[논문]MC3T3-E1 골아세포에서 발효 다시마 추출물에 의한 조골세포 분화의 촉진

정나라; 최영현

doi:10.15433/ksmb.2023.15.1.024

MC3T3-E1 골아세포에서 발효 다시마 추출물에 의한 조골세포 분화의 촉진
Fermented sea tangle (Laminaria japonica Aresch) Accelerates Osteoblast Differentiation in murine osteoblastic MC3T3-E1 Cells 원문보기

한국해양바이오학회지 = Journal of marine bioscience and biotechnology, v.15 no.1, 2023년, pp.24 - 32

Abstract ▼ AI-Helper

The Laminaria japonica Aresch (Sea tangle) belongs to the brown algae and has a long history as a food material in Asia, including Korea. Recent studies have found that the fermented Sea tangle extract (FST) inhibited the differentiation of osteoclasts and protected osteoblasts from oxidative damage. This study aims to explore the possibility that FST can induce the differentiation of osteoblasts and identify the responsible mechanism. According to our results, FST induced differentiation into osteogenic cells in the presence of osteoblastic MC3T3-E1 cells under non-toxic conditions.. This finding was confirmed by phalloidin staining, increased alkaline phosphatase activity, and calcium deposition. Additionally, it was found that this process was achieved by increasing the expression of key factors involved in osteoblast differentiation, such as runt-related transcription factor-2, osterix, β-catenin, and bone morphogenetic protein-2. Moreover, FST increased autophagy, which may contribute to the maintenance of the bone formation homeostasis, and is associated with the activation of the phosphatidylinositol 3-kinase/Akt and mitogen-activated protein kinase signaling pathways. Although further research about the bioactive substances contained in FST and the tests of their efficacy are required, the results of this study indicate that FST has incredible applicability as a functional material for maintaining the bone homeostasis.

주제어

표/그림 (5)

그림 Figure 1. Effects of FST on the cell viability and cytoskeleton morphology in MC3T3-E1 preosteoblasts. (A) Cells were cultured for 72 h in FST-containing α-MEM. (A) Cell viability was assessed by MTT assay. Data were presente
그림 Figure 2. Effects of FST on the ALP activity and calcification in MC3T3-E1 preosteoblasts. After culturing the cells in FST-containing medium α-MEM or OGM alone for 7 day s, the cells were stained for ALP activity using a TRACP & ALP Double-stain Kit (A and B) and for calcification using Alizarin Red S solution (C and D) according to the manufacturer's instructions. (A and C) Representative images were captured using a fluorescence microscope. (B) For quantification of ALP activity, cell lysates were separated from cells, incubated with alkaline phosphatase yellow (pNPP) liquid substrate, measured at 405 nm with a microplate reader, and presented as fold activity of contr ol cells. (D) Calcium deposition was quantified based on Alizarin Red S staining and the results were calculated as fold induction of control cells. Data were presented as mean ± SD (^***P < 0.001 relative to the α-MEM contro l).
그림 Figure 3. Effects of FST on the expression of osteogenic diffe rentiation-related proteins in MC3T3-E1 preosteoblasts. After culturing the cells in FST-containing α-MEM for 7 days, the levels of Runx2, β-catenin and BMP-2 protein expression were detected by Western blot analysis (A) and their relative expression was normalized to glyceralde hyde 3-phosphate dehydrogenase (GAPDH) as an internal control (B). Data were presented as mean ± SD (^***P < 0.001 relative to the α-MEM control).
그림 Figure 4. Effects of FST on the expression of autophagy-relat ed proteins in MC3T3-E1 preosteoblasts. (A) After culturi ng the cells in FST-containing α-MEM for 7 days, immun ofluorescence staining was performed using the LC3 antib ody and representative images obtained under a fluoresce nce microscope are presented. (B and C) The levels of LC3, p62 and Becline 1 protein expression were detected by Western blot analysis (B) and their relative expression was normalized to GAPDH as an internal control (C). Data were presented as mean ± SD (^***P < 0.001 relative to the α-MEM control).
그림 Figure 5. Effects of FST on the expression of PI3K/Akt and MAPKs-related proteins in MC3T3-E1 preosteoblasts. Af ter culturing the cells in FST-containing α-MEM for 7 days, Western blot analysis was used to detect levels of total and phosphorylated proteins using antibodies against the indicated proteins (A and C) and the relative expressio n of the phosphorylated form to total protein of the presen ted proteins was quantified (B and C). Data were presente d as mean ± SD (^***P < 0.001 relative to the α-MEM control).

