摘要
目的观察Wnt和Notch信号通路在大鼠创面愈合模型中的表达及作用。方法取25只SD大鼠幼鼠,早期用溴脱氧尿苷(BrdU)标记表皮干细胞,注射BrdU 60 d后建立全层皮肤缺损创面模型。于大鼠致伤后0 d、7 d、14 d、21 d、30 d五个时间点分别各断颈处死5只取标本,利用免疫印迹、免疫组化和免疫荧光双标法观察创面愈合过程中Wnt1、β-catenin、c-Myc、Jagged1、Notch1、Hes1和Brdu的表达情况。结果免疫印迹结果显示Wnt和Notch信号通路成员在伤后表达上调,于伤后7 d达高峰,持续表达至伤后30 d。免疫组化结果显示,β-catenin开始在细胞膜低表达,伤后逐渐转变为表皮全层表达,且部分呈现异常核表达;伤后,Notch1在表皮全层表达逐渐上调,且在基底层表达上调显著。免疫荧光提示,BrdU/c-Myc和BrdU/Hes1双染阳性细胞率在伤后上升,于伤后7 d达高峰,持续表达至伤后30 d。结论 Wnt和Notch双信号通路可能通过调控表皮干细胞的增殖分化参与大鼠创面愈合过程。
Objective To study the expression and influence of Wnt and Notch signaling pathways in wound healing. Methods 25 new born SD rats were labeled by BrdU to identify epidermalstemcells. The full-thickness skin wound models were established in rats. Samples were collected at 0 d,7 d,14 d,21 d,30 d post-scalding. Western blotting,immunohistochemistry and immunofluorescence double staining were employed to examine the expression of Wnt1、β-catenin、c-Myc、Jagged1、Notch1、Hes1 and BrdU in wound healing. Results Western blotting revealed that the expression of the Notch and Wnt signaling components up-regulated after wound,peaked on the 7th day and down-regulated in the following days. Trough immunohistochemistry,the expression of β-catenin was detected at nucleus of full-thickness skin after the injury,compared with the membrane expression of β-catenin in normal skin; and the expression of Notch1elevated at full-thickness skin after the injury,especially in the stratum basale. Immunofluorescent staining indicated that the proportions of BrdU /c-Myc-positive cells and BrdU /Hes1-positive cells increased during wound healing,peaked on the 7th day and then decreased in the following days. Conclusions Wnt and Notch signaling pathways could promote cell migration,re-epithelization and epidermalstemcells proliferation,accelerating wound healing.
出处
《中华损伤与修复杂志（电子版）》
CAS
2014年第2期31-34,共4页
Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)
基金
国家自然基金(81071557
81372062)
