Protective and Osteogenic Effects of Crude Water Extract from Cuscuta japonica Choisy at Gene Expression Level in Human Gingival Cells

Abstract

This study focused on observing change of gene expression after longterm treatment with high dose of crude water extract from seeds of Cuscuta japonica Choisy (Japanese dodder). The aim of this study was to study the protective and osteogenic effects of dodder seed extract at gene expression level in human gingival cells. In this research, dodder seeds were blended and boiled in water before freeze-drying to preserve as dried powder. Total phenolic content and antioxidant assay were performed. MTT assay was performed with human gingival cells. The concentrations of 100, 250 and 500 µg / mL had no effect on cell viability when cultivated for 48 hours. The extract at the concentration of 250 µg / mL was chosen to treat human gingival cells for many time points to observe the expressions of osteogenic, inflammatory, antioxidant and cancer gene markers by real-time PCR. The results showed that dodder seed water extract could increase the expressions of osteogenic markers; OPN, OCN and Col-I genes. Moreover, dodder seed water extract induced mineralization of human gingival cells cultured in 3D structure. Change of BCL2, CAS3 and LC3 expression indicated involvement in apoptotic process of dodder seed water extract. Significant changes in gene expression of antioxidant markers (GST1, SOD1 and TXNRD1), inflammatory marker (Cox-2) and epithelial cell marker (α-SMA) without change of important cancer genes were observed. These evidences suggested that seeds of Cuscuta japonica Choisy could benefit the application in regenerative medic ine and alternative medic ine.