地塞米松致子代大鼠海马轴突发育损伤

doi:10.19586/j.2095-2341.2021.0193

生物技术进展 ›› 2022, Vol. 12 ›› Issue (3): 467-472.DOI: 10.19586/j.2095-2341.2021.0193

地塞米松致子代大鼠海马轴突发育损伤

谢露露(), 姚宝珍()

武汉大学人民医院儿科，武汉 430000

收稿日期:2021-12-14 接受日期:2022-01-24 出版日期:2022-05-25 发布日期:2022-05-26
通讯作者: 姚宝珍
作者简介:谢露露E-mail：1318569752@qq.com；
基金资助:
武汉大学医学腾飞计划(TFLC2018001)

Dexamethasone-induced Hippocampal Axon Developmental Damage in Offspring Rats

Lulu XIE(), Baozhen YAO()

Department of Pediatrics，Renmin Hospital of Wuhan University，Wuhan 430000，China

Received:2021-12-14 Accepted:2022-01-24 Online:2022-05-25 Published:2022-05-26
Contact: Baozhen YAO

摘要/Abstract

摘要：

为了探讨地塞米松对子代大鼠海马轴突的影响，建立了孕期地塞米松暴露（prenatal dexamethasone exposure, PDE）模型。Wistar大鼠于孕中晚期皮下注射地塞米松（0.2 mg·kg^-1·d^-1），部分子代于孕20天（GD20）、出生后12周（PW12）处死取海马样本，检测海马糖皮质激素受体（glucocorticoid receptor, GR）活化指标以及轴突损伤指标。PDE子代胎鼠海马GR活化，GR、糖皮质激素调节激酶1（glucocorticoid-regulated kinase 1, SGK1）和FK506结合蛋白（FK506 binding protein 5, FKBP5）表达显著增加。轴突损伤指标包括生长相关蛋白43（growth associated protein-43,GAP43）、信号素3A（semaphorin 3A, SEMA3A）和集聚蛋白（agrin）表达明显升高。而PDE成年子代大鼠海马GR无明显活化，轴突损伤指标GAP43、SEMA3A和AGRIN表达明显升高。研究结果证实PDE通过活化胎海马GR引起轴突发育损伤，且轴突损伤可延续至出生后。

关键词: 糖皮质激素受体, 海马, 轴突损伤, 神经疾病

Abstract:

In order to explore the effect of dexamethasone on the hippocampal axons of offspring rats， a prenatal dexamethasone exposure （PDE） model was established. Wistar rats were subcutaneously injected with dexamethasone （0.2 mg·kg^-1·d^-1） in the second and third trimesters， some offspring were sacrificed during the gestational days of 20 （GD20） and at postnatal week of 12 （PW12）， and the hippocampal glucocorticoid receptors （GR） activation index and axon damage index were detected. GR was activated in the hippocampus of PDE offspring， and the expressions of GR， glucocorticoid-regulated kinase 1 （SGK1） and FK506 binding protein 5 （FKBP5） were significantly increased. Axon damage indicators including growth-associated protein-43 （GAP43）， semaphorin 3A （SEMA3A） and agrin expression were significantly increased. However， the hippocampal GR of PDE adult offspring rats was not significantly activated， and the expressions of axonal injury indicators GAP43， SEMA3A and AGRIN were significantly increased. The results comfirmed that PDE induces axonal developmental damage by activating fetal hippocampal GR， and axonal damage can persist after birth.

Key words: glucocorticoid receptor, hippocampus, axon injury, neurological disease

中图分类号:

R742.8

谢露露, 姚宝珍. 地塞米松致子代大鼠海马轴突发育损伤[J]. 生物技术进展, 2022, 12(3): 467-472.

Lulu XIE, Baozhen YAO. Dexamethasone-induced Hippocampal Axon Developmental Damage in Offspring Rats[J]. Current Biotechnology, 2022, 12(3): 467-472.

