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田七总皂苷对PCV-2体外诱导RAW264.7细胞组蛋白乙酰化修饰的调节作用

发表时间:2017-02-16  浏览量:1178  下载量:302
全部作者: 尹 丹,杨 剑,郝祝兵,韦英益,胡庭俊*
作者单位: 广西大学动物科学技术学院
摘 要: 以田七总皂苷(Pannax notoginseng saponins,PNS)处理PCV-2感染的小鼠单核巨噬细胞系RAW264.7,探讨PNS对PCV-2感染RAW264.7细胞组蛋白乙酰化修饰的调节作用。实验设细胞对照组、PCV-2感染组、PNS对照组,PCV-2+PNS组(治疗组)和PNS+PCV-2组(预防组),以200 mg/L PNS溶液作用于正常或PCV-2感染的RAW264.7细胞,分别通过化学发光法、ELISA法、Western blotting法检测12 h PNS对PCV-2病毒感染RAW264.7细胞抗自由基能力及细胞内过氧化氢水平、组蛋白乙酰化相关酶[组蛋白乙酰化酶(histone acetyltransferase,HAT)和组蛋白去乙酰化酶(histone deacetylase,HDAC)]活性及组蛋白H3、H4乙酰化水平的影响。结果显示,预先使用PNS处理能拮抗PCV-2感染引起的RAW264.7细胞抑制羟自由基能力的下降,而对抗超氧阴离子活力的下降无拮抗作用;PNS处理PCV-2感染细胞能显著提高感染细胞抗超氧阴离子活力,显著增强感染细胞抑制羟自由基的能力。PCV-2感染RAW264.7细胞后,引起细胞HAT活性的显著升高及HDAC活性的显著降低,导致细胞趋于组蛋白乙酰化修饰的状态;200 mg/L PNS处理PCV-2感染后的细胞或在PCV-2感染细胞前作用于细胞,均能拮抗PCV-2感染引起的细胞HDAC水平的降低,但对HAT酶活性没有显著影响。研究还发现,PCV-2感染RAW264.7细胞后,引起组蛋白H3乙酰化水平显著升高,对组蛋白H4乙酰化水平没有显著影响;加入PNS处理能抑制PCV-2感染引起的细胞组蛋白H3乙酰化水平的升高,但对细胞组蛋白H4乙酰化水平无显著影响。总之,PNS通过调节RAW264.7细胞HAT、HDAC两种酶活性及组蛋白H3、H4乙酰化水平从而调节其乙酰化修饰,进而维持细胞稳态,有利于缓解PCV-2感染RAW264.7细胞引起的氧化损伤。
关 键 词: 中兽医学;田七总皂苷;PCV-2;RAW264.7细胞;组蛋白乙酰化
Title: Effects of Pannax notoginseng saponins on histone acetylation induced by PCV-2 infection in RAW264.7 cells
Author: YIN Dan, YANG Jian, HAO Zhubing, WEI Yingyi, HU Tingjun
Organization: College of Animal Science and Technology, Guangxi University
Abstract: The objective of the current study is to investigate the effects of Pannax notoginseng saponins (PNS) on histone acetylation induced by PCV-2 infection in RAW264.7 cells. It was assigned to 4 groups which were negative control group, PCV-2 group, PNS control group and PCV-2+PNS group (treatment group) and PNS+PCV-2 group (prevention group). And then RAW264.7 cells or PCV-2 infected RAW264.7 cells were treated with 200 mg/L PNS. In this experiment, anti free radicals ability, hydrogen peroxide level, histone acetyltransferase (HAT) activity and histone deacetylase (HDAC) activity, H3 and H4 acetylation levels of PCV-2 infected RAW264.7 cells, treated with PNS for 12 h, were tested by chemical luminescence method, ELISA, Western blotting, respectively. The results showed that pretreatment with PNS could antagonize the decrease of hydroxyl free radical inhibition activity of PCV-2 infected RAW264.7 cells, but not the decline of anti superoxide anion activity. Treatment with PNS significantly increased anti superoxide anion activity and hydroxyl free radical inhibition activity of PCV-2 infected RAW264.7 cells. And also, noticeable increase and decrease of HAT activity and HDAC activity were observed, respectively. This may result to histone acetylation of cells. The decline of HDAC activity were inhibited following 200 mg/L PNS treatment both before or after PCV-2 infection, but not HAT activity. A significantly elevation of H3 acetylation level was detected after PCV-2 infection. It had no significant effect on H4 acetylation level. However, the elevation of H3 acetylation levels of PCV-2 infected RAW264.7 cells was suppressed after PNS treatment but did not have a significant effect on H4 acetylation level. Results demonstrate that PNS effects RAW264.7 cells acetylation modification by regulating HAT and HDAC activities, H3, H4 acetylation level, which is favourable for cellular homeostasis and relief of oxidative injury in RAW264.7 cells induced by PCV-2 infection.
Key words: traditional Chinese veterinary science; Panax notoginseng saponins; PCV-2; RAW264.7 cells; histone acetylation
发表期数: 2017年2月第3期
引用格式: 尹 丹,杨 剑,郝祝兵,等. 田七总皂苷对PCV-2体外诱导RAW264.7细胞组蛋白乙酰化修饰的调节作用[J]. 中国科技论文在线精品论文,2017,10(3):346-355.
 
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