促皮肤创面修复和虫体断肢再生的美洲大蠊活性多肽研究

美洲大蠊（俗称蟑螂）素有“小强”之称，有极强的伤口愈合和断肢再生能力；干燥虫体提取物制备的中成药“康复新液”含活性多肽有效成分，可促进创面修复，治疗金创和烧烫伤具有良效。两者均涉及到活性多肽促进细胞分裂、迁移和分化，但其多肽成分、作用机制、内在联系均不清楚。申请人实验室已测定了美洲大蠊的基因组，验证了其伤口愈合和断肢再生能力。本项目拟（1）以人皮肤细胞系和表皮干细胞为生测对象，结合HPLC分离、质谱分析和基因组比对鉴定活性多肽，并在小鼠创面修复模型中验证；（2）RNA干扰该活性多肽编码基因观察是否降低美洲大蠊伤口愈合和断肢再生的能力；（3）在哺乳动物和美洲大蠊中确定该活性多肽是否通过JNK, Grainy head, Wnt, Notch, TGF-beta和 Hedgehog 等信号通路诱导细胞分裂和分化。研究成果不仅有助于揭示“小强”的生命奥秘，还对我国传统昆虫中药具有弘扬和推动作用。

Cockroaches, including the American cockroach Periplaneta Americana, are known as “little toughness” with extremely strong capacity of wound healing and amputation regeneration. The Chinese medicine named “rehabilitated new liquid” is made of dry extracts from P. Americana and clinically used for healing incised wounds and burns. The repair process in both insect and human could involve cell division and differentiation promoted by active polypeptides. However, the active ingredients and their functional mechanism as well as the internal link from cockroach to human are nearly unknown. Our lab has sequenced the whole genome from P. Americana, and verified its ability of wound healing and limb regeneration. This project will be focused on the following items: (1) To identify the active polypeptides in vitro with the injured human skin cell lines and skin stem cells for bioassay by combing HPLC separation, mass spectrometry assay, and genome blast, and further confirm their roles in vivo by using the skin-injured mouse model. (2) To observe if RNAi knockdown of the polypeptide genes could decrease wound healing and limb regeneration in the cockroach. (3) To assess if the polypeptides activate cell division and differentiation via the JNK, Grainy head, Wnt, Notch, TGF-beta, and Hedgehog signal transduction pathways in both mammals and cockroach. This study will help unravel the life mystery of cockroach and drive the development of traditional Chinese medicine.