Effect of Erxian Decoction-containing serum on H_2O_2-induced proliferation and osteogenic differentiation of MC3T3-E1 cells via BK channels.
10.19540/j.cnki.cjcmm.20230103.703
- Author:
Ming-Shi REN
1
;
Yu DING
1
;
Zi-Han LI
1
;
Yu-Meng WU
1
;
Si-Min HUANG
1
;
Lan-Lan LUO
1
;
Yu-Jing ZHANG
1
;
Min SHI
2
;
Xun-Li XIA
2
;
Bo LIU
1
Author Information
1. School of Pharmacy,Jiangxi University of Chinese Medicine Nanchang 330004,China Key Laboratory of Traditional Chinese Medicine Prevention and Treatment of Senile Disease, Jiangxi Provincial Administration of Traditional Chinese Medicine Nanchang 330004,China.
2. College of Life Sciences,Jiangxi University of Chinese Medicine Nanchang 330004,China.
- Publication Type:Journal Article
- Keywords:
BK channels;
Erxian Decoction(EXD);
bone formation;
osteoblast;
osteogenic differentiation;
oxidative stress
- MeSH:
Osteogenesis;
Core Binding Factor Alpha 1 Subunit/pharmacology*;
Large-Conductance Calcium-Activated Potassium Channels/pharmacology*;
Proto-Oncogene Proteins c-akt/metabolism*;
Calcium/metabolism*;
Cell Differentiation;
RNA, Messenger/metabolism*;
Cell Proliferation;
Osteoblasts
- From:
China Journal of Chinese Materia Medica
2023;48(9):2522-2529
- CountryChina
- Language:Chinese
-
Abstract:
This study aimed to investigate the effects of Erxian Decoction(EXD)-containing serum on the proliferation and osteogenic differentiation of MC3T3-E1 cells under oxidative stress through BK channels. The oxidative stress model was induced in MC3T3-E1 cells by H_2O_2, and 3 mmol·L~(-1) tetraethylammonium(TEA) chloride was used to block the BK channels in MC3T3-E1 cells. MC3T3-E1 cells were divided into a control group, a model group, an EXD group, a TEA group, and a TEA+EXD group. After MC3T3-E1 cells were treated with corresponding drugs for 2 days, 700 μmol·L~(-1) H_2O_2 was added for treatment for another 2 hours. CCK-8 assay was used to detect cell proliferation activity. The alkaline phosphatase(ALP) assay kit was used to detect the ALP activity of cells. Western blot and real-time fluorescence-based quantitative PCR(RT-qPCR) were used to detect protein and mRNA expression, respectively. Alizarin red staining was used to detect the mineralization area of osteoblasts. The results showed that compared with the control group, the model group showed significantly blunted cell proliferation activity and ALP activity, reduced expression of BK channel α subunit(BKα), collagen Ⅰ(COL1), bone morphogenetic protein 2(BMP2), osteoprotegerin(OPG), and phosphorylated Akt, decreased mRNA expression levels of Runt-related transcription factor 2(RUNX2), BMP2, and OPG, and declining area of calcium nodules. EXD-containing serum could significantly potentiate the cell proliferation activity and ALP activity, up-regulate the protein expression of BKα, COL1, BMP2, OPG, and phosphorylated Akt, and forkhead box protein O1(FoxO1), promote the mRNA expression of RUNX2, BMP2, and OPG, and enlarge the area of calcium nodules. However, BK channel blockage by TEA reversed the effects of EXD-containing serum in promoting the protein expression of BKα, COL1, BMP2, OPG, and phosphorylated Akt and FoxO1, increasing the mRNA expression of RUNX2, BMP2, and OPG, and enlarging the area of calcium nodules. EXD-containing serum could improve the proliferation activity, osteogenic differentiation, and mineralization ability of MC3T3-E1 cells under oxidative stress, which might be related to the regulation of BK channels and downstream Akt/FoxO1 signaling pathway.
