| 摘要: |
| 目的探讨蛛网膜下腔注射SRT1720对慢性坐骨神经结扎（CCI）大鼠脊髓沉默信息调节因子2相关酶1（SIRT1）表达和活性的影响，以及对NF-κBp65去乙酰化的调节作用。方法将120只雄性SD大鼠采用随机数表法分成5组，每组24只，即假手术组（S）组、坐骨神经结扎组（CCI组）、二甲亚砜（DMSO）组、坐骨神经结扎+低剂量SRT1720治疗组（CCI+LD组）和坐骨神经结扎+高剂量SRT1720治疗组（CCI+HD组）。其中CCI、DMSO、CCI+LD和CCI+HD组大鼠提前8d蛛网膜下腔预置给药导管后建立CCI模型。DMSO组从手术日起，每天通过蛛网膜下腔预置细管给予10滋lDMSO，而CCI+LD组和CCI+HD组每天分别给予SRT172010滋g和20滋g（溶于10滋lDMSO）。在CCI前1d、CCI后1、3和7d各组分别处死大鼠6只，取L4~5段手术侧脊髓组织，采用荧光法测定沉默信息调节因子2相关酶1（SIRT1）活性，Westernblot法测定SIRT1、NF-κB、乙酰化-NF-κBp65（Lys310）水平，并计算各组NF-κB的乙酰化水平（乙酰化-NF-κBp65/NF-κB）。结果与S组比较，CCI和DMSO组在CCI后各时点SIRT1水平和酶活性均显著下降（均P＜0.01），相反，NF-κB、乙酰化-NF-κBp65水平和NF-κB乙酰化水平均显著升高（均P＜0.01）；而与CCI组比较，CCI+LD和CCI+HD组在CCI后3d、7dSIRT1水平和酶活性均较高（均P＜0.01），NF-κB、乙酰化-NF-κBp65（Lys310）水平则较低。其中SRT1720高剂量组在CCI后7d，SIRT1水平和酶活性显著高于低剂量组（P＜0.01），而NF-κB的乙酰化水平则更低（P＜0.01）。结论SRT1720蛛网膜下腔给药可改善CCI导致的脊髓SIRT1水平和酶活性的降低，抑制NF-κBp65的过高表达和乙酰化。 |
| 关键词: SRT1720 慢性坐骨神经结扎 沉默信息调节因子 2 相关酶 1 NF-κB 去乙酰化 |
| DOI： |
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| 基金项目:国家自然科学基金青年科学基金项目；浙江省医药卫生科技计划项目 |
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| Effects of intrathecal SRT1720 injection on expressions of SIRT1 and deacetylation of NF-κB in spinal cord in rats with chronic constriction injury |
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LV Chen, GUO Xiaowen, ZHENG Hui, YE Ling, MA Qian, ZHANG Juan
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Zhejiang Hospital of Traditional Chinese Medicine
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| Abstract: |
| Objective To investigate the effect of intrathecal SRT1720 injection on expression and activity of SIRT1, and deacetylation of NF-κBp65 in rats with chronic constriction injury (CCI). Methods One hundred and twenty male SD rats were randomly divided into five groups with 24 animals in each group: Sham group (S group), CCI group, dimethylsulfoxide (DMSO)
group, CCI+LD group and CCI+HD group. Lumbosacral intrathecal (IT) catheter was placed 8d before CCI operation in DMSO, CCI+LD and CCI+HD groups. CCI was induced in all animals except those in S group. 10滋l DMSO was intrathecally injected q.d via the catheter in DMSO group; 10滋g and 20滋g SRT1720 dissolved in 10滋l DMSO was given to rats in CCI+LD and CCI+HD groups. Six rats were sacrificed randomly before and at 1, 3, 7 d after CCI or sham surgery in each group. The L4~5 spinal cord segments were removed for detection of deacetylase activity of SIRT1 by fluorometric kit. The proteins levels of (SIRT1) and NF-κB/Ac-NF-κB p65 were determined by Western blot analysis. Results The deacetylase activity and protein levels of SIRT1 were significantly decreased after operation in CCI and DMSO group at postoperative d1, 3 and 7, and the proteins level of NF-κB/Ac-NF-κB p65 were significantly increased(P＜0.01), compared with S group(P＜0.01). Compared with CCI group, the deacetylase activity and protein level of SIRT1 increased, and NF-κB/Ac-NF-κB p65 protein level decreased both in CCI+LD
group and CCI+HD group at postoperative d3 and d7 (P＜0.01). Furthermore, the activity and protein levels of SIRT1 were higher in CCI+HD group and the proteins levels of Ac-NF-κB p65 were lower (P＜0.01) than those in CCI+LD group at postoperative d7 (P＜0.01). Conclusion SRT1720 intrathecal injection can improve the down-regulation of spinal SIRT1 deacetylase activity and protein levels in CCI rats, furthermore, it also can inhibit over-expression and acetylation of NF-κB p65. |
| Key words: SRT1720 Chronic constriction injury Silentmating type information regulation 2 homolog 1 NF-κB Deacetylation |
