硫酸镁对先兆子痫大鼠病理模型子代海马神经细胞的保护作用

doi:10.3877/cma.j.issn.1673-5250.2015.01.007

目的

探讨硫酸镁(MgSO₄)对先兆子痫(PE)大鼠病理模型子代(胎鼠)海马神经细胞的保护作用。

方法

选择确定妊娠的18只健康Sprague-Dawley(SD)大鼠为研究对象，按照不同处理方式，随机将其分为3组：模型组(n＝6，空白对照)、治疗组(n＝6，使用MgSO₄治疗2 d的PE病理模型大鼠)、对照组(n＝6，正常妊娠大鼠)。模型组和治疗组大鼠均于孕龄为14 d时，采用微量输液泵经尾静脉缓慢注入1.0 g/kg脂多糖溶液诱导建立PE大鼠病理模型，治疗组大鼠于终止妊娠前2 d使用MgSO₄治疗，60～120 mg/(kg·d)×2 d，模型组大鼠不接受任何治疗。对照组大鼠仅注射2 mL生理盐水。3组大鼠均于孕龄为20 d时行剖宫产。采用TUNEL法检测胎鼠脑组织神经细胞凋亡情况，实时荧光定量(RT)-聚合酶链反应(PCR)和免疫组化法测定海马组织中半胱氨酸天冬氨酸蛋白酶(caspase)-3的相对表达水平。

结果

①3组大鼠于孕龄为13 d时收缩压比较，差异无统计学意义(F＝2.876，P>0.05)。模型组和治疗组大鼠孕龄为15 d时收缩压较对照组升高，并且差异有统计学意义(P<0.05)。孕龄为19 d时治疗组收缩压较模型组降低，并且差异有统计学意义(P<0.05)。②3组胎鼠脑海马组织CA1区神经细胞凋亡比较，差异有统计学意义(F＝1 931.4，P<0.05)。模型组和治疗组胎鼠脑海马组织CA1区凋亡神经细胞数较对照组明显增高，并且差异均有统计学意义(P<0.05)；治疗组凋亡神经细胞数较模型组减少，并且差异有统计学意义(P<0.05)。③3组胎鼠脑海马组织中caspase-3 mRNA相对表达水平比较，差异有统计学意义(F＝1 687.13，P＝0.000)。模型组和治疗组胎鼠海马组织中caspase-3 mRNA相对表达水平均较对照组升高，并且差异有统计学意义(P<0.05)，治疗组caspase-3 mRNA相对表达水平较模型组下降，并且差异有统计学意义(P<0.05)。

结论

常规剂量MgSO₄在PE胎鼠脑组织中发挥神经保护作用，其作用机制可能与胎鼠大脑caspase-3活性调控相关。

关键词: 先兆子痫, 海马, 凋亡, MgSO₄, 半胱氨酸天冬氨酸蛋白酶3

Objective

To explore the effects of magnesium sulfate (MgSO₄) on the hippocampal neurons of fetal rats which were delivered by pre-eclampsia (PE) experimental model rats.

Methods

A total of 18 pregnant Sprague-Dawley (SD) rats were included into this study. They were divided into 3 groups according to different treatment methods: model group (n=6, PE experimental model group without any treatment), treatment group (n=6, 2-day MgSO₄ treatment of PE experimental model rats) and control group (normal pregnant rats). Model group and treatment group were given low dose lipopolysaccharides (1 μg/kg) to induce experimental rats model of PE. Treatment group were managed by MgSO₄ 60~120 (mg/kg·d) for 2 days before the caesarean birth at the 20th day of pregnancy. Hippocampal neurons in CA1 region were analyzed for apoptosis by TUNEL assay. Relative expression levels of cysteine aspartic proteases (caspase)-3 of hippocampus in fetal rats were detected by real-time (RT)-PCR and immunohistochemistry.

Results

①There was no significant difference on systolic blood pressure at 13th day during pregnancy among three groups (F=2.876, P>0.05). The systolic blood pressure of model group and treatment group were higher than that of control group on the 15th day during the pregnancy, with significant differences (P<0.05), and the systolic blood pressure of treatment group was lower than than of model group on 19th day during the pregnancy with significant difference (P<0.05). ②There was no significant difference on neuronal apoptosis of CA1 region of hippocampus in rat brains among three groups (F=1 931.4, P<0.05). The number of neuronal apoptosis of CA1 region of neuronal apoptosis in rat brains of model group and treatment group were higher than that of control group (P<0.05), and the number of neuronal apoptosis of CA1 region of neuronal apoptosis in rat brains of treatment group was lower than that of model group (P<0.05). ③ There was no significant difference on the relative expression levels of caspase-3 mRNA of hippocampus in rat brains among three groups (F=1 687.13, P=0.000). The relative expression of caspase-3 mRNA of hippocampus in rat brains of model group and treatment group were higher than that of control group (P<0.05), and the relative expression of caspase-3 mRNA of hippocampus in rat brains of treatment group was lower than that of control group (P<0.05).

