Stimulation mechanism of osteoblast proliferation and differentiation by Duzhong Decoction-containing serum through L-VGCCs.
10.19540/j.cnki.cjcmm.20250221.401
- Author:
Ze-Bin CHEN
1
;
Lan-Lan LUO
2
;
Xin-Yi SHI
2
;
Rui-Tong ZHAO
2
;
Cai-Xian HU
2
;
Yun-Ying FU
2
;
Su-Zhen CHAO
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 Diseases,Jiangxi Administration of Traditional Chinese Medicine Nanchang 330004, China Jiangxi Province Key Laboratory of Pharmacology of Traditional Chinese Medicine Nanchang 330004, China.
2. School of Pharmacy, Jiangxi University of Chinese Medicine Nanchang 330004, China Key Laboratory of Traditional Chinese Medicine Prevention and Treatment of Senile Diseases,Jiangxi Administration of Traditional Chinese Medicine Nanchang 330004, China.
- Publication Type:Journal Article
- Keywords:
Duzhong Decoction;
L-VGCCs;
bone formation;
osteoblast;
osteoporosis
- MeSH:
Animals;
Osteoblasts/metabolism*;
Cell Proliferation/drug effects*;
Cell Differentiation/drug effects*;
Mice;
Drugs, Chinese Herbal/pharmacology*;
Calcium Channels, L-Type/genetics*;
Alkaline Phosphatase/genetics*;
Serum/chemistry*;
Cell Line;
Osteogenesis/drug effects*;
Bone Morphogenetic Protein 2/genetics*
- From:
China Journal of Chinese Materia Medica
2025;50(12):3335-3345
- CountryChina
- Language:Chinese
-
Abstract:
This paper aimed to explore the effects of Duzhong Decoction(DZD)-containing serum on the proliferation and osteoblast differentiation of MC3T3-E1 cells through L-type voltage-gated calcium channels(L-VGCCs). L-VGCCs inhibitors, nifedipine and verapamil, were used to block L-VGCCs in osteoblasts. MC3T3-E1 cells were divided into a control group, a low-dose DZD-containing serum(L-DZD) group, a medium-dose DZD-containing serum(M-DZD) group, a high-dose DZD-containing serum(H-DZD) group, a nifedipine group, a H-DZD + nifedipine group, verapamil group, and a H-DZD + verapamil group. The CCK-8 method was used for cell proliferation analysis, alkaline phosphatase(ALP) assay kits for intracellular ALP activity measurement, Western blot for protein expression level in cells, real-time fluorescence quantitative PCR technology for intracellular mRNA expression level determination, fluorescence spectrophotometer for free Ca~(2+) concentration determination in osteoblasts, and alizarin red staining(ARS) for mineralized nodule formation in osteoblasts. The experimental results show that compared to the control group, DZD groups can promote MC3T3-E1 cell proliferation, ALP activity, and mineralized nodule formation, increase intracellular Ca~(2+) concentrations, and upregulate the protein expression of bone morphogenetic protein 2(BMP2), collagen Ⅰ(COL1), α2 subunit protein of L-VGCCs(L-VGCCα2), and the mRNA expression of Runt-related transcription factor 2(RUNX2), and BMP2. After blocking L-VGCCs with nifedipine and verapamil, the intervention effects of DZD-containing serum were inhibited to varying degrees. Both nifedipine and verapamil could inhibit ALP activity, reduce mineralized nodule areas, and downregulate the expression of bone formation-related proteins. Moreover, the effects of DZD-containing serum on increasing MC3T3-E1 cell proliferation, osteoblast differentiation, and Ca~(2+) concentrations, upregulating the mRNA expression of osteoprotegerin(OPG) and protein expression of phosphorylated protein kinase B(p-Akt) and phosphorylated forkhead box protein O1(p-FOXO1), and upregulating phosphatase and tensin homolog(PTEN) expression were reversed by nifedipine. The results indicate that DZD-containing serum can increase the Ca~(2+) concentration in MC3T3-E1 cells to promote bone formation, which may be mediated by L-VGCCs and the PTEN/Akt/FoxO1 signaling pathway, providing a new perspective on the mechanism of DZD in treating osteoporosis.
