基础医学与临床 ›› 2016, Vol. 36 ›› Issue (8): 1080-1086.

• 研究论文 • 上一篇    下一篇

酪酸梭菌培养上清液通过下调TLR4/NF-κB信号通路抑制SW-480细胞增殖

李世超,张德纯,杨金梅,杨晓容,郭亚楠   

  1. 重庆医科大学
  • 收稿日期:2016-03-28 修回日期:2016-05-24 出版日期:2016-08-05 发布日期:2016-07-13
  • 通讯作者: 郭亚楠 E-mail:272055848@qq.com
  • 基金资助:
    国家自然科学基金(NSFC31000075)

Clostridium butyricumculture supernatant inhibits proliferation of SW-480 cells through inhibition of TLR4/NF-κB pathway

  • Received:2016-03-28 Revised:2016-05-24 Online:2016-08-05 Published:2016-07-13

摘要: 目的 研究酪酸梭菌培养上清液(C.butyricum culture supernatant; C.b.cs)对结肠癌SW-480细胞增殖的抑制作用,并探讨C.b.cs中可能的活性成分及相关分子机制。方法 用C.b.cs处理后SW-480细胞, CCK-8及FCM检测细胞的增殖和凋亡。HPLC检测C.b.cs中主要的有机酸含量。Western blot及qRT-PCR检测经过C.b.cs和丁酸处理后的TLR4的表达。用TLR4配体脂多糖(LPS)激活TLR4,再用C.b.cs和丁酸处理细胞,观察TLR4和NF-κB的表达。结果 C.b.cs呈浓度依赖性地抑制SW-480增殖,并能诱导凋亡和G0/G1期阻滞。HPLC检测C.b.cs中乙酸和丁酸的含量分别为9.27g/L和4.53g/L。C.b.cs和丁酸可下调TLR4的表达(p<0.01),并下调TLR4的下游基因NF-κB的表达(p<0.01)。LPS激活TLR4的表达(p<0.01)后,C.b.cs和丁酸下调TLR4和NF-κB的表达(p<0.05)。结论 C.b.cs通过下调 TLR4/NF-κB通路抑制SW-480细胞增殖,丁酸可能是发挥抑制作用的活性成分之一。

关键词: 酪酸梭菌, 结肠癌, 凋亡, Toll样受体4, 丁酸

Abstract: ObjectiveTo study the effects of proliferation inhibitionof C.butyricum culture supernatant (C.b.cs) on colon cancer SW-480 cells,and to explore the possible active ingredient and related molecular mechanism. MethodsCCK-8 and FCM wereused to test the cell proliferation and apoptosis.Furthermore, high efficiency liquid chromatography (HPLC) was used to analyze the concentrations of the main organic acid in C.b.cs. After the stimulation of lipopolysaccharide (LPS), the expression of TLR4 was detected by Western blot and qRT-PCR after the treatment of C.b.cs andbutyrate.ResultsC.b.cs inhibits the proliferation of SW-480cells in dose-dependent manner, which can also induce apoptosis and G0/G1 phase arrest. HPLC analysis showed that the concentrations of acetic acid and butyric acid were 9.27g/L and 4.53g/Lin C.b.cs respectively. The expression of TLR4 was inhibited after the treatment of C.b.cs and butyrate(p<0.01),as the downstream gene of TLR4, the expression of NF-κBwas also inhibited(p<0.01).Furtherly, the expression of TLR4 was regulated by LPS(p<0.01), and C.b.cs and butyrateattenuatedthe upregulation(p<0.05). ConclusionsC.b.cs and butyrate inhibit SW-480 cell proliferation by downregulation the expression of TLR4/NF-κB pathway.

Key words: C. butyricum, colon cancer, apoptosis, Toll-like receptor 4, butyrate

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