广西医科大学学报2024,Vol.41Issue(2) :218-224.DOI:10.16190/j.cnki.45-1211/r.2024.02.008

牡荆素对脊髓损伤大鼠运动功能和炎症反应的影响

Effect of vitexin on motor function and inflammation in rats with spinal cord injury

谢佩雅 粟叶萍 韦入菲 曾颖虹 陈欣颖 宗少晖 曾高峰
广西医科大学学报2024,Vol.41Issue(2) :218-224.DOI:10.16190/j.cnki.45-1211/r.2024.02.008

牡荆素对脊髓损伤大鼠运动功能和炎症反应的影响

Effect of vitexin on motor function and inflammation in rats with spinal cord injury

谢佩雅 1粟叶萍 2韦入菲 2曾颖虹 2陈欣颖 2宗少晖 3曾高峰1
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作者信息

  • 1. 广西医科大学药学院,南宁 530021
  • 2. 广西医科大学公共卫生学院,南宁 530021
  • 3. 广西医科大学第一附属医院脊柱骨病外科,南宁 530021
  • 折叠

摘要

目的:探究牡荆素(VT)对脊髓损伤(SCI)大鼠和脂多糖(LPS)诱导的BV2小胶质细胞的影响.方法:体内实验:将SD大鼠分为假手术组(sham组)、模型SCI组(SCI组)和VT低剂量组、VT中剂量组和VT高剂量组.采用Basso-Beattie-Bresnah-an(BBB)评分评估大鼠的运动功能;实时荧光定量PCR(RT-qPCR)和酶联免疫吸附实验(ELISA)检测大鼠炎症因子基因和蛋白表达;苏木精—伊红(HE)染色观察脊髓组织结构.体外实验:将BV2小胶质细胞分为正常对照组(control组)、LPS组、VT低、中、高、剂量组和VT高剂量单独给药组,LPS诱导BV2小胶质细胞活化构建体外神经炎症模型;细胞计数试剂盒(CCK-8)法筛选VT安全浓度范围;RT-qPCR和蛋白质免疫印迹法(western blotting)检测细胞炎症基因和蛋白表达水平.结果:体内实验:与SCI组比较,VT中、高剂量组的BBB评分明显升高,损伤第7天,VT中剂量组评分显著高于VT高剂量组(均P<0.05);VT中剂量组TNF-α mRNA表达较SCI组降低,VT中、高剂量组IL-10 mRNA表达较SCI组显著升高(均P<0.05);VT低剂量组、VT中剂量组和VT高剂量组的TNF-α蛋白水平显著降低(均P<0.05),VT各剂量组的IL-10蛋白水平显著升高(均P<0.05);HE结果显示,sham组脊髓组织结构完整,神经元数量较多、排列较为规则整齐;SCI组脊髓组织炎症细胞浸润,出现较多空洞,神经元减少萎缩且排列不整齐;与SCI组比较,VT治疗组损伤更小,炎症浸润减轻较为明显.体外实验:与LPS组相比,VT各剂量组的TNF-α mRNA表达显著降低(均P<0.05),且成剂量依赖性降低;VT各剂量组TNF-α蛋白表达量降低(均P<0.001).结论:VT通过抑制大鼠炎症反应,减轻脊髓组织病理学损伤,改善损伤后脊髓局部微环境,从而保护受损脊髓神经元,达到促进运动功能恢复的作用.

Abstract

Objective:To investigate the effect of vitexin(VT)on spinal cord injury(SCI)rats and lipopolysac-charide(LPS)-induced BV2 microglia.Methods:In vivo experiments:SD rats were divided into sham-operated group(sham group),model SCI group(model group)and VT low-dose,VT medium-dose and VT high-dose groups.Basso-Beattie-Bresnahan(BBB)score was used to evaluate the motor function of rats;reverse transcrip-tion-quantitative PCR(RT-qPCR)and enzyme-linked immunoassay(ELISA)were used to detect the gene and protein expression of inflammatory factors in rats;hematoxylin-eosin(HE)staining was used to observe the spi-nal cord tissue structure.In vitro experiments:BV2 microglia were divided into normal control group(control group),LPS group,VT low-dose,medium-dose and high-dose groups and VT high-dose alone administration group;LPS-induced BV2 microglia activation was used to construct in vitro neuroinflammation model;cell counting kit-8(CCK-8)method was used to screen the range of safe concentration of VT;RT-qPCR and western blotting were used to detect the levels of cellular inflammatory gene and protein expression.Results:In vivo ex-periments:compared with the SCI group,the BBB scores of the VT medium-and high-dose groups were signifi-cantly higher,and the score of the VT medium-dose group was significantly higher than that of the high-dose group on the 7th day of the injury(both P<0.05);the expression of TNF-α mRNA in the VT medium-dose group was lower compared with that of the SCI group,and the expression of IL-10 mRNA in the VT medium-and high-dose groups was significantly higher compared with that in the SCI group(both P<0.05);the protein levels of TNF-α in the VT low-dose group,VT medium-dose group and VT high-dose group were significantly decreased(all P<0.05),and the protein levels of IL-10 in the VT dose groups were significantly increased(all P<0.05);HE results showed that the spinal cord in the sham group was structurally intact,with more neurons and a more regular arrangement;spinal cord tissue in the SCI group was infiltrated with inflammatory cells,showed more cavities,and neurons were reduced and atrophied with irregular arrangement;compared with the SCI group,the damage in the VT treatment group was smaller,and the reduction of inflammatory infiltration was more obvious.In vitro experiments:compared with the LPS group,TNF-α mRNA expression was significantly reduced in each dose group of VT(all P<0.05),and reduced in a dose-dependent manner;TNF-α protein expression was reduced in each group of VT(all P<0.001).Conclusion:VT inhibits the inflammation of rats,reduces the histopathologi-cal damage of the spinal cord,and improves the local microenvironment of the spinal cord after injury,thereby protecting the damaged spinal cord neurons,and promoting the recovery of motor function.

关键词

牡荆素/脊髓损伤/炎症反应/神经保护

Key words

vitexin/spinal cord injury/inflammation/neuroprotection

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基金项目

国家自然科学基金(81860391)

广西自然科学基金(2023GXNSFDA026050)

出版年

2024
广西医科大学学报
广西医科大学

广西医科大学学报

CSTPCD
影响因子:0.788
ISSN:1005-930X
参考文献量25
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