缺氧预处理对颅脑外伤大鼠皮层TrxR2、CHOP表达及神经元细胞超微结构改变的影响

doi:10.3877/cma.j.issn.2095-9141.2015.03.010

目的

探讨缺氧预处理对颅脑外伤大鼠皮层TrxR2、CHOP 表达及神经元细胞超微结构改变的影响。

方法

48 只SD 大鼠随机分成对照(Con)组、缺氧预处理(HPC)组、单纯外伤(TBI)组和缺氧预处理颅脑外伤(HPCT)组。采用改良的Feeney’s自由落体装置制作颅脑损伤大鼠模型，预先在低压氧舱内连续处理3 d(-50 kPa、3 h/d)制作缺氧预处理模型。采用HE染色法和新鲜标本制作超薄切片，分别在光镜和电镜下观察各组大鼠皮层脑组织的大体结构和神经元细胞超微结构改变。qRT-PCR 和Western blotting 法检测挫伤区周围皮层TrxR2、CHOP mRNA 和蛋白的表达变化。

结果

Con组和HPC组病理改变无明显差异，HPCT组较TBI组大鼠病理改变轻，神经元细胞超微结构受损明显减轻。HPCT 与TBI 组比较，TrxR2 mRNA 和蛋白表达显著上调，CHOP mRNA和蛋白表达显著下调，差异均具有统计学意义(均P＜0.05)；Con与HPC组比较，差异无统计学意义(P>0.05)。

结论

缺氧预处理可明显减轻颅脑外伤大鼠皮层神经元细胞形态学改变，这可能与上调TrxR2、下调CHOP 的表达有密切关系，从而对神经元细胞起保护作用。

关键词: 颅脑外伤, 缺氧预处理, TrxR2, CHOP, 超微结构

Objective

To investigate the effects of hypoxic preconditioning on the expression of TrxR2、CHOP and the ultra-structure of the neuronal cells of rats suffered traumatic brain injury.

Methods

Forty eight Sprague Dawley rats were randomly divided into control group (Con), hypoxic preconditioning group (HPC), traumatic brain injury group (TBI) and hypoxic preconditioning traumatic brain injury group (HPCT). The traumatic brain injury model of rats were made by the Feeney’s improved equipment and hypoxic preconditioning model were made by Hypobaric chamber for successive 3 days (-50 kPa、3 h/d). Staining HE and making ultra-thin slicing by fresh specimen to observe the potential change of general structure and ultra-structure of rats’ brain tissue of each group respectively under light microscope and electron microscope.Expressions of TrxR2、CHOP mRNA and proteins were detected in the brain cortex around the contusion area by qRT-PCR and Western blotting.

Results

The pathologic change has no difference between Con gourp and HPC group. HPCT group has a lighter pathologic change than the TBI group.The expressions of TrxR2 mRNA and protein was significantly higher in the brain cortex in the HPCT group than the TBI group, the difference between the two group was significant(P＜0.05),the expressions of CHOP mRNA and protein was significantly lower in the brain cortex in the HPCT group than the TBI group, the difference between the two group was significant (P＜0.05), while the difference between the Con group and the HPC group was not significant (P>0.05).

Conclusion

Hypoxia preconditioning can obviously reduce the craniocerebral trauma rats cortex neuron cell morphology change,which has a close relationship with the upregulated expression of TrxR2 and the downregulated expression of CHOP,in order to protection of neurons.

Key words: Traumatic brain injury, Hypoxic preconditioning, TrxR2, CHOP, Ultra-structure

图1 各处理组大鼠HE染色(×400) A、B为Con组，C、D为TBI组，E、F为HPC组，G、H为HPCT组；A、C、E、G：HE染色×100；B、D、F、H：HE染色×400；A、B和E、F：脑组织形态，结构完整，C、D与G、H：脑组织形态结构不完整，大量炎细胞浸润，细胞核固缩，胞质空泡变；HPCT组较TBI组损伤轻。

图2 各组大鼠挫伤区周围皮层神经元细胞电镜观察(×1万倍) A:假手术（Con）组，神经元细胞微细结构完整，线粒体、内质网结构形态完整；B:缺氧预处(HPC)组，与Con组改变相似；C:单纯外伤组(TBI组)，神经元细胞受损严重，线粒体肿胀、内质网结构形态异常；D：缺氧预处理颅脑外伤(HPC)组，神经元受损较TBI组轻，线粒体、内质网轻度受损