참고문헌 (48)

Suzuki, A., Minamide, M., Iwaya, C., Ogata, K., Iwata,？J. 2020. Role of metabolism in bone development and？homeostasis. Int. J. Mol. Sci. 21, 8992.
Florencio-Silva, R., Sasso, G. R., Sasso-Cerri, E., Simoes,？M. J., Cerri, P. S. 2015. Biology of bone tissue: Structure, function, and factors that influence bone cells. Biomed. Res. Int. 2015, 421746.

상세보기
Song, S., Guo, Y., Yang, Y., Fu, D. 2022. Advances in？pathogenesis and therapeutic strategies for osteoporosis.？Pharmacol. Ther. 237, 108168.

상세보기
Martiniakova, M., Babikova, M., Omelka, R. 2020. Pharm？acological agents and natural compounds: available treat？ments for osteoporosis. J. Physiol. Pharmacol. 71, 307-320.
Arora, S., Cooper, P. R., Ratnayake, J. T., Friedlander,？L. T., Rizwan, S. B., Seo, B., Hussaini, H. M. 2022.？A critical review of in vitro research methodologies used？to study mineralization in human dental pulp cell cultures. Int. Endod. J. 55 (Suppl 1), 3-13.
Patel, U., Macri-Pellizzeri, L., Zakir Hossain, K. M., Scam？mell, B. E., Grant, D. M., Scotchford, C. A., Hannon,？A. C., Kennedy, A. R., Barney, E. R., Ahmed, I., Sottile,？V. 2019. In vitro cellular testing of strontium/calcium？substituted phosphate glass discs and microspheres shows potential for bone regeneration. J. Tissue Eng. Regen.？Med. 13, 396-405.

상세보기
Kobayashi, Y., Uehara, S., Koide, M., Takahashi, N. 2015.？The regulation of osteoclast differentiation by Wnt signals. Bonekey Rep. 4, 713.

상세보기
Kobayashi, Y., Uehara, S., Udagawa, N., Takahashi, N.？2016. Regulation of bone metabolism by Wnt signals.？J. Biochem. 159, 387-392.

상세보기
Yin, X., Zhou, C. C., Li, J. T., Liu, R. K., Shi, B., Yuan,？Q., Zou, S. J. 2019. Autophagy in bone homeostasis and？the onset of osteoporosis. Bone Res. 7, 28.

상세보기
Zhang, K., Liu, F. W., Jin, D. 2019. Autophagy preserves？the osteogenic ability of periodontal ligament stem cells？under high glucose conditions in rats. Arch. Oral. Biol. 101, 172-179.

상세보기
Franceschi, R. T., Ge, C. 2017. Control of the osteoblast？lineage by mitogen-activated protein kinase signaling. Curr. Mol. Biol. Rep. 3, 122-132.
Greenblatt MB, Shim JH, Glimcher LH. Mitogen-activat？ed protein kinase pathways in osteoblasts. Annu Rev Cell Dev Biol. 2013; 29: 63-79.

상세보기
El-Desoky, A. H. H., Tsukamoto, S. 2022. Marine natural products that inhibit osteoclastogenesis and promote？osteoblast differentiation. J. Nat. Med. 76, 575-583.

상세보기
Zhao, X., Patil, S., Xu, F., Lin, X., Qian, A. 2021. Role？of biomolecules in osteoclasts and their therapeutic potential for osteoporosis. Biomolecules 11, 747.

상세보기
Raut, N., Wicks, S. M., Lawal, T. O., Mahady, G. B.？2019. Epigenetic regulation of bone remodeling by natural compounds. Pharmacol. Res. 147, 104350.

상세보기
Jeong, J. W., Ji, S. Y., Lee, H., Hong, S. H., Kim, G.？Y., Park, C., Lee, B. J., Park, E. K., Hyun, J. W., Jeon,？Y. J., Choi, Y. H. 2019. Fermented sea tangle (Laminari a japonica Aresch) suppresses RANKL-induced osteocla？stogenesis by scavenging ROS in RAW 264.7 cells. Foods 8, 290.

