摘要
目的 研究丹酚酸A(SAA)与氧化槐定碱(OSR)协同抑制宫颈上皮内瘤变细胞的作用机制。方法 采用宫颈上皮永生化细胞(H8细胞)作为宫颈上皮内瘤变的细胞模型,设置SAA、OSR及其联合用药3个处理组,并设对照组。观察各组H8细胞形态,通过Hochest33342染色法观察H8细胞凋亡情况,采用流式细胞仪测定H8细胞周期,采用蛋白免疫印迹法检测H8细胞B细胞淋巴相关X蛋白(Bax)、B细胞淋巴瘤-2(Bcl-2)、切割型半胱氨酸天冬氨酸蛋白水解酶-3(Cleaved Caspase-3)蛋白表达情况。结果 与对照组比较,各处理组H8细胞形态均变小、变圆,且黏附力均下降,凋亡细胞数量均增加;OSR组、联合用药组H8细胞G_(2)期细胞数量均明显增加,G_(1)期细胞数量均明显减少,H8细胞被阻滞在G_(2)/M期,SAA组在实验浓度下对H8细胞周期的调控无明显影响,但可协同OSR调控H8细胞周期;SAA、OSR及其联合用药均可抑制Bcl-2蛋白表达,促进Bax、Cleaved Caspase-3蛋白表达,进而促进H8细胞凋亡的发生。结论 SAA、OSR联合用药可通过协同调控H8细胞周期,将H8细胞阻滞在G_(2)/M期,抑制H8细胞增殖;通过协同抑制Bcl-2蛋白表达,促进Bax、Cleaved Caspase-3蛋白表达进而促进H8细胞凋亡。
Objective To study the mechanism of salvianolic acid A(SAA) and oxysophoridine(OSR) synergistic inhibition of cervical intraepithelial neoplasia cells.Methods Cervical epithelial immortarized cells(H8 cells) were used as the cell model of cervical intraepithelial neoplasia.Three treatment groups(SAA,OSR and their combination) were set up, and the control group was set up.The morphology of H8 cells in each group was observed.The apoptosis of H8 cells was observed by Hochest33342 staining.The cell cycle of H8 cells was measured by flow cytometry.Expression of B-cell lymphoid-associated X protein(Bax),B-cell lymphoma-2(Bcl-2) and cleaved cysteine aspartate proteolytic enzyme 3(Cleaved Caspase-3) in H8 cells was detected by western blot.Results Compared with the control group, the morphology of H8 cells in each treatment group became smaller and rounder, and their adhesion decreased, and the number of apoptotic cells increased.The number of H8 cells increased significantly in G_(2)phase while decreased significantly in G_(1)phase in the OSR group and the combination group, and H8 cells were blocked in G_(2)/M phase.The experimental concentration of SAA group had no significant effect on the regulation of H8 cell cycle, but it could cooperate with OSR to regulate H8 cell cycle.SAA,OSR and their combination could inhibit the expression of Bcl-2 protein, promote the expression of Bax and Cleaved Caspase-3 protein, so as to promote H8 cell apoptosis.Conclusion The combination of SAA and OSR can co-regulate H8 cell cycle, block H8 cells in G_(2)/M phase and inhibit the proliferation of H8 cells.By synergistically inhibiting the expression of Bcl-2 protein, it can promote the expression of Bax and Cleaved Caspase-3 protein to promote the apoptosis of H8 cells.
作者
冷雪娇
阚宏飞
吴沁航
李存玉
郑云枫
彭国平
LENG Xuejiao;KAN Hongfei;WU Qinhang;LI Cunyu;ZHENG Yunfeng;PENG Guoping(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing,Jiangsu 210023,China)
出处
《现代医药卫生》
2023年第2期186-191,共6页
Journal of Modern Medicine & Health
基金
国家自然科学基金青年基金项目(82104451)。
关键词
宫颈上皮内瘤变
丹酚酸A
氧化槐定碱
细胞周期
细胞凋亡
Cervical intraepithelial neoplasia
Salvianolic acid A
Oxysophoridine
Cell cycle
Apoptosis
