乌梅总黄酮对MPP^(+)诱导SH-SY5Y细胞线粒体损伤的保护作用  

Protective effect of fructus mume total flavone on mitochondrial damage induced by MPP^(+) in SH-SY5Y cells

作　　者：王春玲 罗宁 文晓东[2] 蒋媛静[2] 刘钰[2] 冯文勇 WANG Chunling;LUO Ning;WEN Xiaodong;JIANG Yuanjing;LIU Yu;FENG Wenyong(School of Pharmacy,Guangxi University of Chinese Medicine,Nanning,Guangxi 530200,China;Department of Neurology,Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning,Guangxi 530011,China)

机构地区：[1]广西中医药大学药学院,南宁530200 [2]广西中医药大学附属瑞康医院神经内科,南宁530011

出　　处：《重庆医学》2022年第12期1981-1986,共6页Chongqing medicine

基　　金：国家自然科学基金项目(82060888);广西中医药管理局立项项目(GZZC2020107);2018广西中医药大学一流学科项目(2018XK089);广西中医药大学2020年校级课题(2020MS049)。

摘　　要：目的探讨乌梅总黄酮(FMF)对神经毒素1-甲基-4-苯基吡啶离子(MPP+)诱导人神经母细胞瘤(SH-SY5Y)细胞建立的帕金森病(PD)细胞模型钙调蛋白激酶(CaMKKβ)/腺苷酸活化蛋白激酶(AMPK)通路的调控作用及对SH-SY5Y细胞线粒体功能的影响。方法250μmol/L MPP+作用于SH-SY5Y细胞24 h后,建立PD体外细胞模型,将细胞分为5组:对照组(只加SH-SY5Y细胞,不加药物)、模型组(250μmol/L MPP+孵育24 h)、FMF低剂量组(MPP^(+)+FMF 10μmol/L)、FMF中剂量组(MPP^(+)+FMF 50μmol/L)和FMF高剂量组(MPP^(+)+FMF 100μmol/L)。CCK-8法检测细胞的存活率,JC-1染色检测线粒体膜电位(ΔΨm);Western blot检测CaMKKβ/AMPK信号通路相关CaMKKβ、AMPK及磷酸化CaMKKβ(p-CaMKK)、磷酸化AMPK(p-AMPK)表达水平。结果CCK-8检测结果显示,FMF预处理24 h后,FMF低、中、高剂量组MPP+损伤细胞的存活率均较模型组明显升高(P<0.05或P<0.01)。ΔΨm检测结果显示,与对照组相比,模型组细胞ΔΨm明显降低(P<0.001);与模型组相比,FMF低、中、高剂量组ΔΨm明显升高(P<0.001)。Western blot检测结果显示,与对照组比较,模型组p-CaMKKβ、p-AMPK表达水平均明显下降(P<0.01);与模型组比较,FMF低、中、高剂量组p-CaMKKβ、p-AMPK表达水平均明显上调(P<0.05或P<0.01)。FMF干预前、后各组间CaMKKβ、AMPK表达水平比较,差异均无统计学意义(P>0.05)。结论FMF可减轻MPP+对SH-SYSY细胞的损伤,这种保护作用可能是通过激活CaMKKβ/AMPK通路,改善线粒体功能实现的。Objective To investigate the effect of fructus mume total flavone(FMF)on the regulation of calmodulin kinase/adenosine monophosphate activated protein kinase(CaMKKβ/AMPK)pathway in human neuroblastoma(SH-SY5Y)cells induced by neurotoxin 1-methyl-4-phenylpyridyl ion(MPP^(+)),a cell model of Parkinson’s disease(PD),and the mitochondrial function in SH-SY5Y cells.Methods SH-SY5Y cells were treated with 250μmol/L MPP^(+)for 24 h,and an in vitro model of PD was established.The cells were divided into 5 groups:the control group(only added SH-SY5Y cells,without any drug),the model group(250μmol/L MPP+incubated for 24 h),and the FMF low-,medium-,and high-dose groups(MPP^(+)+10,50,100μmol/L FMF,respectively).CCK-8 assay was used to detect the cell survival rate,JC-1 staining was usedto detect the change of mitochondrial membrane potential(ΔΨm),Western blot was used to detect the expression levels of CaMKKβ/AMPK signaling pathway relative proteins CaMKKβ,AMPK,phosphorylated CaMKKβ(p-CaMKK)and phosphorylated AMPK(p-AMPK).Results The results of CCK-8 showed that after FMF pretreatment for 24 h,the survival rate of MPP^(+)damaged cells in the FMF low-,medium-,and high-dose groups was significantly higher than that in the model group(P<0.05 orP<0.01).The results ofΔΨm test showed that,compared with the control group,theΔΨm of cells in the model group was decreased significantly(P<0.001).Compared with the model group,theΔΨm of cells in the FMF low-,medium-,and high-dose groups was significantly increased(P<0.001).Western blot analysis of related proteins showed that,compared with the control group,the expression levels of p-CaMKKβand p-AMPK were significantly reduced in the model group(P<0.01).Compared with the model group,the expression levels of p-CaMKKβand p-AMPK were significantly increased in the FMF low-,medium-,and high-dose groups(P<0.05 or P<0.01).There was no significant difference in the expression levels of CaMKKβand AMPK before or after FMF pretreatment(P>0.05).Conclusion FMF can alleviate the d

关 键 词：帕金森病 乌梅总黄酮 1-甲基-4-苯基吡啶离子 钙调蛋白激酶 腺苷酸活化蛋白激酶 线粒体 

分 类 号：R284.141[医药卫生—中药学]