상세보기
Kim, S. Y., Cha, H. J., Hwangbo, H., Park, C., Lee,？H., Song, K. S., Shim, J. H., Noh, J. S., Kim, H. S.,？Lee, B. J., Kim, S., Kim, G. Y., Jeon, Y. J., Choi, Y.？H. 2021. Protection against oxidative stress-induced apoptosis by fermented sea tangle (Laminaria japonica Aresch) in osteoblastic MC3T3-E1 cells through activation？of Nrf2 signaling pathway. Foods 10, 2807.

상세보기
Tao, H., Ge, G., Liang, X., Zhang, W., Sun, H., Li, M.,？Geng, D. 2020. ROS signaling cascades: Dual regulations for osteoclast and osteoblast. Acta. Biochim. Biophys.？Sin. 52, 1055-1062.

상세보기
Domazetovic, V., Marcucci, G., Iantomasi, T., Brandi,？M. L., Vincenzini, M. T. 2017. Oxidative stress in bone？remodeling: Role of antioxidants. Clin. Cases Miner. Bone Metab. 14, 209-216.

상세보기
Agidigbi, T. S., Kim, C. 2019. Reactive oxygen species？in osteoclast differentiation and possible pharmaceutical？targets of ROS-mediated osteoclast diseases. Int. J. Mol.？Sci. 20, 3576.
Choi, W. C., Reid, S. N. S., Ryu, J. K., Kim, Y., Jo,？Y. H., Jeon, B. H. 2016. Effects of γ-aminobutyric acidenriched fermented sea tangle (Laminaria japonica) on？brain derived neurotrophic factor-related muscle growth？and lipolysis in middle aged women. Algae 31, 175-187.
Melak, M., Plessner, M., Grosse, R. 2017. Actin visualization at a glance. J. Cell Sci. 130, 525-530.

상세보기
Khan, A. U., Qu, R., Fan, T., Ouyang, J., Dai, J. 2020.？A glance on the role of actin in osteogenic and adipogenic differentiation of mesenchymal stem cells. Stem Cell？Res. Ther. 11, 283.

상세보기
Rodriguez, J. P., Gonzalez, M., Rios, S., Cambiazo, V.？2004. Cytoskeletal organization of human mesenchymal？stem cells (MSC) changes during their osteogenic differentiation. J. Cell. Biochem. 93, 721-731.

상세보기
Xu, G., Shen, C., Lin, H., Zhou, J., Wang, T., Wan,？B., Binshabaib, M., Forouzanfar, T., Xu, G., Alharbi,？N., Wu, G. 2022. Development, in-vitro characterization？and in-vivo osteoinductive efficacy of a novel biomimetically-precipitated nanocrystalline calcium phosphate with internally-incorporated bone morphogenetic protein-2.？Front. Bioeng. Biotechnol. 10, 920696.
Chen, L., Shi, K., Frary, C. E., Ditzel, N., Hu, H., Qiu,？W., Kassem, M. 2015. Inhibiting actin depolymerization？enhances osteoblast differentiation and bone formation？in human stromal stem cells. Stem Cell Res. 15, 281-289.
Badila, A. E., Radulescu, D. M., Ilie, A., Niculescu, A.？G., Grumezescu, A. M., Radulescu, A. R. 2022. Bone regeneration and oxidative stress: An updated overview.？Antioxidants (Basel) 11, 318.

상세보기
Shahi, M., Peymani, A, 2017. Sahmani M. Regulation？of bone metabolism. Rep. Biochem. Mol. Biol. 5, 73-82.
Nishimura, R., Hata, K., Nakamura, E., Murakami, T.,？Takahata, Y. 2018. Transcriptional network systems in？cartilage development and disease. Histochem. Cell. Biol. 149, 353-363.
Lin, D. P. L., Carnagarin, R., Dharmarajan, A., Dass,？C. R. 2017. Transdifferentiation of myoblasts into osteob？lasts - Possible use for bone therapy. J. Pharm. Pharmacol. 69, 1661-1671.

상세보기
Ryoo, H. M., Lee, M. H., Kim, Y. J. 2006. Critical molecular switches involved in BMP-2-induced osteogenic differentiation of mesenchymal cells. Gene 366, 51-57.

상세보기
Kroemer, G., Marino, G., Levine, B. 2010. Autophagy？and the integrated stress response. Mol. Cell 40, 280-293.
Yoshii, S. R., Mizushima, N. 2017. Monitoring and measuring autophagy. Int. J. Mol. Sci. 18, 1865.
Tanida, I. 2011. Autophagosome formation and molecular mechanism of autophagy. Antioxid. Redox Signal. 14, 2201-2214.

