金银花活性成分的光酶诱导和生物合成机制研究

Lonicera japonica Thunb. has significant antiviral and antioxidant activity, with the efficacy of clearing heat and resolving toxin, coursing wind and dissipating heat, in which caffeoyl guinic acid and iridoid glycoside are the main active ingredients. Our preliminary studies have shown that through enzyme-mediated natural product biosynthesis (EMNPB), the content of caffeoyl guinic acid and iridoid glycoside in Lonicera japonica Thunb. were significantly increased as well as significant improvement of antiviral and antioxidant activity. By comparative proteomic analysis of the EMNPB Lonicera japonica Thunb., 5 proteins related to Lonicera japonica Thunb. secondary metabolism were identified..In order to prove the results of proteomics research and to clarify the molecular mechanism of the improved active ingredients through EMNPB, we intend to apply transcriptome and bioinformatics to functional annotation and analysis of the transcriptome before and after EMNPB, to the discovery of functional genes in metabolic pathways. Finally we verify the results by RT-PCR and Western blot analysis and reveal the molecular mechanism of the active ingredients biosynthesis in the EMNPB Lonicera japonica Thunb. from protein level and gene level. Our study establishes a new method to improve the pharmacological active component content through EMNPB and provides new ideas and methods for the improvement of medicinal plant activity and rational use of traditional Chinese medicine resources.

金银花是常用的清热解毒、凉散风热中药，具有显著的抗病毒和抗氧化作用，咖啡酰奎尼酸和环烯醚萜苷类化合物为其主要活性成分。我们前期研究表明，金银花经过光酶诱导处理后，咖啡酰奎尼酸类及环烯醚萜苷类化合物含量显著增加，抗病毒与抗氧化活性亦有很大提高。通过对光酶诱导前后金银花差异蛋白质组学的研究，鉴定出5个与金银花次生代谢相关的蛋白。.为了印证蛋白质组学的研究结果，并阐明光酶诱导金银花活性成分含量提高的分子机理，我们拟运用转录组学和生物信息学技术，对诱导前后转录组进行功能注释和分析，寻找代谢途径中的功能基因，并应用RT-PCR和Western blot技术加以验证，从蛋白质水平和基因水平探讨光酶诱导金银花活性成分生物合成的分子机制。本研究通过光酶诱导技术提高金银花药理活性成分含量，为提高药用植物活性，合理利用中药资源提供新的思路和研究方法。

金银花是常用的清热解毒、凉散风热中药，具有显著的抗病毒和抗氧化作用。但是其所含的主要活性成分咖啡酰奎尼酸和环烯醚萜类化合物的含量较低。 本研究利用光酶诱导技术提高了金银花中主要活性成分的含量，并且提高了金银花的抗病毒和抗氧化的活性。利用蛋白质组学技术，对光酶诱导前后金银花中活性成分含量提高的分子机制进行了探索，发现光酶诱导导致了咖啡酰奎尼酸和环烯醚萜生物合成途径中关键酶和基因的上调表达，以及氨基酸代谢、逆境、能量代谢相关蛋白的变化。该研究结果对于改善中药资源的质量，为合理有效地利用中药资源提供新的思路。