红树林淡紫拟青霉EPS提取物体外抗HSV-1机制研究

发布时间：2018-01-12 03:20

本文关键词：红树林淡紫拟青霉EPS提取物体外抗HSV-1机制研究　出处：《中国病原生物学杂志》2017年01期 　论文类型：期刊论文

【摘要】：目的探讨红树林淡紫拟青霉胞外多糖(extracellular polysaccharides,EPS)提取物体外抗单纯疱疹病毒Ⅰ型(HSV-1)感染的机制。方法以Vero细胞为病毒感染靶细胞,以不同浓度EPS提取物作用于HSV-1感染过程的各个阶段,MTT法测定提取物对Vero细胞的毒性作用和对HSV-1的直接灭活作用及对病毒吸附和生物合成的影响;采用RT-PCR检测该提取物对病毒gD基因US6和DNA聚合酶基因UL30表达的影响。结果 EPS提取物对Vero细胞无促进增殖作用,其对Vero细胞的半数有毒浓度(CC50)为1 300.17μg/ml,浓度在800μg/ml以下时细胞存活率达90%以上;在25~800μg/ml浓度范围内该提取物可抑制HSV-1吸附和生物合成,并表现出一定量效关系;浓度为800μg/ml时抑制率最高,分别为39.5%和59.7%,与病毒对照组相比差异有统计学意义(P0.01),但并不能达到完全抑制。未发现该提取物对HSV-1有直接灭活作用。RT-PCR显示该提取物可干扰病毒US6和UL30基因转录,灰度扫描分析US6mRNA丰度与病毒对照组相比差异有统计学意义(P0.05);UL30mRNA丰度与病毒对照组相比差异有统计学意义(P0.01)。结论 EPS提取物对Vero细胞毒性较小,使用安全。该提取物具有一定的抗病毒作用,可抑制HSV-1吸附和生物合成,还可抑制US6和UL30基因转录。
[Abstract]:Objective to study extracellular polysaccharides of Penicillium lilacinus in mangroves. Methods Vero cells were used as target cells of HSV-1 infection. Different concentrations of EPS extract were used in different stages of HSV-1 infection. The toxicity of the extract to Vero cells, the direct inactivation of HSV-1 and the effects on virus adsorption and biosynthesis were determined by MTT method. RT-PCR was used to detect the effect of the extract on the expression of GD gene US6 and DNA polymerase gene UL30. Results the EPS extract did not promote the proliferation of Vero cells. The median toxic concentration of CC50 to Vero cells was 1 300.17 渭 g / ml, and the cell survival rate was more than 90% when the concentration was below 800 渭 g / ml. The extract could inhibit HSV-1 adsorption and biosynthesis in the concentration range of 25 渭 g / ml, and showed a dose-effect relationship. When the concentration was 800 渭 g / ml, the inhibition rate was the highest (39.5% and 59.7, respectively, compared with the virus control group, the difference was statistically significant (P 0.01). It was not found that the extract could directly inactivate HSV-1. RT-PCR showed that the extract could interfere with the transcription of US6 and UL30 genes. Compared with the control group, the US6mRNA abundance was significantly different from that of the control group (P 0.05). The abundance of UL30mRNA was significantly higher than that of the control group (P 0.01). Conclusion EPS extract is less toxic to Vero cells. The extract can inhibit HSV-1 adsorption and biosynthesis, as well as US6 and UL30 gene transcription.
【作者单位】：海南医学院热带医学与检验学院;海南医学院临床学院;
【基金】：国家自然科学基金项目(No.81560006) 海南省自然科学基金项目(No.813248) 科研培育基金项目(No.HY2014-015)
【分类号】：R373.11
【正文快照】： ***机体内的多糖除了作为能源和细胞组成成分外,还参与细胞间信息传递、分裂、再生等生命活动,被认为是继蛋白质、核酸之后的第三条生命链。因此,利用结构相似而功能相异的多糖干预细胞代谢和病原体繁殖从而达到抗肿瘤、抗病毒作用是可行的。红树林真菌由于生活在寡营养、弱碱

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