图/表 5

表1 RT-qPCR中大鼠寡核苷酸引物及反应条件

Table 1 Rat oligonucleotide primers and reaction conditions used in RT-qPCR

基因名称	正向引物序列（5′→3′）	反向引物序列（5′→3′）	退火温度/°C
糖皮质激素受体基因（GR）	5′-TGTTGGGGAGACGAGATTTT-3′	5′-TCATGCAACCGAAGTATGAAA-3′	58
糖皮质激素调节激酶1基因（SGK1）	5′-CAATAAGCGACACGGCTCTT-3′	5′-GCGGGTGGCTAACTAGGATA-3′	60
FK506结合蛋白基因（FKBP5）	5′-CCTCGTTGGAGAAGATCAAG-3′	5′-CGTGGTACTCCGTGTGATTG-3′	60
生长相关蛋白43基因（GAP43）	5′-CCAGAGTTAGAGAAGCGTTTAG-3′	5′-CAGGCATTTGCAGCTACT-3′	58
信号素3A基因（SEMA3A）	5′-CACAAGACGACAGGATATAAGG-3′	5′-GCCAATATACCAAGGCTCTC-3′	60
集聚蛋白基因（AGRIN）	5′-ACTTATGAATGCCTGTGTCC-3′	5′-GTGCCTTCTCACTCTCAATC-3′	58
磷酸甘油醛脱氢酶基因（GAPDH）	5′-GGCACAGTCAAGGCTGAGAATG-3′	5′-ATGGTGGTGAAGACGCCAGTA-3′	60

图1 子代胎鼠海马轴突损伤指标表达变化注：**表示与对照组相比较，在P<0.01水平有统计学差异。n=6。

Fig.1 Changes of hippocampal axon injury index in offspring fetal rats

图2 子代成年大鼠海马轴突损伤指标表达变化注：**表示与对照组相比较，在P<0.01水平有统计学差异。n=6。

Fig.2 Expression changes of hippocampal axon injury indexes in offspring adult rats

图3 子代胎鼠海马GR活化指标变化A：子代胎鼠GR的RT-qPCR检测；B：子代胎鼠GR的Western blot定量结果；C：子代胎鼠GR的Western blot检测；D：子代胎鼠SGK1的RT-qPCR检测；E：子代胎鼠FKBP5的RT-qPCR检测。**表示与对照组相比较，在P<0.01水平有统计学差异。n=6。

Fig.3 Changes of hippocampal GR activation in offspring fetal rats

图4 成年子代大鼠海马GR活化指标变化A：子代成年大鼠GR的RT-qPCR检测；B：子代成年大鼠SGK1的RT-qPCR检测；C：子代成年大鼠FKBP5的RT-qPCR检测；D,E：子代成年大鼠GR的Western blot检测。n=6。

