Effect of Astragali Radix Polysaccharide on mRNA and Protein Expression of CYP27B, CYP24A in MC-3T3-E1 Osteoblasts

CHAI Yi-hui; GAO Jie; TIAN Xing-zhong; WU Da-mei; GUAN Lian-cheng; LI Jing; Li Wen; CHEN Yun-zhi

doi:10.13422/j.cnki.syfjx.20181327

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Effect of Astragali Radix Polysaccharide on mRNA and Protein Expression of CYP27B, CYP24A in MC-3T3-E1 Osteoblasts

更新时间：2024-01-04

- Effect of Astragali Radix Polysaccharide on mRNA and Protein Expression of CYP27B, CYP24A in MC-3T3-E1 Osteoblasts
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 13, Pages: 147-151(2018)
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- DOI：10.13422/j.cnki.syfjx.20181327
  CLC： R285.5
- Published：2018
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CHAI Yi-hui, GAO Jie, TIAN Xing-zhong, et al. Effect of Astragali Radix Polysaccharide on mRNA and Protein Expression of CYP27B, CYP24A in MC-3T3-E1 Osteoblasts[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(13): 147-151.
DOI：

CHAI Yi-hui, GAO Jie, TIAN Xing-zhong, et al. Effect of Astragali Radix Polysaccharide on mRNA and Protein Expression of CYP27B, CYP24A in MC-3T3-E1 Osteoblasts[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(13): 147-151. DOI： 10.13422/j.cnki.syfjx.20181327.

摘要

目的：通过研究黄芪多糖对成骨细胞增殖及1α羟化酶（CYP27B），24-羟基化酶（CYP24A）基因与蛋白表达情况的影响对其治疗骨质疏松症（OP）作用机制进行初步探讨。方法：以MC-3T3-E1成骨细胞为研究对象，实验分为5组，分别为正常组，维生素D组（1×10-5 mmol·L-1），黄芪多糖高、中、低剂量组（10，1，0.1 mg·L-1）。用噻唑蓝（MTT）比色法检测不同浓度的黄芪多糖在不同时间（24，36，48 h）对MC-3T3-E1成骨细胞增殖率的影响。实时荧光定量聚合酶链式反应法（Real-time PCR）检测黄芪多糖对MC-3T3-E1成骨细胞CYP24A，CYP27B mRNA的表达情况；蛋白免疫印迹法（Western blot）检测黄芪多糖对MC-3T3-E1成骨细胞CYP24A，CYP27B蛋白的表达情况。结果：MTT比色法显示黄芪多糖的质量浓度在0.1，1，10 mg·L-1时可以提高MC-3T3-E1成骨细胞的增殖率，且在48 h促增殖作用最佳。Real-time PCR，Western blot检测结果显示，与正常组比较，维生素D组MC-3T3-E1成骨细胞CYP27B mRNA表达量、蛋白表达量有显著促进作用（P<0.05）；与维生素D组比较，黄芪多糖在质量浓度为10，1，0.1 mg·L-1时对MC-3T3-E1成骨细胞CYP24A mRNA表达量、蛋白表达量有显著抑制作用（P<0.05）。结论：黄芪多糖能够治疗骨质疏松症其机制与提高成骨细胞活性及调节MC-3T3-E1成骨细胞CYP24A，CYP27B mRNA及蛋白表达水平有关。

Abstract

Objective: To investigate the effects of Astragali Radix polysaccharides on osteoblast proliferation

1α hydroxylase (CYP27B) and 24-hydroxylase (CYP24A) gene and protein expression and preliminarily explore its mechanism in the treatment of osteoporosis (OP). Method: The MC-3T3-E1 osteoblasts were divided into 5 groups:normal group

vitamin D group (1×10-5 mmol·L-1)

Astragali Radix polysaccharide high dose (10 mg·L-1) group

Astragali Radix polysaccharide middle dose (1 mg·L-1) group and Astragali Radix polysaccharide low dose (0.1 mg·L-1) group. The effect of different concentrations of Astragali Radix polysaccharides on the proliferation of MC-3T3-E1 osteoblasts at different time points (24

48 h) was detected by methylthiazoletetrazolium colorimetry method (MTT). The mRNA expression of CYP24A and CYP27B in MC-3T3-E1 osteoblasts was detected by real-time quantitative fluorescence polymerase chain reaction (Real-time PCR) and the protein expression of CYP24A and CYP27B in asthmatic rat MC-3T3-E1 osteoblasts was detected by Western blot. Result: MTT assay showed that the proliferation rate of MC-3T3-E1 osteoblasts could be enhanced by Astragali Radix polysaccharide at the concentrations of 0.1

10 mg·L-1

and the effect was best at 48 h. The results of Real-time PCR and Western blot showed that the mRNA and protein expression levels of CYP27B in MC-3T3-E1 osteoblasts were significantly promoted in vitamin D group as compared with the normal group (P <0.05). As compared with vitamin D group

the Astragali Radix polysaccharides (10

0.1 mg·L-1) significantly inhibited the mRNA and protein expression of CYP24A in MC-3T3-E1 osteoblast (P<0.05). Conclusion: The mechanism of Astragali Radix polysaccharide to treat osteoporosis is related to the improvement of osteoblast activity and the regulation of CYP24A

CYP27B mRNA and protein expression levels in MC-3T3-E1 osteoblast.

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