黄芩苷对白癜风小鼠模型的作用机制研究

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (1): 27-32.

黄芩苷对白癜风小鼠模型的作用机制研究

祝逸平，金嵘，王遂泉，许爱娥

浙江中医药大学附属杭州市第三人民医院皮肤科，杭州 310009，浙江

收稿日期:2016-11-03 修回日期:2016-12-15 出版日期:2017-01-26 发布日期:2017-01-23
通讯作者: 许爱娥，女，本科，主任医师，教授，博士生导师，研究方向：中西医结合防治色素性皮肤病。 Tel: 0571-87827535 E-mail: xuaiehz@man.com
作者简介:祝逸平，女，博士，主治医师，研究方向：中西医结合防治皮肤病。 Tel: 0571-87827535 E-mail: juyeapy@126.com
基金资助:
浙江省自然科学基金（LQ16H290001，LQ15H290006）；国家自然科学基金（81071294，81541084）；市科委重大专项创新项目(20122513A02)；国家临床重点专科建设项目

Effect of baicalin on vitiligo mice induced by monobenzone

ZHU Yiping, JIN Rong,WANG Suiquan, XU Ai'e

Dermatology Department, Third People's Hospital of Hangzhou Affiliated to Zhejiang Chinese Medicine University, Hangzhou 310009, Zhejiang, China

Received:2016-11-03 Revised:2016-12-15 Online:2017-01-26 Published:2017-01-23

摘要/Abstract

摘要：

目的:探讨黄芩苷对莫诺苯宗诱导的白癜风小鼠模型发生发展的影响及其机制。方法: C57BL/6小鼠共40只分为4组，即阴性对照组、模型组、卤米松组及5%黄芩苷组，每组10只。应用40%莫诺苯宗乳膏诱导C57BL/6小鼠脱色，建立白癜风小鼠模型；进行5%黄芩苷对白癜风小鼠的疗效观察及其机制研究等；连续给药50 d，通过肉眼观察小鼠的毛发脱色，反射共聚焦显微镜(RCM)观察皮肤的黑色素和黑素细胞，免疫荧光检测CD+8T细胞。选取与白癜风相关的5个基因（CXCL9，CXCL10，CXCR3，RAB27A，PI3K）进行荧光实时定量PCR检测。结果: 模型组小鼠在用药部位及非用药部位有毛发脱色现象。卤米松组和5%黄芩苷组小鼠脱色减少，较模型组发生率明显降低、脱色出现时间明显延迟，脱色面积减小，且局部淋巴细胞和CD+8T细胞浸润较模型组明显减少。荧光实时定量PCR检测提示卤米松组和5%黄芩苷组CXCL9、CXCL10、CXCR3和RAB27A表达较模型组明显降低，而PI3K则升高明显。卤米松组和5%黄芩苷组之间无明显差异，但卤米松组用药30 d后，用药部位出现皮肤萎缩等不良反应。结论:卤米松组和5%黄芩苷组均具有治疗效果，黄芩苷组更安全有效，可为临床白癜风的治疗提供参考。

关键词: 黄芩苷, 莫诺苯宗, 白癜风

Abstract:

AIM: To study the effect of baicalin on vitiligo mice induced by monobenzone. METHODS: 40 C57BL/6 mice were randomly divided into four groups (n=10), i.e. the negative control group, the model group, the halometasone group and 5% baicalin group. 40% monobenzone cream was applied on C57BL/6 mice to induce vitiligo model. 5% baicalin was used to observe its efficacy on vitiligo and possible mechanism. After treatment for 50 days, hair decolorizing was observed with naked eye, melanin and melanocytes were observed by reflectance confocal microscopy (RCM), and CD+8T cells were tested by immunofluorescence detection. 5 vitiligo-related genes (CXCL9, CXCL10, CXCR3, RAB27A, PI3K) were detected by real-time fluorescence quantitative PCR. RESULTS: Mice in model group showed depigmentation at the monobenzone application and non-application sites.The halometasone group and 5% baicalin group manifested less, delayed, smaller decolorization with fewer infiltrated lymphocytes and CD+8T cells compared with the model group. Expressions of CXCL9, CXCL10, CXCR3 and RAB27A detected by Fluorescence real-time quantitative PCR in the halometasone group and 5% baicalin group was significantly lower than that of the model group, while PI3K increased significantly. No significant difference was observed between the halometasone group and the 5% baicalin group, yet the halometasone group showed incidence of skin atrophy after 30 days' treatment. CONCLUSION: Halometasone and 5% baicalin are both effective for treating vitiligo, while baicalin presented with safer results, which can be referential for clinical application.

