微管蛋白、水通道蛋白4和K+通道蛋白4.1在大鼠脊髓损伤后不同时间的表达变化

doi:10.3969/j.issn.1006-9771.2018.10.007

《中国康复理论与实践》 ›› 2018, Vol. 24 ›› Issue (10): 1151-1158.doi: 10.3969/j.issn.1006-9771.2018.10.007

微管蛋白、水通道蛋白4和K⁺通道蛋白4.1在大鼠脊髓损伤后不同时间的表达变化

陈铁戈^1,2, 郭永强^1,2, 王明^1,2, 张东亮^1,2, 夏亚一^1,2, 汪静², 伍亚民³, 党跃修^1,2, 张海鸿¹

1.兰州大学第二医院骨科,甘肃兰州市 730030;
2.甘肃省骨关节疾病重点实验室,甘肃兰州市 730030;
3.中国人民解放军陆军军医大学大坪医院野战外科研究所三室,创伤、烧伤与复合伤国家重点实验室,重庆市 400042

收稿日期:2018-06-25 出版日期:2018-10-25 发布日期:2018-10-30
通讯作者: 张海鸿、党跃修。E-mail: zhh1968jzwk@sina.com (张海鸿); dangyuexiu@sina.com (党跃修)
作者简介:陈铁戈(1989-),男,汉族,陕西西安市人,硕士研究生,主要研究方向：脊柱脊髓损伤。
基金资助:
甘肃省自然科学基金项目(No. 1506RJZA245)

Expression and Change of Microtubule, Aquaporins-4 and K⁺ Ion Channel Protein-4.1 after Spinal Cord Injury in Rats

CHEN Tie-ge^1,2, GUO Yong-qiang^1,2, WANG Ming^1,2, ZHANG Dong-liang^1,2, XIA Ya-yi^1,2, WANG Jing², WU Ya-min³, DANG Yue-xiu^1,2, ZHANG Hai-hong^1,2

1. Department of Orthopedics, Second Clinical Medical College of Lanzhou University, Lanzhou, Gansu 730030, China;
2. Key Laboratory of Orthopedics of Gansu Province, Lanzhou, Gansu 730030, China;
3. State Key Laboratory of Trauma, Burns and Combined Injury, the Third Department of Research Institute of Surgery, Daping Hospital, Army Medical University, Chongqing 400042, China

Received:2018-06-25 Published:2018-10-25 Online:2018-10-30
Contact: ZHANG Hai-hong, DANG Yue-xiu. E-mail: zhh1968jzwk@sina.com (ZHANG Hai-hong); dangyuexiu@sina.com (DANG Yue-xiu)
Supported by:
Natural Science Foundation of Gansu Province (No. 1506RJZA245)

摘要/Abstract

摘要： 目的探究微管骨架、水通道蛋白4(AQP4)和K⁺通道4.1(Kir4.1)在大鼠急性脊髓损伤后的时间变化规律。方法 90只成年雌性Sprague-Dawley大鼠,随机分组为假手术组(n=30)和损伤组(n=60),损伤组设6 h、1 d、3 d、5 d、7 d,每个亚组12只大鼠。用Allen打击法制备T₁₀打击损伤模型。损伤后各时间点各组行脊髓含水量检测、脊髓HE染色,观察急性脊髓损伤后的病理变化;行微管蛋白、AQP4和Kir4.1免疫组织化学染色和Western blotting检测,观察急性损伤后三组蛋白的表达变化及相对灰度值分析。结果损伤组各时间点脊髓含水量均高于假手术组(P<0.05),且5 d时最高。HE染色显示,损伤后6 h,灰质主要以出血为主;损伤后1 d,灰质出血严重,神经元肿胀加重;损伤后3 d,灰质坏死面积加大,水肿现象明显;损伤后5 d、7 d,灰质坏死更加明显,水肿现象更加严重。Western blotting和免疫组织化学染色结果显示,损伤后AQP4在灰质表达逐渐增高,且5 d表达为高峰,微管蛋白和Kir4.1表达趋势基本相同,为损伤后表达逐渐下降,5 d表达最低。结论脊髓损伤后微管蛋白表达与Kir4.1表达趋势相似,与AQP4表达趋势相反,可能共同参与水肿形成。

