【作者】 卢葳；

【导师】 李惠；

【作者基本信息】 重庆医科大学 ， 皮肤病与性病学， 2010， 硕士

【摘要】 研究目的:本课题基于前期研究发现TLR7受体在毛囊隆突部高表达并且给予TLR7激动剂咪喹莫特后小鼠毛囊数量增加及生长周期延长的发现拟通过RT-PCR的方法验证前期由基因芯片筛选出的可能的调控基因Fos,并通过使用JNK通路阻断剂SP600125在动物试验水平上进一步证实TLR7信号通路作用于毛囊干细胞的可能靶位基因,以期对TLR7信号通路对毛囊增殖调控的机制有初步的探讨。方法:6周大C57雌性小鼠10只,随机分为使用咪喹莫特实验组与使用生理盐水对照组。4周后小鼠背部皮肤取材,将所取皮肤组织TRIZOL法提取RNA,RT-PCR方法分析两组样品Fos基因mRNA的表达差异。再将C57雌性小鼠20只,随机分为咪喹莫特处理组、DMSO对照组、SP00125组和咪喹莫特及SP600125混合组,处理4周后各组小鼠背部皮肤H&E染色观察组织学改变并行磷酸化组蛋白H3检测增殖情况。结果:(1) Fos mRNA在咪喹莫特实验组的表达率显著高于对照组,差异有统计学意义(P<0.05)(2)阻断JNK信号通路后,再使用TLR7激动剂咪喹莫特,小鼠毛囊的增值水平较咪喹莫特处理组明显降低,但仍高于DMSO对照组。结论:(1)咪喹莫特可能通过激活TLR7使JNK通路下游基因FOS的活化来达到促进毛囊增值的作用。(2)TLR7激活后可能通过JNK、NF-κB和IRF等多条信号通路共同作用达到促进毛囊增殖分化的效果。更多还原

【Abstract】 Research objectives: Our previous research found that TLR7 highly expressed in the hair follicle bulge. After using TLR7 agonist imiquimod, the number of hair follicles increased in mice, and extend the growth cycle. The basis of this subject in the preliminary work, further by RT-PCR method verified the possible regulation of gene Fos, and by using the SP600125 (JNK pathway inhibitor) in animal levels studies to further confirm and explore the role of TLR7 signaling pathway in hair follicle stem cells in the target gene view of the TLR7 signaling pathway on proliferation and regulation of hair follicle mechanism was discussed.Methods: 6-week-old C57 female mice, 10 mice were randomly divided into the use of imiquimod in the experimental group and control group using saline. Back skin of mice after 4 weeks were sacrificed to the TRIZOL extraction from skin tissue RNA, RT-PCR to analyze two samples of mRNA expression of Fos gene differences.Then 20 C57 female mice were randomly divided into treatment group imiquimod, DMSO control group, SP00125 group and imiquimod &SP600125 mixed group.4 weeks after treatment the skin of mice in each group H & E staining of histological change the parallel detection of phosphorylated histone H3 proliferation.Results: (1) Fos mRNA in the experimental group imiquimod significantly higher expression rate, the difference was statistically significant (P <0.05) (2) Block the JNK signaling pathway, then use the TLR7 agonist imiquimod, the proliferation level of mouse hair follicles than imiquimod treatment were significantly lower, but still higher than the DMSO control group.Conclusions: (1) imiquimod may activates TLR7 to enhance the proliferation of the follicle progenitor via Fos activities. (2) TLR7 may activates JNK、NF-κB and IRF several other channels to promote joint action of the effect of proliferation and differentiation of hair follicles.更多还原