梅花针叩刺增强咪喹莫特治疗SKH-1小鼠皮肤鳞状细胞癌的疗效  

Plum- blossom needle tapping enhances the efficacy of topical imiquimod immunotherapy for cutaneous sqnamous cell carcinoma in SKH-1 mice

作　　者：张付贺 石磊[2] 罗敏[2] 刘沛[2] 周忠霞[2] 张国龙[2] 王秀丽[1] 

机构地区：[1]安徽医科大学上海皮肤病临床学院,上海200443 [2]上海市皮肤病医院同济大学医学院光医学研究所

出　　处：《中华皮肤科杂志》2017年第4期273-278,共6页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81601601）;上海市卫计委科研课题（201640016）;上海市青年科技英才扬帆计划项目（15YF1410700）;上海申康医院发展中心临床科技创新项目（SHDC12015123）

摘　　要：目的 评估咪喹莫特联合梅花针治疗SKH-1小鼠皮肤鳞状细胞癌（鳞癌）的疗效，探讨其免疫学机制。方法 将紫外线诱导成瘤的40只SKH-1皮肤鳞癌小鼠随机分4组，每组10只：对照组无处理；梅花针组：每天梅花针叩刺所有瘤体1次；咪喹莫特组：每天外涂5%咪喹莫特乳膏1次，剂量为1.2 g/kg；联合组：先梅花针叩刺所有瘤体，止血后再外涂5%咪喹莫特乳膏1次，剂量同前。各组小鼠连续治疗30 d。每日观察并拍照记录各组小鼠肿瘤形态学变化，每3 d测量1次瘤体大小，比较4组瘤体体积变化及小鼠生存率变化。治疗结束后取各组小鼠瘤体，比较4组瘤体组织病理学变化。实时荧光定量PCR检测各组小鼠瘤体内干扰素（IFN）-α、IFN-β、白细胞介素1β（IL-1β）、肿瘤坏死因子α（TNF-α）及IL-12 mRNA表达。结果 联合组小鼠背部瘤体生长缓慢，部分小的瘤体有消退现象；对照组、梅花针组和咪喹莫特组瘤体一直在增长，但梅花针组、咪喹莫特组生长速度较对照组慢。治疗前，4组小鼠瘤体体积差异无统计学意义（F = 0.90，P 〉 0.05）。治疗24 d后，4组小鼠瘤体体积差异有统计学意义（F = 5.16，P 〈 0.05），LSD-t检验示，联合组瘤体体积显著低于对照组（P 〈 0.01），余各组间差异均无统计学意义（P 〉 0.05）。经Log-rank检验，4组小鼠生存率曲线分布不同（χ2 = 8.32，P 〈 0.05），联合组生存情况优于对照组（χ2 = 4.62，P = 0.03），而梅花针组、咪喹莫特组与对照组、联合组比较，小鼠生存率差异均无统计学意义（P 〉 0.05）。组织病理学检查显示，对照组、梅花针组细胞异形性明显，排列密集，可见大量的肿瘤细胞，部分可见角化珠，咪喹莫特组肿瘤细胞少许死亡；联合组肿瘤细胞大量死亡，核异形不明显，角化增多。实时荧光定量PCR显示，联合组IFN-α、IFN-β、IL-12、IL-1Objective To assess the therapeutic effect of plum-blossom needle tapping combined with topical imiquimod immunotherapy on cutaneous squamous cell carcinoma （SCC） in SKH-1 mice, and to explore the immunological mechanism. Methods A total of 40 SKH-1 mice with ultraviolet light-induced cutaneous SCC were randomly and equally divided into 4 groups： control group receiving no treatment, plum-blossom needle group receiving plum-blossom needle tapping on all the tumors once a day,imiquimod group topically treated with imiquimod 5% cream at a dose of 1.2 g/kg once a day, combination group firstly treated with plum-blossom needle tapping on all the tumors, and after the stop of bleeding topically treated with imiquimod 5% cream at the same dose as the imiquimod group once a day. All the mice were treated for 30 days. Morphological changes of tumors in all groups were photographed and recorded every day. The tumor size was measured once every three days, and changes of total tumor volume and survival rate of the mice were compared among the 4 groups. At the end of treatment, tumor tissues were resected, and histopathological changes were compared among the 4 groups. Real-time fluorescence-based quantitative PCR （qRT-PCR） was performed to measure the mRNA of interferon-α （IFN-α）, IFN-β, interleukin-1β （IL-1β）, tumor necrosis factor-α （TNF-α） and IL-12 in tumor tissues. Results In the combination group, tumors on the back of mice grew slowly, and some even regressed. However, tumors grew fast in the control group, plum-blossom needle group and imiquimod group, and grew more slowly in the plum-blossom needle group and imiquimod group than in the control group. Before the treatment, there was no significant difference in the total tumor volume among the 4 groups （F = 0.90, P 〉 0.05）. After 24-day treatment, the total tumor volume significantly differed among the 4 groups （F = 5.16, P 〈 0.05）. The LSD-t test showed that the total tumor volume significantly decreased in the combination

关 键 词：肿瘤 鳞状细胞 梅花针疗法 疾病模型 动物 小鼠 咪喹莫特 

分 类 号：R739.5[医药卫生—肿瘤]