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表儿茶酸对脑损伤小鼠的神经修复作用及机制探讨

The neural function repair and mechanism of (-)-epicatechin on murine traumatic brain injury

摘要:

目的 探讨表儿茶酸[(-)-epicatechin,EC]对脑损伤(traumatic brain injury,TBI)小鼠的神经修复作用及机制.方法 建立小鼠自由落体脑外伤动物模型,将50只C57BL/6小鼠随机(随机数字法)分为脑损伤+生理盐水(Veh)组,脑损伤+表儿茶酸(EC)组(EC组和Veh组各死亡1只,每组24只).术后3d,采用ELISA方法检测炎症因子IL-1β和TNF-α的表达,PI染色检测分析损伤区坏死细胞数;水迷宫实验观察术后28 d小鼠空间学习记忆能力,qRT-PCR检测神经营养因子BDNF和NGF的表达,免疫荧光染色检测NeuN的表达,EdU染色观察SGZ区神经再生.结果 与Veh组相比较,EC治疗组明显改善小鼠的空间学习记忆能力,平台象限停留时间增加[24 d:(26.333±5.037)% vs (26.583±5.802)%,P=0.938;25 d:(33.300±4.724)% vs (29.767±3.347)%,P=0.166;26 d:(41.017±7.246)% vs (32.800±8.145)%,P=0.0951;27 d:(48.017±7.424)% vs(35.267±6.748)%,P=0.011,28 d:(51.617±9.017)% vs (41.116±6.467)%,P=0.043], 逃逸潜伏期时间[24 d:(62.967±5.494)s vs (63.917±7.027)s,P=0.800,25 d:(50.533±10.305)s vs(57.217±13.085)s,P=0.349,26 d:(40.333±10.526)s vs (50.133±11.039)s,P=0.147,27 d:(28.717±4.137)s vs (44.533±7.181)s,P=0.001,28 d:(21.950±6.889)s vs (37.567±5.974)s,P=0.002],炎症因子IL-1β和TNF-α表达量降低[IL-1β(42.690±3.057) ng/mL vs (750.167±51.941)ng/mL,TNF-α(71.670±4.996) ng/mL vs(1 085.167±68.535) ng/mL,P=0.000 6,0.003)],神经营养因子BDNF和神经生长因子NGF的表达增加,损伤区域坏死细胞数显著降低[(54.833±5.486)个vs (74.000±5.323)个,P=0.031],成熟的神经元数量增多[(76.667±6.386)个vs (42.167±5.237)个,P=0.002],SGZ区新生细胞数增多[(12.667±0.760)个vs (7.500±1.258)个,P=0.031].结论 表儿茶酸促进脑损伤小鼠神经功能重塑,与抑制炎性反应、促进神经营养因子分泌和促神经再生密切相关.

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abstracts:

Objective To investigate whether (-)-epicatechin plays a role in neurological repair on traumatic brain injury in mice.Methods The mice model of traumatic brain injury was established by modified weight drop method.Experimental mice were randomly (random number) divided into the injury+ (Veh) group and the injury + (-)-epicatechin (EC) group.At 3 days after operation,the expression of IL-1β and TNF-α were detected.The number of necrotic cells of the lesion area was detected by PI staining.At 28 days after SCI,Morris Water Maze test was performed to observe the ability of spatial learning and memory in mice.The expression of neurotrophic factors BDNF and NGF were examined by qRT-PCR.The expression of NeuN was detected by immunofluorescence staining.EdU staining was used to observe the neurogenesis in the SGZ region.Results Compared with the Veh group,EC treated group showed better spatial learning and memory ability in time spent in correct quadrant at day 27 and 28 [24 d:(26.333±5.037)% vs (26.583±5.802)%,P=0.938;25 d (33.300±4.724)% vs (29.767±3.347)%.P=0.166;26 d:(41.017±7.246)% vs (32.800±8.145)%,P=0.095;27 d:(48.017±7.424)% vs (35.267±6.748)%,P=0.011;28 d:(51.617±9.017)% vs (41.116±6.467)%,P=0.043] and in latency to platform at day 27 and 28 [24 d:(62.967±5.494) s vs (63.917±7.027) s,P=0.800;25 d:(50.533±10.305) s vs (57.217±13.085) s,P=0.349;26 d:(40.333± 10.526) s vs (50.133±11.039) s,P=0.147;27 d:(28.717±4.137) s vs (44.533±7.181) s,P=0.001;28 d:(21.950±6.889) s vs (37.567±5.974) s,P=0.002].There was a decreased expression of IL-lβ and TNF-α and increased level of neurotrophic factor BDNF and NGF after EC treatment in EC treatment group,compared to the veh treatment group [IL-1β and TNF-α:(42.690±3.057) ng/mL and (750.167±51.941) ng/mL vs (71.670±4.996) ng/mL and (1 085.167±68.535) ng/mL,P=0.000 6 and 0.003;BDNF and NGF:0.543±0.033 and 0.334±0.041 vs 0.756±0.088 and 0.514±0.047,P=0.048 and 0.017)].EC decreased the cell death near injury area (54.833±5.486 vs 74.000±5.323,P=0.031),increased NeuN positive cells (76.667±6.386 vs 42.167±5.237,P=0.002),and increased neurogenesis in SGZ area (12.667±0.760 vs 7.500±1.258,P=0.031).Conclusions (-)-Epicatechin plays an important role in functional recovery after traumatic brain injury in mice.The underlying mechanisms are closely related to inhibited inflammation,enhanced neurotrophic factors and improved neurogenesis.

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作者: 程田 [1] 刘雯雯 [2] 马珊珊 [2] 孙同文 [3] 关方霞 [4]
作者单位: 郑州大学第一附属医院骨科 450052 [1] 郑州大学生命科学院 450000 [2] 郑州大学第一附属医院 ICU 450052 [3] 郑州大学生命科学院 450000;郑州大学第一附属医院 ICU 450052 [4]
期刊: 《中华急诊医学杂志》2019年28卷5期 567-573页 ISTICPKUCSCD
栏目名称: 创伤
DOI: 10.3760/cma.j.issn.1671-0282.2019.05.006
发布时间: 2019-06-11
基金项目:
国家自然科学青年基金 河南省科技厅科技攻关项目 河南省高等学校重点科研项目 郑州大学第一附属医院基金(YNQN2017042)Youth National Natural Science Foundation of China Science and Technology Research Project of Henan Province Key Scientific Research Projects of High School in Henan Province The Grant of the First Affiliated Hospital of Zhengzhou University
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