Conclusions

Administration of regular dose of MgSO₄ treatment may play a neuroprotective role in the brain of fetal rats who were delivered by experimental rats model of PE and its mechanism may be related to the regulation of fetal brain caspase-3.

Key words: Pre-eclampsia, Hippocampus, Apoptosis, Magnesium sulfate, Caspase 3

表1 3组大鼠孕期各时间点收缩压变化情况比较(mmHg, ±s)

Table 1 Comparison of the blood pressure change at different time points during pregnancy among three groups (mmHg, ±s)

组别	例数	13 d	15 d	19 d
模型组	6	111.4±4.9	141.2±8.2^a	139.2±9.1^c，d
治疗组	6	109.1±3.9	139.9±7.5^b	119.9±6.8
对照组	6	112.3±4.3	108.5±8.2	110.5±8.2
F值	?	2.876	26.758	165.040
P值	?	0.068	0.000	0.000

表1 3组大鼠孕期各时间点收缩压变化情况比较(mmHg, ±s)

Table 1 Comparison of the blood pressure change at different time points during pregnancy among three groups (mmHg, ±s)

组别	例数	13 d	15 d	19 d
模型组	6	111.4±4.9	141.2±8.2^a	139.2±9.1^c，d
治疗组	6	109.1±3.9	139.9±7.5^b	119.9±6.8
对照组	6	112.3±4.3	108.5±8.2	110.5±8.2
F值	?	2.876	26.758	165.040
P值	?	0.068	0.000	0.000

图1 胎鼠脑海马CA1区TUNEL染色，胎鼠脑海马CA1区神经细胞均出现不同程度的凋亡（TUNEL染色，×400；A：模型组；B：治疗组；C：对照组）

Figure 1 Fetal rat hippocampal CA1 area by TUNEL staining (TUNEL staining, ×400; A: model group; B: treatment group; C: control group)

表2 各组胎鼠脑海马组织CA1区神经细胞凋亡情况比较(个/高倍视野)

Table 2 Comparison of hippocampus of fetal rat(per high power field)

组别	例数	凋亡细胞数
模型组	6	67.9±3.3^{a b}
治疗组	6	30.6±1.6^c
对照组	6	9.6±1.8
F值	?	1 931.4
P值	?	0.000

表2 各组胎鼠脑海马组织CA1区神经细胞凋亡情况比较(个/高倍视野)

Table 2 Comparison of hippocampus of fetal rat(per high power field)

组别	例数	凋亡细胞数
模型组	6	67.9±3.3^{a b}
治疗组	6	30.6±1.6^c
对照组	6	9.6±1.8
F值	?	1 931.4
P值	?	0.000

表3 各组间胎脑海马组织中caspase-3 mRNA相对表达水平比较

Table 3 Comparison of caspase-3 mRNA relative expression levels of hippocampus among three groups

组别	例数	caspase-3 mRNA
模型组	6	2.87±0.32^{a b}
治疗组	6	1.69±0.41^c
对照组	6	1.00±0.22
F值	?	1 687.13
P值	?	0.000

表3 各组间胎脑海马组织中caspase-3 mRNA相对表达水平比较

Table 3 Comparison of caspase-3 mRNA relative expression levels of hippocampus among three groups

组别	例数	caspase-3 mRNA
模型组	6	2.87±0.32^{a b}
治疗组	6	1.69±0.41^c
对照组	6	1.00±0.22
F值	?	1 687.13
P值	?	0.000

图2 胎鼠脑海马CA1区caspase-3免疫组化检测（TAB染色，×400；A：模型组；B：治疗组；对照组）

Figure 2 Immunohistochemistry of caspase-3 of fetal rat brain hippocampal CA1 region (TAB staining, ×400; A：model group; B：treatment group; C:control group)

表4 各组间胎鼠脑海马组织CA1区caspase-3蛋白比较

Table 4 Comparison of caspase-3 protein of hippocampus CA1 area among three groups

组别	样本量	－	＋	＋＋	＋＋＋
模型组	24	0	4	13	7
治疗组	24	0	9	14	1
对照组	24	2	22	0	0
χ²值	?	?	36.057	?	?
P值	?	?	0.000	?	?

