79, SPD mRNA (2-&Delta;&Delta;CT) 3.38±0.25 vs. 5.27±0.28, pro-SPC/β-actin 2.48±0.17 vs. 4.93±1.18, proliferation rate (80.00±7.27)% vs. (98.91±4.36)%, all P < 0.05]. The ability of horizontal migration [wound healing (27.17±3.53)% vs. (52.45±6.52)%, P < 0.05] and homing BMSCs to conditioned medium of ARDS lung tissue [cell count (fold, relative to control) 2.77±0.21 vs. 1.90±0.19, P < 0.05] were increased after activation of Hippo pathway by 2-DG treatment, but those effects were reversed after inhibition of Hippo pathway by 9E1 treatment [wound healing (79.89±8.42)% vs. (52.45±6.52)%, cell count (fold, relative to control) 1.69±0.13 vs. 1.90±0.19, both P < 0.05]. Conclusion Activation of Hippo pathway could enhance differentiation of BMSCs to AECⅡ, promote proliferation and ability of horizontal migration and homing BMSCs to conditioned medium of ARDS lung tissue in vitro.

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Idioma: Chinês Revista: Chinese Critical Care Medicine Ano de publicação: 2019 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Idioma: Chinês Revista: Chinese Critical Care Medicine Ano de publicação: 2019 Tipo de documento: Artigo