儿黄散白及胶双层膜对宫颈癌HeLa细胞增殖和凋亡作用及毒性、刺激性研究

Effect of Erhuangsan gum-bletilla double-film on proliferation and apoptosis of cervical cancer HeLa cells and its toxicity and irritation

目的通过儿黄散白及胶双层膜(EHGBF)对人宫颈癌HeLa细胞增殖抑制及凋亡作用研究和急性毒性试验、皮肤刺激性试验、安全性试验,探讨其有效性和安全性。方法采用MTT法考察不同浓度(0.1、1.0、5.0、10.0 mg/mL)的EHGBF、无白及胶儿黄散双层膜、白及胶双层膜及空白基质膜对HeLa细胞的增殖抑制作用;流式细胞术检测EHGBF对Hela细胞凋亡的影响;通过膜剂阴道给药大鼠急性毒性试验,对家兔完整皮肤及破损皮肤的刺激性试验,及对用药后家兔阴道取材肉眼观察及石蜡切片病理学观察,研究EHGBF对阴道刺激性。结果空白基质膜对HeLa细胞无增殖抑制作用,EHGBF、白及胶双层膜、无白及胶儿黄散双层膜对HeLa细胞均发挥显著增殖抑制作用(P<0.05),作用强弱为EHGBF>白及胶双层膜>无白及胶儿黄散双层膜;经过1、5、10 mg/mL浓度的EHGBF处理后,HeLa细胞凋亡率与对照组比较均显著升高(P<0.05);EHGBF在大剂量使用下无致死性,对皮肤及阴道几乎没有刺激性。结论EHGBF可抑制人宫颈癌HeLa细胞增殖并促进其凋亡,无急性毒性,对皮肤无刺激性,对阴道黏膜几乎没有刺激性,安全性较好。

Objective To investigate the efficacy and safety of skin medication for human cervical cancer with HeLa cell proliferation inhibition and apoptosis by Erhuangsan gum-bletilla double-film(EHGBF).Methods MTT assay was used to investigate the effects of different concentrations(0.1,1.0,5.0,10.0 mg/mL)of EHGBF,Erhuangsan double-film without gum-bletilla,gum-bletilla double-film and blank matrix film on cell proliferation inhibition.The effects of apoptosis were detected by flow cytometry.The vaginal irritancy of the film was investigated by the vaginal acute toxicity test of the film,the irritating test of the intact skin and the damaged skin of the rabbit,and the visual observation of the vaginal material of the rabbit after the administration and the pathological observation of the paraffin section.Result The blank matrix membrane had no inhibitory effect on HeLa cell proliferation,while EHGBF,gum-bletilla double-film,and erhuangsan double-film without gum-bletilla had significant inhibitory effect on HeLa cell proliferation(P<0.05).The inhibitory effect on the proliferation of HeLa cells was that EHGBF>gum-bletilla double-film>erhuangsan double-film without gum-bletilla.The apoptosis rate of HeLa cells treated with EHGBF was significantly higher than that of the control group(P<0.05).EHGBF was not lethal at high doses,and had little irritation to the skin and vagina.Conclusion EHGBF can inhibit the proliferation and promote apoptosis of human cervical cancer HeLa cells,no acute toxicity and irritation to the skin,almost no irritating to vaginal mucosa,and good safety.