复方硝酸溶液处理尖锐湿疣皮损的实验研究

The experimental study on the treatment of condyloma acuminatum with compond nitric acid solution

目的:探讨复方硝酸溶液(商品名:思可得)治疗尖锐湿疣的作用机制,及其对体外培养角质形成细胞(HaCaT细胞)的作用。方法:20例尖锐湿疣患者皮损制成标本用思可得涂抹,并以鬼臼毒素作对照,分别检测2种药物首次用药和重复用药后,皮损内HPV6/11DNA的检出率,并行组织病理学检查。四甲基偶氮唑蓝(MTT)还原法检测思可得对HaCaT细胞存活率的影响,流式细胞仪分析细胞周期。结果:单次用药后,思可得组HPV6/11DNA的检出率为20%,鬼臼毒素组的检出率为70%。重复用药后,思可得组HPV-DNA阳性率为5%,鬼臼毒素组为40%。用药后组织病理学检查显示标本呈凝固性坏死。HaCaT细胞经0.1～5.0μl/mL思可得处理后,细胞存活率降低,并与药物浓度呈正相关。经1.0～3.0μl/mL思可得处理48h的细胞,流式细胞仪检测到凋亡峰。结论:思可得治疗尖锐湿疣,除化学性破坏组织外,尚有抗病毒作用,重复用药作用更强,有利于减少尖锐湿疣的复发。思可得对体外培养的角质形成细胞生长有抑制作用,可促进其凋亡。

Objective： To investigate the mechanism of compond nitric acid solution （i.e. solcoderm） treating condyloma acuminatum, and relation between treatment frequency and occurrence of HPV DNA. To study the effect of solcoderm on cultured HaCaT cells. Methods： The lesion specimens taken from 20 patients with condyloma acuminatum were treated once or repeatedly with solcoderm. And the specimens were also treated with podophyllotoxin as control. After that the DNA frequencies of HPV6 and HPVll were detected in the lesions by polymerase chain reaction （PCR）. Meanwhile pathological changes of treated samples were observed. In addition, the viability of HaCaT cells treated with solcoderm was measured by MTT [3-（4, 5-dimethyl-2-thiazolyl）-2, 5-diphenyl-2H-tetrazolium bromide] assay. The cells growth phase were analyzed by flow cytometry （FCM）. Results： PCR results showed that the positive reaction to HPV6, 11 was found in 4 （20%） and 14 （70%）of 20 specimens once-treated with solcoderm or podophyllotoxin respectively. For the samples repeatedly treated with solcoderm or podophyllotoxin, the positivity rates were reduced to 5% and 40% respectively. The pathological examination showed that the treated tissues present coagulation necrosis. A time- and dose-dependent decrease in cell viability was induced in HaCaT cells after treatment with solcolderm at the concentration of 0.1 to 5 μl/mL. The flow cytometry showed that apoptotic peak present after treatment with 1 to 3μ/mL of solcolderm for 48 h. Conclusions： Not only does chemical destruction of solcoderm give contribution to treating condyloma acuminatum, but also its anti-virus effectiveness. Retreatment with solcoderm may decrease the recurrence of condyloma acuminatum. Solcoderm could inhibit keratinocyte growth in vitro and induce the cells apoptosis.