表1 各组大鼠皮层TrxR2、CHOP mRNA和蛋白表达(n=6，±s)

分组	鼠数(例)	TrxR2		CHOP
分组	鼠数(例)	mRNA	蛋白	mRNA	蛋白
Con组	12	1.01±0.01	0.19±0.00	0.42±0.08	0.35±0.01
HPC组	12	1.02±0.01	0.19±0.00	0.38±0.08	0.35±0.01
TBI组	12	3.92±0.12^a	0.73±0.01^a	1.02±0.01^a	0.67±0.01^a
HPCT组	12	4.03±0.14^ab	0.78±0.03^ab	0.95±0.02^ab	0.66±0.01^ab
F值		2136.000	2129.000	220.500	2209.000
P值		0.000	0.000	0.000	0.000

表1 各组大鼠皮层TrxR2、CHOP mRNA和蛋白表达(n=6，±s)

分组	鼠数(例)	TrxR2		CHOP
分组	鼠数(例)	mRNA	蛋白	mRNA	蛋白
Con组	12	1.01±0.01	0.19±0.00	0.42±0.08	0.35±0.01
HPC组	12	1.02±0.01	0.19±0.00	0.38±0.08	0.35±0.01
TBI组	12	3.92±0.12^a	0.73±0.01^a	1.02±0.01^a	0.67±0.01^a
HPCT组	12	4.03±0.14^ab	0.78±0.03^ab	0.95±0.02^ab	0.66±0.01^ab
F值		2136.000	2129.000	220.500	2209.000
P值		0.000	0.000	0.000	0.000

图3 qRT-PCR检测结果 TBI与Con组，HPCT与HPC组比较，^aP＜0.05；与TBI组比较，^bP＜0.05。

图4 TrxR2、CHOP蛋白电泳条带

[1]	Hemphill MA, Dauth S, Yu CJ, et al. Traumatic Brain Injury and the Neuronal Microenvironment: A Potential Role for Neuropathological Mechanotransduction[J]. Neuron, 2015, 85(6):1177-1192.
[2]	Yokobori S, Mazzeo AT, Hosein K, et al. Preconditioning for traumatic brain injury[J].Transl Stroke Res,2013,4(1):25-39.
[3]	疏龙飞,刘家传,王金标,等.硫氧还蛋白还原酶TrxR2 在颅脑损伤大鼠皮质的表达变化[J].中国微侵袭神经外科杂志,2014,19(11):517-519.
[4]	Oyadomari S, Mori M. Roles of CHOP/GADD153 in endoplasmicreticulumstress[J].CellDeathand Differentiation,2004,11:381-389.
[5]	武孝刚,刘家传,杨艳艳,等.缺氧预处理对创伤性脑损伤大鼠缺氧诱导因子-1α、葡萄糖转运体3型及神经元核蛋白表达的影响[J].中华神经创伤外科电子杂志,2015,4(1):11-15.
[6]	Meyer Y, Buchanan BB, Vignols F, et al. Thioredoxins and glutare-doxins: unifying elements in redox biology[J]. Annual Review of Genetics,2009,43:335-367.
[7]	Seiji Tajiria, Shigetoshi Yano, Motohiro Morioka, et al. CHOP is involved in neuronal apoptosis induced by neurotrophic factor deprivation[J].FEBS Letters,2006,580:3462-3468.
[8]	Liu Y, Sun Z, Sun S, et al. Effects of hypoxic preconditioning on synaptic ultrastructure in mice[J]. Synapse, 2015, 69(1):7-14.
[9]	Galle A, N.M.Jones. The neuroprotective actions of hypoxic preconditioning and postconditioning in a neonatal rat model of hypoxic-ischemic brain injury[J].brain Res,2013,1498:1-8.
[10]	Liamina NP, Karpova ES, Kotel'nikova EV, et al.Adaptation to hypoxia and ischemic preconditioning: from basic research to clinical practice[J].Klin Med,2014,92(2):23-29.

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Effects of hypoxic preconditioning on cortical expression of TrxR2 and CHOP of rats with craniocerebral injury and ultramicrostructural changes of neurons

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Effects of hypoxic preconditioning on cortical expression of TrxR2 and CHOP of rats with craniocerebral injury and ultramicrostructural changes of neurons