表4 各组间胎鼠脑海马组织CA1区caspase-3蛋白比较

Table 4 Comparison of caspase-3 protein of hippocampus CA1 area among three groups

组别	样本量	－	＋	＋＋	＋＋＋
模型组	24	0	4	13	7
治疗组	24	0	9	14	1
对照组	24	2	22	0	0
χ²值	?	?	36.057	?	?
P值	?	?	0.000	?	?

[1]	Tagin M, Shah PS, Lee KS. Magnesium for newborns with hypoxic-ischemic encephalopathy: a systematic review and meta-analysis. J Perinatol. 2013. 33(9): 663–669.
[2]	Parrish AB, Freel CD, Kornbluth S. Cellular mechanisms controlling caspase activation and function[J]. Cold Spring Harb Perspect Biol, 2013, 5(6): a008672.
[3]	Broughton BR, Reutens DC, Sobey CG. Apoptotic mechanisms after cerebral ischemia[J]. Stroke, 2009, 40(5): e331–e339.
[4]	Snigdha S, Smith ED, Prieto GA, et al. Caspase-3 activation as a bifurcation point between plasticity and cell death[J]. Neurosci Bull, 2012, 28(1): 14–24.
[5]	李静，唐卉. 低剂量内毒素诱导构建先兆子痫大鼠模型[J]. 山东医药，2014，58 (19): 26–28.
[6]	Kulp A, Kuehn MJ. Biological functions and biogenesis of secreted bacterial outer membrane vesicles[J]. Annu Rev Microbiol, 2010, 64: 163–184.
[7]	García-Quintanilla M, Pulido MR, Pachón J, et al. Immunization with lipopolysaccharide-deficient whole cells provides protective immunity in an experimental mouse model of acinetobacter baumannii infection[J]. PLoS One, 2014, 9(12): e114410.
[8]	Faas MM, Schuiling GA, Baller JF, et al. A new animal model for human preeclampsia: ultra-low-dose endotoxin infusion in pregnant rats[J]. Am J Obstet Gynecol, 1994, 171(1): 158–164.
[9]	李娜，王晓燕，刘春长，等. caspase-3基因在染铝小鼠神经细胞凋亡中的作用[J]. 环境与职业医学，2011, 28(3): 137–140.
[10]	Neuman KM, Molina-Campos E, Musial TF, et al. Evidence for Alzheimer's disease-linked synapse loss and compensation in mouse and human hippocampal CA1 pyramidal neurons[J]. Brain Struct Funct, 2014 Jul 17. [Epub ahead of print]
[11]	Kondo M, Kitajima T, Fujii S, et al. Modulation of synaptic plasticity by the coactivation of spatially distinct synaptic inputs in rat hippocampal CA1 apical dendrites[J]. Brain Res, 2013，1526: 1–14.
[12]	Goñi-de-Cerio F, Alvarez A, Lara-Celador I, et al. Magnesium sulfate treatment decreases the initial brain damage alterations produced after perinatal asphyxia in fetal lambs[J]. J Neurosci Res, 2012, 90(10): 1932–1940.
[13]	Gathwala G, Khera A, Singh J, et al. Magnesium for neuroprotection in birth asphyxia[J]. J Pediatr Neurosci, 2010, 5(2): 102–104.
[14]	Nakamura T, Lipton SA. Preventing Ca^2＋-mediated nitrosative stress in neurodegenerative diseases: possible pharmacological strategies[J]. Cell Calcium, 2010, 47(2): 190–197.
[15]	Rayssiguier Y, Libako P, Nowacki W, et al. Magnesium deficiency and metabolic syndrome: stress and inflammation may reflect calcium activation[J]. Magnes Res, 2010, 23(2): 73–80.
[16]	Creagh EM. Caspase crosstalk: integration of apoptotic and innate immune signalling pathways[J]. Trends Immunol, 2014, 35(12): 631–640.
[17]	Niemi NM, MacKeigan JP. Mitochondrial phosphorylation in apoptosis: flipping the death switch[J]. Antioxid Redox Signal, 2013, 19(6): 572–582.
[18]	MacKenzie SH, Clark AC. Death by caspase dimerization[J]. Adv Exp Med Biol, 2012, 747: 55–73.
[19]	Murray J, Renslo AR. Modulating caspase activity: beyond the active site[J]. Curr Opin Struct Biol, 2013, 23(6): 812–819.
[20]	McIlwain DR, Berger T, Mak TW. Caspase functions in cell death and disease[J]. Cold Spring Harb Perspect Biol, 2013, 5(4): a008656.

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Effects of magnesium sulfate on the neuroprotection of hippocampus of fetal rats delivered by experimental rats model of preeclampsia

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Effects of magnesium sulfate on the neuroprotection of hippocampus of fetal rats delivered by experimental rats model of preeclampsia