黄芩茎叶黄酮对大鼠急性脊髓损伤后的神经保护作用及对HMGB1/TLR4/NF-κB信号通路的影响
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摘要
目的探究黄芩茎叶黄酮(SSF)对急性脊髓损伤(ASCI)大鼠神经的保护作用,及其潜在的分子作用机制。方法将大鼠分为假手术组、模型组、甲强龙组、SSF低剂量组、SSF中剂量组、SSF高剂量组,每组12只。采用脊髓运动功能(BBB)评分法评定大鼠脊髓损伤情况,HE染色检测脊髓组织病理学变化,TUNEL检测脊髓损伤组织细胞凋亡率,蛋白质印迹法(Western blotting)检测HMGB1、Toll样受体4(TLR4)、核因子κB(NF-κB)的表达水平。结果模型组脊髓坏死严重,出现很多空腔,灰质中神经元结构破坏,细胞核固缩;与模型组相比,甲强龙组和SSF组因坏死形成的空腔减少。甲强龙组和SSF组BBB评分显著高于模型组(P<0.05);甲强龙组和SSF组脊髓细胞凋亡率以及脊髓组织中HMGB1、TLR4、NF-κB表达水平显著低于模型组(P<0.05);SSF高剂量组BBB评分、脊髓细胞凋亡率以及脊髓组织中HMGB1、TLR4、NF-κB表达水平与甲强龙组相比,差异无统计学意义(P>0.05)。结论 SSF对ASCI大鼠神经具有保护作用,这一作用可能是通过调控HMGB1/TLR4/NF-κB信号通路实现的,为ASCI的临床治疗提供一定的理论基础。
Objective: To explore the protective effect of flavonoids from stem leaf of scutellaria baicalensis georgi(SSF) on the nerve of rats with acute spinal cord injury(ASCI) and its potential molecular mechanism.Methods: Rats were divided into the sham operation group,the model group,Methylprednisolone group,SSF low dose group,SSF medium dose group and SSF high dose group,with 12 rats in each group. The spinal cord motor function(BBB) score was used to evaluate the spinal cord injury in rats;the pathological changes of spinal cord tissue were detected by HE staining;TUNEL was used to detect the apoptosis rate of tissue cells in spinal cord injury;the expression levels of HMGB1,TLR4 and NF-κB were detected by Western blotting(WB). Results: In the model group,the spinal cord necrosis was severe;there were many cavities,and the neuron structure in gray matter was destroyed,with nuclear pyknosis. Compared with the model group,the cavity caused by necrosis was reduced in methylprednisolone group and SSF group. The BBB scores in methylprednisolone group and SSF group were significantly higher than those in the model group(P<0.05). The apoptosis rate of spinal cord and the expression level of HMGB1,TLR4 and NF-κB in spinal cord tissue in methylprednisolone group and SSF group were significantly lower than those in model group(P<0.05). The BBB score,the apoptosis rate of spinal cord cells and the expression level of HMGB1,TLR4 and NF-κB in the spinal cord of high dose group of SSF were not statistically significant,compared with those in methylprednisolone group(P>0.05). Conclusion: SSF has a protective effect on the nerve of ASCI rats,which may be achieved by regulating the HMGB1/TLR4/NF-κB signaling pathway,and it provides a theoretical basis for the clinical treatment of ASCI.
