| 王义宁 张钦 杨娅婷 曾振华 陈仲清.EGb761 对LPS诱导THP-1 细胞释放HMGB1 蛋白表达的调节[J].,2015,15(20):3809-3812 |
| EGb761 对LPS诱导THP-1 细胞释放HMGB1 蛋白表达的调节 |
| The effect of EGb761 on the Expression of HMGB1 in LPS-induced THP-1 |
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| DOI： |
| 中文关键词: LPS THP-1 细胞 EGB761 HMGB1 NF-kB（P-65） |
| 英文关键词: Lipopolysaccharides Mononuclear epithelial cells Extract of Ginkgobiloba761 High mobility group box-1 protein Nuclear factor kappa B（p65） |
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| 中文摘要: |
| 目的：探讨EGb761 对LPS诱导THP-1 细胞释放HMGB1 蛋白表达的调节，为EGb761 的临床运用提供可行的依据。方法：
LPS（1 ug/mL）诱导不同时间后，western blotting检测THP-1 细胞上清液中HMGB1 蛋白含量变化及不同浓度EGb761 对LPS 诱
导THP-1 细胞释放HMGB1 蛋白的表达和NF-kB 的活性；酶联免疫吸附法（ELISA）检测细胞中IL-1beta、IL-6 、TNF-alpha的含量。共聚
焦显微镜观察EGb761 对LPS诱导THP-1细胞释放HMGB1 蛋白核转位变化。结果：（1）LPS 组IL-1beta、IL-6 、TNF-alpha的含量在刺
激6-12 h后明显高于空白对照组，而EGb761+LPS 组IL-1 beta、IL-6 、TNF-alpha的含量均显著低于LPS 组（P<0.05）。（2）EGb761 处理
LPS诱导THP-1细胞6 h 后细胞上清液NF-资B 活性表达量较空白对照组低，随着处理时间延长至12 h，NF-资B 的活性表达量呈
明显下降趋势（P<0.05）。（3）LPS诱导THP-1 细胞18 h后，细胞上清液中HMGB1蛋白含量呈明显升高趋势（P<0.05）。(4)不同浓度
EGb761 对LPS 诱导THP-1 细胞18 h 后，HMGB1 蛋白含量较空白对照组有下降趋势，HMGB1 蛋白含量随着EGB761 浓度增加
至100 ug/mL呈下降趋势并呈浓度依赖效应（P<0.05）。（5）LPS诱导THP-1 细胞后，在共聚焦显微镜下可见胞浆中大量HMGB1
蛋白标记分布，而EGb761+LPS共同诱导THP-1 细胞后胞浆中可见少量HMGB1 蛋白分布。结论：LPS 可诱导THP-1 细胞IL-1
茁、IL-6 、TNF-alpha表达增多及NF-kB 活化，导致HMGB1 蛋白表达增多及核转位，而EGB761 能抑制THP-1 细胞IL-1beta、IL-6 、
TNF-alpha表达及NF-kB 活化，调节HMGB1 蛋白的表达及核转位。 |
| 英文摘要: |
| Objective:To investigate the effect of EGb761 on the expression of HMGB1 in LPS-induced THP-1 cells. We expect
this study could provide a viable basis for the clinical application of EGb761.Methods:THP-1 cells were induced by LPS(1 ug/mL) for a
certain period of time, then the amount of HMGB1 in the THP-1 cell supernatant was measured by a Western blot. EGB761 with
different concentrations was added to the LPS-induced THP-1 and then the amount of HMGB1 and the level of phosphorylated NF-kB
were tested. ELISA was used to test the content of IL-1beta, IL-6 and TNF-alpha. A confocal microscope was used to observe the nuclear
translocation of HMGB1 in the LPS induced THP-1 cells to evaluate the effect of EGb761 in this progress.Results:(1) The amount of
IL-1beta, IL-6 and TNF-alpha in the LPS-induced group was significantly increased when compared with the control group after induced for
6-12 h, while the amount of those makers in the EGb761+LPS group was slightly increased compared to the control group but
significantly less than that in the control group (P<0. 05). (2) The amount of NF-kB in the EGb761+LPS group was less than that in the
control group at 6 h, and showed a significantly downward trend at 12 h (P<0.05). (3) The amount of HMGB1 in the THP-1 cell
supernatant increased significantly after the procedure of LPS induction for 18 hours (P<0.05). (4) The amount of HMGB1 in the
EGb761+LPS group decreased significantly when compared with the control group after induced for 18 h, and showed a concentrationdependent
effect when the concentration of EGb761 was increased to 100 ug/mL (P<0.05). (5) We found a large amount of marked
HMGB1 under the confocal microscope in the LPS group, while a relatively small amount of that in the EGb761+LPS group.Conclusion:LPS may induce the expression of IL-1beta, IL-6, TNF-alpha and the activation of NF-kB in the LPS-induced THP-1 cells to
promote the expression and nuclear translocation of HMGB1, while EGb761 may inhibit the progress of expression and activation
mentioned above, thus affect the expression and the progress of nuclear translocation of HMGB1. |
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