天然产物研究与开发 ›› 2022, Vol. 34 ›› Issue (7): 1204-1212.doi: 10.16333/j.1001-6880.2022.7.014

• 开发研究 • 上一篇    下一篇

禾谷镰刀菌胞外多糖诱导SGC-7901细胞凋亡作用及其机制研究

王姣姣1,2,3,袁平川1,2,3,许寒池1,陈   颖1,张斯佳1,田梦云1,余子琪1,柳春燕1,2,3*   

  1. 1皖南医学院安徽省多糖药物工程技术研究中心 皖南医学院药学院药物研发中心;2活性生物大分子安徽省重点实验室;3安徽省皖南地区植物药活性筛选与再评价工程实验室,芜湖 241000
  • 出版日期:2022-07-28 发布日期:2022-07-25
  • 基金资助:
    国家级/省级大学生创新创业训练计划(202110368058,202010368025,S202010368047);安徽省高校自然科学研究重大项目(KJ2021ZD0101)

Mechanism of apoptosis induced by exopolysaccharides from Fusarium graminearum on SGC-7901 cells#br#
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WANG Jiao-jiao1,2,3,YUAN Ping-chuan1,2,3,XU Han-chi1,CHEN Ying1,ZHANG Si-jia1,TIAN Meng-yun1,YU Zi-qi1,LIU Chun-yan1,2,3*#br#
  

  1. 1Wannan Medical College Drug Research & Development Center,School of Pharmacy,Anhui Provincial Engineering Research Center for Polysaccharide Drugs,Wannan Medical College; 2Anhui Province Key Laboratory of Active Biological Macro-Molecules;3Provincial Engineering Laboratory for Screening and Re-evaluation of Active Compounds of Herbal Medicines in Southern Anhui,Wuhu 241000,China
  • Online:2022-07-28 Published:2022-07-25

摘要:

为了探究禾谷镰刀菌胞外多糖(exopolysaccharides from Fusarium graminearum,FGEPS)抗人胃癌细胞SGC-7901作用及其可能的机制,本研究利用液体培养基对禾谷镰刀菌进行发酵培养,通过醇沉、脱色、脱蛋白和透析分离得到胞外多糖,并通过高效凝胶渗透色谱、红外光谱和紫外光谱对其进行分析。采用CCK-8法分别检测FGEPS对人胃癌细胞SGC-7901、BGC-823和MGC-803的增殖抑制作用,通过AnnexinV-FITC/PI、Hoechst 33258及JC-1染色分析FGEPS对筛选出的SGC-7901细胞凋亡的影响,进一步通过Western blot法检测促凋亡蛋白Bax、Cleaved Caspase-3、Cleaved Caspase-9和抑凋亡蛋白Bcl-2的表达水平。结果表明从禾谷镰刀菌发酵液中分离出的胞外多糖具有多糖特征吸收峰,不含有多肽核酸类物质,液相色谱显示含有4个多糖组分。CCK-8结果显示FGEPS对SGC-7901细胞的抑制效果最为显著,能显著降低SGC-7901细胞线粒体膜电位,诱导细胞核染色质浓缩,并产生凋亡小体(P<0.05或P<0.01)。FGEPS能上调SGC-7901细胞中促凋亡蛋白Bax、Cleaved Caspase-3和Cleaved Caspase-9表达(P<0.01或P<0.001),下调抑凋亡蛋白Bcl-2表达(P<0.001)。综上所述,我们从禾谷镰刀菌发酵液中首次分离出的FGEPS能通过线粒体途径诱导SGC-7901细胞发生凋亡。

关键词: 禾谷镰刀菌胞外多糖, SGC-7901, 细胞凋亡, 线粒体通路

Abstract:

To investigate the effect and mechanism of exopolysaccharides from Fusarium graminearum (FGEPS) on human gastric cancer SGC-7901 cells.Fusarium graminearum was fermented in liquid medium.The exopolysaccharides were separated by alcohol precipitation,decolorization,deproteinization and dialysis,and analyzed by high performance gel permeation chromatography (HPGPC),infrared (IR) and ultraviolet (UV) spectroscopy.CCK-8 method was used to detect the proliferation inhibition activity of SGC-7901,BGC-823,and MGC-803 cells.Effects of FGEPS on apoptosis of SGC-7901 cells were analyzed by Annexin V-FITC/PI,JC-1 and Hoechst 33258 staining.The expressions of pro-apoptotic proteins Bax,Cleaved Caspase-3,Cleaved Caspase-9 and anti-apoptotic protein Bcl-2 were detected by Western blot.FGEPS was isolated from the fermentation broth of Fusarium graminearum,which had the characteristic absorption of polysaccharides and contained four components. In vitro anti-tumor activity assay showed that FGEPS inhibited SGC-7901 cells more significantly than BGC-823 and MGC-803 cells.FGEPS could significantly reduce the mitochondrial membrane potential of SGC-7901 cells,induce the concentration of nuclear chromatin,and produce apoptotic bodies (P<0.05 or P<0.01).FGEPS could raise the expression of pro-apoptotic proteins Bax,Cleaved Caspase-3 and Cleaved Caspase-9 in SGC-7901 cells (P<0.01 or P<0.001),and reduce the expression of anti-apoptotic protein Bcl-2 (P<0.001).In summary,FGEPS isolated from the fermentation broth of Fusarium graminearum could induce apoptosis of SGC-7901 cells through mitochondrial pathway.

Key words: exopolysaccharides from Fusarium graminearum, SGC-7901, apoptosis, mitochondrial pathway

中图分类号:  R285.5