夹脊电针对急性脊髓损伤大鼠运动功能及脊髓组织NogoA、RhoA、ROCK Ⅱ蛋白表达的影响
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摘要
目的:观察夹脊电针对急性脊髓损伤模型大鼠肢体运动功能和脊髓组织髓鞘相关抑制因子勿动蛋白A(Nogo-A)、Ras同源基因家族成员A(RhoA)和Rho激酶Ⅱ(ROCKⅡ)蛋白表达的影响。方法:将18只成年雌性Wistar大鼠随机分为假手术组、模型组和夹脊电针组,每组6只。采用NYU打击器制备急性脊髓损伤模型,夹脊电针组给予损伤节段上下一对夹脊穴电针治疗。治疗1天、3天、7天、14天、28天后采用BBB评分法对大鼠急性脊髓损伤后运动功能进行评估;治疗28天后采用HE染色观察脊髓组织病理形态学变化;采用Western blot法检测脊髓组织Nogo-A、RhoA和ROCKⅡ蛋白表达。结果:与假手术组比较,模型组和夹脊电针组在各时间点BBB评分显著降低(P<0.05);与模型组比较,夹脊电针组在治疗3天后BBB评分显著升高(P<0.05)。与假手术组比较,模型组Nogo-A、RhoA和ROCKⅡ蛋白表达均显著升高(P<0.05);与模型组比较,夹脊电针组Nogo-A、RhoA和ROCKⅡ蛋白表达均显著降低(P<0.05)。结论:夹脊电针治疗能够显著改善急性脊髓损伤模型大鼠肢体运动功能和脊髓组织病理形态学变化,可能与其下调脊髓损伤微环境中Nogo-A、RhoA、ROCKⅡ蛋白表达有关。
Objective: To observe the effect of Jiaji electro-acupuncture on limb motor function and protein expressions of myelin-related inhibitory factor Nogo-A,as well as homologous genes of RhoA and ROCKⅡin rats with acute spinal cord injury( ASCI). Methods: 18 Wistar rats were randomly divided into the sham operation group( group A),the model group( group B),and the Jiaji electro-acupuncture group( group C),with 6 rats in each group. The model of ASCI was made with NYU striker. The electro-acupuncture group was treated with Jiaji points of one section upper and lower respectively of the injury segment. The motor function was evaluated by BBB score after 1 d,3 d,7 d,14 d and 28 d of the treatment; the pathological changes of spinal cord were observed by HE staining after 28 days of the treatment; protein expressions of Nogo-A,RhoA,and ROCKⅡ were detected with Western blot method. Results: In terms of BBB score,it was decreased significantly both in group B and group C when compared to group A at each time point( P < 0. 05); it was significantly increased in group C after 3 days of the treatment when compared to group B( P < 0. 05). In terms of protein expressions of Nogo-A,RhoA and ROCKⅡ,they were significantly increased in group B compared to group B( P < 0. 05); they were all lower in group C compared to those in group B( P < 0. 05). Conclusion: The treatment of Jiaji electro-acupuncture can significantly improve the limb motor function and pathological changes of spinal cord in rats with ASCI,which may be related to the down-regulation of the protein expressions of Nogo-A,RhoA and ROCKII in micro-environment of spinal cord injury.
