<strong>Effects of Low-dose Methotrexate on Spinal Cord Contusion-Induced Neural Cell Apoptisis in Rats</strong>

Abstract

Abstract:Objective：This study examined the effect of low-dose methotrexate on spinal cord contusion–induced neural cell apoptosis in rats and its potential protective mechanism.Methods:Rat spinal cord contusion model was established by Pinpoint Precision Cortical Impactor™ apparatus and then methotrexate (0.3 mg • kg^-1• BW) was subcutaneously administrated daily for 2 weeks starting from 30 min post-injury. Immunohistochemical staining and TUNEL method were used to examine the expression of neuronal nuclear antigen (NeuN) and neural cell apoptosis at damaged area, respectively.Results:One day after the traumatic injury, gray and white matter structures were unaltered and neuronal necrosis was only noticeable at the epicenter. TUNEL-positive cells were primarily located in the posterior horn of gray matter. 3～7 days after injury, the TUNEL-positive cells were increased and the location of these cells spreaded to the ventral and diffused periphery sections of the epicenter. Meanwhile, the number of NeuN immunoreactive neurons gradually decreased (P<0.05) from 0 mm to ±3 mm rostral and candual to the epicenter. Fourteen days after injury, the gray matter structure was no longer identifiable at the epicenter while the TUNEL-positive cells and NeuN immunoreactive neurons in the remaining anatomical positions remained at high levels. In rats that received methotrexate treatment, the number of NeuN immunoreactive neurons was significantly higher than those in model group (P<0.05,P<0.01).Conclusion:Low-dose methotrexate may decrease the nerve cell apoptosis via its metabolic product by exerting a protective effect on secondary injury of spinal cord.

Key words:methotrexate,low-dose,traumatic spinal cord injury,apoptosis,neuronal nuclear antigen

CLC Number:

R965.1

ZHANG Si, WANG Shuo-yu, GU Bing, LI Hua-nan, ZHANG Guo-fu, ZHANG Shui-yin.Effects of Low-dose Methotrexate on Spinal Cord Contusion-Induced Neural Cell Apoptisis in Rats[J]. Acta Neuropharmacologica, 2013, 3(6): 1-8.

References

[1] Alok Sharma. Pharmacological management of acute spinal cord injury [J]. J Assoc Physicians India, 2012, 60(Supp l): 13-18.

[2] Jefferson R Wilson, Nicole Forgione, Michael G Fehlings. Emerging therapies for acute traumatic spinal cord injury [J]. CMAJ, 2013, 185(6): 485-492.

[3] Lars Olson. Combinatory treatments needed for spinal cord injury [J]. Exp Neurol, 2013, 248: 309-315.

[4] 肖寒贫，王烁宇，李华南，等. 脊髓损伤治疗药物的研究现状 [J]. 神经药理学报, 2012，2(2): 32-36.

[5] Becker D, McDonald JW 3rd. Approaches to repairing the damaged spinal cord: overview [J]. Handb Clin Neurol, 2012, 109: 445-461.

[6] M P Keith, J D Edison, W R Gilliland. Progress toward personalized treatment of rheumatoid arthritis [J]. Clin Pharmacol Ther, 2012, 92(4): 440-442.

[7] Yukihiko Saeki, Toshihiro Matsui, Koichiro Saisho, et al. Current treatments of rheumatoid arthritis: from the 'NinJa' registry [J]. Expert Rev Clin Immunol, 2012, 8(5): 455-465.

[8] Edwin S Chan, Bruce N Cronstein. Methotrexate--how does it really work? [J]. Nat Rev Rheumatol, 2010, 6(3): 175-178.

[9] Ahmet Metin Sanli, Gokhan Serbes, Mustafa F Sargon, et al. Methothrexate attenuates early neutrophil infiltration and the associated lipid peroxidation in the injured spinal cord but does not induce neurotoxicity in the uninjured spinal cord in rats [J]. Acta Neurochir (Wien), 2012, 154(6): 1045-1054.

[10] Bulent Bakar, Emine A Köse, Sebnem Kupana Ayva, et al. Effects of low-dose methotrexate in spinal cord injury in rats [J]. Ulus Travma Acil Cerrahi Derg, 2013, 19(4): 285-293.

