홍화, 홍화씨 추출물이 MC3T3E1 세포의 골분화 과정에 미치는 영향 = Effect of Safflower and Safflower Seed Extract on Osteogenic Differentiation of MC3T3E1 Cells

다국어 초록 (Multilingual Abstract)

Objectives This study investigated the effect of purified safflower (Carthamus tinctorius Linne) and safflower seed (Carthamus tinctorius L. seed; CS) extract, using hot water and ethanol extract methods , on the osteogenic differentiation of MC3T3E1 cells.Methods The safflower and safflower seed were extracted with hot water and ethanol. The samples were concentrated by a rotary evaporator and then freeze-dried using a freeze-dryer. The MC3T3E1 cells were propagated and maintained in DMEM (Gibco) containing 10% FBS and a 1% antibiotic antimycotic solution. To induce osteogenic differentiation, the cells were treated for 14 days with DMEM with 10 mM β-glycerophosphate and 50 μM ascorbic acid. Extract doses were confirmed by the results of an MTT assay, and treatment of the extracts was performed in a differentiation medium every two days. The ALP staining and activity were tested after osteogenic differentiation for five days, and after 14 days, osteogenic differentiation was determined by alizarin red S staining. The mRNA expressions of osteogenic-related genes were quantified using quantitative real-time PCR.Results In the results of the MTT assay, all concentrations of safflower extracts had no toxicity in the MC3T3El cells. But in the groups of 100 ng/ml and 200 ng/ml concentrations of safflower seed extracts, the cell viability was significantly reduced by up to 40-50%. So we fixed the treatment concentration of the extract at 50 ng/ml. In the ALP and alizarin red S staining, all extract groups increased osteogenic differentiation compared with the control group. The water-safflower extract group showed the highest mRNA level of Alp, Runx2, and Dlx5 genes. The mRNA level of Ocn, an osteogenic gene related to late-stage differentiation, in the ethanol-safflower extract group increased the mineralization more significantly than in other groups.Conclusions These data suggest that the extract of safflower increases the osteoblastic differentiation activates of MC3T3E1 cells like the extract of safflower seed. The water-extract and ethanol-extract of safflower have effects on different stages of osteogenesis in MC3T3El. Not only safflower seed but also safflower will be useful therapeutic reagents for age-associated chronic diseases such as osteoporosis.

참고문헌 (Reference)

1 최철한, "홍화씨(잇꽃, Carthamus tinctorius L.) 연구 동향에 대한 고찰" 대한한의학원전학회 24 (24): 63-90, 2011

2 김덕규, "홍화씨 추출물의 국소투여가 백서 두개골결손부 재생에 미치는 영향" 29 (29): 297-310, 1999

3 김기영, "홍화씨 Yoghurt 급여가 난소절제 Rat의 골다공증에 미치는 영향" 한국동물자원과학회 46 (46): 69-76, 2004

4 이지원, "미역취뿌리 추출물이 성장기 흰쥐의 골대사에 미치는 영향" 한국생명과학회 15 (15): 236-241, 2005

5 김광진, "난소 절제 흰쥐의 골대사에 미치는 골쇄보의 영향" 22 (22): 175-181, 2001

6 문준성, "골다공증의 진단과 치료" 25 (25): 19-30, 2008

7 김정연, "골다공증에 대한 동서의학적고찰" 6 : 293-315, 1996

8 Oh HJ, "Therapy of osteoporosis in climacteric" 21 : 20-27, 2005

9 Boonen S, "The prevention of treatment of age-related osteoporosis in the elderly by systemic recombinant growth factor therapy(rhIGF-I or rhRGF-β): a perspective" 242 (242): 285-290, 1997

10 Hawker GA, "The epidemiology of osteoporosis" 45 : 2-5, 1996

11 Cho SH, "The effects of 17-β etradiol, medroxyprogesterone acetate and parathyroid hormone on the differentiation of osteoblast cell" 39 (39): 1497-1506, 1996

12 Ryoo HM, "Stage-specific expression of Dlx-5 during osteoblast differentiation involvement in regulation of osteocalcin gene expression" 11 (11): 1681-1694, 1997

13 Choi EM, "Soybean ethanol extract increases the funtion of osteoblastic MC3T3-E1 cells" 56 : 733-739, 2001

14 Hansen MA, "Role of peak bone mass and bone loss in postmenopausal osteoporosis: 12year study" 303 : 961-964, 1991

15 Aldercreutz H, "Phyto-estrogens in relation to cancer and other human health risks" 55 : 399-417, 1996

16 Jung HS, "Peroxisome proliferator-activated receptor gamma/singal transducers and activators of transcription 5a pathway plays a key factor in adipogenesis of human bone marrow-derived stromal cells and 3T3-L1 preadipocytes" 21 : 465-475, 2012

17 Tanaka Y, "Osteoblasts and osteoclasts in bone remodeling and inflammation" 4 (4): 325-328, 2005

18 Pole HAP, "Multinational placebo-controlled, randomised trial of the effects of alendronate on bone density and fracture risk in postmenopausal women with law bone mass: Results of the FOSIT" 9 : 461-468, 1999

19 Kutsuna H, "Identification and determination of platelet aggregation inhibitor from safflower (Carthamus tinctorius Linne)" 108 : 1101-1103, 1988

20 Soo-miKim, "Effects of Safflower Seeds on the Serum Levels of Insulin-like Growth Factors, Insulin-like Growth Factor Binding Protein-3 and BALP in Osteoporosis Induced-ovariectomized Rats" 한국임상수의학회 20 (20): 263-273, 2003

21 Lee YS, "Effect of isoflavones on proliferation and oxidatives stress of MC3T3-E1 osteoblastic like cells" 18 : 35-42, 2001

22 Hwang JS, "Differential Expression of STATs and Its Function in Osteogenesis of Mesenchymal Stem Cells" 16 (16): 1-9, 2013

23 Lee RH, "Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue" 14 : 311-324, 2004

24 Kim HJ, "Bone-protecting effect of safflower seeds in ovariectomized rats" 71 (71): 88-94, 2002

25 Kim YJ, "Bone morphogenetic protein-2-induced alkaline phosphatase expression is stimulated by Dlx5 and repressed by Msx2" 279 : 50773-50780, 2004

26 Ott SM, "Attainment of peak bone mass" 71 (71): 1082A-1082C, 1990