目的 通過兒黃散白及膠雙層膜（EHGBF）對人宮頸癌HeLa細胞增殖抑制及凋亡作用研究和急性毒性試驗、皮膚刺激性試驗、安全性試驗，探討其有效性和安全性。方法 采用MTT法考察不同濃度（0.1、1.0、5.0、10.0 mg/mL）的EHGBF、無白及膠兒黃散雙層膜、白及膠雙層膜及空白基質(zhì)膜對HeLa細胞的增殖抑制作用；流式細胞術檢測EHGBF對Hela細胞凋亡的影響；通過膜劑陰道給藥大鼠急性毒性試驗，對家兔完整皮膚及破損皮膚的刺激性試驗，及對用藥后家兔陰道取材肉眼觀察及石蠟切片病理學觀察，研究EHGBF對陰道刺激性。結(jié)果 空白基質(zhì)膜對HeLa細胞無增殖抑制作用，EHGBF、白及膠雙層膜、無白及膠兒黃散雙層膜對HeLa細胞均發(fā)揮顯著增殖抑制作用（P<0.05），作用強弱為EHGBF > 白及膠雙層膜 > 無白及膠兒黃散雙層膜；經(jīng)過1、5、10 mg/mL濃度的EHGBF處理后，HeLa細胞凋亡率與對照組比較均顯著升高（P<0.05）；EHGBF在大劑量使用下無致死性，對皮膚及陰道幾乎沒有刺激性。結(jié)論 EHGBF可抑制人宮頸癌HeLa細胞增殖并促進其凋亡，無急性毒性，對皮膚無刺激性，對陰道黏膜幾乎沒有刺激性，安全性較好。

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.

R285.5

哈爾濱市應用技術研究與開發(fā)項目（2016RAXXJ047）；黑龍江中醫(yī)藥大學研究生創(chuàng)新科研項目（YJSCX2017058）