Objective: To explore the protective effect of flavonoids from stem leaf of scutellaria baicalensis georgi(SSF) on the nerve of rats with acute spinal cord injury(ASCI) and its potential molecular mechanism.Methods: Rats were divided into the sham operation group,the model group,Methylprednisolone group,SSF low dose group,SSF medium dose group and SSF high dose group,with 12 rats in each group. The spinal cord motor function(BBB) score was used to evaluate the spinal cord injury in rats;the pathological changes of spinal cord tissue were detected by HE staining;TUNEL was used to detect the apoptosis rate of tissue cells in spinal cord injury;the expression levels of HMGB1,TLR4 and NF-κB were detected by Western blotting(WB). Results: In the model group,the spinal cord necrosis was severe;there were many cavities,and the neuron structure in gray matter was destroyed,with nuclear pyknosis. Compared with the model group,the cavity caused by necrosis was reduced in methylprednisolone group and SSF group. The BBB scores in methylprednisolone group and SSF group were significantly higher than those in the model group(P<0.05). The apoptosis rate of spinal cord and the expression level of HMGB1,TLR4 and NF-κB in spinal cord tissue in methylprednisolone group and SSF group were significantly lower than those in model group(P<0.05). The BBB score,the apoptosis rate of spinal cord cells and the expression level of HMGB1,TLR4 and NF-κB in the spinal cord of high dose group of SSF were not statistically significant,compared with those in methylprednisolone group(P>0.05). Conclusion: SSF has a protective effect on the nerve of ASCI rats,which may be achieved by regulating the HMGB1/TLR4/NF-κB signaling pathway,and it provides a theoretical basis for the clinical treatment of ASCI.
引文
[1]曹鹏,石长贵,袁文.甲基强的松龙在急性脊髓损伤中的应用进展[J].中国脊柱脊髓杂志,2015,25(2):174-178.
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[4]白江涛,龚理,赵淑敏,等.黄芩茎叶总黄酮对缺血再灌注大脑皮质神经元和突触超微结构的保护作用[J].中华老年心脑血管病杂志,2015,17(10):1091-1093.
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[7]孔维,苗光新,纪海茹,等.黄芩茎叶总黄酮预处理对脑缺血再灌注脊髓突触超微结构和突触密度的影响[J].解剖学杂志,2015,38(1):41-44.
[8]Basso DM,Beattie MS,Bresnahan JC.Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection[J].Experimental Neurology,1996,139(2):244-256.
[9]贺琴琴,王廷华,罗朝志.微RNA在脊髓损伤中的作用及研究进展[J].华西医学,2016,35(10):1782-1789.
[10]黄学英,王丽华,柳尧花.MOTOmed运动训练在颈髓损伤四肢瘫痪患者康复中的应用[J].海军医学杂志,2015,36(6):581-583.
[11]朱双龙,段会全,刘英富,等.柴胡皂苷a对大鼠急性脊髓损伤的神经保护作用与机制研究[J].中国修复重建外科杂志,2017,31(7):825-829.
[12]陈博来.活血通督汤治疗急性脊髓损伤及其与血清内皮素关系的实验研究[D].广州:广州中医药大学,2001.
[13]乔若飞,王凤英,李良业,等.益气活血汤治疗急性脊髓损伤及对SOD、MDA的影响[J].光明中医,2012,27(2):272-273.
[14]程建军,赵泓翔,郭可,等.黄芩茎叶黄酮减轻复合Aβ所致大鼠记忆障碍及抑制神经细胞凋亡的调节机制[J].中国新药杂志,2016,25(22):2627-2636.
[15]王玉梅,刘永平,曹凯,等.黄芩茎叶黄酮对大鼠缺血性记忆障碍和神经炎症的作用[J].中国药理学与毒理学杂志,2011,25(2):135-140.
[16]王玉梅,刘永平,曹凯,等.黄芩茎叶黄酮对慢性脑缺血大鼠脑内NMDA受体和VEGF表达的影响[J].中国病理生理杂志,2012,28(2):353-357.
[17]Lu B,Tang Y,Zhao X,et al.Regulation of post-translational modifications of HMGB1 during immune responses[J].Antioxidants&Redox Signaling,2015,24(12):620-634.
[18]杨晶,王国忠,刘雪华,等.高压氧对大鼠脊髓损伤后脊髓组织高迁移率族蛋白B1表达的影响[J].中华航海医学与高气压医学杂志,2012,19(4):193-197.
[19]张立才.芍药苷通过TLR4炎症通路和Nrf2氧化应激通路减轻大鼠脊髓损伤的实验研究[J].海南医学院学报,2017,23(8):1030-1033.
