黑果枸杞花色苷联合葡萄糖酸锌对急性脊髓损伤模型大鼠脊髓组织病理形态及炎症因子的影响

Effects of Lycium ruthenicum Murr. anthocyanin combined with zinc gluconate on histopathology and inflammatory factors of spinal cord in rats model with acute spinal cord injury

目的研究黑果枸杞花色苷联合葡萄糖酸锌对急性脊髓损伤(ASCI)模型大鼠神经功能的保护作用及其作用机制。方法将120只SD大鼠按照随机数字表法分为6组,即假手术组、模型组、醋酸泼尼松组、黑果枸杞花色苷低剂量+葡萄糖酸锌组(简称'低剂量组')、黑果枸杞花色苷中剂量+葡萄糖酸锌组(简称'中剂量组')、黑果枸杞花色苷高剂量+葡萄糖酸锌组(简称'高剂量组'),每组20只。假手术组仅给予手术暴露,其余各组均采用改良的Allen’s法建立ASCI大鼠模型,造模成功后各组分别给予相应药物灌胃,每日1次,连续灌胃14 d。分别于给药12 h、1 d、4 d、7 d、14 d比较各组大鼠脊髓损伤行为学(BBB)评分,采用ELISA法测定各组大鼠血清肿瘤坏死因子-α(TNF-α)和白细胞介素(IL)-6含量,观察脊髓组织病理形态。结果与假手术组比较,模型组各时间点BBB评分明显降低,血清TNF-α、IL-6含量明显升高,差异均有统计学意义(P<0.05或P<0.01)。与模型组比较,醋酸泼尼松组、中剂量组、高剂量组给药3,7,14 d BBB评分明显升高,低剂量组给药7,14 d BBB评分明显升高,差异均有统计学意义(P<0.05或P<0.01);醋酸泼尼松组、高剂量组给药1,3,7,14 d血清TNF-α、IL-6含量明显降低,低剂量组给药14 d血清IL-6含量明显降低,中剂量组给药3 d血清TNF-α含量明显降低,给药1,3,7,14 d血清IL-6含量明显降低,差异均有统计学意义(P<0.05或P<0.01)。模型组大鼠脊髓可见多个出血点,大量的炎性细胞浸润,神经元细胞水肿且数量减少,灰质和白质之间结构紊乱;黑果枸杞花色苷联合葡萄糖酸锌能明显改善上述脊髓组织的病理改变。结论黑果枸杞花色苷联合葡萄糖酸锌能明显改善ASCI大鼠脊髓组织的病理改变,降低血清炎症因子TNF-α、IL-6含量,从而保护大鼠急性脊髓损伤后的神经功能。

Objective To investigate the protective effect and mechanism of Lycium ruthenicum Murr.anthocyanin combined with zinc gluconate on nerve function in rats models with acute spinal cord injury(ASCI).Methods Totally 120 SD rats were divided into 6 groups according to random number table,including sham operation group,model group,prednisone acetate group,Lycium ruthenicum Murr.anthocyanin low dose plus zinc gluconate group(abbreviated as'low dose group'),Lycium ruthenicum Murr.anthocyanin medium dose plus zinc gluconate group(abbreviated as'medium dose group'),Lycium ruthenicum Murr.anthocyanin high dose plus zinc gluconate group(abbreviated as'high dose group'),with 20 rats in each group.Only the sham operation group was exposed to surgery,the other groups used modified Allen’s method to establish the ASCI rats model.After successful establishment of the model,each group was given corresponding drugs by gavage once a day for consecutive 14 days.The Basso,Beattie and Bresnahan(BBB)scores of spinal cord injury were compared at 12 h,1 day,4 days,7 days and 14 days after administration.The serum levels of TNF-αand IL-6 were measured by ELISA,and the pathological morphology of spinal cord tissue was observed.Results Compared with the sham operation group,BBB scores decreased significantly at each time point in the model group,serum levels of TNF-αand IL-6 increased significantly,the difference was statistically significant(P<0.05 or P<0.01).Compared with the model group,BBB scores increased significantly in prednisone acetate group,medium dose group and high dose group after administration for 3,7 and 14 days,and BBB scores increased significantly in the low dose group after administration for 7 and 14 days,the difference was statistically significant(P<0.05 or P<0.01).The serum levels of TNF-αand IL-6 decreased significantly after administration for 1,3,7 and 14 days in prednisone acetate group and high dose group,the serum level of IL-6 decreased significantly after administration for 14 days in the low dose group,the serum level of TNF-αdecreased significantly after administration for 3 days in the medium dose group,and the serum level of IL-6 decreased significantly after administration for 1,3,7 and 14 days,the difference was statistically significant(P<0.05 or P<0.01).In the model group,there were many hemorrhagic spots in the spinal cord,a large number of inflammatory cells infiltration,neuronal cell edema and decrease in number,and structural disorder between gray matter and white matter.It can markedly improve the pathological change of the above spinal cord tissues to use Lycium ruthenicum Murr.anthocyanin combined with zinc gluconate.Conclusion Lycium ruthenicum Murr.anthocyanin combined with zinc gluconate can significantly improve the pathological change of spinal cord tissue in ASCI rats,reduce the levels of serum inflammatory factors TNF-αand IL-6,thus protecting the neurological function of rats after acute spinal cord injury.