雌激素受体介导的柚皮苷抗Aβ_(25-35)损伤PC12细胞凋亡作用研究  

Effects of Estrogen Receptor-mediated Naringin on Apoptosis of Aβ_(25-35)Injured PC12 Cells

作　　者：王媛 武凤 熊辉[1] 徐艳明[2] 雷霞[1] 徐红丹[2] 孙慧峰[1] 张宁[1,2] 杨波[1] 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)

机构地区：[1]黑龙江中医药大学,哈尔滨黑龙江150040 [2]黑龙江中医药大学佳木斯学院,佳木斯黑龙江154007

出　　处：《辽宁中医药大学学报》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)。

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.

关 键 词：柚皮苷 Β淀粉样蛋白25-35 雌激素受体 抗凋亡作用 

分 类 号：R285.5[医药卫生—中药学]