香萱解郁方对血清剥夺诱导HT22细胞损伤模型的神经保护作用  

Effect of Xiangxuan Jieyu decoction on serum deprivation-induced HT22 cells injury

作　　者：高圆媛 沙中玮 薛纯纯[1] 徐建[1] 李欧 GAO Yuanyuan;SHA Zhongwei;XUE Chunchun;XU Jian;LI Ou(Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China)

机构地区：[1]上海中医药大学附属市中医医院,上海200071

出　　处：《西部医学》2023年第6期793-798,共6页Medical Journal of West China

基　　金：上海市青年科技英才扬帆计划项目(20YF1446100);上海市加强公共卫生体系建设三年行动计划重点学科建设项目(GWV-10.1-XK20)。

摘　　要：目的探讨香萱解郁方含药血清对血清剥夺致小鼠海马神经元HT22细胞损伤模型的影响。方法采用血清剥夺培养HT22细胞建立神经损伤体外模型,实验分为对照组、模型组和中药组[中药组A(含药血清15%浓度)、中药组B(含药血清20%浓度)],各组血清剥夺12 h后,通过CCK8法检测各组细胞活力,确定15%浓度含药血清干预后细胞的存活率最高,设定为后续实验中药组的药物浓度。免疫荧光染色法检测神经元特异性微管蛋白(β-TubulinⅢ)在各组中的表达,蛋白质免疫印迹法及qPCR法检测脑源性神经营养因子(BDNF)蛋白及其mRNA在各组中的表达。结果在加药后培养12 h,中药组的细胞活力明显高于对照组与模型组(P<0.001)。培养24 h,模型组细胞活力较对照组明显下降(P<0.001),中药组较模型组明显提高(P<0.001)。培养36 h,模型组细胞活力较对照组显著下降(P<0.001),中药组较模型组明显提高(P<0.001)。模型组BDNF蛋白含量及BDNF mRNA表达量较对照组显著降低(P<0.05,P<0.001),模型组少数神经元长出突起;中药组BDNF蛋白含量及BDNF mRNA表达量较模型组显著增加(P<0.05),中药组HT22细胞轴突突起长度和分支增加,β-TubulinⅢ阳性表达明显增多。结论香萱解郁方含药血清对血清剥夺诱导小鼠海马神经元HT22细胞损伤具有神经保护作用,可能与促进BDNF蛋白表达有关。Objective To investigate the effect of Xiangxuan Jieyu decoction on serum deprivation induced HT22 cells injury.Methods In this study,serum deprivation culture was used to establish a cell injury model,the CCK8 assay was used to examine the effect of the drug-containing serum on cell viability.Based on viability results,test concentrations for subsequent experiments were set at 15%Xiangxuan Jieyu decoction drug-containing serum,as the TCM group.Then the experiment was divided into control group,model group and TCM group.Serum was deprived for 12 hours,then immunofluorescence staining was performed to detect the neuron-specificβ-tubulin III expression.Western blot and qPCR analysis was used to detect the expression of trophic factors brain-derived neurotrophic factor(BDNF)and BDNF mRNA.Results Compared with control group and model group,cell viability was increased significantly in TCM group at 12h(P<0.001).Cell viability was decreased in model group compared with control group(P<0.001),which was increased in TCM group compared with model group(P<0.001)at 24h.Compared with control group,cell viability was decreased in model group(P<0.001),and increased in TCM group compared with model group(P<0.001)at 36h.BDNF protein(P<0.05)and BDNF mRNA expression(P<0.001)was significantly decreased in the model group,and a fewβ-tubulin III-positive neurites compared with control group.BDNF protein and BDNF mRNA expression(P<0.05)was significantly increased in the TCM group,the neurite branch and neurite length in the HT22 cell neurons was increased,and moreβ-tubulin III-positive neurites compared with the model group.Conclusion Xiangxuan Jieyu decoction drug-containing serum could exert the neuroprotective effect against serum deprivation-induced cell injury in HT22 hippocampal neuronal cells by promoting BDNF expression.

关 键 词：含药血清 血清剥夺 HT22细胞 神经保护 

分 类 号：R285.5[医药卫生—中药学]