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黑加仑提取物促进MC3T3-E1前成骨细胞的分化。

Blackcurrant extract promotes differentiation of MC3T3‑E1 pre‑osteoblasts.

作者信息

Nanashima Naoki, Horie Kayo, Oey Indrawati

机构信息

Department of Nutrition, Faculty of Health Science, Aomori University of Health and Welfare, Aomori 030-8505, Japan.

Department of Bioscience and Laboratory Medicine, Hirosaki University Graduate School of Health Sciences, Aomori 036-8564, Japan.

出版信息

Biomed Rep. 2024 Jun 19;21(2):121. doi: 10.3892/br.2024.1810. eCollection 2024 Aug.

Abstract

Osteoporosis risk increases in menopausal individuals owing to the decrease in estrogen secretion. Blackcurrant extract (BCE) ameliorates osteoporosis; however, the underlying mechanisms are unclear. Furthermore, although BCE has phytoestrogenic activity, its effects on osteoblasts are unknown. In the present study, we investigated BCE-mediated attenuation of osteoporosis using mouse MC3T3-E1 pre-osteoblasts, with a focus on osteogenesis. After treating MC3T3-E1 cells with BCE for 48 h, cell proliferation was assessed using Cell Counting Kit-8. Levels of osteoblast differentiation markers, namely alkaline phosphatase activity and total collagen content in the cells, were evaluated after 3 and 14 days of BCE treatment, respectively. The expression of genes encoding osteoblast differentiation markers, including collagen type I (), alkaline phosphatase (), bone γ-carboxyglutamate protein (), and runt-related transcription factor 2 (), was evaluated using reverse transcription-quantitative polymerase chain reaction. Mineralization of the cells was evaluated using Alizarin Red staining. Femoral tissues of ovariectomized (OVX) rats with or without 3% BCE were stained using ALP to evaluate osteogenic differentiation in femoral tissue. After treating MC3T3-E1 cells with BCE, cell proliferation had increased. BCE treatment increased Alp activity and total collagen content. Moreover, the expression of , , , and increased in BCE-treated cells. Furthermore, when MC3T3-E1 cells were treated with BCE for 21 days, the levels of calcified nodules increased. Alp staining intensity was stronger in the epiphyses on femoral tissue of OVX rats treated with 3% BCE than in those of untreated OVX rats. The results suggest that BCE may promote osteogenesis by inducing osteoblast differentiation.

摘要

由于雌激素分泌减少,绝经后个体患骨质疏松症的风险增加。黑加仑提取物(BCE)可改善骨质疏松症;然而,其潜在机制尚不清楚。此外,尽管BCE具有植物雌激素活性,但其对成骨细胞的影响尚不清楚。在本研究中,我们使用小鼠MC3T3-E1前成骨细胞研究了BCE介导的骨质疏松症缓解作用,重点是骨生成。用BCE处理MC3T3-E1细胞48小时后,使用细胞计数试剂盒-8评估细胞增殖。分别在BCE处理3天和14天后,评估细胞中成骨细胞分化标志物的水平,即碱性磷酸酶活性和总胶原蛋白含量。使用逆转录-定量聚合酶链反应评估编码成骨细胞分化标志物的基因的表达,包括I型胶原蛋白()、碱性磷酸酶()、骨γ-羧基谷氨酸蛋白()和 runt相关转录因子2()。使用茜素红染色评估细胞的矿化情况。对有或没有3%BCE的去卵巢(OVX)大鼠的股骨组织进行碱性磷酸酶染色,以评估股骨组织中的成骨分化。用BCE处理MC3T3-E1细胞后,细胞增殖增加。BCE处理增加了碱性磷酸酶活性和总胶原蛋白含量。此外,在BCE处理的细胞中,、、和的表达增加。此外,当用BCE处理MC3T3-E1细胞21天时,钙化结节的水平增加。在用3%BCE处理的OVX大鼠股骨组织的骨骺中,碱性磷酸酶染色强度比未处理的OVX大鼠更强。结果表明,BCE可能通过诱导成骨细胞分化来促进骨生成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7076/11229392/0af3fd93bd80/br-21-02-01810-g00.jpg

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