[1]潘晓彦,张明娇,曾晓云,等.狼毒大戟提取物通过NF-κB通路激活潜伏HIV[J].南方医科大学学报,2015,(11):1614.
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狼毒大戟提取物通过NF-κB通路激活潜伏HIV()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2015年11期
页码:
1614
栏目:
出版日期:
2015-11-15

文章信息/Info

Title:
Euphorbia fischeriana extract reactivates latent HIV through nuclear factor-κB pathway
作者:
潘晓彦张明娇曾晓云林健李琳李珉珉赵伟
关键词:
NF-κBprostratin潜伏HIV狼毒大戟
Keywords:
nuclear factor-κB prostratin latent HIV Euphorbia fischeriana
摘要:
目的研究狼毒大戟提取物对潜伏HIV激活的影响,探讨其激活潜伏HIV可能的机制以及清除HIV的作用。方法取新鲜
狼毒大戟植物根部组织,液氮速冻并粉碎成末,冷冻干燥3 d后称取少量,采用丙酮一步法提取并浓缩。以甲醇-水为流动相,采
用HPLC反相C18柱分离,并通过MS鉴定后,得到狼毒大戟提取物(EFE)。以潜伏HIV细胞系J-Lat 10.6为激活模型,以TNF-α
(10 ng/mL)作为阳性对照,EFE(50 μg/mL)处理24 h后,采用FACS分析GFP阳性率以观测激活活性。采用不同浓度NF-κB抑
制剂(Bay 11-7082)抑制EFE活性,并用WB检测2 h 内p65 入核水平,以及24 h 内HIV蛋白p24 随时间点变化情况。结果丙
酮一步法可简便快速从狼毒大戟植物中提取得到EFE,经鉴定含prostratin 及其类似物,分离后测得其中prostratin 浓度为
0.53 mmol/L。EFE(50 μg/mL)作用24 h可产生约50%激活潜伏HIV活性,同时HIV蛋白p24水平随时间显著增加。进一步研
究发现NF-κB通路抑制剂(Bay 11-7082)可对EFE活性产生浓度依赖抑制作用,且2 h 内胞核p65 水平在EFE作用下增加明
显。结论狼毒大戟提取物EFE含有效成分prostratin及其类似物,可产生较强激活潜伏HIV活性,其机制为通过激活NF-κB
通路促进p65入核从而发挥作用。
Abstract:
Objective To investigate the effect of Euphorbia fischeriana extract on latent HIV reactivation and the pathway involved
in this process and discuss the value of Euphorbia fischeriana extract in eliminating HIV. Methods Fresh tissues of Euphorbia
fischeriana root were crushed into powder after quick freezing with liquid nitrogen and extracted with acetone followed by a
three-day vacuum freeze-drying for dehydration of the extract. The extract (EFE) was separated using RP-C18 column with
high-performance liquid chromatography (HPLC) and identified with mass spectrometry (MS). The activity of reactivated latent
HIV was analyzed by fluorescence-activated cell sorting in a J-Lat 10.6 cell model treated with EFE (50 μg/mL) for 24 h, using
TNF-α (10 ng/mL) as the positive control. The effect of a NF-κB pathway inhibitor (Bay 11-7082) on EFE activity was tested. The
changes in P65 expression in the cell nuclei within 2 h and HIV protein p24 expression within 24 h were analyzed by Western
blotting in cells treated with EFE. Results EFE was obtained by one-step acetone extraction, and the concentration of prostratin
in the extract was around 0.53 mmol/L. About 50% of the cells showed HIV reactivation after treatment with 50 μg/mL EFE for
24 h accompanied by a significantly increased p24 expression. The activity of EFE in reactivating latent HIV was inhibited by
Bay 11-7082 in a concentration-dependent manner, and p65 accumulation was detected in the cell nuclei within 2 h. Conclusion
EFE we obtained contains the active compounds of prostratin and its analogues and shows a strong capacity to reactivate latent
HIV through classical NF-κB pathway.

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更新日期/Last Update: 1900-01-01