白藜芦醇对惊厥持续状态大鼠海马Toll样受体4、核因子-κB、Caspase-3表达的影响

周琴* 张勤 李光乾

解剖学报 ›› 2014, Vol. 45 ›› Issue (3) : 321-327.

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解剖学报 ›› 2014, Vol. 45 ›› Issue (3) : 321-327. DOI: 10.3969/j.issn.0529-1356.2014.03.005
神经生物学

白藜芦醇对惊厥持续状态大鼠海马Toll样受体4、核因子-κB、Caspase-3表达的影响

  • 周琴1*  张勤1 李光乾2
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Effects of resveratrol on the expression of Toll-like receptor 4, NF-кB and Caspase-3 in the hippocampus of the status convulsion rat

  • ZHOU Qin 1*  ZHANG Qin1  LI Guang-qian2
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摘要

目的 观察大鼠惊厥持续状态(SC)后脑组织海马中Toll样受体4(TLR4)、核因子-κB(NF-κB)和半胱氨酸门冬氨酸特异性蛋白酶3(Caspase-3)的表达及神经细胞凋亡的变化,并探讨白藜芦醇(Res)对三者的影响。 方法 106只SD大鼠随机分为对照组(A)、SE组(B)和白藜芦醇干预组(C),其中B组再随机分为B1~B4组(分别于惊厥后4h、24h、48h和72h处死),B组和C组采用氯化锂-匹罗卡品法制作大鼠SC模型,C组在大鼠惊厥终止后30min,给予30 mg/kg白藜芦醇腹腔注射,每天1次,连用3d,于惊厥后72 h处死。免疫组织化学法检测海马TLR4、NF-κB/p65蛋白的表达变化;RT-PCR技术检测海马TLR4、Caspase-3 mRNA表达的动态改变;TUNEL法检测神经细胞凋亡数的变化。 结果 免疫组织化学显示,B组各时间点TLR4蛋白的表达均明显高于A组(P<0.05),并随时间延长明显增高;C组TLR4蛋白表达较B4组显著降低(P<0.01)。B组可见NF-кB/p65蛋白在胞核内有不同程度表达,与A组比较差异有统计学意义(P<0.05)。C组与B4组比较,NF-κB/p65蛋白表达水平明显降低(P<0.01)。RT-PCR显示,B组各时间点TLR4、Caspase-3 mRNA的表达均较A组高(P<0.05),且随时间延长逐步升高,惊厥后72h达高峰;C组海马TLR4、Caspase-3mRNA的表达明显低于B4组(P<0.05)。B组在惊厥24h后海马CA1区TUNEL阳性细胞数已显著高于A组(P<0.01),72h达高峰,而C组TUNEL阳性细胞数较B4组显著下降(P<0.01)。 结论 SC后大鼠海马TLR4、NF-κB/p65和Caspase-3的表达增强。白藜芦醇可下调匹罗卡品致惊厥持续状态大鼠海马TLR4、NF-κB/p65和Caspase-3的表达,并使神经细胞凋亡减少;提示白藜芦醇对SC引起的脑损伤可能有保护作用。

Abstract

Objective To study the expression of toll-like receptor 4 (TLR4), nuclear factor κB (NF-кB) and Caspase-3 and the change of neuron apoptosis of the hippocampus in the status convulsion rat, and to explore the effect of resveratrol on them.Methods A total of 106 male Sprague-Dawley(SD)rats were randomly divided into the control (A), convulsion (B) and resveratrol treatment (C) groups. Group B was further randomly divided into four subset groups (B1-B4) which were executed at 4h, 24h, 48h, 72h after convulsion discontinued. Continuous epilepticus was induced by injecting lithium chloride and pilocarpine, and group C was daily injected with 30mg/kg resveratrol 30minutes after convulsion stopped for 3 days. TLR4 and NF-κB/p65 proteins were detected by immunohistochemistry(IHC); the expressions of TLR4 and Caspase-3 mRNA were detected by RT-PCR. The neuron apoptosis was observed by TUNEL. Results The IHC staining of TLR4 protein in group B was significantly higher than in group A (P<0.05), and that in group C was much lower than in B4 group (P<0.01). The expression of NF-кB/p65 showed that hippocampal neurons had positive expression in cell nuclei in group B compared with group A (P<0.05), and the expression of NF-кB/p65 protein in group C was much lower than that in group B4 (P<0.01). The mRNA expressions of TLR4 and Caspase-3 in the rat hippocampus of group B were significantly elevated than that in group A (P<0.05), and the tendency was increased gradually, reaching the peak at 72 hours after seizure, and that in group C was much lower than that in B4 group (P<0.01). The TUNEL positive cells in hippocampus CAl of B group were more than that of group A after the SC 24hours(P<0.01),reached the highest level at the 72nd hour, and that in group C the TUNEL positive cells were lower than that in B4 group (P<0.01). Conclusion The expression of TLR4, NF-кB and Caspase-3 increased after SC. Resveratrol can down regulate the expression of TLR4, NF-кB and Caspase-3 in the hippocampus with pilocarpine-induced seizures, reduced the number of neuronal apoptosis.These results suggest that resveratrol may have a protective effect against the hippocampus damage caused by status convulsion.

