Effects of astragalus polysaccharides on CYP17A1,CYP19A1 mRNA and protein expres-sion in MC-3T3-E1 osteoblasts induced bv dexamethasone
Objective To investigate the effects of Astragalus polysaccharide on the proliferation of MC-3T3-E1 osteoblasts and the expression of CYP17A1 and CYP19A1 genes and proteins,and to explore the mechanism of Astragalus polysaccharide in the treatment of glucocorticoid osteoporosis(GIOP).Methods MC-3T3-El osteoblasts were studied and divided into 6 groups:normal group,model group,vitamin D3 group(1 ×10-5 mmol·L-1),and Astragalus polysaccharide high-dose,medi-um-dose and low-dose groups(1,0.1,0.01mg·L-1).The effect of Astragalus polysaccharide on the proliferation inhibition rate of MC-3T3-E1 osteoblasts induced by dexamethasone was determined by MTT method.The effect of Astragalus polysac-charide on calcium content in dexamethasone-induced supernatant of MC-3T3-E1 osteoblasts was determined by colorimetry.The effect of Astragalus polysaccharide on the content of 25(OH)D3 in the supernatant of MC-3T3-E1 osteoblasts induced by dexamethasone was determined by ELISA.The effects of Astragalus polysaccharide on the relative expression levels of CYP17A1 and CYP19A1 mRNA and protein in dexamethasone-induced MC-3T3-E1 osteoblasts were detected by RT-PCR and West-ern blot.Results Compared with the normal group,the proliferation inhibition rate of MC-3T3-E1 osteoblasts in the model group was significantly increased at 24,36,48 h(P＜0.01).Compared with model group,the proliferation inhibition rate of MC-3T3-E1 osteoblasts in Astragalus polysaccharide high,medium and low dose groups and vitamin D3 groups was significant-ly decreased at 24,36,48 h(P＜0.01).Colorimetric test results showed that calcium content of MC-3T3-E1 osteoblasts in model group was significantly decreased compared with normal group(P＜0.01).Compared with model group,calcium content of MC-3T3-E1 osteoblasts in APS high-dose group was significantly increased(P＜0.01).ELISA results showed that com-pared with normal group,the content of MC-3T3-E1 osteoblast 25(OH)D3 in model group was significantly decreased(P＜0.01).Compared with model group,the content of MC-3T3-E1 osteoblast 25(OH)D3 in APS high-dose and medium-dose groups was significantly increased(P＜0.01).RT-PCR and Western-Blot detection results showed that compared with normal group,the relative expression levels of CYP17A1 and CYP19A1 mRNA and protein in MC-3T3-E1 osteoblasts in model group were significantly decreased(P＜0.01).Compared with model group,mRNA and protein relative expressions of CYP17A1 and CYP19A1 in MC-3T3-El osteoblasts in APS high-dose group were significantly increased(P＜0.01)and were positively correlated with drug concentration.Conclusion Astragalus polysaccharide can improve glucocorticoid-related indexes of osteopo-rosis,and its possible mechanism is related to increasing the activity of MC-3T3-E1 osteoblasts and the mRNA and protein ex-pression levels of CYP17A1 and CYP19A1.
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