低剂量环磷酰胺联合胸腺肽疗法对尖锐湿疣角质形成细胞增殖及凋亡的影响

Effect of Low-dose Cyclophosphamide Combined with Thymosin on Proliferation and Apoptosis of Keratinocyte in Condyloma Acuminatum

目的探讨低剂量环磷酰胺联合胸腺肽疗法对尖锐湿疣(CA)角质形成细胞(KCs)增殖及凋亡的影响。方法观察华中科技大学同济医学院附属同济医院皮肤科55例完成该疗法治疗1个月的CA患者的疗效,从有效病例中随机抽取20例,用免疫组化ABC法检测治疗前后疣体中KCs增殖指标Ki-67及凋亡指标Cleaved Caspase-3的表达情况;同时用原位杂交技术、实时荧光定量聚合酶链反应(PCR)检测其治疗前后疣体HPV阳性细胞表达及HPV6/11-DNA载量变化。结果20例有效患者中治疗前Ki-67阳性表达率为80%(16/20),治疗后Ki-67阳性表达率45%(9/20);治疗前Cleaved Caspase-3阳性表达率35%(7/20),治疗后Cleaved Caspase-3阳性表达率75%(15/20);治疗后HPV6/11-DNA拷贝数量级(106)较治疗前(109)明显下降(P<0.01)。结论低剂量环磷酰胺联合胸腺肽治疗1个月后,患者疣体内KCs增殖能力下降、凋亡能力增强;患者疣体HPV病毒载量消减达103个数量级。

Objective To investigate the effect of low-dose cyclophosphamide combined with thymosin treatment on the proliferation and apoptosis of keratinocytes(KCs)in condyloma acuminatum(CA).Methods To observe the efficacy of 55 CA patients who received the treatment for 1 month,and 20 cases were selected randomly from the effective cases.Ki-67 and cleaved Caspase-3,as proliferation and apoptosis markers respectively,were evaluated by immunohistochemistry with ABC chromogenic method.Meanwhile,the changes of HPV positive expression and HPV6/11-DNA copies in warts before and after treatment were detected by in situ hybridization and real-time fluorescence quantitative polymerase chain reaction(PCR).Results Among the 20 patients with effective therapy,the Ki-67 positive expression rate was 80%(16/20)before treatment,while decreased to 45%(9/20)after treatment.The positive cell expression rate of cleaved Caspase-3 was 35%(7/20)before treatment,and increased to 75%(15/20)after treatment.HPV6/11-DNA copy number(106)after treatment was significantly lower than that before treatment(109)(P<0.01).Conclusion Treatment of low-dose cyclophosphamide combined with thymosin for one month contributes to decreased KCs proliferation and increased apoptotic capacity in warts.Moreover,the HPV copies in the warts are reduced by 1000 times.