Key words: baicalin, monobenzone, vitiligo

中图分类号:

R965.2

祝逸平，金嵘，王遂泉，许爱娥. 黄芩苷对白癜风小鼠模型的作用机制研究[J]. 中国临床药理学与治疗学, 2017, 22(1): 27-32.

ZHU Yiping, JIN Rong,WANG Suiquan, XU Ai'e. Effect of baicalin on vitiligo mice induced by monobenzone[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(1): 27-32.

参考文献

［1］刘栋，朱文元，谭城. 皮质类固醇对培养Cloudman S91黑素瘤细胞株黑素合成的影响［J］. 临床皮肤科杂志，2003，32(1)：3-6.
［2］张海湃，余昕，贲艺雯，等. 黄芩防晒作用的研究［J］. 长春中医药大学学报，2014，（2）：224-226.
［3］Wu RL, Feng S,Wang JD. Stay on the inhibition effect of baicalin and baicalin metallic coordination［J］. Sci Technol Rev, 2006, 24(2): 212.
［4］刘璋，胡佑伦，韩瑞玲. 黑素生成过程中黄芩的调节作用［J］. 武汉大学学报（医学版）,2005，26(1)：66-67.
［5］Zhu YP, Wang SQ, Xu A. A mouse model of vitiligo induced by monobenzone［J］. Exp Dermatol, 2013, 22(7): 499-501.
［6］祝逸平，王遂泉，李阳，等. 局部针刺对莫诺苯宗诱导小鼠白癜风样模型的影响［J］. 中华皮肤科杂志，2014，47(1)：25-28.
［7］Harris JE, Harris TH, Weninger W, et al. A mouse model of vitiligo with focused epidermal depigmentation requires IFN-γ for autoreactive CD+8T-cell accumulation in the skin［J］. J Invest Dermatol,2012, 132(7): 1869-1876.
［8］McGuire J, Hendee J. Biochemical basis for depigmentation of skin by phenolic germicides［J］. J Invest Dermatol, 1971, 57: 256-261.
［9］张兰兰，闫明，刘晓东，等. 两种化学脱色剂对两种动物模拟白癜风作用比较［J］. 医药导报，2009，28(6)：690-692.
［10］Jasper G, van den Boom JG, Konijnenberg D, et al. Autoimmune destruction of skin melanocytes by perilesional T cells from vitiligo patients［J］. Invest Dermatol, 2009, 129: 2220-2232.
［11］Wankowicz-Kalinska A, van den Wijngaard RM, Tigges BJ, et al. Immunopolarization of CD+4 and CD+8 T cells to Type-1-like is associated with melanocyte loss in human vitiligo［J］. Lab Invest, 2003, 83: 683-695.
［12］Harris JE, Harris TH, Weninger W, et al. A mouse model of vitiligo with focused epidermal depigmentation requires IFN-γ for autoreactive CD+8T-cell accumulation in the skin［J］. J Invest Dermatol, 2012, 132(7): 1869-1876.
［13］Rashighi M, Agarwal P, Richmond JM, et al. CXCL10 is critical for the progression and maintenance of depigmentation in a mouse model of vitiligo［J］. Sci Transl Med, 2014, 6(223): 223ra23.
［14］Zhai Y, Shen XD, Gao F, et al. CXCL10 regulates liver innate immune response against ischemia and reperfusion injury［J］. Hepatology,2008, 47(1): 207-214.
［15］Wang XX, Wang QQ, Wu JQ, et al. Increased expression of CXCR3 and its ligands in patients with vitiligo and CXCL10 as a potential clinical marker for vitiligo［J］. Br J Dermatol,2016, 174(6): 1318-1326.
［16］Griscelli C, Durandy A, Guy-Grand D, et al. A syndrome associating partial albinism and immunodeficiency［J］. Am J Med, 1978, 65(4): 691-702.
［17］Stromberg S, Bjorklund MG, Asplund A, et al. Transcriptional profiling of melanocytes from patients with vitiligo vulgaris［J］. Pigment Cell Melanoma Res, 2008, 21:162-171.
［18］Janes SM, Ofstad TA, Campbell DH, et al. Transient activation of FOXN1 in keratinocytes induces a transcriptional programme that promotes terminal differentiation: contrasting roles of FOXN1 and Akt［J］. J Cell Sci, 2004, 117: 4157-4168.
［19］Kim NH, Jeon S, Lee HJ, et al. Impaired PI3K/Akt activation-mediated NF-kappaB inactivation under elevated TNF-alpha is more vulnerable to apoptosis in vitiliginous keratinocytes［J］. J Invest Dermatol, 2007, 127: 2612-2617.

