Effect of Pinoresinol Diglucoside and Pinoresinol from Eucommiae Cortex on Expression of OPG and RANKL in Osteoblasts

HU Qian-ying; YIN Rui-lin; WANG Yi-fei; LIU Xue-jing; DING Yan-xia

doi:10.13422/j.cnki.syfjx.20180908

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Effect of Pinoresinol Diglucoside and Pinoresinol from Eucommiae Cortex on Expression of OPG and RANKL in Osteoblasts

更新时间：2024-01-04

- Effect of Pinoresinol Diglucoside and Pinoresinol from Eucommiae Cortex on Expression of OPG and RANKL in Osteoblasts
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 10, Pages: 181-186(2018)
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- DOI：10.13422/j.cnki.syfjx.20180908
  CLC： R285
- Published：2018
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HU Qian-ying, YIN Rui-lin, WANG Yi-fei, et al. Effect of Pinoresinol Diglucoside and Pinoresinol from Eucommiae Cortex on Expression of OPG and RANKL in Osteoblasts[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(10): 181-186.
DOI：

HU Qian-ying, YIN Rui-lin, WANG Yi-fei, et al. Effect of Pinoresinol Diglucoside and Pinoresinol from Eucommiae Cortex on Expression of OPG and RANKL in Osteoblasts[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(10): 181-186. DOI： 10.13422/j.cnki.syfjx.20180908.

摘要

目的：观察杜仲中松脂素二葡萄糖苷及其苷元松脂素对体外培养的小鼠胚胎成骨细胞前体细胞MC3T3-E1的影响。方法：采用质量浓度为1×10-7~1×103 μg·L-1的松脂素二葡萄糖苷与松脂素体外干预MC3T3-E1成骨细胞48，72 h，采用噻唑蓝法检测细胞增殖情况，碱性磷酸酶（ALP）试剂盒检测细胞分化情况，酶联免疫吸附测定法（ELISA）和蛋白质免疫印迹法（Western blot）检测细胞骨保护素（OPG）与核转录因子-κB受体活化因子配体（RANKL）蛋白表达的水平。结果：与空白组相比，培养48 h，1×10-4~100 μg·L-1松脂素二葡萄糖苷及1×10-4~1 μg·L-1松脂素均能促进成骨细胞增殖（P<0.05，P<0.01）；培养72 h，1×10-3~100 μg·L-1松脂素二葡萄糖苷及0.1 μg·L-1松脂素能促进成骨细胞增殖（P<0.05，P<0.01）。与空白组相比，培养48 h时，质量浓度为1×10-4~1×103 μg·L-1的松脂素二葡萄糖苷、松脂素均能够显著促进成骨细胞ALP分泌（P<0.01）；培养72 h时，质量浓度为0.1~1×103 μg·L-1的松脂素二葡萄糖苷和1×10-4~1×103 μg·L-1松脂素均能显著促进成骨细胞ALP分泌（P<0.05，P<0.01）。与空白组相比，培养48 h，1×10-5~10 μg·L-1的松脂素二葡萄糖苷和松脂素均能促进细OPG蛋白分泌（P<0.05，P<0.01）；只有松脂素在1×10-5，1×10-3 μg·L-1时能明显抑制RANKL蛋白的分泌（P<0.01），在1×10-5~0.1 μg·L-1时提高OPG/RANKL（P<0.01）。结论：松脂素二葡萄糖苷与松脂素均能通过促进成骨细胞的增殖和分化达到抗骨质疏松作用，但其作用机制不同，松脂素二葡萄糖苷主要通过促进OPG分泌来发挥，而松脂素则既能通过促进OPG分泌又能通过抑制RANKL表达来发挥作用。

Abstract

Objective: To investigate the effect of pinoresinol diglucoside (PDG) and pinoresinol from Eucommiae Cortex on osteoblastic-like cell-line of MC3T3-E1. Method: MC3T3-E1 osteoblasts were cultured with 1×10-7 -1×103 μg·L-1 of PDG and pinoresinol for 48

72 h

respectively.The proliferation of cell was detected by methyl thiazolyl tetrazolium (MTT) assay

the differentiation of osteoblasts was detected by alkaline phosphatase (ALP) kit

the protein expression of osteoprogerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were examined by Western blot and enzyme linked immunosorbent assay (ELISA). Result: Compared to the control group

when cultivating 48 h

1×10-4 -100 μg·L-1 of PDG and 1×10-4-1 μg·L-1 of pinoresinol stimulated the proliferation of cell (P<0.05

P<0.01);at 72 h treatment time point

1×10-3-100 μg·L-1 of PDG and 0.1 μg·L-1 of pinoresinol stimulated the proliferation of cell (P<0.05

P<0.01).Compared to the control group

at 48 h treatment time point

1×10-4-1×103 μg·L-1 of PDG and pinoresinol significantly promoted the secretion of ALP in osteoblasts (P<0.01);at 72 h treatment time point

0.1-1×103 μg·L-1 of PDG and 1×10-4 -1×103 μg·L-1 of pinoresinol significantly promoted the secretion of ALP in osteoblasts (P<0.05

P<0.01).Compared to the control group

at 48 h treatment time point

1×10-5-10 μg·L-1 of PDG and pinoresinol promoted the secretion of OPG (P<0.05

P<0.01);1×10-5

1×10-3 μg·L-1 of pinoresinol inhibited the secretion of RANKL (P<0.01) and 1×10-5-0.1 μg·L-1 of pinoresinol raised the OPG/RANKL ratio (P<0.01). Conclusion: PDG and pinoresinol can promote anti-osteoporosis by promoting the proliferation and differentiation of osteoblasts

but their mechanisms of action are different.PDG mainly through the promotion of OPG secretion

but pinoresinol can play a role not only by promoting the secretion of OPG but also inhibiting the expression of RANKL.

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