[11] T Brett Reece, David O Okonkwo, Peter I Ellman, et al. The evolution of ischemic spinal cord injury in function, cytoarchitecture，and inflammation and the effects of adenosine A2A receptor activation [J]. J Thorac Cardiovasc Surg, 2004, 128(6): 925-932.

[12] Hayri Kertmen, Bora Gürer, Erdal Resit Y?lmaz, et al. The protective effect of low-dose methotrexate on ischemia-reperfusion injury of the rabbit spinal cord [J]. Eur J Pharmacol, 2013, 714(1-3): 148-156.

[13] 肖寒贫，王烁宇，李华南，等. TreadScan步态分析系统在评价大鼠脊髓挫伤模型中的应用 [J]. 神经药理学报，2012, 2(3)：in press.?

[14] Bruno Benetti Torres, Fatima Maria Caldeira, Mardelene Geisa Gomes, et al. Effects of dantrolene on apoptosis and immunohistochemical expression of NeuN in the spinal cord after traumatic injury in rats [J]. Int J Exp Pathol, 2010, 91(6): 530-536.

[15] Ma Yong-jie, Wang Bing-bing, Li Wen-liang, et al. Reduction of intersectin1-s induced apoptosis of human glioblastoma cells [J]. Brain Res, 2010, 1351: 222-228.

[16] Venkata Ramesh Dasari, Daniel G Spomar, Li L, et al. Umbilical cord blood stem cell mediated down regulation of fas improves functional recovery of rats after spinal cord injury [J]. Neurochem Res, 2008, 33(1): 134-149.

[17] Michael G Fehlings, Darryl C Baptise. Current status of clinical trials for acute spinal cord injury [J]. J Injury, 2005, 36(Suppl 2): B113-B122.

[18] Jose E Pereira, Luis M Costa, Antonio M Cabrita, et al. Methylprednisolone fails to improve functional and histological outcome following spinal cord injury in rats [J]. Exp Neurol, 2009, 220(1): 71-81.

[19] L Jackson Roberts 2nd, John A Oates, M F Linton, et al. The relationship between dose of vitamin E and suppression of oxidative stress in humans [J]. Free Radic Biol Med, 2007, 43(10): 1388-1393.

[20] Anna-Leena Sirén, Theresa Fasshauer, Claudia Bartels, et al. Therapeutic potential of erythropoietin and its structural or functional variants in the nervous system [J]. Neurotherapeutics, 2009, 6(1): 108-127.

[21] Darry C Baptiste, Michael G Fehlings. Update on the treatment of spinal cord injury [J]. Prog Brain Res, 2007, 161: 217-133.

[22] Tim D Hewitson, Teresa Bisucci, Ian A Darby. Histochemical localization of apoptosis with in situ labeling of fragmented DNA [J]. Methods Mol Biol, 2006, 326: 227-234.

[23] Joachim Scholz, Andrea Abele, Claudiu Marian, et al. Low-dose methotrexate reduces peripheral nerve injury-evoked spinal microglial activation and neuropathic pain behavior in rats [J]. Pain, 2008, 138(1): 130-142.

[24] Ayromlou Hormoz, Hajipour Babak, Hossenian Mohammad Mehdi, et al. Oxidative effect of methotrexate administration in spinal cord of rabbits [J]. J Pak Med Assoc, 2011, 61(11): 1096-1099.

[25] Kobayashi T, Okamoto K, Kobata T, et al. Apomodulation as a novel therapeutic concept for the regulation of apoptosis in rheumatoid synoviocytes [J]. Curr Opin Rheumatol, 1999, 11(3): 188-193.

[26] Genestier L, Paillot R, Quemeneur L, et al. Mechanisms of action of methotrexate [J]. Immunopharmacology, 2000, 47(2-3): 247-257.

[27] Wang Zhi-gang, Chen Zhe, Yang S, et al. (1)H NMR-based metabolomic analysis for identifying serum biomarkers to evaluate methotrexate treatment in patients with early rheumatoid arthritis[J]. Exp Ther Med, 2012, 4(1):165-71.

[28] Aoyama K, Matsumura N, Watabe M, et al. Caffeine and uric acid mediate glutathione synthesis for neuroprotection [J]. Neuroscience, 2011, 181: 206-15.