Objective: To observe the effect of Jiaji electro-acupuncture on limb motor function and protein expressions of myelin-related inhibitory factor Nogo-A,as well as homologous genes of RhoA and ROCKⅡin rats with acute spinal cord injury( ASCI). Methods: 18 Wistar rats were randomly divided into the sham operation group( group A),the model group( group B),and the Jiaji electro-acupuncture group( group C),with 6 rats in each group. The model of ASCI was made with NYU striker. The electro-acupuncture group was treated with Jiaji points of one section upper and lower respectively of the injury segment. The motor function was evaluated by BBB score after 1 d,3 d,7 d,14 d and 28 d of the treatment; the pathological changes of spinal cord were observed by HE staining after 28 days of the treatment; protein expressions of Nogo-A,RhoA,and ROCKⅡ were detected with Western blot method. Results: In terms of BBB score,it was decreased significantly both in group B and group C when compared to group A at each time point( P < 0. 05); it was significantly increased in group C after 3 days of the treatment when compared to group B( P < 0. 05). In terms of protein expressions of Nogo-A,RhoA and ROCKⅡ,they were significantly increased in group B compared to group B( P < 0. 05); they were all lower in group C compared to those in group B( P < 0. 05). Conclusion: The treatment of Jiaji electro-acupuncture can significantly improve the limb motor function and pathological changes of spinal cord in rats with ASCI,which may be related to the down-regulation of the protein expressions of Nogo-A,RhoA and ROCKII in micro-environment of spinal cord injury.
引文
[1]吕威,李冰,景泉凯,等.电针“大椎”“命门”对脊髓损伤大鼠神经元细胞凋亡及JNK信号通路相关蛋白表达的影响[J].针刺研究,2017,42(1):14-19.
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[3]李华南,张海明,顾兵,等.针刺促进脊髓损伤后神经功能恢复的机制及相关信号通路的作用[J].中国康复理论与实践,2017,23(6):641-644.
[4]Fujita Y,Yamashita T.Axon growth inhibition by Rho A/ROCK in the central nervous system[J].Front Neurosci,2014,8(338):1-12.
[5]吴锋,李怀斌,赵健,等.电针结合天麻素对局灶性脑缺血大鼠神经功能、额叶皮质勿动蛋白A及其受体表达的影响[J].针刺研究,2016,41(1):65-69.
[6]Monnier PP,Sierra A,Schwab JM,et al.The Rho/ROCK pathway mediates neurite growth-inhibitory activity associated with the chondroitin sulfate proteoglycans of the CNS glial scar[J].Mol Cell Neurosci,2003,22(3):319-330.
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[9]李晓宁,张良,杨庆红,等.夹脊电针对脊髓损伤后大鼠c-fosmRNA及BNIP3 mRNA表达影响的实验研究[J].针灸临床杂志,2015,31(7):83-86.
[10]Basso DM,Beattie MS,Bresnahan JC.Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection[J].Exp Neurol,1996,139(2):244-256.
[11]李晓宁,吴磊,迟蕾,等.不同治疗周期夹脊电针对急性脊髓损伤大鼠运动功能及细胞凋亡的影响[J].针刺研究,2016,41(6):492-496.
[12]李晓宁,吴磊,梅继林,等.夹脊电针联合甲强龙对急性脊髓损伤大鼠肢体运动功能及尼氏小体影响的实验研究[J].针灸临床杂志,2016,32(12):61-64,91.
[13]胡华辉,黄小龙,刘飞,等.电针夹脊穴治疗脊髓损伤机制的实验研究进展[J].上海针灸杂志,2016,35(12):1480-1483.
[14]霍会霞.头电针配合夹脊电针治疗脊髓损伤后下肢运动功能障碍的临床观察[D].哈尔滨:黑龙江中医药大学,2014.
[15]SCHWAB ME.Repairing the injured spinal cord[J].Science,2002,295(5557):1029-1031.
[16]尹明锡,时素华,宋萌.电针对大鼠脊髓损伤后Nogo-A表达的实验研究[J].陕西中医,2010,31(3):371-373.
[17]朱枫,鲁秀敏,王永堂.中枢神经再生抑制因子及免疫治疗研究[J].现代生物医学进展,2008,8(9):1743-1745.
[18]Wu BQ,Bi ZG,Qi Q.Inactivation of the Rho-ROCK signaling pathway to promote neurologic recovery after spinal cord injuries in rats[J].Chin Med J(Engl),2013,126(19):3723-3727.
