摘要
目的观察芍药苷对垂体瘤细胞增殖凋亡的影响及作用机制。方法取新鲜垂体瘤组织消化分离培养垂体瘤细胞,分别采用不同浓度芍药苷处理48 h后,采用四唑盐(MTT)法检测细胞存活率,选择半数细胞存活率对应的芍药苷浓度进行实验。分别采用芍药苷、信号传导及转录激活因子3(STAT3)信号通路激动剂与芍药苷+激动剂处理垂体瘤细胞,MTT法检测细胞存活率,流式细胞技术检测细胞凋亡率,实时荧光定量聚合酶链式反应(RT-qPCR)检测Janus激酶2(JAK2)、STAT3、B淋巴细胞瘤-2基因(Bcl-2)和Bcl-2相关X蛋白(Bax)mRNA水平,免疫印迹法(Western Blot)检测磷酸化JAK2(p-JAK2)、磷酸化STAT3(p-STAT3)、JAK2、STAT3、Bcl-2、Bax蛋白表达。结果芍药苷对垂体瘤细胞生长有明显的抑制作用并促进凋亡,这种作用呈剂量依赖性。RT-qPCR和Western Blot结果显示:芍药苷对STAT3信号通路有抑制作用,可抑制Bcl-2转录及表达,促进Bax转录及表达。结论芍药苷可能通过抑制STAT3信号通路,调控细胞凋亡相关基因Bcl-2、Bax表达,从而抑制垂体瘤细胞增殖,诱导其凋亡。
Objective To investigate the effect of paeoniflorin on proliferation and apoptosis of pituitary tumor cells and its mechanism.Methods Pituitary tumor cells were digested,separated,and cultured from fresh pituitary tumor tissue.After treatment with different concentrations of paeoniflorin for 48 h,the cell survival rate was detected by tetrazolium salt(MTT)method,and the concentration of paeoniflorin corresponding to half of the cell survival rate was selected for the experiment.Pituitary tumor cells were treated with paeoniflorin,signal transducer and activator of transcription 3(STAT3)signaling pathway agonist and paeoniflorin+agonist,respectively.The cell survival rate was detected by MTT assay,the cell apoptosis rate was detected by flow cytometry.The mRNA levels of Janus kinase 2(JAK2),STAT3,B cell lymphocyte-2(Bcl-2),and Bcl-2-associated X protein(Bax)were detected by real-time quantitative polymerase chain reaction(RT-qPCR),and the expressions of p-JAK2,p-STAT3,JAK2,STAT3,Bcl-2,and Bax protein were detected by Western Blot.Results Paeoniflorin significantly inhibited the growth of pituitary tumor cells and promoted apoptosis in a dose-dependent manner.RT-qPCR and Western Blot results showed that paeoniflorin possessed clear inhibitory effect on the STAT3 signaling pathway.Paeoniflorin could inhibit the transcription and expression of Bcl-2 and promote the transcription and expression of Bax.Conclusion Paeoniflorin may inhibit the proliferation of pituitary tumor cells and induce their apoptosis by inhibiting the STAT3 signaling pathway and regulating the expression of apoptosis-related genes Bcl-2 and Bax.
作者
徐大伟
刁玉领
惠磊
周文科
汲乾坤
XU Dawei;DIAO Yuling;HUI Lei;ZHOU Wenke;JI Qiankun(The First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100,Henan,China)
出处
《中西医结合心脑血管病杂志》
2022年第16期2930-2935,共6页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
基金
2019年河南省医学科技攻关计划联合共建项目(No.LHGJ20190445)。
关键词
垂体瘤细胞
芍药苷
增殖
凋亡
信号传导及转录激活因子3信号通路
实验研究
pituitary tumor cells
paeoniflorin
proliferation
apoptosis
signal transducer and activator of transcription 3 signaling pathway
experimental study