关键词: 脊髓水肿, 微管, 水通道蛋白4, K⁺通道4.1, 大鼠

Abstract: Objective To explore the expression and the changes of microtubule, aquaporin-4 (AQP4) and potassium ion channel 4.1 (Kir4.1) after spinal cord injury in rats.Methods Ninety female adult Sprague-Dawley rats were randomly divided into sham operation group (n=30) and injury group (n=60). The injury group was divided into six hours, one day, three days, five days and seven days subgroups, with twelve rats in each subgroup. Spinal cord injury at T₁₀ was established with modified Allen's method (20 g×25 mm) in the injury group. The water content of spinal cord was measured at each time point after injury. Then, the pathology was observed with HE staining, the expression of α-Tubulin, AQP4 and Kir4.1 was detected and analyzed with immunohistochemical staining and Western blotting.Results The water content of the spinal cord was higher in the injured group than in the sham operation group (P<0.05), and was highest on the fifth day. HE staining showed that the gray matter hemorrhage at six hours after injury; one day after injury, the gray matter bled seriously, and neuron swelling was aggravated; three days after injury, the area of gray matter necrosis increased, and the edema phenomenon was obvious; five days and seven days after injury, the gray matter necrosis and the edema phenomenon were more serious. Western blotting and immunohistochemistry showed that the expression of AQP4 gradually increased after injury, and raised at peak on the fifth day; the expression of α-Tubulin and Kir4.1 was similar, and the expression gradually decreased after injury, especially on the fifth day.Conclusion The expression of α-Tubulin and Kir4.1 is similar after spinal cord injury, and is contrary to the expression of AQP4. α-Tubulin, AQP4 and Kir4.1 may be related after injury and may participate in the formation of spinal cord edema.

Key words: spinal cord edema, microtubule, aquaporin 4, potassium ion channel 4.1, rats

中图分类号:

R651.2

陈铁戈, 郭永强, 王明, 张东亮, 夏亚一, 汪静, 伍亚民, 党跃修, 张海鸿. 微管蛋白、水通道蛋白4和K⁺通道蛋白4.1在大鼠脊髓损伤后不同时间的表达变化[J]. 《中国康复理论与实践》, 2018, 24(10): 1151-1158.

CHEN Tie-ge, GUO Yong-qiang, WANG Ming, ZHANG Dong-liang, XIA Ya-yi, WANG Jing, WU Ya-min, DANG Yue-xiu, ZHANG Hai-hong. Expression and Change of Microtubule, Aquaporins-4 and K⁺ Ion Channel Protein-4.1 after Spinal Cord Injury in Rats[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2018, 24(10): 1151-1158.

