ATP6V1H facilitates osteogenic differentiation in MC3T3-E1 cells via Akt/GSK3β signaling pathway

Fusong Jiang; Haojie Shan; Chenhao Pan; Zubin Zhou; Keze Cui; Yuanliang Chen; Haibo Zhong; Zhibin Lin; Nan Wang; Liang Yan; Xiaowei Yu

doi:10.1080/15476278.2019.1633869

ATP6V1H facilitates osteogenic differentiation in MC3T3-E1 cells via Akt/GSK3β signaling pathway

Organogenesis. 2019;15(2):43-54. doi: 10.1080/15476278.2019.1633869. Epub 2019 Jul 4.

Authors

Fusong Jiang¹, Haojie Shan², Chenhao Pan², Zubin Zhou², Keze Cui³, Yuanliang Chen³, Haibo Zhong³, Zhibin Lin³, Nan Wang⁴, Liang Yan⁵, Xiaowei Yu²

Affiliations

¹ a Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Clinical Center for Diabetes , Shanghai , China.
² b Department of Orthopaedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital , Shanghai , China.
³ c Department of Orthopaedic Surgery, Haikou Orthopedics and Diabetes Hospital of Shanghai Sixth People's Hospital , Haikou , China.
⁴ d Department of Emergency, The First Affiliated Hospital of Zhengzhou University , Zhengzhou , Henan , China.
⁵ e Department of Ophthalmology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital , Shanghai , China.

Abstract

Type 2 diabetes mellitus (T2DM) accounts for approximately 90% of all diabetic patients, and osteoporosis is one of the complications during T2DM process. ATP6V1H (V-type proton ATPase subunit H) displays crucial roles in inhibiting bone loss, but its role in osteogenic differentiation remains unknown. Therefore in this study, we aimed to explore the biological role of ATP6V1H in osteogenic differentiation. OM (osteogenic medium) and HG (high glucose and free fatty acids) were used to induce the MC3T3-E1 cells into osteogenic differentiation in a T2DM simulating environment. CCK8 assay was used to detect cell viability. Alizarin Red staining was used to detect the influence of ATP6V1H on osteogenic differentiation. ATP6V1H expression increased in OM-MC3T3-E1 cells, while decreased in OM+HG-MC3T3-E1 cells. ATP6V1H promoted osteogenic differentiation of OM+HG-MC3T3-E1 cells. Overexpression of ATP6V1H inhibited Akt/GSK3β signaling pathway, while knockdown of ATP6V1H promoted Akt/GSK3β signaling pathway. ATP6V1H overexpression promoted osteogenic differentiation of OM+HG-MC3T3-E1 cells. The role of ATP6V1H in osteogenic differentiation in a T2DM simulating environment involved in Akt/GSK3β signaling pathway. These data demonstrated that ATP6V1H could serve as a potential target for osteogenic differentiation in a T2DM simulating environment.

Keywords: ATP6V1H; Akt/GSK3β; MC3T3-E1; Osteogenic Differentiation; T2DM.

MeSH terms

3T3 Cells
Animals
Cell Differentiation
Cell Survival
Gene Expression Regulation*
Glycogen Synthase Kinase 3 beta / metabolism*
Mice
Osteoblasts / cytology
Osteogenesis*
Proto-Oncogene Proteins c-akt / metabolism*
Signal Transduction
Vacuolar Proton-Translocating ATPases / metabolism*

Substances

Akt1 protein, mouse
Glycogen Synthase Kinase 3 beta
Gsk3b protein, mouse
Proto-Oncogene Proteins c-akt
ATP6V1H protein, mouse
Vacuolar Proton-Translocating ATPases

Grants and funding

This work was supported by the National Natural Science Foundation of China (No. 81873993, No. 81572155); Municipal Human Resources Development Program for Outstanding Leaders in Medical Disciplines in Shanghai (No. 2018BR38); the Special Funds of the Key Clinical Research Project (No. DYZD201803); Shanghai Clinical Ability Construction of the Three Grade Hospital (No. SHDC12015904).