[1]	祝逸平，金嵘，林福全，周妙妮，王遂泉，许爱娥. 黄芩苷对H₂O₂氧化损伤人黑素细胞的保护机制研究[J]. 中国临床药理学与治疗学, 2018, 23(4): 377-382.
[2]	韩超，杨柳，张秋玲，潘竞锵，徐俊. 黄芩苷对哮喘大鼠气道重塑作用的实验研究[J]. 中国临床药理学与治疗学, 2017, 22(7): 749-754.
[3]	杨迪，袁婷婷，干舒蕾，崔亚娇，丁茹，吴玉林，杨劲. 静脉注射黄芩苷、黄芩素在大鼠体内的药代规律比较研究[J]. 中国临床药理学与治疗学, 2017, 22(4): 394-400.
[4]	薛薇, 李敏, 庞宏贤, 李扬, 胡欣, 史爱欣. LC-MS/MS法测定粪便样品中百可利和黄芩苷的浓度及在粪便排泄中的应用[J]. 中国临床药理学与治疗学, 2016, 21(12): 1372-1377.
[5]	黄浩, 付雷, 易艳东. 黄芩苷下调沙眼衣原体感染小鼠TLR2和TLR4基因表达的研究[J]. 中国临床药理学与治疗学, 2011, 16(6): 606-610.
[6]	沈红燕, 吴晓冬. 黄芩苷对大鼠脂肪细胞胰岛素抵抗的改善作用[J]. 中国临床药理学与治疗学, 2011, 16(3): 263-269.
[7]	祝诚诚, 李自强, 刘志东, 高国义, HAJI Juma, 舒乐欣, 何新. 药物溶出仿生系统研究黄芩苷固体制剂的释放规律[J]. 中国临床药理学与治疗学, 2010, 15(1): 11-20.
[8]	马珺, 吴晓冬. 黄芩苷对动脉粥样硬化家兔的保护作用及其机制[J]. 中国临床药理学与治疗学, 2008, 13(2): 188-194.
[9]	张晓丹, 吴晓冬. 黄芩苷对离体大鼠心肌缺血的保护作用[J]. 中国临床药理学与治疗学, 2007, 12(2): 177-181.
[10]	宋珏, 路通, 谢林, 王广基, 刘晓东. 黄芩苷及其含药血清在大鼠体内的药动学、药效学相关性研究[J]. 中国临床药理学与治疗学, 2006, 11(12): 1350-1354.
[11]	吕健, 吴晓冬. 黄芩苷对异丙肾上腺素诱导大鼠心肌缺血的治疗作用[J]. 中国临床药理学与治疗学, 2005, 10(9): 992-995.
[12]	冯正权, 沈敏鹤, 吴良村. 黄芩苷体外调控乳腺癌细胞表达TIMP2的实验研究[J]. 中国临床药理学与治疗学, 2005, 10(6): 705-708.
[13]	戴德. 黄芩苷对庆大霉素致耳蜗螺旋神经节细胞损伤作用的影响[J]. 中国临床药理学与治疗学, 2004, 9(8): 939-941.