Carbenoxolone inhibits mechanical stress-induced osteogenic differentiation of mesenchymal stem cells by regulating p38 MAPK phosphorylation

Shenglong Li; Jing Wang; Yudi Han; Xiaoteng Li; Changjian Liu; Zhengshuai Lv; Xiuhui Wang; Xin Tang; Zhe Wang

doi:10.3892/etm.2018.5757

Carbenoxolone inhibits mechanical stress-induced osteogenic differentiation of mesenchymal stem cells by regulating p38 MAPK phosphorylation

Exp Ther Med. 2018 Mar;15(3):2798-2803. doi: 10.3892/etm.2018.5757. Epub 2018 Jan 17.

Authors

Shenglong Li¹, Jing Wang², Yudi Han³, Xiaoteng Li⁴, Changjian Liu⁴, Zhengshuai Lv⁴, Xiuhui Wang⁵, Xin Tang⁴, Zhe Wang⁴

Affiliations

¹ Department of Bone and Soft Tissue Tumor Surgery, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, Liaoning 110042, P.R. China.
² Department of Plastic and Cosmetic Surgery, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
³ Department of Plastic and Reconstructive Surgery, General Hospital of Chinese PLA, Beijing 100853, P.R. China.
⁴ Department of Orthopedic Trauma, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
⁵ Department of Orthopedics, Shanghai Zhoupu Hospital, Shanghai 201318, P.R. China.

Abstract

The aim of the present study was to explore the effects of pannexin1 (Px1) protein channels on osteogenic differentiation of mesenchymal stem cells (MSCs) under mechanical stress stimulation. MSCs were isolated from Sprague Dawley rats (3 weeks old, weighing 100-120 g) and cultured in vitro. A safe concentration of carbenoxolone was determined (CBX, an inhibitor of Px1 channels; 100 µM) on MSCs using the Cell Counting Kit-8 (CCK8) method. MSCs were divided into 6 groups: Control, stress (4,000 µ strain), and stress following 3, 6, 12, and 24 h pretreatment with CBX. Stress groups were stimulated with mechanical stress for 15 min. Alkaline phosphatase (ALP) activity, type I collagen expression, intracellular calcium ion (Ca²⁺) concentration, Px1 expression, p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated phosphorylation were determined. ALP activity was increased in the stress group, and this was prevented by pretreatment with CBX. Similarly, stress-induced increases in type I collagen expression, Ca²⁺ concentration, Px1 expression, and p38 MAPK phosphorylation decreased in the presence of CBX. ERK phosphorylation was decreased by stress, however was not affected by CBX treatment. Altogether, the results suggest that mechanical stress promoted the osteogenic differentiation of MSCs, and this promotion was inhibited by pretreatment with CBX, possibly through regulating the phosphorylation of p38 MAPK.

Keywords: gap conjunction; mechanical stress; mesenchymal stem cells; osteogenic differentiation; p38 mitogen-activated protein kinase; phosphorylation.