[20]李咪咪,刘芳,许昌声,等.甲氨蝶呤对急性脊髓损伤大鼠核因子κB及炎性因子的影响[J].中国临床药理学杂志,2016,32(17):1587-1590.
[2]吴毛,冯骅.中西医结合治疗急性脊髓损伤后便秘48例[J].现代中西医结合杂志,2008,17(35):5483-5484.
[3]Shang YZ,Gong MY,Zhou XX,et al.Improving effects of SSF on memory deficits and pathological changes of neural and immunological systems in senescent mice[J].Acta pharmacologica Sinica,2001,22(12):1078-1083.
[4]白江涛,龚理,赵淑敏,等.黄芩茎叶总黄酮对缺血再灌注大脑皮质神经元和突触超微结构的保护作用[J].中华老年心脑血管病杂志,2015,17(10):1091-1093.
[5]Allen AR.Surgery of experimental lesion of spinal cord equivalent to crush injury of fracture dislocation of spinal column[J].Journal of the American Medical Association,1911,57(7):878-880.
[6]张南文,叶剑,蔡燕亮,等.姜黄素预处理保护急性脊髓损伤小鼠神经功能的实验研究[J].中国临床药理学杂志,2015,31(11):950-953.
[7]孔维,苗光新,纪海茹,等.黄芩茎叶总黄酮预处理对脑缺血再灌注脊髓突触超微结构和突触密度的影响[J].解剖学杂志,2015,38(1):41-44.
[8]Basso DM,Beattie MS,Bresnahan JC.Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection[J].Experimental Neurology,1996,139(2):244-256.
[9]贺琴琴,王廷华,罗朝志.微RNA在脊髓损伤中的作用及研究进展[J].华西医学,2016,35(10):1782-1789.
[10]黄学英,王丽华,柳尧花.MOTOmed运动训练在颈髓损伤四肢瘫痪患者康复中的应用[J].海军医学杂志,2015,36(6):581-583.
[11]朱双龙,段会全,刘英富,等.柴胡皂苷a对大鼠急性脊髓损伤的神经保护作用与机制研究[J].中国修复重建外科杂志,2017,31(7):825-829.
[12]陈博来.活血通督汤治疗急性脊髓损伤及其与血清内皮素关系的实验研究[D].广州:广州中医药大学,2001.
[13]乔若飞,王凤英,李良业,等.益气活血汤治疗急性脊髓损伤及对SOD、MDA的影响[J].光明中医,2012,27(2):272-273.
[14]程建军,赵泓翔,郭可,等.黄芩茎叶黄酮减轻复合Aβ所致大鼠记忆障碍及抑制神经细胞凋亡的调节机制[J].中国新药杂志,2016,25(22):2627-2636.
[15]王玉梅,刘永平,曹凯,等.黄芩茎叶黄酮对大鼠缺血性记忆障碍和神经炎症的作用[J].中国药理学与毒理学杂志,2011,25(2):135-140.
[16]王玉梅,刘永平,曹凯,等.黄芩茎叶黄酮对慢性脑缺血大鼠脑内NMDA受体和VEGF表达的影响[J].中国病理生理杂志,2012,28(2):353-357.
[17]Lu B,Tang Y,Zhao X,et al.Regulation of post-translational modifications of HMGB1 during immune responses[J].Antioxidants&Redox Signaling,2015,24(12):620-634.
[18]杨晶,王国忠,刘雪华,等.高压氧对大鼠脊髓损伤后脊髓组织高迁移率族蛋白B1表达的影响[J].中华航海医学与高气压医学杂志,2012,19(4):193-197.
[19]张立才.芍药苷通过TLR4炎症通路和Nrf2氧化应激通路减轻大鼠脊髓损伤的实验研究[J].海南医学院学报,2017,23(8):1030-1033.
[20]李咪咪,刘芳,许昌声,等.甲氨蝶呤对急性脊髓损伤大鼠核因子κB及炎性因子的影响[J].中国临床药理学杂志,2016,32(17):1587-1590.