[19]尤武林,张亚峰,陈剑峰,等.中药脊髓康对脊髓损伤大鼠NogoNgR基因表达的影响[J].中国骨伤,2015,28(3):235-239.
[20]姚年伟.丹参酮IIA对大鼠急性脊髓损伤保护作用及其机制研究[D].武汉:湖北中医药大学,2016.
[2]Jain NB,Ayers GD,Peterson EN,et al.Traumatic spinal cord injury in the United States,1993-2012[J].JAMA,2015,313(22):2236-2343.
[3]李华南,张海明,顾兵,等.针刺促进脊髓损伤后神经功能恢复的机制及相关信号通路的作用[J].中国康复理论与实践,2017,23(6):641-644.
[4]Fujita Y,Yamashita T.Axon growth inhibition by Rho A/ROCK in the central nervous system[J].Front Neurosci,2014,8(338):1-12.
[5]吴锋,李怀斌,赵健,等.电针结合天麻素对局灶性脑缺血大鼠神经功能、额叶皮质勿动蛋白A及其受体表达的影响[J].针刺研究,2016,41(1):65-69.
[6]Monnier PP,Sierra A,Schwab JM,et al.The Rho/ROCK pathway mediates neurite growth-inhibitory activity associated with the chondroitin sulfate proteoglycans of the CNS glial scar[J].Mol Cell Neurosci,2003,22(3):319-330.
[7]Lehmann M,Fournier A,Selles-Navarro I,et al.Inactiviation of Rho signaling pathway promotes CNS axon regeneration[J].J Neurosci,1999,19(17):7535-7547.
[8]李晓宁,迟蕾.夹脊配合督脉电针治疗脊髓损伤后功能障碍临床观察[J].上海针灸杂志,2015,34(10):972-975.
[9]李晓宁,张良,杨庆红,等.夹脊电针对脊髓损伤后大鼠c-fosmRNA及BNIP3 mRNA表达影响的实验研究[J].针灸临床杂志,2015,31(7):83-86.
[10]Basso DM,Beattie MS,Bresnahan JC.Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection[J].Exp Neurol,1996,139(2):244-256.
[11]李晓宁,吴磊,迟蕾,等.不同治疗周期夹脊电针对急性脊髓损伤大鼠运动功能及细胞凋亡的影响[J].针刺研究,2016,41(6):492-496.
[12]李晓宁,吴磊,梅继林,等.夹脊电针联合甲强龙对急性脊髓损伤大鼠肢体运动功能及尼氏小体影响的实验研究[J].针灸临床杂志,2016,32(12):61-64,91.
[13]胡华辉,黄小龙,刘飞,等.电针夹脊穴治疗脊髓损伤机制的实验研究进展[J].上海针灸杂志,2016,35(12):1480-1483.
[14]霍会霞.头电针配合夹脊电针治疗脊髓损伤后下肢运动功能障碍的临床观察[D].哈尔滨:黑龙江中医药大学,2014.
[15]SCHWAB ME.Repairing the injured spinal cord[J].Science,2002,295(5557):1029-1031.
[16]尹明锡,时素华,宋萌.电针对大鼠脊髓损伤后Nogo-A表达的实验研究[J].陕西中医,2010,31(3):371-373.
[17]朱枫,鲁秀敏,王永堂.中枢神经再生抑制因子及免疫治疗研究[J].现代生物医学进展,2008,8(9):1743-1745.
[18]Wu BQ,Bi ZG,Qi Q.Inactivation of the Rho-ROCK signaling pathway to promote neurologic recovery after spinal cord injuries in rats[J].Chin Med J(Engl),2013,126(19):3723-3727.
[19]尤武林,张亚峰,陈剑峰,等.中药脊髓康对脊髓损伤大鼠NogoNgR基因表达的影响[J].中国骨伤,2015,28(3):235-239.
[20]姚年伟.丹参酮IIA对大鼠急性脊髓损伤保护作用及其机制研究[D].武汉:湖北中医药大学,2016.