丹酚酸B对鼻息肉成纤维细胞的增殖抑制及诱导凋亡的研究

罗丹; 赵艳霞; 张彦惠; 周媛; 丁健; 孙臻峰

doi:10.13201/j.issn.1001-1781.2018.03.005

丹酚酸B对鼻息肉成纤维细胞的增殖抑制及诱导凋亡的研究

- 上海交通大学附属第一人民医院耳鼻咽喉头颈外科(上海, 200080)
基金项目:
上海市自然科学基金（No：122R1424700）

详细信息

通讯作者: 孙臻峰,E-mail:sun_zhenf@aliyun.com

中图分类号: R765.25

收稿日期: 2017-11-30

Effect of salvianolic acid B on proliferation and apoptosis of human nasal polyps fibroblasts

- Department of Otolaryngology Head and Neck Surgery, Shanghai General Hospital, Shanghai Jiaotong University, Shanghai, 200080, China

More Information

Corresponding author: SUN Zhenfeng, E-mail: sun_zhenf@aliyun.com

Received Date: 30 November 2017

摘要

摘要: 目的：探讨丹酚酸B对人鼻息肉成纤维细胞（NPFs）增殖及凋亡的影响及其分子机制。方法：采用5、10、20、40 μmol/L不同浓度丹酚酸B处理体外培养的鼻息肉来源的NPFs，CCK-8法确定药物丹酚酸B最佳作用浓度，将实验分为对照组、TGF-β1刺激组、丹酚酸B组、TGF-β1和丹酚酸B联合处理组。MTT法检测各组细胞增殖的情况，Annexin V-FITC/PI双标法检测细胞凋亡率，Western Blot法检测凋亡相关蛋白Bcl-2、Bax表达，分光光度法检测Caspase-3活性。结果：CCK-8结果显示丹酚酸B对NPFs增殖具有抑制作用，抑制率与丹酚酸B浓度呈正相关（P<0.05），选取丹酚酸B浓度为10 μmol/L进行后续实验，MTT结果显示丹酚酸B能抑制TGF-β1对NPFs的促增殖作用（P<0.01）；Annexin V-FITC/PI双标法结果显示丹酚酸B促进NPFs凋亡（P<0.05）；经丹酚酸B作用后，Caspase-3的活性升高（P<0.05）；Western Blot结果显示丹酚酸B能使Bcl-2蛋白表达下降，Bax蛋白表达升高，Bax/Bcl-2比值增大，差异有统计学意义（P<0.05）。结论：丹酚酸B对NPFs有抑制增殖和诱导凋亡的作用，其机制可能与下调Bcl-2蛋白、上调Bax蛋白表达、升高Caspase-3活性有关。
- 鼻息肉成纤维细胞 /
- 丹酚酸B /
- 细胞增殖 /
- 细胞凋亡 /
- Bcl-2 /
- Bax /
- Caspase-3
Abstract: Objective: The aim of this study is to discuss the effects and mechanism of salvianolic acid B (SAB) on the proliferation and apoptosis of human nasal polyps fibroblasts (NPFs).Method: The NPFs were treated with SAB at different concentrations (5, 10, 20, 40 μmol/L). CCK-8 assay was used to determine the inhibition of SAB on the proliferation of NPFs and determined the optimum concentration for the following experiments. The NPFs were divided into four groups:a) control group:the cells were cultured in DMEM without TGF-β1 or salvianolic acid B; b) the TGF-β1 group:the cells were cultured in DMEM containing 5 ng/ml TGF-β1; c) the SAB group:the cells were cultured in DMEM containing SAB at the optimum concentration by CCK-8 assay; d) the TGF-β1+SAB treatment groups:the cells were cultured in DMEM containing 5 ng/ml TGF-β1 and SAB at the optimum concentration by CCK-8 assay. MTT assay was used to determine the proliferation of NPFs in four groups. The apoptosis rate was analyzed by Annexin-V FITC/PI double staining. The activities of Caspase-3 were detected with spectrophotometric method. Apoptosis related proteins Bcl-2 and Bax were measured by western blot.Result: CCK-8 assay showed that SAB could inhibite NPFs proliferation in a dose-dependent pattern (P<0.05). The following experiments were carried out with the concentration of salvianolic acid B at 10 μmol/L. MTT results showed that SAB could inhibit the proliferation of NPFs stimulated by TGF-β1 (P<0.01). Annexin-V FITC/PI double staining showed that SAB could induce apoptosis of NPFs (P<0.05). The activities of Caspase-3 were increased with SAB (P<0.05). Western blot results showed that SAB reduced the Bcl-2 protein expression, and increased the Bax protein expression and the ratio of Bax/Bcl-2 (P<0.05).Conclusion: Salvianolic acid B could inhibit proliferation and induce apoptosis of NPFs. Its mechanism may be associated with increasing the activities of Caspase-3 by inhibiting the expression of Bcl-2 protein and promoting the expression of Bax protein.
- human nasal polyps fibroblasts /
- alvianolic acid B /
- proliferation /
- apoptosis /
- Bcl-2 /
- Bax /
- Caspase-3

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