摘要
目的探讨人参皂苷Rh2(ginsenoside Rh2,G-Rh2)对小鼠黑色素瘤B16细胞的促凋亡作用及其作用机制。方法培养黑色素瘤B16细胞,将黑色素瘤B16细胞随机分为6组:control、10 mg/mL、20 mg/mL、30 mg/mL、40 mg/mL、50 mg/mL,培养24 h、48 h、72 h后,采用CCK-8法检测不同浓度G-Rh2对B16细胞增殖的影响;采用Annexin V-FITC/PI双染,流式细胞仪检测细胞凋亡率和细胞周期分布情况;采用分光光度法检测Caspase-9、Caspase-3蛋白的活性。结果不同浓度G-Rh2(10 mg/mL、20 mg/mL、30 mg/mL、40 mg/mL、50 mg/mL)作用于B16细胞24 h、48 h和72 h后,与空白对照组比,随给药浓度升高和药物干预时间延长对B16细胞的抑制率均有显著升高,并且呈浓度依赖性和时间依赖性(P<0.05);10 mg/mL、20 mg/mL、30 mg/mL、40 mg/mL、50 mg/mL的G-Rh2作用B16细胞48 h后,凋亡率分别为(9.43±0.23)%、(17.50±0.11)%、(24.14±0.38)%、(38.93±0.59)%,与空白对照组(6.67±0.10)%相比,随着G-Rh2的浓度增加,B16细胞凋亡率可显著增加(P<0.05);细胞周期研究结果表明,与空白对照组相比,G0/G1期细胞增多,由空白对照组(60.64±0.68)增加至40 mg/mL时的(82.88±0.17)%,G2/M期细胞减少,由空白对照组(16.31±0.11)%下降至40 mg/mL时的(7.49±0.69)%,说明G-Rh2可诱导B16细胞周期阻滞于G0/G1期;分光光度法研究表明,G-Rh2可以明显增加B16细胞内Caspase-9、Caspase-3蛋白的活性(P<0.05)。结论人参皂苷Rh2可抑制黑色素瘤B16细胞增殖,诱导其凋亡,其凋亡机制可能与上调Caspase-9、Caspase-3,激活Caspase家族有关。
To investigate the apoptotic effect of ginsenoside Rh2(G-Rh2)on mouse melanoma B16 cells and its mechanism.Methods Melanoma B16 cells were cultured and randomly divided into 6 groups:Control,10 mg/mL,20 mg/mL,30 mg/mL,40 mg/mL,and 50 mg/mL.Cells were cultured for 24 h,48 h and 72 h.CCK-8 assay was used to detect the effects of different concentrations of G-Rh2 on the proliferation of B16 cells.Annexin V-FITC/PI double staining was used to detect the apoptosis rate and cell cycle distribution by flow cytometry.Spectrophotometry was used to detect the protein activities of Caspase-9 and Caspase-3.Results Different concentrations of G-Rh2(10 mg/mL,20 mg/mL,30 mg/mL,40 mg/mL,and 50 mg/mL)treated B16 cells for 24 h,48 h and 72 h,compared with the blank control group,the inhibition rate of B16 cells increased significantly with the increase of the concentration of G-Rh2 and the extension of the intervention time.It was concentration-dependent and time-dependent(P<0.05).B16 cells were treated with 10 mg/mL,20 mg/mL,30 mg/mL,40 mg/mL,and 50 mg/mL G-Rh2 for 48h.The apoptosis rates were(9.43±0.23)%,(17.50±0.11)%,(24.14±0.38)%and(38.93±0.59)%,respectively.Compared with the blank control group(6.67±0.10)%,the apoptosis rate of B16 cells was significantly increased with the increase of G-Rh2 concentration(P<0.05).Compared with the blank control group,cells in G0/G1 phase increased from(60.64±0.68)%to(82.88±0.17)%at 40 mg/mL,and cells in G2/M phase decreased.It decreased from(16.31±0.11)%in the blank control group to(7.49±0.69)%at 40 mg/mL,suggesting that G-Rh2 could induce B16 cell cycle arrest in G0/G1 phase.Spectrophotometric study showed that G-Rh2 could significantly increase the activity of caspase-9 and caspase-3 proteins in B16 cells(P<0.05).Conclusion Ginsenoside Rh2 can inhibit the proliferation of melanoma B16 cells and induce their apoptosis,which may be related to the up-regulation of caspase-9 and caspase-3 and activation of the Caspase family.
作者
任煜
刘婷婷
祁冰洁
REN Yu(Anqing Medical College, Anqing 246052, China)
出处
《牡丹江医学院学报》
2021年第6期1-5,共5页
Journal of Mudanjiang Medical University
基金
安徽省高校自然科学研究重点项目(KJ2018A0936)
安庆医药高等专科学校自然科学研究项目(2020ZR007)。