Effects of Low-dose Methotrexate on Spinal Cord Contusion-Induced Neural Cell Apoptisis in Rats

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles15

Recommended Articles

Metrics

[1]	ZHANG Yu-rong，YU Jie，ZHANG Meng-di，SUN Hong-liu.Xenon Exerts Antiepileptic Effects via Increased Autophagy on Kainic Acid-Induced Acute Generalized Seizures in Rats[J]. Acta Neuropharmacologica, 2018, 8(4): 1-3.
[2]	LIU Cai-hong，WU Xian，TANG Su-su，HONG Hao*.Involvement of TGR5 in Aβ-Induced Neurotoxicity in Vivo[J]. Acta Neuropharmacologica, 2018, 8(4): 11-12.
[3]	WANG Jia-Yue，DUAN Yan-Hong，Wang Xin-He，Zhang Xu-Liang，Xu Mei-Chen, Cao Xiao-Hua *.The Effect of PHA-543613 on Memory Disorders in Presenilin1 and Presenilin2 Conditional Double Knockout Mice[J]. Acta Neuropharmacologica, 2018, 8(4): 52-53.
[4]	WU Xian, LV Yang-ge, TANG Su-Su, HONG Hao.Involvement of TGR5 in Aβ-induced Neurotoxicity in Vivo[J]. Acta Neuropharmacologica, 2018, 8(4): 53-54.
[5]	ZHANG Li-na，ZHANG Xin，XUE Juan，ZHANG Dan-shen.Advances Achievements of Traditional Chinese Medicine on the Signaling Pathways in Central Nervous Degenerative Diseases[J]. Acta Neuropharmacologica, 2017, 7(3): 33-42.
[6]	LIU Jun-ju，FU Dong-jun，NIU Jin-bo，ZHANG Qi，ZHANG Yan-bing.Antiproliferation Effects of 2’-bro-2-hydroxy-4,6,4’,5’-tetramethoxychalcone on Human Neuroblastoma SK-N-SH Cells[J]. Acta Neuropharmacologica, 2016, 6(1): 18-24.
[7]	HU Bao-ling, GUO Chun-yan.Advances Achievements about Neuroprotective Mechanisms of Paeoniflorin[J]. Acta Neuropharmacologica, 2015, 5(6): 51-56.
[8]	YANG Wen-liang, SONG Yu, ZHAO Can-can, XIE Xin-mei, PANG Xiao-bin, DU Guan-hua.Protective Effect of Astragaloside Ⅳ on PC12 Cells Injured by Oxygen-glucose Deprivation[J]. Acta Neuropharmacologica, 2015, 5(1): 10-14.
[9]	YAN Juan，ZHENG Mao-dong.Protective Effects and Mechanisms of L-carnitine in the Nervous System[J]. Acta Neuropharmacologica, 2015, 5(1): 45-50.
[10]	ZHAO Wei, LI Fang-jiang, WANG Shu.Progress in Understanding the Mechanism of Protective Effects of Cerebral Ischemia Injury[J]. Acta Neuropharmacologica, 2014, 4(6): 55-64.
[11]	YAN Juan，ZHENG Mao-Dong.Protective Effects and Mechanisms of Breviscapine on the Nervous System[J]. Acta Neuropharmacologica, 2014, 4(3): 31-38.
[12]	ZHANG Si, GU Bing, WANG Shuo-yu, LI Hua-nan, ZHANG Guo-fu, ZHANG Shui-yin.Effects of Methotrexate on Acute Phrase of Spinal Cord Contusion-Induced Lipid Peroxidation in Rats[J]. Acta Neuropharmacologica, 2014, 4(1): 11-16.
[13]	YAN Juan，ZHANG Dan-shen.Protective Effects and Mechanisms of Chrysophanol in the Nervous System[J]. Acta Neuropharmacologica, 2014, 4(1): 52-57.
[14]	WU Miao-miao，YUAN Yu-he，CHEN Nai-hong.The Effect of Polygalasaponin F on Rotenone-induced PC12 Cells and the Expression of CREB[J]. Acta Neuropharmacologica, 2013, 3(1): 12-16.
[15]	DONG Xiao-hua, MENG Xian-yong, ZHANG Li, YANG Jian-ming.Effects of Osthole on Neuronal Apoptosis and Cell Cycle in AD Rats[J]. Acta Neuropharmacologica, 2012, 2(3): 7-14.