| 刘钢 侯晓峰 于晨 刘佳 孙天胜.和厚朴酚抑制线粒体可溶蛋白诱导小胶质细胞活化的体外实验研究[J].,2014,14(13):2433-2436 |
| 和厚朴酚抑制线粒体可溶蛋白诱导小胶质细胞活化的体外实验研究 |
| Honokiol Inhibiting Microglia Activation Induced byMDP in vitroExperimental Study |
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| DOI： |
| 中文关键词: 小胶质细胞活化 线粒体可溶蛋白 Klf-4 和厚朴酚 |
| 英文关键词: Microglia activation MSP Klf4 Honokiol |
| 基金项目:国家自然科学基金项目（81150020） |
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| 中文摘要: |
| 目的：探讨线粒体可溶蛋白诱导小胶质细胞活化作用及和厚朴酚对小胶质细胞活化的影响。方法：培养BV2 小胶质细胞，分
空白对照组（control）、线粒体可溶蛋白组(MDP) 、和厚朴酚干预组(HNK)。ELISA 检测不同浓度线粒体可溶蛋白（MSP）刺激不同
时间的细胞上清液中IL-6 和TNF-α含量；RT-PCR 半定量法测定各组转录因子Klf4 的表达情况；倒置相差显微镜观察各组细胞
形态变化。结果：1.ELISA:MSP（10 ug/ml，100 ug/ml）处理7h 时，IL-6 和TNF-α含量与control 组比较显著升高（P<0.05）; 100
ug/ml MSP 处理1、3、5、7h 时，IL-6 和TNF-α表达与control 组比较明显升高（P<0.05），且7h 时HNK组含量明显低于MDP 组
（P<0.05）。2.RT-PCR 结果显示MDP组KLP4 的表达显著高于空白对照组和HNK 组（P<0.05）。3.倒置相差显微镜观察示空白组
胶质细胞形态呈静息状态；MDP 组小胶质细胞的胞体变圆或者椭圆，突起消失，呈“阿米巴”状；HNK组活化状态的小胶质细胞明
显减少。结论：线粒体可溶蛋白可以激活小胶质细胞，促进白细胞介素IL-6 和TNF-α的释放，和厚朴酚能够有效地抑制线粒体可
溶蛋白诱导的小胶质细胞活化,其机制可能是通过下调Klf4 的表达发挥作用。 |
| 英文摘要: |
| Objective:To investigate the activation of microglia induced by MSP and the activated process affected by Honokiol.Methods:Mice BV2 microglia in logarithmic growth phase were randomly divided into 3 groups. (1)control group: the cells were
cultured with serum free medium; (2)MDP group:the cells were treated with MDP after cultured for 24 h． (3)HNK group: after
pretreated with Honokiol for 30 min, MDP was added in the cells. ELISA: The concentration of IL-6 and TNF-αin culture supernatant of
these group were determined; Semi-quantity RT-PCR method was used to analyze the dynamic expression of transcription factor Klf4 in
different groups; Cellular morphological changes were observed under phase-contrast microscope.Results:ELISA: MSP (10 ug/ml, 100
ug/ml) treated for 7h. Compared with the control group, Il-6 and TNF-αcontent in cell supernatant have statistical significance (P<
0.05), suggesting microglia activation have certain concentration dependent manner; MSP (100 滋g/ml) treated for 1,3,5,7 h. Compared
with control group, Il-6 and TNF-αcontent in cell supernatant have statistical significance at each time point (P< 0.05), suggesting
microglia activation have certain time dependent manner; MPS (100 ug/ml) treated for 7 h. Compared with the HNK group, Il-6 and
TNF-αcontent in cell supernatant have statistical significance (P< 0.05), suggesting Honokiol could decrease the expression of Il-6 and
TNF-α. (2)RT-PCR：Klf4 expression level raised obviously after stimulating of microglia by MSP, and honokiol could downregulate Klf4
expression. (3)Morphological observation: Under phase-contrast microscope, microglia in the control group were in resting state;
microglia in the MDP group, the cell body became round, neurites disappeared, presenting "Amoeba" shape; The activation of microglia
in HNK group was reduced obviously.Conclusion:1.MSP can induce microglia activation, and promote Il-6 and TNF-αrelease, and its
activation effect is in the manner of concentration and time dependence. 2. Honokiol can effectively inhibit the microglia activation
induced by MSP, the possible mechanismis to down-regulate Klf4 expression of the microglia. |
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