摘要
目的探究红花黄素(SY)对脊髓损伤(SCI)的保护作用及其作用机制。方法将SCI模型大鼠分为模型(Model)组、重组高迁移率组蛋白(HMG)B1处理组(HMGB1组,8μg/kg)、SY干预组(SY组,40 mg/kg)、HMGB1+SY组(8μg/kg HMGB1+40 mg/kg SY),另设假手术(Control组),各组均12只,各组连续给药28 d。行为学运动(BBB)评分法评定运动行为;获取脊髓组织,采用苏木素-伊红(HE)染色观察脊髓组织病理形态学变化;免疫印迹法(WB)检测脊髓组织中HMG B1、Toll样受体(TLR)4、核转录因子(NF)-κB蛋白表达;制备脊髓组织匀浆液,采用酶联免疫吸附试验检测超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSHP)、过氧化氢酶(CAT)、丙二醛(MDA)水平;Tunel染色观察脊髓组织中神经元细胞凋亡情况;Western印迹法检测B淋巴细胞瘤-2蛋白(Bcl-2)、Cleaved-caspase3、Bcl-2相关X蛋白(Bax)蛋白表达。结果与Control组相比,Model组BBB评分显著降低,SOD、GSHP、CAT水平显著降低,MDA水平显著升高,HMGB1、TLR4、NF-κB蛋白表达显著升高,脊髓组织神经细胞凋亡率显著升高,Bax、Cleaved-caspase3蛋白表达显著升高,Bcl-2蛋白表达显著降低(P<0.05)。与Model组相比,HMGB1组BBB评分显著降低,SOD、GSHP、CAT水平显著降低,MDA水平显著升高,HMGB1、TLR4、NF-κB蛋白表达显著升高,脊髓组织神经细胞凋亡率显著升高,Bax、Cleaved-caspase3蛋白表达显著升高,Bcl-2蛋白表达显著降低(P<0.05);SY组、HMGB1+SY组与Model组及HMGB1组比较BBB评分显著升高,SOD、GSHP、CAT水平显著升高,MDA水平显著降低,HMGB1、TLR4、NF-κB蛋白表达显著降低,脊髓组织神经细胞凋亡率显著降低,Bax、Cleaved-caspase3蛋白表达显著降低,Bcl-2蛋白表达升高(P<0.05)。结论 SY可能通过抑制HMGB1/TLR4/NF-κB信号通路,缓解SCI后氧化应激水平,减低脊髓组织神经元细胞凋亡,缓解脊髓组织、运动功能损伤。
Objective To investigate the protective effect of Safflor Yellow(SY) on spinal cord injury(SCI) rats and explore its mechanism.Methods SCI model rats were divided into model, recombinant high-mobility group box(HMGB)1 treatment(HMGB1 group, 8 μg/kg), SY intervention(40 mg/kg), HMGB1+SY treatment(8 μg/kg HMGB1+40 mg/kg SY), and sham(control group) groups, with 12 rats in each group. Rats in each group were administered continuously for 28 days. BBB score was used to evaluate the motor behavior of rats. The spinal cord tissues of rats were obtained. HE staining was used to observe the histopathological changes of spinal cord tissues. Western blot was used to detect the expressions of HMGB1, Toll-like receptor(TLR)4, and Nuclear factor kappa(NF-κ)B in spinal cord tissues. Homogenate of spinal cord tissue was prepared, the levels of superoxide dismutase(SOD), glutathione peroxidase(GSHP), catalase(CAT) and malondialdehyde(MDA) were detected by ELISA. Tunel staining was used to observe neuronal apoptosis in spinal cord. Western blot was used to detect the expressions of Bcl-2 protein, Cleaved-caspase 3, and Bcl-2 related X protein(Bax).Results Compared with control group, BBB score decreased, SOD, GSHP, CAT levels decreased, MDA levels increased, HMGB1, TLR4, NF-κB protein expression increased, apoptotic rate of spinal cord neurons increased, Bax, Cleaved-caspase 3 protein expression increased, Bcl-2 protein expression decreased in model group(P<0.05). Compared with model group, BBB score decreased, SOD, GSHP, CAT level decreased, MDA level increased, HMGB1, TLR4, NF-κB protein expression increased, apoptotic rate of spinal cord neurons increased, Bax, Cleaved-caspase 3 protein expression increased, Bcl-2 protein expression decreased in HMGB1 group(P<0.05);BBB score increased, SOD, GSHP and CAT levels increased, MDA levels decreased, HMGB1, TLR4, NF-κB protein expression decreased, apoptotic rate of spinal cord neurons decreased, Bax, Cleaved-caspase 3 protein expression decreased and Bcl-2 protein expression increased in SY group and HMGB1+SY group(P<0.05).Conclusions SY might alleviate oxidative stress after spinal cord injury, reduce apoptosis of neurons in spinal cord tissue, and alleviate spinal cord tissues injury and motor function injury by inhibiting HMGB1/TLR4/NF-κB signaling pathway.
作者
王大伟
胡培
吴迎爽
赵春桃
WANG Da-Wei;HU Pei;WU Ying-Shuang(Department of Orthopedics and Arthrology,Zhangjiakou First Hospital,Zhangjiakou 075000,Hebei,China)
出处
《中国老年学杂志》
CAS
北大核心
2020年第6期1313-1319,共7页
Chinese Journal of Gerontology
基金
河北省医学科学研究重点课题计划项目(20160249)
