摘要
目的:研究七福饮含药血清对高糖诱导作用下小鼠海马神经元细胞(HT22细胞)的损伤保护作用。方法:确定高糖培养液最适造模浓度;将雄性大鼠预留10只作为正常对照组,其余20只灌胃七福饮8.6 g/kg。灌胃7 d后大鼠腹主动脉采血,制备浓度为5%、10%、15%七福饮含药血清;将各浓度含药血清作用于HT22细胞48 h后以细胞计数试剂(CCK-8)法检测细胞活力;以CCK-8法检测七福饮含药血清对高糖诱导下HT22细胞的活力;以流式细胞术Annexin-V PI细胞双染法检测HT22细胞凋亡率;提取HT22细胞蛋白,聚氰基丙烯酸正丁酯(BCA)试剂盒测定蛋白浓度;蛋白免疫印迹(Western Blot)法检测高糖诱导下HT22细胞相关蛋白表达。结果:选择75 mmol/L高糖干预细胞48 h作为最适造模浓度。与正常对照组相比,模型对照组细胞活力显著降低,细胞密度显著降低,细胞凋亡率显著升高,磷酸化JNK、P38蛋白、BAX、Caspase-3蛋白表达显著上调,BCL2蛋白表达显著下调(P<0.01);与模型对照组相比,5%、10%七福饮含药血清组细胞活力均显著升高,细胞密度显著升高,细胞凋亡率显著下降(P<0.01);半胱氨酸蛋白酶-3(Caspase-3)、BCL-2相关X蛋白(BAX)、磷酸化P38蛋白(p-P38)、磷酸化c-Jun氨基末端激酶(p-JNK)的表达明显下调(P<0.05或P<0.01),B淋巴细胞瘤-2基因(BCL-2)蛋白表达显著上调(P<0.01)。结论:七福饮可通过调节P38/c-Jun氨基末端激酶通路(JNK)信号通路,从而改善高糖诱导的HT22细胞损伤和凋亡。
Objective:To study the protective effect of Qifu Decoction(七福饮)-containing serum on the injury induced by high glucose in mouse hippocampal neurons(HT22 cells).Methods:The optimal concentration of glucose in the high sugar culture medium for modeling was determined.Ten male rats were selected as the normal control group,and the remaining 20 rats were administrated with 8.6 g/kg Qifu Decoction by gavage.After 7 days of administration,blood was collected from the abdominal aorta of rats,and the sera containing 5%,10%,and 15%Qifu Decoction were prepared.After HT22 cells were treated with the sera containing different concentrations of Qifu Decoction for 48 h,the cell counting kit-8(CCK-8)was employed to measure cell viability.Flow cytometry with Annexin V-FITC/PI double staining was employed to examine the apoptosis rate of HT22 cells.The concentration of protein extracted from HT22 cells was measured by the polybutylcyanoacrylate(BCA)protein assay kit.Western blotting was used to determine the expression levels of proteins in HT22 cells exposed to high glucose.Results:The optimal modeling scheme was determined as intervention with 75 mmol/L high glucose for 48 h.Compared with the normal control group,the model group showed decreased cell viability and density,increased apoptosis rate,up-regulated expression of phosphorylated c-Jun amino terminal kinase(p-JNK),phosphorylated P38 protein(p-P38),cysteine aspartate-specific protease-3(Caspase-3),and BCL-2-associated X protein(BAX),and down-regulated expression of B-cell lymphoma 2(BCL-2)(P<0.01).The apoptotic rate of HT22 Compared with the normal control group,the model control group showed decreased cell viability and cell density(P<0.01).Compared with the model control group,the 5%and 10%Qifu Decoction-containing serum groups showed increased cell viability and density,decreased apoptosis rate(P<0.01),down-regulated expression of Caspase-3,BAX,p-P38,and p-JNK(P<0.05 or P<0.01),and up-regulated expression of BCL-2(P<0.01).Conclusion:Qifu Decoction can alleviate
作者
史磊磊
张静文
王欣
李溪
张雪航
韩朝军
王斌
卫昊
刘继平
SHI Leilei;ZHANG Jingwen;WANG Xin;LI Xi;ZHANG Xuehang;HAN Chaojun;WANG Bin;WEI Hao;LIU Jiping(College of Pharmacy,Shaanxi University of Chinese Medicine,Xianyang 712046;Key Laboratory of Pharmacodynamic Mechanism and Material Basis of Traditional Chinese Medicine,Shaanxi Administration of Traditional Chinese Medicine,Xianyang 712046)
出处
《中药药理与临床》
CSCD
北大核心
2023年第10期48-53,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
陕西省重点研发计划项目(编号:2021SF-072)
陕西省教育厅服务地方专项科研计划项目(编号:22JC029)
陕西中医药大学学科创新团队项目(编号:2019-YL13)
陕西省中医药管理局中医药科研项目(编号:2021-ZZ-JC003)。
