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[木豆提取物对人成骨样TE85细胞的细胞功能及破骨样细胞衍生的影响]

[Effects of the extracts of Cajanus cajan L. on cell functions in human osteoblast-like TE85 cells and the derivation of osteoclast-like cells].

作者信息

Zheng Yuan-yuan, Yang Jing, Chen Di-hua, Sun Lan

机构信息

Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China.

出版信息

Yao Xue Xue Bao. 2007 Apr;42(4):386-91.

Abstract

The cajanine (longistylin A-2-carboxylic acid) is isolated and identified from extracts of Cajanus cajan L. (ECC) , which structure is similar to diethylstilbestrol. The regulation properties of the cajanine and other four extracts of Cajanus cajan L. (32-1, 35-1, 35-2, and 35-3) were tested in human osteoblast-like (HOS) TE85 cells and marrow-derived osteoclast-like cells. By using MTT assay to test the change of cell proliferation, 3H-proline incorporation to investigate the formation of collagen, and by measuring alkaline phosphatase (ALP) activity, bone formation in HOS TE85 cell was evaluated after pretreated for 48 hours. Bone marrow cells were cultured to examine the derivation of osteoclast cells (OLCs), which were stained with tartrate-resistant acid phosphatase (TRAP). The long term effect (pretreated for 18 days) on promoting mineralized bone-like tissue formation was tested by Alizarin red S staining in HOS TE85 cells. After the treatment with cajanine (1 x 10(-8) g x mL(-1)) for 48 hours, cell number increased significantly (57.7%). 3H-Proline incorporation also statistically increased (98.5%) in those cells. Significant change of ALP activity was also found (P < 0.01) in 35-1 and 35-3 treated cells (they were 66.2% and 82.4% in the concentration of 1 x 10(-8) g x mL(-1), respectively). The long term (18 days) effects of 32-1 and 35-3 on promoting mineralized bone-like tissue formation in HOS TE85 cell were obvious. There were much more red blots over the field of vision compared with that of control group. After the treatment of cajanine, derived-osteoclast cells appeared later and much less compared with control. The inhibition of cajanine was 22.8% while it was 37.9% in 32-1 treated cells in the dose of 1 x 10(-7) g x mL(-1). It is obvious that cajanine and ECCs promoted the osteoblast cells proliferation and mineralized bone-like tissue formation in HOS TE85 cells, while inhibited derivation of osteoclast cells. All of these suggested that cajanine has the estrogen-like action on osteoblast and osteoclast, which could be developed as anti-osteoporosis drugs.

摘要

从木豆提取物中分离并鉴定出木豆宁(长柱豆素A - 2 - 羧酸),其结构与己烯雌酚相似。对木豆宁以及木豆的其他四种提取物(32 - 1、35 - 1、35 - 2和35 - 3)在人成骨样(HOS)TE85细胞和骨髓来源的破骨样细胞中进行调节特性测试。通过MTT法检测细胞增殖变化,采用3H - 脯氨酸掺入法研究胶原蛋白的形成,并通过测量碱性磷酸酶(ALP)活性,在预处理48小时后评估HOS TE85细胞中的骨形成。培养骨髓细胞以检查破骨细胞(OLCs)的来源,并用抗酒石酸酸性磷酸酶(TRAP)染色。通过茜素红S染色在HOS TE85细胞中测试(预处理18天)促进矿化骨样组织形成的长期效果。用木豆宁(1×10(-8)g×mL(-1))处理48小时后,细胞数量显著增加(57.7%)。这些细胞中的3H - 脯氨酸掺入也有统计学意义的增加(98.5%)。在35 - 1和35 - 3处理的细胞中也发现ALP活性有显著变化(P < 0.01)(在1×10(-8)g×mL(-1)浓度下分别为66.2%和82.4%)。32 - 1和35 - 3对促进HOS TE85细胞中矿化骨样组织形成的长期(18天)效果明显。与对照组相比,视野中有更多红色斑点。用木豆宁处理后,破骨样细胞出现较晚且数量比对照组少得多。在1×10(-7)g×mL(-1)剂量下,木豆宁的抑制率为22.8%,而32 - 1处理的细胞中为37.9%。显然,木豆宁和木豆提取物促进了HOS TE85细胞中成骨细胞的增殖和矿化骨样组织的形成,同时抑制了破骨细胞的来源。所有这些表明木豆宁对成骨细胞和破骨细胞具有雌激素样作用,有望开发成为抗骨质疏松药物。

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