丁苯酞对脂肪源性神经元细胞凋亡的作用  

The role of butylphthalide on apoptosis of neurons derived from adipose-derived stromal cells

作　　者：陆艳卉 李斌[1] 王彦[1] 周桂娟[1] 纪茹英[1] 元小冬[2] LU Yanhui;LI Bin;WANG Yan;ZHOU Guijuan;JI Ruying;YUAN Xiaodong(Tangshan People's Hoepital,Tangshan 063000,China)

机构地区：[1]河北省唐山市人民医院神经内科,063000 [2]开滦总医院神经内科,河北省神经生物机能重点实验室

出　　处：《中国煤炭工业医学杂志》2023年第4期337-345,共9页Chinese Journal of Coal Industry Medicine

基　　金：河北省医学科学研究课题计划(编号:20190172)。

摘　　要：目的观察丁苯酞对成人脂肪基质细胞(ADSC)来源神经元细胞凋亡的影响。方法本研究时间为2019年4月—2020年12月。脂肪基质细胞在β-巯基乙醇诱导下分化为神经元,诱导后细胞分为未加丁苯酞组和加丁苯酞组,每组记录诱导1h、3h、5h、8h四个时间点。未加丁苯酞组与加丁苯酞组均通过倒置相差显微镜、透射电子显微镜观察细胞形态变化,免疫组化法检测NSE、NF-200、GFAP、caspase-3,MTT检测神经元细胞的生长曲线,TUNEL法检测细胞凋亡。结果未加丁苯酞组及加丁苯酞组诱导后ADSC均呈典型的神经元形态,电镜下一些神经元细胞的胞体回缩,胞核表面不光滑,核膜回缩、细胞核碎裂,可见凋亡小体,线粒体出现肿胀、嵴断裂、线粒体膜断裂。两组神经元细胞NSE、NF-200表达为阳性。未加丁苯酞组NF-200及NSE随诱导时间延长阳性率逐渐增高,诱导5h达高峰,NSE及NF-200诱导8h与5h相比P>0.05,其余时间点两两比较P<0.05。加丁苯酞组NF-200及NSE诱导8h达高峰,NSE诱导8h与5h相比P>0.05,其余时间点两两比较P<0.05;加丁苯酞组NF-200各个时间点两两比较P<0.05。NSE及NF-200四个时间点两组之间P<0.05。GFAP表达为阴性。Caspase-3在二组中,随诱导时间延长阳性率逐渐升高,8h达高峰,且各个时间点两两比较和相同时间点两组之间比较差异均有统计学意义(P<0.05)。二组TUNEL随诱导时间延长阳性表达率逐渐增高,8h达高峰,两组中各个时间点两两比较P<0.05,诱导1h时两组之间比较P>0.05,诱导3h、5h、8h两组之间比较P<0.05。二组MTT随诱导时间的延长细胞数量逐渐减少;未加丁苯酞组诱导3h与5h相比P>0.05,其余时间点两两比较P<0.05。加入丁苯酞组诱导后各时间点细胞数量差异有统计学意义(P<0.05),各时间点两组之间比较差异有统计学意义(P<0.05)。结论细胞凋亡是脂肪基质细胞诱导分化为神经元过程中发生死亡的主要原因,丁苯酞对�Objective To observe the effect of butphthalide on apoptosis of adult adipose stromal cells(ADSC)-derived neurons. Methods This study was conducted from April 2019 to December 2020.Adipose stromal cells differentiated into neurons induced by β-mercaptoethanol, and the cells were divided into untetrabenzphthalide group and butrimphthalide group, and four time points of induction 1h, 3h, 5h, and 8h were recorded in each group.The untetrabenzphthalide group was observed by inverted phase contrast microscopy, NSE,NF-200,GFAP,caspase-3 were detected by immunohistochemistry, the growth curve of neuron-like cells was detected by MTT,and apoptosis was detected by TUNEL method.Gatephthalide group immunohistochemistry to detect Caspase-3,MTT to detect the growth curve of neuron-like cells, and TUNEL to detect apoptosis. Results ADSC in non-butylphthalide group with the addition of butylphthalide group showed typical neuronal morphology.Under electron microscope, some neuronal cells showed cell body retraction, nucleus surface was not smooth, nuclear membrane retraction, nucleus fragmentation, apoptotic bodies, mitochondrial swelling, crest fracture and mitochondrial membrane fracture.The expressions of NSE and NF-200 in neurons of two groups were positive.The positive rates of NF-200 and NSE in non-butylphthalide group increased with extension of induction time, and reached the peak in 5h induction group.NSE and NF-200 in unbutylphthalide group were induced at 8h compared with 5h P<0.05,and pairings at other time points P<0.05.NF-200 and NSE induced 8h group reached the peak.In addition to butylphthalide group, NSE induction at 8h was P<0.05 compared with that at 5h, and pairwise comparison at other time points was P<0.05.Pair-to-pair comparison of NF-200 at each time point P<0.05.There were significant differences in NSE and NF-200 between two groups at four time points(P<0.05).GFAP expression was negative.In both groups, the positive rate of Caspase-3 increased gradually with extension of induction time, and reached the

关 键 词：丁苯酞 成人ADSC 诱导分化 神经元 

分 类 号：R349.65[医药卫生—基础医学]