参考文献

[1] Jin BJ, Hua Z, Binder DK, et al.Aquaporin-4-dependent K+ and water transport modeled in brain extracellular space following neuroexcitation[J]. J Gen Physiol, 2013, 141(1): 119-132.
[2] Zeng XN, Sun XL, Gao L, et al.Aquaporin-4 deficiency down-regulates glutamate uptake and GLT-1 expression in astrocytes[J]. Mol Cell Neurosci, 2007, 34(1): 34-39.
[3] Hubbard JA, Binder DK.Unaltered glutamate transporter-1 protein levels in aquaporin-4 knockout mice[J]. ASN Neuro, 2017, 9(1): 1759091416687846.
[4] Janke C, Bulinski JC.Post-translational regulation of the microtubule cytoskeleton: mechanisms and functions[J]. Nat Rev Mol Cell Biol, 2011, 12(12): 773-786.
[5] Ingber DE. Tensegrity I.Cell structure and hierarchical systems biology[J]. J Cell Sci, 2003, 116(Pt 7): 1157-1173.
[6] Yan X, Liu J, Wang X, et al.Pretreatment with AQP4 and NKCC1 inhibitors concurrently attenuated spinal cord edema and tissue damage after spinal cord injury in rats[J]. Front Physiol, 2018, 9: 6.
[7] Li XQ, Fang B, Tan WF, et al.miR-320a affects spinal cord edema through negatively regulating aquaporin-1 of blood-spinal cord barrier during bimodal stage after ischemia reperfusion injury in rats[J]. BMC Neurosci, 2016, 17: 10.
[8] Borgens RB, Liusnyder P.Understanding secondary injury[J]. Q Rev Biol, 2012, 87(2): 89-127.
[9] Tran LV.Understanding the pathophysiology of traumatic brain injury and the mechanisms of action of neuroprotective interventions[J]. J Trauma Nurs, 2014, 21(1): 30-35.
[10] Hasegawa H, Ma T, Skach W, et al.Molecular cloning of a mercurial-insensitive water channel expressed in selected water-transporting tissues[J]. J Biol Chem, 1994, 269(8): 5497-5500.
[11] Hubbard JA, Hsu MS, Seldin MM, et al.Expression of the astrocyte water channel aquaporin-4 in the mouse brain[J]. Asn Neuro, 2015, 7(5): 1759091415605486.
[12] Nagelhus EA, Mathiisen TM, Ottersen OP.Aquaporin-4 in the central nervous system: cellular and subcellular distribution and coexpression with KIR4.1[J]. Neuroscience, 2004, 129(4): 905-913.
[13] Saadoun S, Papadopoulos MC.Aquaporin-4 in brain and spinal cord oedema[J]. Neuroscience, 2010, 168(4): 1036-1046.
[14] Oklinski MK, Skowronski MT, Skowronska A, et al.Aquaporins in the spinal cord[J]. Int J Mol Sci, 2016, 17(12): E2050.
[15] Olsen ML, Campbell SC, Mcferrin MB, et al.Spinal cord injury causes a wide-spread, persistent loss of Kir4.1 and glutamate transporter 1: benefit of 17ß-oestradiol treatment[J]. Brain , 2010, 133(4):1013-1025.
[16] Najafi E, Stoodley MA, Bilston LE, et al.Inwardly rectifying potassium channel 4.1 expression in post-traumatic syringomyelia[J]. Neuroscience, 2016, 317: 23-35.
[17] Janmey PA.The cytoskeleton and cell signaling: component localization and mechanical coupling[J]. Physiol Rev, 1998, 78(3): 763-781.
[18] Ruschel J, Hellal F, Flynn KC, et al.Axonal regeneration. Systemic administration of epothilone B promotes axon regeneration after spinal cord injury[J]. Science, 2015, 348(6232): 347-352.
[19] Ruschel J, Bradke F.Systemic administration of epothilone D improves functional recovery of walking after rat spinal cord contusion injury[J]. Exp Neurol, 2018, 306: 243-249.
[20] Crunkhorn S.CNS injury: microtubule stabilizer repairs spinal cord injury[J]. Nat Rev Drug Discov, 2015, 14(5): 310.
[21] Zelinski B, Müller N, Kierfeld J.Dynamics and length distribution of microtubules under force and confinement[J]. Phys Rev E Stat Nonlin Soft Matter Phys, 2012, 86(4 Pt 1): 041918.
[22] Akhmanova A, Steinmetz MO.Tracking the ends: a dynamic protein network controls the fate of microtubule tips[J]. Nat Rev Mol Cell Biol, 2008, 9(4):309-322.
[23] Zhao H, Yao X, Wang TX, et al.PKCαregulates vasopressin-induced aquaporin-2 trafficking in mouse kidney collecting duct cells in vitro via altering microtubule assembly[J]. Acta Pharmacol Sin, 2012, 33(2): 230-236.
[24] Conner MT, Conner AC, Brown JE, et al.Membrane trafficking of aquaporin 1 is mediated by protein kinase C via microtubules and regulated by tonicity[J]. Biochemistry, 2010, 49(5): 821-823.
[25] Vossenkämper A, Nedvetsky PI, Wiesner B, et al.Microtubules are needed for the perinuclear positioning of aquaporin-2 after its endocytic retrieval in renal principal cells[J]. Am J Physiol Cell Physiol, 2007, 293(3): 1129-1138.
[26] Sunshine JE, Dagal A, Burns SP, et al.Methylprednisolone therapy in acute traumatic spinal cord injury: analysis of a regional spinal cord model systems database[J]. Anesth Analg, 2017, 124(4): 1200-1205.

微管蛋白、水通道蛋白4和K⁺通道蛋白4.1在大鼠脊髓损伤后不同时间的表达变化

Expression and Change of Microtubule, Aquaporins-4 and K⁺ Ion Channel Protein-4.1 after Spinal Cord Injury in Rats

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微管蛋白、水通道蛋白4和K+通道蛋白4.1在大鼠脊髓损伤后不同时间的表达变化

Expression and Change of Microtubule, Aquaporins-4 and K+ Ion Channel Protein-4.1 after Spinal Cord Injury in Rats

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微管蛋白、水通道蛋白4和K⁺通道蛋白4.1在大鼠脊髓损伤后不同时间的表达变化

Expression and Change of Microtubule, Aquaporins-4 and K⁺ Ion Channel Protein-4.1 after Spinal Cord Injury in Rats