椿乳凝胶调节宫颈病变细胞内人乳头瘤病毒表达与阴道菌群失调的机制研究  

Mechanism of Chunru gel in regulating HPV expression in cervical lesion cells and imbalance of vaginal flora

作　　者：罗桂芳 朱金鹏 张鹏 龚云 张宗利 LUO Gui-fang;ZHU Jin-peng;ZHANG Peng;GONG Yun;ZHANG Zong-li(Hunan Center for Drug Safety Evaluation,Changsha 410331;Hunan Key Laboratory for Drug Pharmacodynamics and Safety Evaluation,Changsha 410331;Zhuzhou Qianjin Pharmaceutical Co.Ltd,Zhuzhou Hunan 412000)

机构地区：[1]湖南省药物安全评价研究中心,长沙410331 [2]新药药效与安全性评价湖南省重点实验室,长沙410331 [3]株洲千金药业股份有限公司,湖南株洲412000

出　　处：《中南药学》2020年第2期222-227,共6页Central South Pharmacy

摘　　要：目的研究椿乳凝胶对宫颈上皮细胞损伤的修复和对人宫颈癌细胞内人乳头瘤病毒(HPV)阳性的清除作用及其作用机制。方法采用人宫颈上皮细胞(H8)、人宫颈癌细胞CaSki、Hela(分别为HPV 16阳性和HPV 18阳性),药物干预后,CCK-8法检测细胞增殖抑制率,采用流式细胞术检测H8细胞周期和CaSki、Hela细胞凋亡,Western Blot法检测E6/E7蛋白表达。采用加德纳菌和白假丝酵母菌,检测椿乳凝胶对两种菌的最小抑菌浓度(MIC)。结果椿乳凝胶对H8、CaSki和Hela细胞的IC50的计算值均>10 mg·mL^-1,TC0均>5 mg·mL^-1;可促进H8细胞由G0/G1期进入S期和G2/M期,减少G0/G1期细胞,促进细胞分裂增殖;可诱导宫颈癌细胞凋亡,且具有浓度-效应关系;可抑制HPV16E6/E7、HPV18 E6/E7蛋白的表达,且具有浓度-效应关系;可抑制加德纳菌和白假丝酵母菌的生长,其MIC分别为62.5和125 mg·mL^-1。结论在本实验条件下,椿乳凝胶可抑制CaSki和Hela细胞增殖,调节HPV16/18 E6/E7蛋白表达,促进CaSki和Hela细胞凋亡,调节H8细胞周期。可抑制宫颈炎菌群失衡中的条件致病菌,两种机制作用下共同修复受损宫颈组织。Objective To determine the effect of Chunru gel on the repair of cervical epithelial cell injury and the clearance of human papillomavirus(HPV)in human cervical cancer cells,and to explore its mechanism.Methods Human cervical epithelial cells(H8)and human cervical cancer cells CaSki(HPV 16 positive)and Hela(HPV 18 positive)were treated with Chunru gel.The cell proliferation inhibition rate was detected with CCK-8,the H8 cell cycle and the apoptosis of CaSki and Hela cells were detected with flow cytometry,and the expression of E6/E7 protein by Western Blot.The minimal inhibitory concentration(MIC)of Chunru gel against the two strains was determined by Gardner and Candida albicans.Results IC50 values of Chunru gel for H8,CaSki and Hela cells were all higher than 10 mg·mL^-1,and TC0 were higher 5 mg·mL^-1.Chunru gel promoted H8 cells from G0/G1 phase to S phase and G2/M phase,reduced G0/G1 phase cells,and promoted cell division and proliferation.Chunru gel induced the apoptosis of cervical cancer cells in a concentration-effect manner.The expression of HPV16 E6/E7 and HPV18 E6/E7 was inhibited in a concentration-effect manner.The MIC values of Gardner and Candida albicans were 62.5 and 125 mg·mL^-1,respectively.Conclusion Under the conditions of this experiment,Chunru gel inhibits the proliferation of CaSki and Hela cells,regulates the expression of HPV16/18 E6/E7 protein,promote the apoptosis of CaSki and Hela cells,and regulate the H8 cell cycle.It also inhibits the conditional pathogenic bacteria in the imbalance of cervical inflammatory flora and repairs the damaged cervical tissue under the two mechanisms.

关 键 词：椿乳凝胶 人乳头瘤病毒E6/E7阳性 人宫颈癌细胞 细胞增殖 阴道菌群失调 

分 类 号：R286[医药卫生—中药学]