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人参皂苷 Rb1 不能阻止去卵巢大鼠的骨质疏松性骨丢失。

Ginsenoside Rb1 does not halt osteoporotic bone loss in ovariectomized rats.

机构信息

Department of Pharmacology, Guangdong Medical University, Zhanjiang, China.

Guangdong Key Laboratory for Research and Development of Natural Drugs, Guangdong Medical University, Zhanjiang, China.

出版信息

PLoS One. 2018 Sep 13;13(9):e0202885. doi: 10.1371/journal.pone.0202885. eCollection 2018.

Abstract

Osteoporosis (OP) is a systemic skeletal disorder, manifesting with a reduction in bone mass and deterioration of the microarchitecture. Mesenchymal stem cells (MSCs) have an innate ability to differentiate into several cell types, including osteoblasts (OB). Ginsenoside Rb1 (GRb1) is an ethanol extract from ginseng and contains a highly concentrated form of ginsenoside. GRb1 shows extensive beneficial health effects such as anti-oxidative and anti-inflammatory functions, modulating the immune system and inhibiting osteoclastogenesis. We hypothesized that GRb1 can promote MSC differentiation into OBs and inhibit bone loss. In the present study, we aimed to address two questions: (1) Will GRb1 have a positive effect on osteogenic differentiation of MSCs? and (2) Will GRb1 halt bone loss in ovariectomized (OVX) rats? We investigated the effects of GRb1 on viability and osteogenic differentiation of rat mesenchymal stem cells (rMSCs). Our results showed that GRb1 at concentrations of 10-8 M and 10-6 M can increase alkaline phosphatase activity, mineralization and the expression of osteogenic related proteins, such as osteopontin and osteoprotegerin, while incubating rMSCs with osteogenic induction medium and GRb1. Adding GRb1 into the medium can prevent rMSCs from Oxidative damage at the concentration of 25μM H2O2. Furthermore, 40 4-month-old rats were assigned to 5 groups(8 rats per group): the basal group, the sham group, the OVX group, the high dose of GRb1 group (6 mg/kg/day) and the low dose of GRb1 group (3 mg/kg/day). Rats recrived treatment 3days after surgery and last for 14 weeks. Examinations included serum analysis, mechanical testing, Masson-Goldner trichrome staining and bone histomorphometry analysis. The results showed that OVX can lead to dyslipidemia and excessive oxidative stress, whereas GRb1 cannot significantly halt dyslipidemia and excessive oxidative stress in OVX rats. In addition, the bone density of the lumbar vertebra and femur were decreased significantly in the OVX rats, and GRb1 could not inhibit bone loss. Bone histomorphometry analysis showed that the number and width of bone trabecula of the tibia were reduced in OVX rats, and GRb1 could not prevent their occurrence. A bone biomechanics assay showed that GRb1 cannot improve the ability of bone structure to resist fracture of the femur in OVX rats. The current study demonstrated that GRb1 has an obvious effect on osteogenic differentiation in rMSCs but no obvious effect on bone loss in OVX rats. These findings indicate GRb1 has a positive effect on rMSCs but does not have an effect on bone loss in OVX rats at the concentration we used.

摘要

骨质疏松症(OP)是一种全身性骨骼疾病,表现为骨量减少和微结构恶化。间充质干细胞(MSCs)具有分化为多种细胞类型的内在能力,包括成骨细胞(OB)。人参皂苷 Rb1(GRb1)是从人参中提取的乙醇提取物,含有高度浓缩的人参皂苷。GRb1 具有广泛的有益健康作用,如抗氧化和抗炎功能、调节免疫系统和抑制破骨细胞生成。我们假设 GRb1 可以促进 MSC 分化为 OB 并抑制骨丢失。在本研究中,我们旨在解决两个问题:(1)GRb1 是否会对 MSC 的成骨分化产生积极影响?(2)GRb1 是否会阻止去卵巢(OVX)大鼠的骨丢失?我们研究了 GRb1 对大鼠间充质干细胞(rMSCs)的活力和成骨分化的影响。我们的结果表明,GRb1 在 10-8 M 和 10-6 M 浓度下可以增加碱性磷酸酶活性、矿化和骨形成相关蛋白的表达,如骨桥蛋白和护骨素,同时孵育 rMSCs 与成骨诱导培养基和 GRb1。在 25μM H2O2 的浓度下,将 GRb1 添加到培养基中可以防止 rMSCs 发生氧化损伤。此外,将 40 只 4 月龄大鼠分为 5 组(每组 8 只):基础组、假手术组、OVX 组、高剂量 GRb1 组(6 mg/kg/天)和低剂量 GRb1 组(3 mg/kg/天)。大鼠在手术后 3 天开始接受治疗,持续 14 周。检查包括血清分析、机械测试、Masson-Goldner 三色染色和骨组织形态计量学分析。结果表明,OVX 可导致血脂异常和过度氧化应激,而 GRb1 不能显著抑制 OVX 大鼠的血脂异常和过度氧化应激。此外,OVX 大鼠的腰椎和股骨骨密度明显降低,GRb1 不能抑制骨丢失。骨组织形态计量学分析显示,OVX 大鼠胫骨骨小梁的数量和宽度减少,GRb1 不能预防其发生。骨生物力学检测表明,GRb1 不能改善 OVX 大鼠股骨结构抵抗骨折的能力。本研究表明,GRb1 对 rMSCs 的成骨分化有明显作用,但对 OVX 大鼠的骨丢失没有明显作用。这些发现表明,GRb1 对 rMSCs 有积极作用,但在我们使用的浓度下,对 OVX 大鼠的骨丢失没有作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/80c3/6136715/c89b04207b6c/pone.0202885.g001.jpg

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