氯倍他索对新生大鼠脑白质损伤的治疗作用研究

doi:10.3877/cma.j.issn.2095-5820.2023.01.002

目的

探讨氯倍他索对新生大鼠脑白质损伤（WMI）模型是否具有治疗作用。

方法

选取新生3 d龄SD大鼠，结扎右侧颈总动脉，采用使大鼠缺氧80 min的方法构建新生WMI大鼠模型。采用2 mg·kg^-1·d^-1和5 mg·kg^-1·d^-1剂量的氯倍他索连续腹腔注射WMI大鼠模型5 d。HE染色观察WMI大鼠模型脑组织病理损伤；免疫荧光染色观察髓鞘碱性蛋白（MBP）及 CC1表达水平；免疫印迹检测诱导型一氧化氮合酶（iNOS）表达水平。

结果

经2 mg·kg^-1·d^-1和5 mg·kg^-1·d^-1的氯倍他索处理后，WMI大鼠脑白质结构疏松缓解，细胞肿胀减轻，髓鞘增多，脑白质病理损伤缓解。氯倍他索可以通过上调MBP及CC1表达水平促进WMI大鼠髓鞘形成。同时，氯倍他索可以通过恢复iNOS表达来缓解WMI大鼠髓鞘损伤。

结论

氯倍他索可通过增加MBP、CC1及iNOS的表达，对新生大鼠WMI病理损伤具有缓解作用，为氯倍他索用于早产儿WMI的治疗提供了理论依据。

关键词: 氯倍他索, 早产儿脑白质损伤, 髓鞘碱性蛋白

Objective

To investigate the therapeutic effects of clobetasol on premature white matter injury (WMI) in rats.

Methods

Newborn 3-day-old SD rats were selected, and the WMI model was made by ligation of the right common carotid artery and hypoxia for 80 minutes. WMI rats were intraperitoneally injected with clobetasol (2 mg·kg^-1·d^-1 and 5 mg·kg^-1·d^-1) for 5 days. The pathological damage of brain tissue in WMI rats was observed by Hematoxylin and Eeosin (HE) staining. The expression levels of myelin base protein (MBP) and CC1 were observed by immunofluorescence. The expression of iNOS was detected by Western blotting.

Results

After treated with clobetasol at 2 mg·kg^-1·d^-1 and 5 mg·kg^-1·d^-1, the loose of white matter structure was relieved, cell swelling was reduced, myelin sheath was increased, and the pathological damage of white matter was relieved. Clobetasol can promote myelination in WMI rats by up-regulating the expression levels of MBP and CC1. In addition, clobetasol alleviated myelin sheath injury in WMI rats by restoring iNOS expression.

Conclusions

Clobetasol can alleviate the pathological injury of neonatal white matter injury by increasing the expressions of MBP, CC1 and iNOS, which providing a theoretical basis for the treatment of premature WMI with clobetasol.

