局部针刺对莫诺苯宗诱导C57BL／6小鼠白癜风样模型的影响

Effect of local acupuncture on monobenzone-induced vitiligo-like depigmentation in mice

目的探讨外伤对莫诺苯宗诱导白癜风小鼠脱色的影响。方法将40只C57BI）6小鼠随机分为对照组、模型组、针刺组和针刺配合莫诺苯宗组4个组，每组10只。模型组应用40％莫诺苯宗乳膏诱导C57BL／6小鼠脱色，建立白癜风动物模型；针刺配合莫诺苯宗组通过针刺配合40％莫诺苯宗乳膏的方法，研究针刺对小鼠脱色的影响。实验过程中肉眼观察毛发颜色以及共聚焦激光扫描显微镜（RCM）观察皮肤脱色。实验结束后麻醉小鼠，取非用药部位脱色皮肤行组织学检查，HE染色检测淋巴细胞浸润情况，免疫荧光染色检测CD8＋T细胞表达。结果模型组小鼠在用药部位及非用药部位均有不同程度脱色。针刺配合莫诺苯宗组小鼠同样具有以上特点，且其脱色出现时间早，面积大并稳定。实验第65天，模型组和针刺配合莫诺苯宗组用药部位的脱色面积指数分别为3．45±0．17和3．90±0．25，两组比较差异有统计学意义，t=7．433，19〈0．05；非用药部位分别为1．90±0．12和2．85±0．27，两组比较差异有统计学意义，t=7．529，19〈0．05。免疫荧光显示非用药部位脱色区CD8＋T细胞荧光强度模型组为175．528±10．711，针刺配合莫诺苯宗组为645．928±12．652，两者比较差异有统计学意义，t=8．105，P〈0．05。针刺配合莫诺苯宗组在非用药部位脱色斑局部淋巴细胞和CD8’T细胞浸润更明显。结论局部皮肤屏障破坏可以促进莫诺苯宗诱导的白癜风小鼠脱色。

Objective To evaluate the effect of injuries on monobenzone-induced vitiligo-like depigmentation in mice. Methods Forty C57BL/6 mice were randomly and equally divided into four groups： negative control group topically treated with vaseline cream, model group induced by topical monobenzone （40%） cream, acupuncture group receiving acupuncture treatment （15 times） once every three days, and acupuncture combined with monobenzone group receiving both monobenzone induction and acupuncture treatment. The treatment lasted 50 days and mice were sacrificed 15 days after the end of treatment. Hair decolorization was observed with naked eyes, and skin decolorization with reflectance confocal microscopy （RCM） on a daily basis. Tissue specimens were obtained from depigmented skin at monobenzone-uninduced sites, and subjected to hematoxylin and eosin （HE） staining for the cvaluation of lymphocytic infiltration as well as immunofluorescence staining for the detection of CD8~ T cell expression. Statistical analysis was done by t test. Results Varying degrees of depigmentation were observed in both monobenzone-induced and -uninduced sites in both the model group and acupuncture combined with monobenzone group, and the latter group showed earlier, larger and more stable depigmentation than the former group. At 15 days after the end of treatment, the decolorization area index in the model group and acupuncture combined with monobenzone group was 3.45 ＋ 0.17 and 3.90 ± 0.25 at monobenzone-induced sites respectively（t = 7.433, P 〈 0.05）, 1.90 ± 0.12 and 2.85 ± 0.27 at monobenzone-uninduced sites respectively （t = 7.529, P 〈 0.05）. Significant differences were observed in the fluorescence intensity of CD8 ＋ T cells at monobenzone-uninduced depigmented sites between the model group and acupuncture combined with monobenzone group （175.528 ± 10.711 vs. 645.928±12.652, t = 8.105, P 〈 0.05）, and there was a more evident infiltrate with lymphocytes and CD8＋ T cells in the monobenzone-uninduced depigmented sites in the acupuncture combined with monobenzone group. Conclusion Local destruction of skin barrier may promote monobenzone-induced vitiligo-like decolorization in