丁苯酞对溴化乙锭诱导的脱髓鞘大鼠胼胝体Bcl-2和Bax表达的影响
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摘要
目的观察丁苯酞(Dl-3-n-butylphthalide,NBP)对溴化乙锭(ehidium bromide,EB)诱导的脱髓鞘大鼠脑胼胝体凋亡因子B淋巴细胞瘤2基因(Bcl-2)和Bcl-2相关X蛋白(Bax)表达水平的影响,并探讨NBP对脱髓鞘大鼠脑白质保护作用的可能机制。方法将18只SD大鼠随机分为模型组、治疗组、对照组,每组各6只,给予模型组及治疗组大鼠侧脑室注射EB制备脱髓鞘模型,对照组以相同方法注射等体积生理盐水。造模后,治疗组每天经腹腔注射NBP注射液,模型组及对照组腹腔注射等体积生理盐水,10 d后取脑组织制作石蜡切片,行HE染色、LFB髓鞘染色观察大鼠侧脑室周围胼胝体脱髓鞘情况,免疫组化法检测侧脑室周围胼胝体Bcl-2、Bax表达的情况。结果模型组HE染色示胼胝体结构不清、完整性破坏、大量空泡形成,LFB染色示大片区域髓鞘未被蓝染(提示脱髓鞘),与模型组相比,治疗组胼胝体空泡形成较少,髓鞘脱失率明显降低(17.36%vs. 42.05%,P<0.05);免疫组化结果显示,治疗组胼胝体Bcl-2阳性表达率较模型组明显增高(69.47%vs. 29.95%,P<0.05),Bax阳性细胞表达率明显降低(33.52%vs. 68.61%,P<0.05)。结论 NBP可对脱髓鞘大鼠脑白质产生保护作用,减少髓鞘的脱失,其机制可能与上调脑Bcl-2、下调Bax的表达有关。
Objective To observe the effect of Dl-3-n-butylphthalide(NBP) on the expression of apoptosis factors bcl-2 and Bax in corpus callosum of Ethidium bromide(EB) induced demyelinating rats, and to investigate possible mechanisms of protective effect of NBP on white matter in demyelinated rats. Methods 18 SD rats were randomly divided into three groups, namely, a model, a treated, and a control group. All the rats except those from the control group received intracerebroventricular injection with EB, whereas the control group was injected with the same volume of normal saline. NBP was injected intraperitoneally every day in the treated group. The brain tissue was harvested 10 days later, and demyelination of corpus callosum around the lateral ventricle of rats was observed by HE staining and LFB myelin staining, the levels of Bcl-2 and Bax were determined using immunohistochemistry. Results Compared with the model group, the rate of demyelination in the treatment group was significantly decreased(17.36% vs. 42.05%, P<0.05), and immunohistochemical results showed that the positive expression rate of bcl-2 in the corpus callosum of the treatment group was significantly increased(69.47% vs. 29.95%, P<0.05), and the expression rate of Bax was significantly decreased(33.52% vs. 68.61%, P<0.05). Conclusions NBP can protect the white matter of demyelinating rats and reduce the loss of myelin sheath, which may be related to the up-regulation of bcl-2 and down-regulation of Bax.
Objective To observe the effect of Dl-3-n-butylphthalide(NBP) on the expression of apoptosis factors bcl-2 and Bax in corpus callosum of Ethidium bromide(EB) induced demyelinating rats, and to investigate possible mechanisms of protective effect of NBP on white matter in demyelinated rats. Methods 18 SD rats were randomly divided into three groups, namely, a model, a treated, and a control group. All the rats except those from the control group received intracerebroventricular injection with EB, whereas the control group was injected with the same volume of normal saline. NBP was injected intraperitoneally every day in the treated group. The brain tissue was harvested 10 days later, and demyelination of corpus callosum around the lateral ventricle of rats was observed by HE staining and LFB myelin staining, the levels of Bcl-2 and Bax were determined using immunohistochemistry. Results Compared with the model group, the rate of demyelination in the treatment group was significantly decreased(17.36% vs. 42.05%, P<0.05), and immunohistochemical results showed that the positive expression rate of bcl-2 in the corpus callosum of the treatment group was significantly increased(69.47% vs. 29.95%, P<0.05), and the expression rate of Bax was significantly decreased(33.52% vs. 68.61%, P<0.05). Conclusions NBP can protect the white matter of demyelinating rats and reduce the loss of myelin sheath, which may be related to the up-regulation of bcl-2 and down-regulation of Bax.
引文
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[13] 董雅洁, 高维娟. Bcl-2、bax、caspase-3在细胞凋亡中的作用及其关系[J]. 中国老年学杂志, 2012, 32(21):4828-4830.
[2] Wang W, Cha XX , Reiner J, et al. Synthesis and biological activity of n-butylphthalide derivatives[J]. Eur J Med Chem, 2010, 45(5):1941-1946.
[3] Ali AI, Jing GY. A review of recent advances in neuroprotective potential of 3-N-butylphthalide and its derivatives[J]. Biomed Res Int, 2016,2016:5012341.
[4] Wang YG, Li Y, Wang CY, et al. L-3-n-butylphthalide protects rats' cardiomyocytes from ischaemia/reperfusion-induced apoptosis by affecting the mitochondrial apoptosis pathway[J]. Acta Physiologica, 2014, 210(3):524-533.
[5] Li J, Li Y, Ogle M, et al. dl-3-n-Butylphthalide prevents neuronal cell death after focal cerebral ischemia in mice via the JNK pathway[J]. Brain Res, 2010,1359:216-226.
[6] Wu Y, Huang Q, Liu X, et al. Dl-3-n-butylphthalide is effective for demyelination:A case-combined study[J]. Clin Neurol Neurosurg, 2015,137:83-88.
[7] 包新民, 舒斯云.大鼠脑立体定位图谱[M].北京:人民卫生出版社,1991:1-35.
[8] Brück W, Stadelmann C. The spectrum of multiple sclerosis:new lessons from pathology[J]. Curr Opin Neurol, 2005, 18(3):221-224.
[9] Caprariello AV, Mangla S, Miller RH, et al. Apoptosis of oligodendrocytes in the central nervous system results in rapid focal demyelination[J]. Ann Neurol, 2012, 72(3):395.
[10] 吕静波, 陈强. 少突胶质细胞凋亡与脑白质脱髓鞘病变发病机制关系的研究进展[J]. 浙江医学, 2014, 36(23):1965-1968.
[11] 许茸茸, 李应东. Bcl-2家族与线粒体凋亡通路相互作用研究进展[J]. 中国老年学杂志, 2013, 33(12):2977-2979.
[12] Lindsay J , Esposti MD , Gilmore AP . Bcl-2 proteins and mitochondria—Specificity in membrane targeting for death[J]. BBA-Mol Cell Res, 2011, 1813(4):532-539.
[13] 董雅洁, 高维娟. Bcl-2、bax、caspase-3在细胞凋亡中的作用及其关系[J]. 中国老年学杂志, 2012, 32(21):4828-4830.