PDF(2932 KB)
PDF(2932 KB)
PDF(2932 KB)
组蛋白乙酰化调控对新生大鼠大脑皮层损伤的保护作用
Protective effect of histone acetylation against cortical injury in neonatal rats
目的 研究组蛋白乙酰化调控对新生大鼠缺氧缺血大脑皮层损伤的保护作用。方法 将90只3日龄新生大鼠分为假手术组、缺氧缺血模型组以及组蛋白去乙酰化酶抑制剂丁酸钠组。模型组及丁酸钠组大鼠腹腔注射LPS（0.05 mg/kg）2 h后行右侧颈总动脉结扎并置于低氧舱中（6.5%氧浓度）90 min；假手术组腹腔注射生理盐水后仅分离暴露右侧颈总动脉，但不予结扎和低氧处理；丁酸钠组在建模后立即腹腔注射丁酸钠（300 mg/kg），每天1次，共治疗7 d，假手术组及模型组注射等体积生理盐水。模型建立后第7天，采用Western blot方法检测各组大脑皮层组蛋白H3（HH3），乙酰化组蛋白H3（AH3），B细胞淋巴瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）、活化型半胱天冬酶-3（CC3），以及脑源性神经营养因子（BDNF）蛋白表达；采用免疫荧光法检测皮层细胞增殖指标BrdU表达。结果 丁酸钠组HH3/AH3比值显著低于模型组（P < 0.05），提示丁酸钠组HH3乙酰化程度增加；丁酸钠组凋亡相关蛋白Bcl-2/Bax比值较模型组显著增高（P < 0.05），CC3表达显著减少（P < 0.05），BDNF表达显著增加（P < 0.05）；丁酸钠组较模型组皮层BrdU阳性细胞数显著增加（P < 0.05），主要表达在神经元。结论 组蛋白乙酰化水平增加可通过减少细胞凋亡及促进神经元再生保护新生大鼠大脑皮层损伤，其机制可能与BDNF表达增加相关。
Objective To investigate the protective effect of histone acetylation against hypoxic-ischemic cortical injury in neonatal rats. Methods A total of 90 neonatal rats aged 3 days were divided into three groups: sham-operation, cortical injury model, and sodium butyrate (a histone deacetylase inhibitor) treatment. The rats in the model and the sodium butyrate treatment groups were intraperitoneally injected with lipopolysaccharide (0.05 mg/kg), and then right common carotid artery ligation was performed 2 hours later and the rats were put in a hypoxic chamber (oxygen concentration 6.5%) for 90 minutes. The rats in the sham-operation group were intraperitoneally injected with normal saline and the right common carotid artery was only separated and exposed without ligation or hypoxic treatment. The rats in the sodium butyrate treatment group were intraperitoneally injected with sodium butyrate (300 mg/kg) immediately after establishment of the cortical injury model once a day for 7 days. Those in the sham-operation and the model groups were injected with the same volume of normal saline. At 7 days after establishment of the model, Western blot was used to measure the protein expression of histone H3 (HH3), acetylated histone H3 (AH3), B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated X protein (BAX), cleaved caspase-3 (CC3), and brain-derived neurotrophic factor (BDNF). Immunofluorescence assay was used to measure the expression of 5-bromo-2'-deoxyuridine (BrdU) as the cortex cell proliferation index. Results The sodium butyrate treatment group had a significantly lower HH3/AH3 ratio than the model group (P < 0.05), which suggested that the sodium butyrate treatment group had increased acetylation of HH3. Compared with the model group, the sodium butyrate treatment group had a significant increase in Bcl-2/Bax ratio, a significant reduction in CC3 expression, and a significant increase in BDNF expression (P < 0.05). The sodium butyrate treatment group had a significant increase in the number of BrdU-positive cells in the cortex compared with the model group (P < 0.05), and BrdU was mainly expressed in the neurons. Conclusions Increased histone acetylation may protect neonatal rats against cortical injury by reducing apoptosis and promoting regeneration of neurons. The mechanism may be associated with increased expression of BDNF.
组蛋白乙酰化调控 / 组蛋白去乙酰化酶 / 脑源性神经营养因子 / 新生大鼠
Histone acetylation / Histone deacetylase / Brain-derived neurotrophic factor / Neonatal rats
[1] Volpe JJ. Brain injury in the premature infant:overview of clinical aspects, neuropathology, and pathogenesis[J]. Semin Pediatr Neurol, 1998, 5(3):135-151.
[2] Eaton-Rosen Z, Melbourne A, Orasanu E, et al. Longitudinal measurement of the developing grey matter in preterm subjects using multi-modal MRI[J]. Neuroimage, 2015, 111:580-589.
[3] Zhang H, Shang YP, Chen HY, et al. Histone deacetylases function as novel potential therapeutic target for cancer[J]. Hepatol Res, 2016.[Epub ahead of print].
