天麻素对镉诱导的星形胶质细胞损伤的保护效应  

Protective Effect of Gastrodin on Cadmium-induced Injury of Astrocytes of Mice

作　　者：周翠红 薛姗姗 刘江正 王化宁 顾婷婷 彭正午 ZHOU Cui-hong;XUE Shan-shan;LIU Jiang-zheng;WANG Hua-ning;GU Ting-ting;PENG Zheng-wu(Department of Psychosomatic,Xijing Hospital,Fourth Military Medical University,Xi'an,Shaanxi,710032,China;Department of toxicology,Fourth Military Medical University,Xi'an,Shaanxi,710032,China;Department of Anesthesia,Xijing Hospital,Fourth Military Medical University,Xi'an,Shaanxi,710032,China)

机构地区：[1]空军军医大学西京医院心身科,陕西西安710032 [2]空军军医大学毒理教研室,陕西西安710032 [3]空军军医大学西京医院麻醉科,陕西西安710032

出　　处：《现代生物医学进展》2020年第6期1015-1021,共7页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81630032,81401109);国家科技支撑计划项目(2016YFC1307100)。

摘　　要：目的:探讨氯化镉(CdCl2)和天麻素(GAS)对小鼠星形胶质细胞活力及神经营养因子GDNF和抗氧化基因Nrf2,HO-1,SOD-1表达的影响。方法:首先,给予体外培养的小鼠星形胶质细胞不同浓度的CdCl2(Con,2.5μM,5μM,10μM,20μM)处理24 h或48 h,随后检测细胞活力筛选出造成星形胶质细胞损伤的CdCl2浓度和时间。然后使用上述筛选的CdCl2作用浓度(5μM)构建星形胶质细胞损伤的同时再给予不同浓度的天麻素(0,20μg/mL,30μg/mL,40μg/mL,50μg/mL)处理24 h或48 h,随后检测细胞活力并提取细胞RNA检测其caspase3,GDNF(胶质源性神经营养因子Glial cell-derived neurotrophic factor,GDNF)和Nrf2(Nuclear factor erythroid2-related factor2),HO-1(Heme oxygenase 1),SOD-1(superoxide dismutase 1)等抗氧化基因的mRNA表达的变化。结果:(1)2.5μM CdCl2处理24 h后星形胶质细胞活力已经有明显下降(P<0.05),5μM CdCl2处理24 h后,星形胶质细胞活力显著下降(P<0.01);(2)CdCl2浓度越大,细胞损伤严重;(3)一定浓度的天麻素处理可以缓解CdCl2造成的星形胶质损伤,恢复其细胞活力,下调caspase3 mRNA水平;(4)CdCl2下调了星形胶质细胞的GDNF,Nrf2,HO-1和SOD-1的mRNA水平,天麻素可以抑制CdCl2对上述基因的mRNA水平的调节作用,且浓度越高调节作用越强。结论:天麻素可能通过调节小鼠星形胶质细胞的GDNF,Nrf2,HO-1和SOD-1基因表达缓解CdCl2导致的细胞损伤。Objective:To investigate the effect of cadmium and gastrodin on cell viability and the effect on the expression level of GDNF,Nrf2,HO-1 and SOD-1 of astrocytes from the hippocampus of mice.Methods:First,cultured hippocampal astrocytes from new-born(one day old)mice were divided into control and cadmium groups:control group was treated with medium(5%FBS+DMEM)and cadmium treatment groups were treated with 2.5,5,10 or 20μM CdCl2 for 24 or 48 hours respectively.Then cell viability was measured by CCK-8 and the appropriate concentration and time point(5μM,24 h)of CdCl2 to induce cell injury were determined.After that,cultured astrocytes were divided into control,sham and GAS groups:control group was treated with medium(5%FBS+DMEM),sham group was treated with 5μM CdCl2 for 24 h,GAS groups were treated with 5μM CdCl2 and 20μg/m L,30μg/m L,40μg/m L,50μg/m L gastrodin for 24 h respectively at the same time.Then cell viability was measured by CCK-8 and total RNA was extracted from cells and the m RNA levels of GDNF,Nrf2,HO-1 and SOD-1 were measured by Real-time PCR.Results:(1)Compared with the control,CdCl2 significantly decreased the cell viability of astrocytes;(2)The effect of isoflurane revealed a dose effect---compared with 2.5μM group,cells treated with 5.0-20μM of CdCl2 suffered more influence;(3)5μM CdCl2 treated 24 h significantly down-regulated the m RNA levels of GDNF,Nrf2,HO-1 and SOD-1 m RNA level in the astrocytes;(4)Gastrodin administration restored the cell viability and expression of caspase3,GDNF,Nrf2,HO-1 and SOD-1 in the astrocytes post-CdCl2 exposures.Conclusion:Gastrodin administration restored the decreased cell viability and reversed the disturbed expression of GDNF,Nrf2,HO-1 and SOD-1 in astrocytes exposed to CdCl2.

关 键 词：镉 星形胶质细胞 GDNF Nrf2 HO-1 SOD-1 

分 类 号：R-33[医药卫生] R285.5