海带多糖抑制平滑肌细胞增殖及抗氧化活性研究

彭臻菲; 李泳宁; 魏碧娜; 林碧珠

doi:10.19303/j.issn.1008-0384.2019.12.014

海带多糖抑制平滑肌细胞增殖及抗氧化活性研究

doi: 10.19303/j.issn.1008-0384.2019.12.014

福建卫生职业技术学院，福建　福州　350101

基金项目: 福建省医学创新基金项目（2015-CXB-27）；福建卫生职业技术学院科研基金项目（2014-1-2）

详细信息

作者简介:
彭臻菲（1978−），女，博士，讲师，研究方向：生物大分子活性表征（E-mail：Zhenfei2001@163.com）

通讯作者:
李泳宁（1979−），男，博士，副教授，研究方向：生物制药（E-mail：yongningli@163.com）

中图分类号: P 745
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出版历程
- 收稿日期: 2019-10-28
- 修回日期: 2019-12-09
- 刊出日期: 2019-12-01

Inhibitory Effects of Laminaria Japonica Polysaccharides on Proliferation and Antioxidant Activities of Vascular Smooth Muscle Cells

Fujian Health College, Fuzhou, Fujian　350101, China

摘要

摘要: 目的研究海带多糖对H₂O₂诱导血管平滑肌细胞(vascular smooth muscle cells, VSMC)增殖及胞内脂质过氧化物生成量的影响，为阐明海带多糖对VSMC的作用机制奠定基础。方法以H₂O₂为诱导剂建立体外VSMC增殖模型，通过四甲基偶氮唑盐法和细胞形态观察评价海带多糖对H₂O₂诱导VSMC生长和增殖的影响，同时以丙二醛为指标考察海带多糖对H₂O₂诱导VSMC胞内脂质过氧化物生成量的影响。结果以50 μmol·L⁻¹ H₂O₂为诱导剂建立VSMC体外增殖模型。四甲基偶氮唑盐法测定结果表明海带多糖对H₂O₂诱导VSMC增殖具有显著抑制活性，最大增殖抑制率达到73.56%。细胞形态学观察结果表明海带多糖作用下H₂O₂诱导的VSMC生长状态发生改变且细胞数量显著减少。丙二醛检测结果表明海带多糖作用下VSMC胞内脂质过氧化物量显著减少。结论海带多糖能够抑制H₂O₂诱导的VSMC增殖，且显著降低VSMC胞内脂质过氧化物生成量。
- 海带多糖 /
- 血管平滑肌细胞 /
- H₂O₂诱导 /
- 细胞增殖 /
- 脂质过氧化物
Abstract: Objective Effects of Laminaria japonicapolysaccharides on the proliferation and lipid peroxide production of H₂O₂-induced vascular smooth muscle cells (VSMCs) were examined in vitro for further understanding on the metabolic mechanism. Method Proliferation of VSMCs was artificially induced by using 50 µmol·L⁻¹ H₂O₂. Inhibitory rate on the proliferation by the L. japonica polysaccharides were determined by a methyl thiazolyl tetrazolium (MTT) assay. Morphological changes on VSMCs were observed under a microscope. Malondialdehyde (MDA) was employed as an indicator of the lipid peroxide production in VSMCs. Result A significant inhibition effect of the polysaccharides on the H₂O₂-induced VSMC proliferation with the greatest rate of 73.56% was shown by MTT. Morphologically, the number of VSMCs significantly decreased after an exposure to the polysaccharide. The MDA analysis indicated that the lipid peroxide production by VSMCs significantly declined after the treatment with L. japonica polysaccharides. Conclusion The L. japonica polysaccharides significantly inhibited the proliferation and lipid peroxide production in the H₂O₂-induced VSMCs.
- Laminaria japonica polysaccharides /
- vascular smooth muscle cell /
- H₂O₂-induced model /
- cellular proliferation /
- lipid peroxide

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图 1 海带多糖对H₂O₂诱导VSMC增殖抑制率

注：a: 相同海带多糖预处理时间下，与0.1 mg·mL⁻¹海带多糖试验组相比，P＜0.05；b：相同海带多糖预处理浓度下，与预处理时间12 h海带多糖试验组相比，P＜0.05。

Figure 1. Effects of L. japonica polysaccharides on H₂O₂-induced VSMC proliferation

Note: a: during the same pretreatment time of polysaccharide, indicated significant differences compared with the group of 0.1 mg·mL⁻¹ polysaccharide, P＜0.05; b: under the same concentration of polysaccharide pretreatment, indicated significant differences compared with the group of 12 h treatment with polysaccharide, P＜0.05.

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图 2 VSMC形态（×100）

注：A₁：对照组，A₂ ：H₂O₂模型组，A₃ ：0.5 mg·mL⁻¹ 海带多糖+H₂O₂处理组，A₄：1 mg ·mL⁻¹海带多糖+H₂O₂处理组。

Figure 2. Morphology of VSMC (×100)

Note: A₁: Control group; A₂: H₂O₂ model group; A₃: the group treatment with 0.5 mg·mL⁻¹ polysaccharide and H₂O₂; A₄: the group treatment with 1 mg·mL⁻¹ polysaccharide and H₂O₂.

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表 1 MTT测定VSMC增殖率

Table 1. Viability of VSMCs determined by MTT assay

H₂O₂浓度 H₂O₂Concentration/ （µmol·L⁻¹）	12 h	24 h	36 h
10	102.11±5.22	112.23±3.15	108.74±4.42
20	112.08±2.15	127.65±1.55	117.24±5.68
50	129.56±2.43 a	142.54±2.91 a	133.25±3.48 a
100	132.87±3.56 a	144.29±3.68 a	138.62±2.84 a
150	107.23±5.27	114.87±4.35	96.48±2.57
注：a: 与相同作用时间下H₂O₂浓度为10 µmol·L⁻¹试验组比，P＜0.05。 Note: a: during the same pretreatment time, indicated significant differences compared with the group of 10 µmol·L⁻¹ H₂O₂, P＜0.05.

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表 2 VSMC胞内MDA生成量

Table 2. MDA concentration in VSMC

样品Samples	丙二醛MDA/（µmol·L⁻¹）
对照组 Control	1.94±0.05
H₂O₂模型组 H₂O₂ Model Group	3.21±0.09 a
0.1 mg·mL⁻¹海带多糖+H₂O₂ 0.1 mg·mL⁻¹Laminaria japonica polysaccharide+H₂O₂	2.91±0.04
0.5 mg·mL⁻¹海带多糖+H₂O₂ 0.5mg·mL⁻¹Laminaria japonica polysaccharide+H₂O₂	1.75±0.08 b
1.0 mg·mL⁻¹ 海带多糖+H₂O₂ 1.0 mg·mL⁻¹Laminaria japonica polysaccharide+H₂O₂	1.27±0.05 b
注：a:与对照组相比，P＜0.01；b:与H₂O₂模型组相比，P＜ 0.01。 Note: a: indicated significant differences compared with the control group, P＜0.01; b: indicated significant differences compared with the H₂O₂ model group, P＜0.01.

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