하엽(荷葉)이 파골세포의 분화 및 뼈 흡수 관련 유전자의 조절과 조골세포 의 분화에 미치는 영향

다국어 초록 (Multilingual Abstract)

Background and Purpose: Osteoporosis is a disease characterized by low bone mass and micro-architectural deterioration of bone tissue, leading to increased fracture risk. This study aims to investigate the potential therapeutic effects of leaves of Nelumbo nucifera Gaertner ethanol extract (NG_E) on osteoporosis, focusing on its impact on osteoclast differentiation and osteoblast activity.

Materials and methods: NG_E and water extracts of leaves of Nelumbo nucifera Gaertner (NG_W) were prepared and tested on RAW 264.7 cells induced by RANKL to evaluate osteoclast differentiation. The formation of F-actin rings and the expression of NFATc1 and c-Fos proteins were assessed at RAW 264.7 cells in which NG_E was induced with RANKL. Additionally, the effects on TRAP, ATP6v0d2, MMP-9, CTK, and CA2 mRNA expression were measured. For osteoblast activity, MC3T3-E1 cells were used to evaluate the promotion of osteoblast differentiation through Alizarin red S staining.

Results: NG_E significantly inhibited the formation of F-acting rings and reduced NFATc1 and c-Fos protein expression in RANKL-induced RAW 264.7 cells. Furthermore, it significantly inhibited the mRNA expression of TRAP, ATP6v0d2, MMP-9, CTK, and CA2. However, NG_W did not show significant effects on osteoclast differentiation or activity. In terms of osteoblast activity, NG_E significantly promoted the differentiation of MC3T3-E1 cells into osteoblasts, as shown by increased Alizarin red S staining.

Conclusions: The results suggest that NG_E has the potential to serve as a therapeutic agent for osteoporosis by inhibiting osteoclast differentiation and promoting osteoblast activity. Further studies, including in vivo and clinical trials, are warranted to explore its clinical applications.

목차 (Table of Contents)

• I. 서론 1
• II. 재료 및 방법 4
• 2.1. 시약 4
• 2.2. 荷葉 에탄올(NG_E) 및 열수 추출물(NG_W)의 제조 4
• 2.3. RAW 264.7 세포 및 MC3T3-E1 세포 배양 5
• 2.4. 세포 독성 검증 5
• 2.5. 파골세포 분화 및 TRAP 염색 5
• 2.6. F-actin ring 형성 분석 6
• 2.7. Western blot 분석 6
• 2.8. Reverse transcription (RT)-polymerase chain reaction (PCR) 분석 7
• 2.9. 조골세포 분화 및 Alizarin red S 염색 8
• 2.10. 통계 분석 9
• III. 결과 10
• 3.1. NG_E와 NG_W이 RAW 264.7 세포 생존률에 미치는 영향 10
• 3.2. NG_E가 RANKL로 유도된 RAW 264.7 세포에서 파골세포 분화에 미치는 영향 12
• 3.3. NG_W가 RANKL로 유도된 RAW 264.7 세포에서 파골세포 분화 및 활성 억제에 미치는 영향 14
• 3.4. NG_E가 RANKL로 유도된 RAW 264.7 세포에서 F-actin ring 형성에 미치는 영향 16
• 3.5. NG_E가 RANKL로 유도된 RAW 264.7 세포에서 NFATc1 및 c-Fos 단백질 발현에 미치는 영향 18
• 3.6. NG_E가 RANKL로 유도된 RAW 264.7 세포에서 NFATc1, TRAP 및 ATP6v0d2 mRNA 발현에 미치는 영향 20
• 3.7. NG_E가 RANKL로 유도된 RAW 264.7 세포에서 MMP-9, CTK 및 CA2 mRNA 발현에 미치는 영향 22
• 3.8. NG_E가 MC3T3-E1 세포에서 조골세포 분화에 미치는 영향 24
• Ⅳ. 고 찰 26
• Ⅴ. 결 론 30
• 참 고 문 헌 31
• ABSTRACT 38