Key words: clobetasol, white matter injury, myelin base protein

图1 WMI大鼠模型脑组织病理损伤（HE染色）

图2 免疫荧光观察髓鞘基蛋白及CC1表达水平注：MERGE代表DAPI、MBP及CC1三色叠加。

图3 脑组织iNOS表达水平（免疫印迹）

1	CAYAM-RAND D, GUO T, SYNNES A, et al. Interaction between preterm white matter injury and childhood thalamic growth[J]. Annals of Neurology, 2021, 90(4): 584-594.
2	OPHELDERS D R M G, GUSSENHOVEN R, Klein L, et al. Preterm brain injury, antenatal triggers, and therapeutics: timing is key[J]. Cells, 2020, 9(8): 1871.
3	HE Y, ZHANG Y, LI F, et al. White matter injury in preterm infants: pathogenesis and potential therapy from the aspect of the gut-brain axis[J]. Frontiers in Neuroscience, 2022, 16: 849372.
4	GUILLOT M, MILLER S P. The dimensions of white matter injury in preterm neonates[J]. Seminars in Perinatology, 2021, 45(7): 151469.
5	SCHNEIDER J, MILLER S P. Preterm brain injury: white matter injury[J]. Handbook of Clinical Neurology, 2019, 162: 155-172.
6	MILLER S P, FERRIERO D M, LEONARD C, et al. Early brain injury in premature newborns detected with magnetic resonance imaging is associated with adverse early neurodevelopmental outcome[J]. The Journal of Pediatrics, 2005, 147(5): 609-616.
7	VAN WEZEL-MEIJLER G, STEGGERDA S J, LEIJSER L M. Cranial ultrasonography in neonates: role and limitations[J]. Seminars in Perinatology, 2010, 34(1): 28-38.
8	NAJM F J, MADHAVAN M, ZAREMBA A, et al. Drug-based modulation of endogenous stem cells promotes functional remyelination in vivo[J]. Nature, 2015, 522(7555): 216-220.
9	RAPÔSO C, LUNA R L d A, NUNES A K S, et al. Role of iNOS-NO-cGMP signaling in modulation of inflammatory and myelination processes[J]. Brain Research Bulletin, 2014, 104: 60-73.
10	GOPAGONDANAHALLI K R, Li J, FAHEY M C, et al. Preterm hypoxic-ischemic encephalopathy[J]. Frontiers in Pediatrics, 2016, 4: 114.
11	WANG X, ZANG J, YANG Y, et al. Transplanted human oligodendrocyte progenitor cells restore neurobehavioral deficits in a rat model of preterm white matter injury[J]. Frontiers in Neurology, 2021, 12: 749244.
12	Al-GRIW M A, SALTER M G, WOOD I C. Inhibition of ionotropic GluR signaling preserves Oligodendrocyte lineage and myelination in an ex vivo rat model of white matter ischemic injury[J]. Acta Neurobiologiae Experimentalis, 2021, 81(3): 233-248.
13	BOCCAZZI M, VAN STEENWINCKEL J, SCHANG A L, et al. The immune-inflammatory response of oligodendrocytes in a murine model of preterm white matter injury: the role of TLR3 activation[J]. Cell Death and Disease, 2021, 12(2): 166.
14	Wu Z, XUE H, GAO Q, et al. Effects of early postnatal sevoflurane exposure on oligodendrocyte maturation and myelination in cerebral white matter of the rat[J]. Biomedicine and Pharmacotherapy, 2020, 131: 110733.
15	CORMACK B E, HARDING J E, MILLER S P, et al. The influence of early nutrition on brain growth and neurodevelopment in extremely preterm babies: A narrative review[J]. Nutrients, 2019, 11(9): 2029.
16	QIU H, QIAN T, WU T, et al. Umbilical cord blood cells for the treatment of preterm white matter injury: potential effects and treatment options[J]. Journal of Neuroscience Research, 2021, 99(3): 778-792.
17	AGUT T, ALARCON A, CABAÑAS F, et al. Preterm white matter injury: ultrasound diagnosis and classification[J]. Pediatric Research, 2020, 87(Suppl 1): 37-49.
18	GUO T, CHAU V, PEYVANDI S, et al. White matter injury in term neonates with congenital heart diseases: Topology & comparison with preterm newborns[J]. Neuroimage, 2019, 185: 742-749.
19	DUERDEN E G, HALANI S, NG K, et al. White matter injury predicts disrupted functional connectivity and microstructure in very preterm born neonates[J]. NeuroImage Clinical, 2019, 21: 101596.
20	CHEW L J, DEBOY C A. Pharmacological approaches to intervention in hypomyelinating and demyelinating white matter pathology[J]. Neuropharmacology, 2016, 110(Pt B): 605-625.
21	WARNOCK A, TOOMEY L M, WRIGHT A J, et al. Damage mechanisms to oligodendrocytes and white matter in central nervous system injury: The Australian context[J]. Journal of Neurotrauma, 2020, 37(5):739-769.
22	BANDO Y, NOMURA T, BOCHIMOTO H, et al. Abnormal morphology of myelin and axon pathology in murine models of multiple sclerosis[J]. Neurochemistry International, 2015, 81: 16-27.
23	JEONG I H, CHOI J Y, KIM S H, et al. Comparison of myelin water fraction values in periventricular white matter lesions between multiple sclerosis and neuromyelitis optica spectrum disorder[J]. Multiple Sclerosis: Clinical and Laboratory Research, 2016, 22(12): 1616-1620.
24	HAYNES R L, FOLKERTH R D, KEEFE R J, et al. Nitrosative and oxidative injury to premyelinating oligodendrocytes in periventricular leukomalacia[J]. Journal of Neuropathology and Experimental Neurology, 2003, 62(5): 441-450.
25	荣箫, 周伟. 早产儿脑白质损伤各种动物模型的特点与选择[J]. 中国新生儿科杂志, 2010, 25(5): 310-212.
26	杨印祥, 索磊, 杜庆安, 等. 缺氧时间对脑白质损伤程度影响的实验研究[J]. 临床儿科杂志, 2015, 33(10): 883-886.
27	HAGEN M W, RIDDLE A, MCCLENDON E, et al. Role of recurrent hypoxia-ischemia in preterm white matter injury severity[J]. Public Library of Science One, 2014, 9(11): e112800.
28	BACK S A, LUO N L, BORENSTEIN N S, et al. Late oligodendrocyte progenitors coincide with the developmental window of vulnerability for human perinatal white matter injury[J]. The Journal of Neuroscience: The Official Journal of the Society for Neuroscience, 2001, 21(4): 1302-1312.
29	FENYK-MELODY J E, GARRISON A E, BRUNNERT S R, et al. Experimental autoimmune encephalomyelitis is exacerbated in mice lacking the NOS2 gene[J]. The Journal of Immunology: Official Journal of the American Association of Immunologists, 1998, 160(6): 2940-2946.
30	ARNETT H A, HELLENDALL R P, MATSUSHIMA G K, et al. The protective role of nitric oxide in a neurotoxicant-induced demyelinating model[J]. The Journal of Immunology: Official Journal of The American Association of Immunologists, 2002, 168(1): 427-433.

[1]	阎晨, 刘涛, 宣斐. 超声引导联合全麻对老年肺癌肺叶切除术患者NGF-β、MBP及术后转归分析[J]. 中华肺部疾病杂志(电子版), 2022, 15(02): 192-196.
[2]	邱炜, 黄娜, 郭志旺, 常会民. 亚低温治疗重症高血压脑出血中髓鞘碱性蛋白变化的临床观察[J]. 中华神经创伤外科电子杂志, 2018, 04(02): 103-105.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Therapeutic effect of clobetasol on white matter injury in neonatal rats

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Therapeutic effect of clobetasol on white matter injury in neonatal rats