摘要
【目的】探究辣木籽油对人小脑颗粒细胞γ-氨基丁酸受体(GABAA)细胞膜氯离子通道的影响。【方法】首先利用冷榨法制得辣木籽油,通过连续培养人小脑颗粒细胞(human cerebellar granule cells,HCGC),采用CCK8法检测辣木籽油对细胞活力的影响,14 d后表达功能性GABAA受体,经不同剂量(2、4、8μg/mL)辣木籽油、油酸、β-谷甾醇、豆甾醇处理,空白组给予生理盐水,阳性药给予戊巴比妥钠(10 mmol/L),最后采用N,N二甲基二吖啶硝酸盐(MQAE)荧光探针检测小脑颗粒细胞内的氯离子浓度。【结果】辣木籽油等样品在10μg/mL浓度以下对细胞活力没有任何影响;与空白组相比,辣木籽油(4、8μg/mL)、油酸(4、8μg/mL)、豆甾醇(4、8μg/mL)及β-谷甾醇(8μg/mL)对HCGC细胞以剂量依赖方式激活氯离子通道,且具有统计学差异(P<0.05)。此外,阳性药戊巴比妥钠(10 mmol/L)亦可激活HCGC细胞的氯离子通道,并具有统计学差异(P<0.01)。【结论】证明辣木籽油体外抑制神经元兴奋与GABA神经系统有关。
【Objective】To explore the effect of Moringa oleifera seeds oil(MOO)on chloride ion flow of GABAA receptors in human cerebellar granule cells(HCGC).【Method】The MOO was obtained by the cold pressed method.The HCGC cells were cultured 14 d successively.The CCK8 assay was used to de‐termine cell viability,then the functional GABAA receptor was expressed.The cells were treated with‐MOO,oleic acid,β-sitosterol and stigmasterol at different concentrations i.e.2,4,8μg/mL.The blank control group was given normal saline;the positive group was treated with pentobarbital sodium solution at the concentration of 10 mmol/L.MQAE fluorescence probe was used to detect chloride ion concentration of HCGC cells.【Result】The cytotoxicity assay showed that all treatments at the concentration of 10μg/mL had noncytotoxic to the cells.Compared with blank control group,the treatments of MOO(4,8μg/mL),oleic acid(4,8μg/mL),stigmasterol(4,8μg/mL)andβ-sitosterol(8μg/mL)significantly activated chloride channel of HCGC cells in dose-dependent manner(P<0.05).Moreover,the positive group was dramatically activated the chloride channel(P<0.01).【Conclusion】It is suggested that MOO inhib‐ited neuron excitation is related with GABA nervous system.
作者
刘威良
徐晴芳
高秀
蔡建
LIU Weiliang;XU Qingfang;GAO Xiu;CAI Jian(Qujing Normal University,Yunnan Engineering Research Center of Fruit Wine,Qujing 655011,China)
出处
《甘肃农业大学学报》
CAS
CSCD
2022年第4期177-183,共7页
Journal of Gansu Agricultural University
基金
云南省省校教育合作共建重点项目(曲靖师范学院-中国农业大学云南高原特色水果深加工新产品研发与成果转化平台建设)。
关键词
辣木籽油
冷榨法
人小脑颗粒细胞
Γ-氨基丁酸受体
氯离子通道
Moringa oleifera seed oil
cold pressed method
human cerebellar granulosa cells
GABAA receptor
chloride channels
