摘要
目的探讨雌激素受体介导的柚皮苷(NG)抗β淀粉样蛋白25-35(Aβ_(25-35))诱导大鼠肾上腺嗜铬细胞瘤(PC12)细胞的凋亡作用及其与雌激素受体(ER)信号通路的关系。方法实验分为空白组、Aβ_(25-35)组、E2+Aβ_(25-35)组、NG+Aβ_(25-35)组、ICI182780+E2+Aβ_(25-35)组、ICI182780+NG+Aβ_(25-35)组。实验采用Annexin V-FITC/PI双染法检测细胞凋亡率;蛋白免疫印迹法(WB)检测细胞Bax、Bcl-2和Caspase-3的表达;RT-qPCR法检测细胞凋亡因子mRNA的表达。结果Annexin V-FITC/PI双染色流式细胞术结果显示,与空白组相比,Aβ_(25-35)组细胞凋亡率升高(P<0.01);与Aβ_(25-35)组相比,NG+Aβ_(25-35)组细胞凋亡率降低(P<0.01);与NG+Aβ_(25-35)组相比,ICI182780+NG+Aβ_(25-35)组细胞凋亡率升高(P<0.01)。WB结果显示,与空白组相比,Aβ_(25-35)组细胞Bax和Caspase-3蛋白表达量升高(P<0.01),Bcl-2则相反(P<0.01);与Aβ_(25-35)组相比,E2+Aβ_(25-35)组Bax和Caspase-3蛋白表达量降低(P<0.01),Bcl-2则相反(P<0.01);雌激素受体介导的NG的作用与E2相似,而ICI182780逆转了NG的作用。RT-qPCR结果显示,NG对细胞Caspase-3、Bax和Bcl-2 mRNA表达的影响与蛋白结果一致。说明NG是通过激活ER信号通路发挥抗凋亡作用,作用与E2一致。结论雌激素受体介导的NG有明显的抗凋亡作用,通过提高Bcl-2的正常表达,降低Bax和Caspase-3的正常表达发挥抗凋亡作用,其抗凋亡作用可能经ER介导。
Objective To investigate the effect of estrogen receptor-mediated naringin(NG)on the apoptosis of rat adrenal pheochromocytoma(PC12)cells induced byβ-amyloid protein25-35(Aβ_(25-35))and its relationship with estrogen receptor(ER)signaling pathway.Methods The experiment was divided into blank group,Aβ_(25-35) group,E2+Aβ_(25-35) group,NG+Aβ_(25-35) group,ICI182780+E2+Aβ_(25-35) group,ICI182780+NG+Aβ_(25-35) group.Annexin V-FITC/PI double staining was used to detect the apoptosis rate of cells.The expression of Bax,Bcl-2 and Caspase-3 were detected by western blotting(WB).The mRNA expression of apoptosis factor was detected by RT-PCR.Results Annexin V-FITC/PI double staining flow cytometry showed that the apoptosis rate of Aβ_(25-35) group was higher than that of blank group(P<0.01).Compared with Aβ_(25-35) group,the apoptosis rate of NG+Aβ_(25-35) group was decreased(P<0.01).Compared with NG+Aβ_(25-35) group,the apoptosis rate of ICI182780+NG+Aβ_(25-35) group was increased(P<0.01).WB results showed that compared with blank group,the expression of Bax and Caspase-3 protein in Aβ_(25-35) group was increased(P<0.01),but the expression of Bcl-2 protein was opposite(P<0.01).Compared with Aβ_(25-35) group,the expressions of Bax and Caspase-3 protein in E2+Aβ_(25-35) group were decreased(P<0.01),but the expression of Bcl-2 protein was opposite(P<0.01).The effects of estrogen receptor mediated NG are similar to those of E2,whereas ICI182780 reverses the effects of NG.RT-PCR results showed that NG had the same effect on mRNA expression of caspase-3,Bax and Bcl-2.These results indicate that NG exerts anti-apoptotic effect through activating ER signaling pathway,which is consistent with E2.Conclusion NG mediated by estrogen receptor has obvious anti-apoptotic effect,which may be mediated by ER by increasing the normal expression of Bcl-2 and decreasing the normal expression of Bax and Caspase-3.
作者
王媛
武凤
熊辉
徐艳明
雷霞
徐红丹
孙慧峰
张宁
杨波
WANG Yuan;WU Feng;XIONG Hui;XU Yanming;LEI Xia;XU Hongdan;SUN Huifeng;ZHANG Ning;YANG Bo(Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,Heilongjiang,China;Jiamusi College,Heilongjiang University of Traditional Chinese Medicine,Jiamusi 154007,Heilongjiang,China)
出处
《辽宁中医药大学学报》
CAS
2023年第2期63-67,共5页
Journal of Liaoning University of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81673581)
国家科技重大专项(2018ZX09201009)
黑龙江省属高校科技成果研发、培育项目(TSTAU-C2018020)
黑龙江省卫生健康委科研课题(2020-301,2020-303)
黑龙江中医药大学科研基金项目(2018jkcy05,2018pt04,2019BJP01)。
作者简介
王媛(1997-),女,山东泰安人,硕士在读,研究方向:中药药效物质基础及体内代谢研究;通讯作者:杨波(1972-),男,辽宁铁岭人,副研究员,硕士研究生导师,博士,研究方向:中药药效物质基础及质量评价研究。
