Effects of Sanhua Jiegu San on Wnt/beta-catenin signaling pathway in osteoblasts

doi:10.12307/2023.488

Abstract

Abstract: BACKGROUND: Sanhua Jiegu San is a common drug for clinical fracture and tendon injury, which have good effect. However, there is a lack of relevant experimental evidence, and its mechanism of action remains unclear.
OBJECTIVE: To observe the effect of Sanhua Jiegu San on Wnt/β-catenin signaling pathway in osteoblasts.
METHODS: Firstly, the optimal concentration of Sanhua Jiegu San for osteoblast intervention was screened by pre-experiment. Then, the proliferation of osteoblasts was tested by cell counting kit-8 method. The maturity of osteoblasts was detected by alkaline phosphatase assay. The mRNA expressions of alkaline phosphatase, Runt-related transcription factor 2, osteocalcin, osteopontin, collagenase I, Axin2, Dkk4, Lef-1, and Tcf-1 were detected by PCR. The protein expressions of β-Catenin and P-β-Catenin were detected by western blot method.
RESULTS AND CONCLUSION: After 5 days of osteoblast culture, the number of osteoblasts reached the maximum at the concentration of 10 mg/L. After 21 days of osteoblast culture, the proliferation of cells in different concentration groups of Sanhua Jiegu San increased, which was obvious in the 10 mg/L group, but tended to decrease at 20 mg/L. After 7 days of osteoblast culture, alkaline phosphatase expression was increased in the low, medium and high concentration groups, especially in the high concentration group (20 mg/L). After 7 days of osteoblast culture, mRNA expressions of alkaline phosphatase, Runt-related transcription factor 2, osteocalcin, osteopontin, collagenase I were increased in the low, medium and high concentration groups, especially in the high concentration group (20 mg/L). After 21 days of osteoblast culture, the mRNA expressions of Axin2 and Dkk4 were decreased in the low, medium and high concentration groups, while the mRNA expressions of Lef-1 and Tcf-1 were increased, especially in the high concentration group (20 mg/L). After 21 days of osteoblast culture, the expression of P-β-Catenin protein was decreased in the low, medium and high concentration groups, while the expression of β-Catenin protein was increased, especially in the high concentration group (20 mg/L). To conclude, Sanhua Jiegu San can regulate the Wnt/β-Catenin signaling pathway in osteoblasts, promote the mRNA expression of alkaline phosphatase, Runt-related transcription factor 2, osteocalcin, osteopontin, collagenase I, Lef-1, and Tcf-1 and the protein expression of β-Catenin, and decrease the mRNA expression of Axin2 and Dkk4 and the protein expression of P-β-Catenin, thereby promoting osteoblast proliferation. This may be the mechanism by which Sanhua Jiegu San promotes fracture healing.

Key words: Sanhua Jiegu San, osteoblast, cell proliferation, fracture healing, Wnt/β-catenin signaling pathway, molecular mechanism

CLC Number:

Duan Jiahao, Tan Xuyi, Lu Min, Kuang Gaoyan, Fang Chuanlong, Xiao Man. Effects of Sanhua Jiegu San on Wnt/beta-catenin signaling pathway in osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(20): 3230-3235.

Figures/Tables 8

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