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Study on the Mechanism of Enhancement Effects of Astragalus complanatus Polysaccharides on the
Proliferation of Meniscal Fibrochondrocyte Cells in Rabbits
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SHEN Huarui ，AO Liang ，WANG Lisheng ，WANG Guoyou ，HAO Qi ，LI Ting（1.Dept. of Joint Surgery，the
Affiliated Hospital of Traditional Chinese Medicine of Southwest Medical University，Sichuan Luzhou 646000，
China；2.School of Pharmacy，Southwest Medical University，Sichuan Luzhou 646000，China）
ABSTRACT OBJECTIVE：To study the mechanism of enhancement effects of Astragalus complanatus polysaccharides（ACP）
on the proliferation of meniscal fibrochondrocytes cells in rabbits. METHODS：The meniscal fibrochondrocytes cells were isolated
from 1-month-old New Zealand white rabbits. The meniscal fibrochondrocytes cells were divided into normal control group（PBS），
positive control group （glucosamine sulfate，10 mg/mL） and ACP high-dose，medium-dose and low-dose groups （40，20，10
mg/mL）. The morphology of meniscal fibrochondrocytes cells were observed under microscope. Cell proliferation rate was detected
by MTT assay. Cell cycle was observed with flow cytometry. ELISA assay was used to detect relative expression of medium
collagen type Ⅱ （Col Ⅱ） and alkaline phosphatase protein（ALP） in meniscal fibrochondrocytes cells. RT-qPCR and Western
blotting assay were adopted to detect mRNA and protein expression of transforming growth factor β 1 （TGF-β 1 ） and bone
morphogenetic protein 2（BMP-2）. RESULTS：After cultured for 72 h，meniscal fibrochondrocytes cells were fused into a single
layer，and most of them were slender type in appearance. Compared with normal control group，the proliferation rate of meniscal
fibrochondrocytes cells and the percentage of cells at G1/G0 phase were decreased significantly in positive control group and ACP
high-dose，medium-dose and low-dose groups（P＜0.05）；the percentage of cells at S phase，protein expression of Col Ⅱ and
ALP，mRNA and protein expression of TGF-β 1 and BMP-2 were increased significantly（P＜0.05）. Compared with positive control
group，inhibitory rate of meniscal fibrochondrocytes cells proliferation and the percentage of cells at G1/G0 phase were decreased
significantly in ACP high-dose group（P＜0.05），while the percentage of cells at S phase，protein expression of Col Ⅱ and ALP，
mRNA and protein expression of TGF-β 1 and BMP-2 were increased significantly（P＜0.05）. The inhibitory proliferation rate of
meniscal fibrochondrocytes cells and the percentage of cells at G1/G0 phase were increased significantly in ACP low-dose group（P＜
0.05），while the percentage of cells at S phase，protein expression of Col Ⅱ and ALP，mRNA and protein expression of TGF-β 1
and BMP-2 were decreased significantly（P＜0.05）. There was no statistical significance in above indexes of ACP medium-dose
group. CONCLUSIONS：ACP can promote the proliferation of meniscal fibrochondrocytes cells，reduce the percentage of cells at
G1/G0 phase，promote cell transformation to S phase；the mechanism of which may be related to up-regulating TGF-β 1，BMP-2
mRNA and protein expression，promoting Col Ⅱ and ALP protein expression enhancement.
KEYWORDS Astragalus complanatus polysaccharides；Meniscal fibrochondrocytes cells；Proliferation；Col Ⅱ；ALP；TGF-β 1；
BMP-2；Mechanism

半月板是膝关节的重要组成部分，具有载荷、吸收 白（Col Ⅱ）是软骨细胞外基质的主要成分之一，具有抵
震荡、润滑、增加关节接触面、营养关节软骨等功能，而 抗外界压力、维持软骨结构的作用，其表达水平升高是
[8]
[1]
半月板损伤会导致关节不稳定、甚至出现骨性关节炎 。 软骨细胞增殖的特征性指标 。碱性磷酸酶（ALP）是软
据统计，我国每 10 万人中发生半月板损伤的患者约为 骨发育过程中的关键酶蛋白，其水平升高与软骨细胞的
[9]
60～70人 。半月板损伤患者多接受保留半月板或半月 分化与成熟密切相关 。转化生长因子β（TGF-β）超家
[2]
板重建和移植的治疗办法，但其疗效仍存在较大的提升 族对骨形成、纤维化、伤口愈合具有调节作用 ，其中
[10]
空间 [3-4] 。因此，研究半月板损伤治疗的新途径符合临床 TGF-β 1是骨重建的关键因子，参与软骨细胞的增殖、分
[11]
需求。纤维软骨细胞和细胞外基质构成了半月板外部 化、死亡等过程 。骨形态发生蛋白 2（BMP-2）也是
结构，纤维软骨细胞可从半月板中直接取出并进行体外 TGF-β超家族成员之一，是关节修复过程中必不可少的
培养、传代，是研究半月板损伤修复的重要细胞来源 。 蛋白 。基于此，本文研究沙苑子多糖对软骨细胞增殖
[5]
[12]
沙苑子为豆科植物扁茎黄芪（Astragalus complana- 的促进作用，并检测细胞中Col Ⅱ、ALP蛋白和TGF-β 1、
tus R.Br.）的干燥成熟种子，归肝肾经，具有补肾助阳、养 BMP-2 mRNA及蛋白的表达水平变化，以期深入了解沙
肝明目等功效 。沙苑子多糖为沙苑子的主要有效成 苑子多糖的作用机制。
[6]
分，相关研究表明，沙苑子多糖具有体外促成骨细胞增 1 材料
[7]
殖的作用 。但是，沙苑子多糖对半月板纤维软骨细胞 1.1 仪器
（以下简称“软骨细胞”）的增殖是否有促进作用，目前尚 Gallios型流式细胞仪（美国Beckman Coulter公司）；
未见文献报道。 Multiskan fc 型酶标仪（美国 Thermo Fisher Scientific 公
关节软骨的主要成分是胶原蛋白，其中Ⅱ型胶原蛋 司）；BPN-50CHUV型二氧化碳培养箱（青岛名博环保科

·1098 · China Pharmacy 2020 Vol. 31 No. 9 中国药房 2020年第31卷第9期