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Trimethylpyrazine이 조골세포 분화에 미치는 영향
손효은 ․ 장원구 대구대학교 산업기술연구소 2018 産業技術硏究 Vol.29 No.2
Trimethylpyrazine (TrMP) is known to be derived from cocoa and nuts. In present study, we examined the effects of TrMP on osteoblast differentiation. The mRNA expression of osteogenic genes in MC3T3-E1 cells were determined by RT-PCR. TrMP induced the expression of osteogenic genes such as, inhibitor of differentiation-1 (Id1), distalless related homeobox (Dlx5), runt related transcription factor 2(Runx2), alkaline phosphatase( ALP), osteocalcin (OC). Alizarin red s staining was performed to measure calcium deposition. TrMP increased calcium deposition in MC3T3-E1 cells. These results suggest that TrMP was induces osteoblast differentiation in MC3T3-E1.
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손효은(Hyo-Eun Son),김은아(Eunna Kim),변미순(Misoon Byun),장원구(Won-Gu Jang) 한국식품영양과학회 2019 한국식품영양과학회지 Vol.48 No.3
본 연구에서는 ellagic acid가 조골세포 분화에 미치는 영향을 조사하였다. 먼저 MTT assay를 통하여 세포독성 실험을 하였고 독성이 확인되지 않은 3 μM의 농도에서 실험을 진행하였다. 그리고 조골세포로 분화할 수 있는 중간엽 줄기세포인 C3H10T1/2 세포에 ellagic acid를 처리하여 BMP2, Runx2, ALP의 발현을 확인하였다. 또한 ellagic acid는 BMP2, Dlx5, Runx2의 단백질 발현을 증가시킴을 확인하였다. 그리고 ALP 활성 및 Alizarin Red S 염색을 확인한 결과 ellagic acid 처리에 의해서 ALP 활성이 현저히 증가하고 칼슘 침착 또한 증가하는 것으로 확인하였다. 뿐만 아니라 LPS에 의한 조골세포 분화의 감소가 ellagic acid에 의해 회복됨을 확인하였다. 이러한 결과들을 토대로 천연 폴리페놀 화합물인 ellagic acid는 조골세포 분화를 촉진한다는 것을 알 수 있고, ellagic acid는 골 질환 예방 가능성이 있을 것으로 생각된다. Ellagic acid is a compound found widely in strawberries, raspberries, and green tea and has been reported to have anti-oxidant, anti-inflammatory and anti-cancer activities. In previous reports, anti-oxidants induced osteoblast differentiation via the expression of osteogenic genes. This study examined the effect of ellagic acid on osteoblast differentiation using C3H10T1/2 cells. First, a MTT assay was performed to determine the concentration at which ellagic acid does not exhibit cytotoxicity. The cells were treated at a concentration of 3 μM without cytotoxicity. The effects of ellagic acid on the mRNA expression levels of the osteogenic genes in C3H10T1/2 cells were determined by real-time PCR. Ellagic acid increased the expression of bone morphogenetic proteins 2 (BMP2), runt-related transcription factor 2 (Runx2), and alkaline phosphatase (ALP). In addition, ellagic acid induced the protein expression levels of BMP2, distal-less homeobox 5 (Dlx5), and Runx2 according to a western blots assay. Next, a staining assay was performed to evaluate the ALP activity or mineralization. Ellagic acid enhanced both the ALP activity and mineralization. In addition, ellagic acid recovered lipopolysaccharide-induced osteoblast differentiation. These results suggest that ellagic acid may enhance osteoblast differentiation.
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