白桦脂酸通过阻断Fas-Fasl信号通路抑制异氟醚诱导的大鼠脑神经元损伤  

Inhibitive effect of betulinic acid on isoflurane-induced brain neuron injury by blocking the Fas-Fasl signaling pathway

作　　者：寿琼华[1] 解雅英[1] SHOU Qionghua;XIE Yaying(Department of Anesthesiology,Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,China)

机构地区：[1]内蒙古医科大学附属医院麻醉科

出　　处：《山西医科大学学报》2019年第8期1122-1127,共6页Journal of Shanxi Medical University

基　　金：内蒙古自治区自然科学基金资助项目(2015MS0886)

摘　　要：目的研究白桦脂酸对异氟醚诱导的发育期大鼠脑神经元细胞的效应,探讨其通过阻断Fas-Fasl信号通路抑制神经元损伤的干预机制。方法分离4周龄SD大鼠海马组织,并培养海马神经元细胞,将海马神经元细胞随机分为对照组、异氟醚诱导组、白桦脂酸低剂量组、白桦脂酸中剂量组、白桦脂酸高剂量组。异氟醚诱导组细胞于2.4%异氟醚环境中暴露2 h,白桦脂酸各剂量组分别给相应剂量药物培养后置于2.4%异氟醚环境中暴露2 h,对照组于正常环境中培养。流式细胞仪检测海马神经元细胞的凋亡及线粒体膜电位水平;RT-PCR和Western blot检测脑神经细胞元Fas、Fasl、caspase-8、cyt c、BID、caspase-9、Bcl-2、Bax、caspase-3、PARP表达水平;氧电极法检测大鼠线粒体呼吸水平;紫外分光光度法检测细胞内ATP含量。结果流式细胞分析表明,与异氟醚诱导组比较,白桦脂酸组脑神经元细胞的凋亡均受到抑制,线粒体膜电位水平升高(均P<0.05);与对照组和异氟醚诱导组比较,白桦脂酸能够使Fas、Fasl、caspase-8、cyt c、BID、caspase-9、Bax、caspase-3、PARP表达水平下调(均P<0.05),Bcl-2表达水平上调(P<0.05);氧电极和紫外分光光度检测表明,与对照组和异氟醚诱导组比较,白桦脂酸组大鼠线粒体呼吸水平和细胞内ATP含量提高(P<0.05)。结论一定浓度的白桦脂酸可通过阻断Fas-Fasl信号通路,显著抑制异氟醚诱导发育期大鼠脑神经元的损伤。Objective To observe the effect of betulinic acid on isoflurane-induced neurons in rat brain during the development,and to explore the intervention mechanism of betulinic acid inhibiting neuron injury by blocking the Fas-fasl signaling pathway.Methods The hippocampal neurons of 4-week-old SD rats were isolated and cultured.The hippocampal neurons were randomly divided into blank control group,isoflurane group,low dose group 1(10^-3 mol/L betulinic acid),medium dose group(5×10^-3 mol/L betulinic acid)and high dose group(10^-2 mol/L betulinic acid).Cells in isoflurane group were exposed to 2.4%isoflurane environment for 2 h,cells in betulinic acid groups were exposed 2.4%isoflurane environment for 2 h after different concentrations of betulinic acid,and cells in control group were cultured in normal environment.Flow cytometry was used to detect the apoptosis of hippocampal neurons and the level of mitochondrial membrane potential.RT-PCR and Western blot were used to detect the expression levels of Fas,Fasl,caspase-8,cyt c,BID,caspase-9,Bcl-2,Bax,caspase-3 and PARP.The oxygen electrode method was used to detect the mitochondrial respiratory level in rats,and ultraviolet spectrophotometry was used to detect the content of ATP in cells.Results Flow cytometry analysis showed that the apoptosis was inhibited in betulinic acid groups and the mitochondrial membrane potential was increased compared with isoflurane group(all P<0.05).Compared with control group and isoflurane group,the levels of Fas,Fasl,caspase-8,cyt c,BID,caspase-9,Bax,caspase-3 and PARP were down-regulated in betulinic acid groups,and Bcl-2 levels was up-regulated(all P<0.05).The mitochondrial respiratory level and intracellular ATP content in rats were increased in betulinic acid groups compared with control group and isoflurane group(all P<0.05).Conclusion Betulinic acid can significantly inhibit the injury of brain neurons induced by isoflurane by blocking the Fas-fasl signaling pathway.

关 键 词：白桦脂酸 Fas-Fasl信号通路 异氟醚 脑神经元损伤 

分 类 号：R363[医药卫生—病理学]