鹰嘴豆芽异黄酮对小鼠成骨前体细胞MC3T3-E1增殖 及分化的研究.

Objective To investigate the effects and its mechanism of isoflavones extracted from chickpea sprouts (ICS) on the proliferation and the differentiation of the osteogenic precursor MC3T3-E1 cells. Methods CCK-8 was used to detect the proliferation of MC3T3-E1 cells at different concentrations of ICS (0, 0. 1, 0. 5, 1, 5, 10, 50, 100 mg/L) at 24, 48, 72 h. Different concentrations of ICS (0, 0.5, 1195 mg/L) were added into the osteogenic induction medium to observe its effects on the osteogenic differentiation: Alkaline phosphatase staining (ALP) assay was performed to observe the enzyme activities after 7-day induction; Alizarin red staining was performed on the 21st day to observe the calcium deposition; RT-qPCR was used to detect the mRNA expression levels of runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) and type 1 collagen (COL1) in each group on the 7th day of induction; Western Blot was used to detect the expression levels of RUNX2 and COL1 on the 7th day of induction, and estrogen receptor (ER) inhibitor ICI 182, 780 (1 μmol/L) was also added to detect the effect on the protein expression of the osteogenic differentiation-related proteins RUNX2 and COL1. Results ICS promoted cell proliferation in a certain concentration range (1-10 mg/L), and had some toxicity to cells when the concentration was greater than 10 mg/L (P< 0. 05). Compared with the control group (0 mg/L), ICS at concentrations of 0. 5,1,5 mg/L could promote alkaline phosphatase activity, with significant differences in a dose-dependent manner; ICS could promote calcium deposition, with a significant increase in the number of calcium nodules with increasing dose, with statistical differences; ICS could up-regulate the mRNA transcription levels of osteogenic differentiation-related genes RUNX2, OCN and COLT (P<0. 05), and promote the expression of osteogenic differentiation-related proteins COL1 and RUNX2 (P<0. 05). The estrogen receptor inhibitor ICI 182 780 inhibited the promoting effect of ICS on osteogenic differentiation-related proteins COL1 and RUNX2 (P < 0. 05). Conclusion ICS can promote the proliferation of MC3T3-E1 osteogenic precursor cells in mice, and improve the osteogenic differentiation of MC3T3-E1 cells by up-regulating the expression levels of osteogenic differentiation-related genes and proteins. The use of estrogen receptor inhibitor ICI 182,780 can inhibit the osteogenic differentiation of ICS. It is speculated that ICS promotes osteogenic differentiation through ER-related pathway. [ABSTRACT FROM AUTHOR]