关键词

惊厥 / 凋亡 / Toll样受体4 / Caspase-3 / 白藜芦醇 / 免疫组织化学 / 大鼠

Key words

Convulsion / Apoptosis / Toll like receptor 4 / Caspase-3 / Resveratrol / Immunohistochemistry / Rat

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周琴* 张勤 李光乾. 白藜芦醇对惊厥持续状态大鼠海马Toll样受体4、核因子-κB、Caspase-3表达的影响[J]. 解剖学报. 2014, 45(3): 321-327 https://doi.org/10.3969/j.issn.0529-1356.2014.03.005
ZHOU Qin* ZHANG Qin LI Guang-qian. Effects of resveratrol on the expression of Toll-like receptor 4, NF-кB and Caspase-3 in the hippocampus of the status convulsion rat[J]. Acta Anatomica Sinica. 2014, 45(3): 321-327 https://doi.org/10.3969/j.issn.0529-1356.2014.03.005

参考文献

[1]Li GQ, Wang HP, Jiang ChM. Effects of edravone on the expression of GRP78, Caspase-12 and neuron apoptosis in juvenile rat hippocampus after status convulsivus [J]. Chinese Journal of Pediatrics, 2011, 49(1): 53-59.(in Chinese)
李光乾,王海萍,蒋春明. 惊厥持续状态幼年大鼠海马中葡萄糖调节蛋白78与半胱氨酸门冬氨酸特异性蛋白酶12的表达及依达拉奉对其影响[J]. 中华儿科杂志,2011,49(1): 53-59.
[2]Zhou Q, Li GQ. Role of TLR4/NF-κB signal pathway in the hippocampus injury of status epileptic rats [J]. Chinese Journal of Neurology, 2008, 41(10): 689-694.(in Chinese)
周琴,李光乾.Toll样受体4/核因子-κB信号通路在癫痫持续状态大鼠海马损伤中的作用[J].中华神经科杂志,2008,41(10): 689-694.
[3]Voutsinos-Porche B, Koning E, Kaplan H, et al. Temporal patterns of the cerebral inflammatory response in the rat lithium-pilocarpine model of temporal lobe epilepsy [J]. Neurobiol Dis, 2004, 17(3): 385-402.
[4]Chakravarty S, Herkenham M. Toll-like receptor 4 on nonhematopoietic cells sustains CNS inflammation during endotoxemia, independent of systemic cytokines [J]. J Neurosci, 2005, 25(7): 1788-1796.
[5]Mclaughlin B. The kinder side of killer proteases: caspase activation contributes to neuroprotection and CNS remodeling [J]. Apoptosis, 2004, 9(2): 111-121.
[6]Crespel A, Coubes P, Rousset MC, et al. Inflammatory reaction in human medial temporal lobe epilepsy with hippocampal sclerosis [J]. Brain Res, 2002, 952(2):159-169.
[7]Ruckdeschel K, Pfaffinger G, Haase R, et al. Signaling of apoptosis through TLRs critically involves Toll/IL-1 receptor domain-containing adapter inducing IFN-β but not MyD88, in bacteria-infected murine macrophages [J]. J Immunol, 2004, 173(5): 3320-3328.
[8]Hsu LC, Park JM, Zhang K, et al. The protein kinase PKR is required for macrophage apoptosis after activation of Toll-like receptor 4 [J]. Nature, 2004, 428(6980): 341-345.
[9]Jung DY, Lee H, Jung BY, et al. TLR4, but not TLR2, signals autoregulatory apoptosis of cultured microglia: a critical role of IFN-beta as a decision maker [J]. J Immunol, 2005, 174(10): 6467-6476.
[10]Han YS, Zheng WH, Bastianetto S, et al. Neuroprotective effects of resveratrol against beta-amyloid-induced neurotoxicity in rat hippocampal neurons: involvement of protein kinase C [J]. Br J Pharmacol 2004, 141(6):997-1005.
[11]Li M, Wang QS, Chen Y, et al. Resveratrol inhibits neuronal discharges in rat hippocampal CA1 area [J]. Sheng Li Xue Bao, 2005, 57(3): 355-360. 
[12]Xu J, Wang XY, Yang TF, et al. Protective effect of polydatin on NF-κB expression in rat cortical neurons during cerebral ischem ia-reperfusion injury in vitro [J]. Acta Academiae Medicinae Militaris Tertiae, 2009, 31(2): 113-116. (in Chinese)
徐静,王兴勇,杨廷芳,等. 白藜芦醇苷对体外缺血再灌注脑损伤中NF-κB的保护作用[J]. 第三军医大学学报, 2009, 31(2): 113-116.

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白藜芦醇对惊厥持续状态大鼠海马TLR4/NF-кB信号通路的调控


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