疣毒净通过MAPK信号通路对H8细胞增殖的影响  

Effect of Youdujing on Proliferation of H8 Cells by MAPK Signaling Pathway

作　　者：刘丽华[1] 陈小凤 肖静 LIU Lihua;CHEN Xiaofeng;XIAO Jing(The Second Clinical College,Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong,China;The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong,China)

机构地区：[1]广州中医药大学第二临床医学院,广东广州510006 [2]广州中医药大学第二附属医院,广东广州510006

出　　处：《中华中医药学刊》2020年第3期145-148,I0023,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(81473716)。

摘　　要：目的探讨疣毒净制剂影响宫颈上皮永生化细胞H8细胞增殖作用的可能机制。方法以转染HPV16基因后稳定传代的宫颈上皮永生化细胞H8细胞为研究对象;经0.5、1、2、4、8 g·L-1疣毒净作用于H8细胞,MTT法检测细胞增殖抑制效果;EdU细胞增殖检测法测定各实验组H8细胞增殖变化;流式细胞术检测各实验组H8细胞的周期情况;Western Blot法检测各实验组H8细胞MEK和ERK蛋白表达情况。结果与空白组相比,疣毒净药物干预24、48、72 h后,细胞生长力明显下降(P<0.05),抑制细胞生长率明显增加(P<0.05);EdU免疫荧光染色结果显示,疣毒净组EdU增殖细胞阳性率显著下降(P<0.05);与空白组相比,疣毒净各浓度组H8细胞的周期中,药物将细胞周期阻滞在G0/G1期(P<0.05);与空白组相比,疣毒净组MEK蛋白明显下调(P<0.05)和ERK蛋白明显下调(P<0.05),呈浓度下调趋势,二者下调的趋势一致。结论疣毒净能明显抑制宫颈永生化上皮细胞H8细胞的增殖,这一影响可能是通过MAPK信号通路起作用的。Objective The purpose of this study was designed to investigate the possible mechanism of Youdujing affecting the proliferation of cervical epithelial immortalized H8 cells. Methods H8 cells,which were transfected with HPV-16 gene and passed stably,were chosen to be the object of the study.H8 cells were intervened by 0.5, 1, 2, 4, 8 g·L-1 Youdujing and detected by MTT colorimetry. EdU cell proliferation assay was used to determine the proliferation of H8 cells. Flow cytometry was used to detect the cell cycle of H8 cells in each experimental group. The expression levels of MEK and ERK proteins were detected by WB method. Results The growth of H8 cells intervened by 0.5,1,2,4,8 g·L-1 Youdujing was significantly inhibited in concentration-dependent and time-dependent manners(P<0.05) compared with blank group. EdU immunofluorescence staining results showed that the positive rate of EdU proliferation cells in the Youdujing group was significantly decreased(P<0.05). Compared with the blank group, the cycle of H8 cells in each experimental group, the proportion of cells in G0/G1 phase was increased gradually(P<0.05), and the proportion of cells in S phase and G2/M phase were decreased gradually. Compared with the blank group, the MEK protein in the Youdujing group was significantly down-regulated(P<0.05). And ERK protein was significantly down-regulated(P<0.05). Conclusion Youdujing can significantly inhibit the proliferation of cervical immortalized epithelial cells H8 cells, which may be through the MAPK signaling pathway.

关 键 词：宫颈永生化上皮细胞H8细胞 增殖 疣毒净 MAPK信号通路 

分 类 号：R285.5[医药卫生—中药学]