견우자 추출물이 MC3T3-E1의 조골세포 분화에 미치는 영향 = Effects of Pharbitidis Semen Extract on Osteoblast Differentiation in MC3T3-E1 Cells

다국어 초록 (Multilingual Abstract)

Osteoblasts mineralize bone matrix in the process of bone formation and osteoblastic differentiation is an important step of bone formation. Some drugs have useful effects for bone formation in oriental medicine. In this study, to confirm the anti-inflammation and osteoblast differentiation effect of Pharbitidis Semen extract used in oriental medicine, we investigated cell viability, Alizarin red S assay, and Alkaline phosphatase(ALP) activity with Pharbitidis Semen extract. Also, we studied the inhibition effect of nitric oxide(NO) generation with Pharbitidis Semen extract in RAW 264.7 cell. Our results showed that Pharbitidis Semen extract had almost no cytotoxicity to the cell and it significantly enhanced the induction of ALP activity and Alizarin red S staining for osteogenesis in MC3T3-E1 cell. Also, all Pharbitidis Semen extract inhibited the LPS-induced NO generation in RAW 264.7 cell. Especially, Pharbitidis Semen ethyl acetate extract and Pharbitidis Semen butanol extract showed to be powerful inhibitors of NO generation. In conclusion, Pharbitidis Semen extract increased the ALP activity, Alizarin red S assay activity for bone formation and attenuated NO generation for inflammation. These results suggest that Pharbitidis Semen extract may act as a successful drug for osteogenic differentiation in vitro and could have the potential to be use as a therapeutic agent for bone formation.

참고문헌 (Reference)

1 박중원, "발생중인 생쥐 치아 및 치주조직에서 지추인대-특이 단백질 ; PDLs22의 발현" 대한치주과학회 32 (32): 1-12, 2002

2 K Osawa, "The inhibitory effect of plant extracts on the collagenolytic activity and cytotoxicity of human gingival fibroblasts by Porphyromonas gingivalis crude enzyme" 32 : 1-1, 1991

3 SH Cho, "The effects of 17-$\beta$ estradiol, medroxyprogesteroneacetate and parathyroid hormone on the differentiation ofosteoblast cell" 39 : 1497-1497, 1996

4 EM Choi, "Soybean ethanol extract increases the function of osteo-blastic MC3T3-E1 cells" 56 : 733-733, 2001

5 SJ Kim, "Signal Transduction in the Osteoblast Cells" 7 : 329-329, 1999

6 GS Stein, "Relationship of cellgrowth to the regulation for tissue-specific gene expressionduring osteoblast differentiation" 4 : 3111-3111, 1990

7 MP Sherman, "Pyrrolidine Dithiocarbamate Inhibits Induction of Nitric Oxide Synthase Activity in Rat Alveolar Macrophages" 191 : 1301-1301, 1993

8 Y Goo, "Periodontology" Koonja press 2006

9 N Wada, "Periodontal ligament cells secrete the factor that inhibits osteoclastic differentiation and function:the Factor is osteoprotegrin/osteoclastdogenesis inhibitory factor" 36 : 56-56, 2001

10 MJ Somerman, "Periodontal ligament cells and gingival fibroblasts respond differently to attachment factors in vitro" 60 : 73-73, 1989

11 F Strutz, "Identifiaction and characterization of a fibroblast marker:FSP1" 130 : 393-393, 1995

12 F Strutz, "Identifiaction and characterization of a fibroblast marker:FSP1" 130 : 874-874, 2001

13 SH Ralston, "Human osteoblast-like cells produce nitric oxide and express inducible nitric oxide synthase" 135 : 330-330, 1994

14 SH Raltson, "Human osteoblast-like cells produce nitric oxide and express inducible nitric oxide synthase" 135 : 330-336, 1994

15 M Kassem, "Growth hormon stimulates proliferation of normal human bone marrow stromal osteoblast precursor cells in vitro" 4 : 131-131, 1994

16 PC Schiller, "Gap-junctional communication is required for the maturationprocess of osteoblastic cells in culture" 28 : 362-362, 2001

17 RP Phipps, "Fibroblast heterogeneity in the periodontium and other tissue" 32 : 159-159, 1997

18 MH Helfrich, "Expression of nitric oxide synthase isoforms in bone and bone cell cultures" 12 : 1108-1108, 1997

19 JA Riancho, "Expression and functional role of nitric oxide synthase in osteoblast-like cells" 10 : 439-446, 1995

20 M Ernst, "Estradiol effects on proliferation messenger ribonucleic acid collagen and insulin-like growth factor-I and parathyroid hormone-stimulated adenylate cyclase activity in osteoblastic cells from calvariae and long bones" 125 : 822-822, 1989

21 KS Moon, "Element and Utilization of Herbage" Science and Encyclopedia Publishers 1984

22 EM Choi, "Effects of soybean ethanol extracton the prostaglandin E2 and interleukin-6 production inosteoblastic cells" 35 : 893-893, 2002

23 CH Yang, "Effect of zizyphi fructus extract on the biological activity of gingival fibroblast" 24 : 144-144, 1994

24 YS Lee, "Effect of isoflavones on proliferation and oxidativesstressof MC3T3-E1 osteoblastic like cells" 18 : 35-35, 2001

25 J Lindhe, "Clinical Periodontology and Implant Dentistry" Iowa State Press 2003

26 HJ Verhaar, "A comparison of the action of progestins and estrogen on the growth and differentiation of normal adult human osteoblast-like cells in vitro" 15 : 307-307, 1994

27 BI Seo, "A Philological Study on poisoning of Pharbitidis Semen" 33 : 1-1, 2008