Fig.4 Changes of hippocampal GR activation indexes in adult offspring rats

参考文献 28

1	KHALAF-NAZZAL R, FRANCIS F. Hippocampal development-old and new findings[J]. Neuroscience, 2013, 248: 225-242.
2	QIN S, CHO S, CHEN T, et al.. Hippocampal-neocortical functional reorganization underlies children's cognitive development[J]. Nat. Neurosci., 2014, 17(9): 1263-1269.
3	BOCK J, WAINSTOCK T, BRAUN K, et al.. Stress in utero: prenatal programming of brain plasticity and cognition[J]. Biol. Psychiatry, 2015, 78(5): 315-326.
4	WITCHER M R, ELLIS T L, Astroglial networks and implications for therapeutic neuromodulation of epilepsy[J/OL]. Front. Comput. Neurosci., 2012(6): 61[2022-01-22]. .
5	NOORLANDER C W, VISSER G H A, RAMAKERS G M J, et al.. Prenatal corticosteroid exposure affects hippocampal plasticity and reduces lifespan[J]. Dev. Neurobiol., 2008. 68(2): 237-246.
6	ANDREWS M H, KOSTAKI A, SETIAWAN E, et al.. Developmental regulation of 5-HT1A receptor mRNA in the fetal limbic system: response to antenatal glucocorticoid[J]. Brain Res. Dev. Brain Res., 2004, 149(1): 39-44.
7	NOORLANDER C W, TIJSSELING D, HESSEL E V S, et al.. Antenatal glucocorticoid treatment affects hippocampal development in mice[J/OL]. PLoS ONE, 2014, 9(1): e85671[2022-01-22]. .
8	UNO H, LOHMILLER L, THIEME C, et al.. Brain damage induced by prenatal exposure to dexamethasone in fetal rhesus macaques. I. Hippocampus[J]. Brain Res. Dev. Brain Res., 1990, 53(2): 157-167.
9	郭艳, 陈筱青, 李红英, 等, 产前不同剂量地塞米松对早产仔鼠脑发育的影响 [J]. 南京医科大学学报(自然科学版), 2008,28(12): 1584-1586.
10	ZHANG C, RASBAND M N. Cytoskeletal control of axon domain assembly and function[J]. Curr. Opin. Neurobiol., 2016, 39: 116-121.
11	ADACHI R, YAMADA R, KUBA H. Plasticity of the axonal trigger zone[J]. Neurosci. Rev. J. Bring. Neurobiol. Neurol. Psychiatry, 2015, 21(3): 255-265.
12	KOLE M H, BRETTE R. The electrical significance of axon location diversity[J]. Curr. Opin. Neurobiol., 2018, 51: 52-59.
13	LI S, YOU M, CHAI W, et al.. Developmental exposure to nonylphenol induced rat axonal injury in vivo and in vitro[J]. Arch. Toxicol., 2019. 93(9): 2673-2687.
14	WANG X, JIAO X, LIU Z, et al.. Crocetin potentiates neurite growth in hippocampal neurons and facilitates functional recovery in rats with spinal cord injury[J]. Neurosci. Bull., 2017, 33(6): 695-702.
15	CROWTHER C A, MCKINLAY C J, MIDDLETON P, et al.. Repeat doses of prenatal corticosteroids for women at risk of preterm birth for improving neonatal health outcomes[J/OL]. Cochrane Dtabase System. Rev., 2015(7): CD003935[2022-01-22]. .
16	PATTANITTUM P, EWENS M R, LAOPAIBOON M, et al.. Use of antenatal corticosteroids prior to preterm birth in four South East Asian countries within the SEA-ORCHID project[J/OL]. BMC Pregn. Childbirth, 2008, 8: 47[2022-01-22]. .
17	RAJADURAI V S, TAN K H. The use and abuse of steroids in perinatal medicine[J]. Ann. Acad. Med. Singap., 2003, 32(3): 324-334.
18	VOGEL J P, SOUZA J P, GULMEZOGLU A M, et al.. Use of antenatal corticosteroids and tocolytic drugs in preterm births in 29 countries: an analysis of the WHO multicountry survey on maternal and newborn health[J]. Lancet, 2014, 384(9957): 1869-1877.
19	MOISIADIS V G, MATTHEWS S G. Glucocorticoids and fetal programming part 1: outcomes. nature reviews[J]. Endocrinology, 2014, 10(7): 391-402.
20	REAGAN-SHAW S, NIHAL M, AHMAD N, Dose translation from animal to human studies revisited[J]. FASEB J., 2008, 22(3): 659-661.
21	CHUNG D, SHUM A, CARAVEO G. GAP-43 and BASP1 in axon regeneration: implications for the treatment of neurodegenerative diseases[J/OL]. Front. Cell Dev. Biol., 2020, 8: 567537[2022-01-22]. .
22	ZIAK J, WEISSOVA R, JEŘÁBKOVÁ K, et al.. CRMP2 mediates Sema3F-dependent axon pruning and dendritic spine remodeling[J/OL]. EMBO. Rep., 2020. 21(3): e48512[2022-01-22]. .
23	OENTARYO M J, TSE A C K, LEE C W. Neuronal MT1-MMP mediates ECM clearance and Lrp4 cleavage for agrin deposition and signaling in presynaptic development[J/OL]. J. Cell Sci., 2020, 133(15):jcs246710 [2022-01-22]. .
24	LIU T T, FENG L, LIU H F, et al.. Altered axon initial segment in hippocampal newborn neurons, associated with recurrence of temporal lobe epilepsy in rats[J]. Mol. Med. Rep., 2017, 16(3): 3169-3178.
25	APRILE D, FRUSCIONEF, BALDASSARIS, et al.. TBC1D24 regulates axonal outgrowth and membrane trafficking at the growth cone in rodent and human neurons[J]. Cell Death Differ., 2019. 26(11): 2464-2478.
26	STOUFFER M A, GOLDEN J A, and FRANCIS F. Neuronal migration disorders: Focus on the cytoskeleton and epilepsy[J]. Neurobiol. Dis., 2016, 92(Pt A): 18-45.
27	LÖSING P, NITURAD C E, HARRER M, et al.. SRF modulates seizure occurrence, activity induced gene transcription and hippocampal circuit reorganization in the mouse pilocarpine epilepsy model[J/OL]. Mol. Brain, 2017, 10(1): 30[2022-01-22]. .
28	SONG X J, HAN W, HE R, et al.. Alterations of hippocampal myelin sheath and axon sprouting by status convulsion and regulating Lingo-1 expression with RNA interference in immature and adult rats[J]. Neurochem. Res., 2018, 43(3): 721-735.

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地塞米松致子代大鼠海马轴突发育损伤

Dexamethasone-induced Hippocampal Axon Developmental Damage in Offspring Rats

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