상세보기
Park, J. M., Huang, S., Wu, T. T., Foster, N. R., Sinicrope, F. A. 2013. Prognostic impact of Beclin 1, p62/sequestosome 1 and LC3 protein expression in colon carcinoma？s from patients receiving 5-fluorouracil as adjuvant chem？otherapy. Cancer Biol. Ther. 14, 100-107.
Li, D. Y., Yu, J. C., Xiao, L., Miao, W., Ji, K., Wang,？S. C., Geng, Y. X. 2017. Autophagy attenuates the oxidative stress-induced apoptosis of Mc3T3-E1 osteoblasts.？Eur. Rev. Med. Pharmacol. Sci. 21, 5548-5556.
Bartolome, A., Lopez-Herradon, A., Portal-Nunez, S.,？Garcia-Aguilar, A., Esbrit, P., Benito, M., Guillen, C.？2013. Autophagy impairment aggravates the inhibitory？effects of high glucose on osteoblast viability and function. Biochem. J. 455, 329-337.

상세보기
Lai, E. H., Hong, C. Y., Kok, S. H., Hou, K. L., Chao,？L. H., Lin, L. D., Chen, M. H., Wu, P. H., Lin, S. K.？2012. Simvastatin alleviates the progression of periapical？lesions by modulating autophagy and apoptosis in osteoblasts. J. Endod. 38, 757-763.

상세보기
Zhang, S., Shen, S., Ma, P., Fan, D. 2022. Biochemical？targets and molecular mechanism of ginsenoside compound K in treating osteoporosis based on network pharmacology. Int. J. Mol. Sci. 23, 13921.
Cheng, Y. H., Dong, J. C., Bian, Q. 2019. Small molecul？es for mesenchymal stem cell fate determination. World？J. Stem Cells 11, 1084-1103.

상세보기
Hipskind, R. A., Bilbe, G. 1998. MAP kinase signaling？cascades and gene expression in osteoblasts. Front. Biosci. 3, d804-816.

상세보기
Takeno, A., Kanazawa, I., Tanaka, K. I., Notsu, M., Sugimoto, T. 2019. Phloretin suppresses bone morphogenetic protein-2-induced osteoblastogenesis and mineralization via inhibition of phosphatidylinositol 3-kinases/Akt pathway. Int. J. Mol. Sci. 20, 2481.
Ghosh-Choudhury, N., Abboud, S. L., Nishimura, R.,？Celeste, A., Mahimainathan, L., Choudhury, G. G. 2002.？Requirement of BMP-2-induced phosphatidylinositol 3-kinase and Akt serine/threonine kinase in osteoblast differentiation and Smad-dependent BMP-2 gene transcription. J. Biol. Chem. 277, 33361-33368.

상세보기
Songjang, W., Nensat, C., Tohtong, R., Suthiphongchai,？T., Phimsen, S., Rattanasinganchan, P., Metheenukul, P.,？Kasekarn, W., Jiraviriyakul, A. 2022. Porcine placenta？extract induced Akt, ERK, and JNK signaling to heighte n the osteogenic activity of human osteoblasts. J. App.？Pharm. Sci. 12, 18-25.
Salasznyk, R. M., Klees, R. F., Hughlock, M. K., Plopper, G. E. 2004. ERK signaling pathways regulate the osteogenic differentiation of human mesenchymal stem cells？on collagen I and vitronectin. Cell Commun. Adhes. 11, 137-153.

상세보기
Reid, S. N. S., Ryu, J. K., Kim, Y., Jeon, B. H. 2018.？GABA-enriched fermented Laminaria japonica improves cognitive impairment and neuroplasticity in scopolamine- and ethanol-induced dementia model mice. Nutr. Res. Pract. 12, 199-207.

원문보기 상세보기
Molagoda, I. M. N., Athapaththu, A. M. G. K., Park,？E. K., Choi, Y. H., Jeon, Y. J., Kim, G. Y. 2022. Fermented oyster (Crassostrea gigas) extract cures and prevents？prednisolone-induced bone resorption by activating osteoblast differentiation. Foods 11, 678.

상세보기
Li, H., Wu, Y., Huang, N., Zhao, Q., Yuan, Q., Shao,？B. 2020. γ-Aminobutyric acid promotes osteogenic differentiation of mesenchymal stem cells by inducing TNFA？IP3. Curr. Gene Ther. 20, 152-161.？

저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증