관동화(款冬花)가 RANKL 유도 뼈파괴세포의 분화 및 뼈흡수 관련 유전자 발현에 미치는 영향

다국어 초록 (Multilingual Abstract)

Background and Purpose: Osteoporosis has long been a socially prominent disease. It is characterized by a decrease in bone mass and a weakening of the bone tissue's microstructure, making bones more fragile and increasing the risk of fractures. This systemic disease has a rising prevalence and socioeconomic cost every year, necessitating caution. In this study, I investigated the effects of Tussilago farfara Linné (TF) on the differentiation of osteoclasts and the expression of bone resorption-related genes to find potential improvement effects on osteoporosis.

Materials and methods: Ethanol extracts of TF (TF-E) at various concentrations (100, 200, 400 µg/ml) were used, and tartrate-resistant acid phosphatase (TRAP) activation was assessed in these groups to evaluate the inhibitory effect on osteoclast differentiation. The formation of the F-actin ring, which is essential for bone resorption, was evaluated, as well as the reduction effect on nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), a key factor in osteoclast differentiation, and its gene suppression effect. The gene suppression effect of ATPase H+ transporting V0 subunit d2 (ATP6v0d2), a gene related to bone resorption, was evaluated, along with the expression of matrix metallopeptidase 9 (MMP-9) and carbonic anhydrase 2 (CA2), which are related to bone resorption. The experiments were evaluated using TRAP assay, Western blot, and RT-PCR.

Results: TF-E treatment reduced the number of osteoclasts and TRAP activity. The formation of the F-actin ring was also inhibited, and NFATc1 decreased significantly in a concentration-dependent manner. The gene expression of NFATc1, TRAP, and ATP6v0d2 was reduced, as well as the gene expression of MMP-9 and CA2. Unfortunately, Cathepsin K (CTK) expression did not decrease.

Conclusions: The results indicate that TF holds potential for treating osteoporosis through its ability to inhibit osteoclast differentiation and the expression of genes related to bone resorption. To fully examine its clinical potential, further comprehensive research is essential.

Key words: Osteoporosis, Osteoclasts, Tussilago farfara L, RANKL, TRAP, NFATc1, MMP-9, CA2, ATP6v0d2, Bone density.

목차 (Table of Contents)

• I. 서론
• II. 재료 및 방법
• 2.1. 시약
• 2.2. 세포 배양
• 2.3. 관동화 에탄올 추출물(TF-E) 및 열수 추출물(TF-W)의 제조
• 2.4. 세포 생존율 분석
• 2.5. TRAP 염색 및 활성도 분석
• 2.6. F-actin ring 형성 분석
• 2.7. Western blot 분석
• 2.8. Reverse transcription polymerase chain reaction (RT-PCR) 분석
• 2.9. 통계 분석
• III. 결과
• 3.1. 용매별 관동화(TF) 추출물의 세포 독성
• 3.2. TF-E의 파골세포 분화 억제
• 3.3. TF-W의 파골세포 분화 억제
• 3.4. TF-E의 F-actin ring 형성 억제
• 3.5. TF-E의 파골세포 분화 필수 전사 인자 억제
• 3.6. TF-E의 파골세포 분화 관련 mRNA 발현 억제
• 3.7. TF-E의 골흡수 관련 mRNA 발현 억제
• Ⅳ. 고 찰
• Ⅴ. 결 론
• 참 고 문 헌
• ABSTRACT