The synergistic action of estradiol and L-ascorbic acid on the osteoblast-like cell line MG-63 / 中华老年医学杂志

ABSTRACT

Objective To observe the synergistic action of 17? estradiol(E 2) and L ascorbic acid on the activity of alkaline phosphatase (ALP), cell proliferation and gene expression in osteosarcoma MG 63 cell line, and to study the mechanism of E 2 in treating osteoporosis. Methods MG 63 cells were treated with 10 8 mol/L E 2 with or without 100 ?g/L L ascorbic acid, and then the ALP activity was determined by using ? nitrophenyl phosphate as a substrate. Cell proliferation was measured by ( 3H) thymidine( 3H TdR) incorporation method. mRNA expression of type I collagen, matrix metalloproteinase 1(MMP 1), MMP 2 and osteoprotegerin(OPG) was detected by semiquantitative RT PCR method. Gelatin zymogram was used for the assay of MMP 2 activity. The synergistic action of E 2 and L ascorbic acid on MG 63 cell lines could be evaluated basing on the above indexes since they stand for separately the cell proliferation, gene expression and cell differentiation. Results The action of combined treatment with E 2 and L ascorbic acid on MG 63 cells was obviously stronger than the action using single E 2 or single L ascorbic acid. E 2 and L ascorbic acid increased the ALP activity to 200% ( P