[4] Wang ZF, Fessler EB, Chuang DM. Beneficial effects of mood stabilizers lithium, valproate and lamotrigine in experimental stroke models[J]. Acta Pharmacol Sin, 2011, 32(12):1433-1445.
[5] Shein NA, Shohami E. Histone deacetylase inhibitors as therapeutic agents for acute central nervous system injuries[J]. Mol Med, 2011, 17(5-6):448-456.
[6] Chuang DM, Leng Y, Marinova Z, et al. Multiple roles of HDAC inhibition in neurodegenerative conditions[J]. Trends Neurosci, 2009, 32(11):591-601.
[7] Langley B, Brochier C, Rivieccio MA. Targeting histone deacetylases as a multifaceted approach to treat the diverse outcomes of stroke[J]. Stroke, 2009, 40(8):2899-2905.
[8] Ziemka-Nalecz M, Zalewska T. Neuroprotective effects of histone deacetylase inhibitors in brain ischemia[J]. Acta Neurobiol Exp (Wars), 2014, 74(4):383-395.
[9] Volpe JJ, Kinney HC, Jensen FE, et al. The developing oligodendrocyte:key cellular target in brain injury in the premature infant[J]. Int J Dev Neurosci, 2011, 29(4):423-440.
[10] Vincer MJ, Allen AC, Allen VM, et al. Trends in the prevalence of cerebral palsy among very preterm infants (< 31 weeks' gestational age)[J]. Paediatr Child Health, 2014, 19(4):185-189.
[11] Chenouard A, Gascoin G, Gras-Le Guen C, et al. Neurodevelop-mental impairment in preterm infants with late-onset infection:not only in extremely preterm infants[J]. Eur J Pediatr, 2014, 173(8):1017-1023.
[12] Yang D, Sun YY, Lin X, et al. Taming neonatal hypoxic-ischemic brain injury by intranasal delivery of plasminogen activator inhibitor-1[J]. Stroke, 2013, 44(9):2623-2627.
[13] Wang X, Svedin P, Nie C, et al. N-acetylcysteine reduces lipopolysaccharide-sensitized hypoxic-ischemic brain injury[J]. Ann Neurol, 2007, 61(3):263-271.
[14] Dean JM, Moravec MD, Grafe M, et al. Strain-specific differences in perinatal rodent oligodendrocyte lineage progression and its correlation with human[J]. Dev Neurosci, 2011, 33(3-4):251-260.
[15] Wang LW, Chang YC, Lin CY, et al. Low-dose lipopolysac-charide selectively sensitizes hypoxic ischemia-induced white matter injury in the immature brain[J]. Pediatr Res, 2010, 68(1):41-47.
[16] Fessler EB, Chibane FL, Wang Z, et al. Potential roles of HDAC inhibitors in mitigating ischemia-induced brain damage and facilitating endogenous regeneration and recovery[J]. Curr Pharm Des, 2013, 19(28):5105-5120.
[17] Yang PM, Tseng HH, Peng CW, et al. Dietary flavonoid fisetin targets caspase-3-deficient human breast cancer MCF-7 cells by induction of caspase-7-associated apoptosis and inhibition of autophagy[J]. Int J Oncol, 2012, 40(2):469-478.
[18] 王雪维, 李燕. Bcl-2、Bax在青光眼神经节细胞凋亡中的研究进展[J]. 医学综述, 2015, 21(22):4077-4079.
[19] Siddiqui WA, Ahad A, Ahsan H. The mystery of BCL2 family:Bcl-2 proteins and apoptosis:an update[J]. Arch Toxicol, 2015, 89(3):289-317.
[20] Kim G, Kim E. The effects of antecedent exercise on motor function recovery and brain-derived neurotrophic factor expression after focal cerebral ischemia in rats[J]. J Phys Ther Sci, 2013, 25(5):553-556.
[21] Wang NQ, Wang LY, Zhao HP, et al. Luoyutong treatment promotes functional recovery and neuronal plasticity after cerebral ischemia-reperfusion injury in rats[J]. Evid Based Complement Alternat Med, 2015, 2015:369021.
[22] Kim HJ, Leeds P, Chuang DM. The HDAC inhibitor, sodium butyrate, stimulates neurogenesis in the ischemic brain[J]. J Neurochem, 2009, 110(4):1226-1240.
[23] Heiser JH, Schuwald AM, Sillani G, et al. TRPC6 channel-mediated neurite outgrowth in PC12 cells and hippocampal neurons involves activation of RAS/MEK/ERK, PI3K, and CAMKIV signaling[J]. J Neurochem, 2013, 127(3):303-313.
国家自然科学基金（81330016；81630038；81270724；81300526）；四川省科技厅基金（2014SZ0149；2016TD0002）；国家临床重点专科（儿科新生儿专业）建设项目（1311200003303）。
