大鼠血清外泌体对皮质酮诱导损伤PC12细胞的影响
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摘要
目的观察大鼠血清外泌体对皮质酮诱导损伤的PC12细胞存活率的影响,并探讨其对胞内5-羟色胺含量的影响。方法试剂盒法提取外泌体; MTT法检测PC12细胞活性;细胞免疫荧光法测定PC12细胞胞内5-羟色胺含量。结果成功提取和鉴定大鼠血清外泌体;皮质酮能浓度依赖性地诱导PC12细胞损伤;大鼠血清外泌体能够抑制皮质酮诱导的细胞损伤;皮质酮诱导损伤细胞中胞内5-羟色胺含量增加,血清外泌体抑制该诱导作用,降低5-羟色胺的含量。结论大鼠血清外泌体能够提高皮质酮诱导损伤的PC12细胞活性,降低损伤细胞胞内5-羟色胺的含量。
Objective To investigate the effect of rat serum-derived exosomes on the survival rate of PC12 cells induced by corticosterone,and observe the changes of intracellular serotonin content.Methods Exosomes were extracted by using exosomes extraction kit.MTT assay was used to determine activity of PC12 cells.The intracellular serotonin content of PC12 cells was determined by immunofluorescence assay.Results The rat serum-derived exosomes were successfully extracted and identified.Corticosterone damaged PC12 cells in a concentration-dependent manner.The rat serum-derived exosomes inhibited corticosteroneinduced PC12 cell injury.The increase of the intracellular serotonin content in PC12 cells was induced by corticosterone,while serum-derived exosomes inhibited the phenomenon and decreased the content of serotonin.Conclusion Rat serum-derived exosomes can increase the activity of injured PC12 cells induced by corticosterone and reduce the content of intracellular serotonin.
Objective To investigate the effect of rat serum-derived exosomes on the survival rate of PC12 cells induced by corticosterone,and observe the changes of intracellular serotonin content.Methods Exosomes were extracted by using exosomes extraction kit.MTT assay was used to determine activity of PC12 cells.The intracellular serotonin content of PC12 cells was determined by immunofluorescence assay.Results The rat serum-derived exosomes were successfully extracted and identified.Corticosterone damaged PC12 cells in a concentration-dependent manner.The rat serum-derived exosomes inhibited corticosteroneinduced PC12 cell injury.The increase of the intracellular serotonin content in PC12 cells was induced by corticosterone,while serum-derived exosomes inhibited the phenomenon and decreased the content of serotonin.Conclusion Rat serum-derived exosomes can increase the activity of injured PC12 cells induced by corticosterone and reduce the content of intracellular serotonin.
引文
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[2]王鑫伟,毛建文.外泌体的功能浅析[J].临床与病理杂志,2016,36(3):290-294.
[3]STEINBICHLERT B,DUDAS J,RIECHLMANN H,et al.The role of exosomes in cancer metastasis[J].Semin Cancer Biol,2017,44:170-181.
[4]ARAUJO FARIAS V,O'VALLE F,SERRANO-SAENZ S,et al.Exosomes derived from mesenchymal stem cells enhance radiotherapy-induced cell death in tumor and metastatic tumor foci[J].Mol Cancer,2018,17(1):122-133.
[5]VICENCIO J M,ZHENG Y,DAS D,et al.Plasma exosomes from rats and humans protect the myocardium from ischemiareperfusion injury[J].Cardiovasc Res,2014,65(15):1525-1536.
[6]TSILIONI I,PANAGIOTIDOU S,THEOHAIDES T C.Exosomes in neurologic and psychiatric disorders[J].Clin Ther,2014,36(6):882-888.
[7]HUANG Shan,GE Xintong,YU Jinwen,et al.Increased miR-124-3p in microglial exosomes following traumatic brain injury inhibits neuronal inflammation and contributes to neurite outgrowth via their transfer into neurons[J].Faseb J,2018,32(1):512-528.
[8]朱维煜,冯霞,王瑾,等.定志小丸对miR-16及5-HT重摄取的影响[J].中国中药杂志,2018,43(17):3513-3518.
[9]NUMAKAWA T,ADCHI N,RICHARDS M,et al.Brain-derived neurotrophic factor and glucocorticoids:reciprocal influence on the central nervous system[J].Neuroscience,2013(23),239:157-172.
[10]黄文超,李云峰,罗质璞.单胺递质对皮质酮所致的PC12细胞损伤的保护作用[J].中国药理学通报,2003,19(1):103-106.
[11]何小燕,陈建丽,向欢,等.谷氨酸和皮质酮诱导的PC12抑郁症细胞模型差异性的~1H NMR代谢组学研究[J].药学学报,2017,52(2):245-252.
[12]ADEOSUN S O,ALBERTl P R,AUSTIN M C,et al.17β-estradiol-induced regulation of the novel 5-HT 1A-related transcription factors NUDR and Freud-1 in SH SY5Y cells[J].Cell Mol Neurobiol,2012,32(4):517-521.
[13]JIANG Baoping,LIU Yamin,LE Liang,et al.Cajaninstilbene acid prevents corticosterone-induced apoptosis in PC12 cells by inhibiting the mitochondrial apoptotic pathway[J].Cell Physiol Biochem,2014,34(3):1015-1026.
[14]QU Mengke,LIN Qing,HUANG Luyi,et al.Dopamine-loaded blood exosomes targeted to brain for better treatment of Parkinson's disease[J].J Control Release,2018,287:156-166.
[15]刘金鹏.两种造模方式致抑郁症大鼠模型的脑PET研究[D].北京:中国人民解放军军事医学科学院,2011.
[16]李永超,彭亮,王高华,等.炎性因子、皮质醇、5-羟色胺与抑郁严重程度关系的初步研究[J].国际精神病学杂志,2015,42(6):6-9.
[17]JASINSKA A J,LOWRY C A,BURMEISTER M.Corrigendum:serotonin transporter gene,stress,and raphe-raphe interactions:a molecular mechanism of depression[J].Trends Neurosci,2012,35(7):454-455.
[18]王静,马强,刘洪涛.5-羟色胺对过量皮质酮影响海马神经元钾电流的干预作用[R].中国生理学会第21届全国代表大会暨学术会议,2002.