姜黄素通过调节AKT/CREB/BDNF通路缓解朊病毒感染的神经细胞损伤  

Curcumin Alleviates Neuronal Cell Injury in Prion Infection by Modulating the AKT/CREB/BDNF Pathway

作　　者：杨文静[1] 赵晓燕[1] 刘淑娇[1] 高瑞娜[1] YANG Wenjing;ZHAO Xiaoyan;LIU Shujiao;GAO Ruina(Shuozhou Vocational Technical College,Shuozhou 036002,China)

机构地区：[1]朔州职业技术学院,朔州036002

出　　处：《病毒学报》2023年第5期1243-1252,共10页Chinese Journal of Virology

摘　　要：探究姜黄素对朊病毒感染模型的蛋白激酶B/环磷酸腺苷反应元件结合蛋白/脑源性神经营养因子(Protein kinase B/CAMP response element binding protein/brain-derived neurotrophic factor,AKT/CREB/BDNF)信号通路的影响。首先摸索姜黄素的安全给药浓度和给药时间,将培养的细胞分为对照组(SMB-PS细胞+DMSO)、对照组+姜黄素(SMB-PS细胞+姜黄素)、模型组(SMB-S15细胞+DMSO)、模型组+姜黄素低剂量组(SMB-S15细胞+姜黄素+DMSO)。采用MTT法检测各组细胞存活率,检测乳酸脱氢酶(Lactic dehydrogenase,LDH)、丙二醛(Malondialdehyde,MDA)和谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px)评估细胞活性,Hoechst 33258染色观察各组细胞凋亡,Western Blot法检测各组磷酸化蛋白激酶B(Phosphorylated protein kinase B,pAKT)/AKT、磷酸化环磷酸腺苷反应元件结合蛋白(Phosphorylated CAMP response element binding protein,pCREB)/CREB和BDNF蛋白表达,实时荧光定量PCR检测AKT/CREB/BDNF mRNA表达。对照组细胞呈锥形,细胞突触显著,长而有光泽。模型组细胞呈悬浮样,突触消失或变圆,皱缩显著,细胞存活率、GSH-px、pAKT/AKT、pCREB/CREB和BDNF的蛋白及mRNA表达下调(P<0.05),LDH、MDA、细胞内的荧光强度、光密度值显著增加(P<0.05)。与模型组比,姜黄素低和高剂量组细胞突触和皱缩显著改善,细胞存活率、GSH-px、pAKT/AKT、pCREB/CREB和BDNF的蛋白及mRNA表达上调(P<0.05),LDH、MDA、细胞内的荧光强度、光密度值显著降低(P<0.05)。模型组NO和谷氨酸表达上调(P<0.01),乙酰胆碱(Acetylcholine,ACh)表达下调(P<0.01);与模型组比,姜黄素干预后的NO和谷氨酸表达下调(P<0.01),ACh表达上调(P<0.01),且均呈姜黄素浓度依赖趋势。姜黄素可调节AKT/CREB/BDNF信号通路缓解朊病毒感染神经细胞损伤,且具有浓度依赖性趋势。To examine the effect of curcumin on the protein kinase B/CAMP response element binding protein/brain-derived neurotrophic factor(AKT/CREB/BDNF)signalling pathway in a prion infection model.The effect of the BDNF signalling pathway on the brain-derived neurotrophic factor(BDNF)was investigated.The cultured cells were divided into SMB-PS cell(control group),SMB-PS cell(control group)+curcumin,SMBS15 cell(model group),SMB-S15 cell(model group)+curcumin.The cell survival rate of each group was measured by MTT,and the cell activity was assessed by lactate dehydrogenase(LDH),malondialdehyde(MDA)and glutathione peroxidase(GSH-Px),and Hoechst 33258 staining for apoptosis and Western Blot for phosphorylated protein kinase B(pAKT)/AKT,phosphorylated CAMP response element binding protein(pCREB),and GSH-Px.The expression of AKT/CREB/BDNF mRNA was measured by real-time fluorescence PCR.The control cells were conical in shape,with obvious synapses,long and shiny.In the model group,the cells were suspension-like,the synapses disappeared or became rounded,and wrinkled significantly.The cell viability,GSH-px,pAKT/AKT,pCREB/CREB and BDNF protein and mRNA were significantly reduced(P<0.05),and LDH,MDA,intracellular fluorescence intensity and optical density values were significantly increased(P<0.05).Compared with the model group,the low and high doses of curcumin significantly improved synapses and wrinkles,increased cell viability,GSH-px,pAKT/AKT,pCREB/CREB and BDNF protein and mRNA(P<0.05),and significantly decreased LDH,MDA,intracellular fluorescence intensity and optical density values(P<0.05).In the model group,NO and glutamate levels increased significantly(P<0.01)and acetylcholine(Ach)release decreased significantly(P<0.01);after curcumin treatment,NO and glutamate release decreased significantly(P<0.01)and ACh release increased significantly(P<0.01)compared to the model group.Curcumin can regulate the AKT/CREB/BDNF signaling pathway to alleviate the damage of prion-infected neuronal cells.

关 键 词：神经细胞 朊病毒 姜黄素 蛋白激酶B/环磷酸腺苷反应元件 蛋白/脑源性神经营养因子 

分 类 号：R373.9[医药卫生—病原生物学] Q421[医药卫生—基础医学]