结合辨病论治探索白发症小鼠动物模型的构建方法  

Construction Method Exploration of Mouse Model of Leukotrichia by Combining Disease Differentiation and Treatment

作　　者：王俊杰 吴涛 谢明君 王晓钰 乔明琦 WANG Junjie;WU Tao;XIE Mingjun;WANG Xiaoyu;QIAO Mingqi(School of Traditional Chinese Medicine,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;School of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Emotional Disease and Syndrome Innovation Research Team,Shandong University of Traditional Chinese Medicine,Jinan 250355,China)

机构地区：[1]山东中医药大学中医学院,山东济南250355 [2]山东中医药大学药学院,山东济南250355 [3]山东中医药大学情志病证创新研究团队,山东济南250355

出　　处：《中医药学报》2023年第7期15-21,共7页Acta Chinese Medicine and Pharmacology

基　　金：国家自然科学基金项目(82204794);山东省自然科学基金项目(ZR2021LZY043)。

摘　　要：目的:探讨白发症小鼠动物模型的构建方法。方法:80只雄性C57BL/6J小鼠背部剃毛后,模型组分别采用不同浓度过氧化氢(H_(2)O_(2))、氢醌背部经皮给药,对照组给予等量生理盐水,连续诱导42 d,分别在第7天(D7)、第14天(D14)、第21天(D21)、第42天(D42)拍照观察小鼠背部毛色的变化,HE染色法观察毛囊形态,黑色素染色切片统计不含黑色素毛囊的个数,qPCR法检测与黑色素相关指标TYR、TRP-1、TRP-2、MITF基因表达水平,Western blot法检测TYR、MITF的蛋白表达水平。结果:不同浓度H_(2)O_(2)诱导组在42 d内未见白毛产生,并且对皮肤刺激性伤害明显。4%氢醌诱导1周后小鼠背部用药区域逐渐开始出现白毛,42 d内均见明显白毛出现。与氢醌对照组小鼠比较,4%氢醌组皮肤组织中黑色素毛囊数量减少(P<0.05)。TYR、TRP-1、TRP-2基因表达水平显著下降(P<0.05,P<0.01),TYR蛋白表达水平显著降低(P<0.05)、MITF蛋白表达减少,但差异无统计学意义(P>0.05)。结论:氢醌造模法能成功构建白发症小鼠模型并具有造模时间短、易操作、对小鼠皮肤刺激性小等优点。Objective:To explore the construction method of leukotrichia mouse model.Methods:80 C57BL/6J male mice were shaved on their back.After that,they were administered with different concentrations of hydrogen peroxide(H 2O 2)and hydroquinone on the skin of their back for 42 consecutive days,and the control group was given an equal amount of normal saline in the same way.The changes of the back hair color of the mice were observed,and photos were taken on day 7(D7),day 14(D14),day 21(D21),and day 42(D42)respectively.HE staining was used to observe the hair follicle morphology,melanin staining section was used to count the number of hair follicles without melanin,qPCR was used to detect the gene expression level of melanin related indicators TYR,TRP-1,TRP-2 and MITF,and Western blot was employed to detect the protein expression level of TYR and MITF.Results:Different concentrations of H_(2)O_(2) induced groups showed no white hair production within 42 days but showed significant skin irritation.One week after 4%hydroquinone induction,white hairs began to appear on the back of the mice,and obvious white hairs appeared within 42 days on all mice.Compared with the hydroquinone control group,the 4%hydroquinone group showed a decrease in the number of melanin hair follicles in the skin tissue(P<0.05),the expression levels of TYR,TRP-1,and TRP-2 genes were significantly reduced(P<0.05,P<0.01),the expression level of TYR protein was significantly lowered and the expression level of MITF protein was reduced.But the difference was not statistically significant(P>0.05).Conclusions:Hydroquinone modeling method can successfully construct leukotrichia mouse models and has the advantages of short modeling time,easy operation and less irritation to the mouse skin.

关 键 词：动物模型 白发症 氢醌 双氧水 毛囊 

分 类 号：R275[医药卫生—中医皮科]