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中国临床药理学与治疗学 ›› 2014, Vol. 19 ›› Issue (10): 1081-1086.

• 基础研究 •    下一篇

丹酚酸B对转化生长因子β1诱导人肺成纤维细胞增殖和分化的影响

张敏, 曹述任   

  1. 暨南大学第四附属医院 广州市红十字会医院 重症监护中心, 广州 510220,广东
  • 收稿日期:2013-12-10 修回日期:2014-07-28 出版日期:2014-10-26 发布日期:2014-10-29
  • 作者简介:张敏,女,博士研究生,主任医师,硕士生导师,主要研究方向为肺纤维化防治。 Tel: 13632462245 E-mail: zhangmincc@163.com
  • 基金资助:
    国家自然科学基金资助项目(81100042); 广东省医学科研基金立项项目(A2012498); 广州市医药卫生科技一般引导项目(20121A011044)

Effect of salvianolic acid B on TGF-β1-induced proliferation and differentiation in lung fibroblasts

ZHANG Min, CAO Shu-ren   

  1. Department of Intensive Care Medicine (ICU), Guangzhou Red Cross Hospital, Jinan University, Guangzhou 510220, Guangdong ,China
  • Received:2013-12-10 Revised:2014-07-28 Online:2014-10-26 Published:2014-10-29

摘要: 目的 观察丹酚酸B(Sal B)对转化生长因子β1(TGF-β1)诱导的人肺成纤维细胞增殖及分化的影响。方法 将人胚肺成纤维细胞(MRC-5)随机分为6组:对照组(未加入TGF-β1或Sal B);1 μmol/L Sal B组;10 μmol/L Sal B组;10 ng/mL TGF-β1组;TGF-β1(10 ng/mL)+1 μmol/L Sal B组;TGF-β1(10 ng/mL)+10 μmol/L Sal B组。采用MTT法观察各组细胞增殖情况;RT-PCR和Western blot 方法分别检测各组细胞α-SMA mRNA和蛋白的表达情况;免疫荧光方法检测细胞内纤维形肌动蛋白(F-actin,Fibrous Actin)重组及观察细胞形态变化。结果 与对照组比较,TGF-β1组细胞增殖率、α-SMA mRNA和蛋白的表达水平均明显增加,细胞形态发生明显改变,胞内可见大量F-actin聚合形成的应力纤维(stress fiber)(P<0.01);与对照组比较,单纯加入Sal B对细胞增殖、α-SMA表达、细胞形态和F-actin重组均未产生影响(P>0.05); 与单纯TGF-β1组比较,TGF-β1+Sal B两组细胞增殖率、α-SMA mRNA和蛋白表达水平均下降,发生形态改变的细胞减少,胞内F-actin聚合形成的stress fibers减少(P<0.05), 且 10 μmol/L Sal B对TGF-β1抑制效应优于 1 μmol/L Sal B,差异均有统计学意义(P<0.05)。结论 Sal B体外能够抑制TGF-β1诱导的人肺成纤维细胞增殖和向肌纤维母细胞分化。

关键词: 丹酚酸B, 肺成纤维细胞, 转化生长因子β1;, 增殖, 分化

Abstract: AIM: To investigate the effect of salvianolic acid B on TGF-β1-induced proliferation and differentiation in lung fibroblasts. METHODS: The human embryonic lung fibroblasts (MRC-5) were randomly divided into six groups as follows: control group (cells cultured with DMEM without TGF-β1 or Sal B); Sal B groups (cells cultured with 1 μmol/L or 10 μmol/L Sal B); TGF-β1 group (cells cultured with 10 ng/mL TGF-β1); TGF-β1 (10 ng/mL) with 1 or 10 μmol/L of Sal B treated groups. The cell proliferation rates were analyzed by MTT assay. The protein levels of α-SMA were detected by Western blot, and the expressions of α-SMA mRNA were evaluated by RT-PCR.The expression of F-actin was detected by immunofluorescence. RESULTS: TGF-β1 treatment of lung fibroblasts increased cell proliferation rates and α-SMA expression, changed cell morphology from a spindle shape to a stellate shape, and induced F-actin reorganization to form abundant long stress fibers (P<0.01 as compared with control). The treatment with only Sal B did not affect the proliferation, α-SMA expression, cell morphology and F-actin reorganization of lung fibroblasts (P>0.05 as compared with control). Sal B was found to inhibit TGF-β1-induced cell proliferation, α-SMA expression, cell morphology change, and F-actin reorganization in lung fibroblasts (P<0.05 as compared with TGF-β1 group). The inhibitory effect of Sal B on TGF-β1-induced proliferation and differentiation in lung fibroblasts was more significant with the treatment of high-dose Sal B (1 vs 10 μmol/L, P<0.05). CONCLUSION: Sal B could inhibit TGF-β1-induced cell proliferation and differentiation in lung fibroblasts in vitro.

Key words: salvianolic acid B, pulmonary fibroblast, transforming growth factor-β1;, proliferation, differentiation

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