Systematic Biological Platforms Applying in Endemic Taiwanese Plants

It pays much attention to develop natural products for treating or preventing human diseases at present. The influence of aging on internal and external, eg: neuron and skin problems, was an aspect of the important researches. The subjects of neuroprotective effect and skin care were emphasized recently. In order to develop Taiwanese abundant species and to study biological diversity of the researches, we have screened a series of Taiwanese plants by using the cellular tyrosinase activities in human epidermal melanocytes (HEMn) in our laboratory. Since 2006, we hold the CCMP's project for “Training in genomic research and core techniques in Chinese medicine.” We already set up the neuron cell model, differentiated PC12 cells, for the application. In our screening systems, Hygrophila pogonocalyx Hayata and Acer albopurpurascens Hayata exhibited the most potent free radical scavenging activity against hydroxyl (IC50 values were 0.8 and 3.1 μg/ml, respectively), superoxide (IC50 values were 12.8 and 5.3 μg/ml, respectively), and 2,2’-azino-bis(3-ethylbenzothiazoline- 6-sulfonic acid (ABTS) (IC50 values were 7.9 and 2.4 μg/ml, respectively) anion radicals. We further evaluated the two extracts in both bioactive platforms, 6-hydroxydopamine (6-OHDA)-treated nerve growth factor (NGF)-differentiated PC 12 cells and human epidermal melanocytes. They also showed the potent activities in the two models when the concentration was 100 μg/ml. Therefore, the active constituents of the plants will be extracted, purified, and identified by bio-guided system. The isolated constituents will further be determined the biological effects and the related mechanism in the project. Specifically, we will pursue the following three aims: The 1st year: The large amount of H. pogonocalyx will be collected and extracted with 95% ethanol. By using bioassay-guide, the active fractions will be done as followed. After passing through the various chromatographic columns, the pure constituents will be isolated. The structures of isolated active constituents will be identified. They will be evaluated both systems, including the neuroprotective effect in 6-OH-treated nerve growth factor (NGF)-differentiated PC12 cells as well as the cellular tyrosinase and matrix metalloproteinase inhibitory activities in human skin cells. The conditions of plant tissue cultures for H. pogonocalyx will also be tested. The 2nd year: The large amount of A. albopurpurascens will be collected and extracted with 95% ethanol. By using bioassay-guide, the active fractions will be done as followed. After passing through the various chromatographic columns, the pure constituents will be isolated. The structures of isolated active constituents will be identified. They will be evaluated both systems, including the neuroprotective effect in 6-OH-treated NGF-differentiated PC12 cells as well as the cellular tyrosinase and matrix metalloproteinase inhibitory activities in human skin cells. Based on the study of the 1st year, we will compare the differences between wild-type and plant tissue cultures of H. pogonocalyx by using chemical and biological analysis. The 3rd year: We will examine the signal transduction mechanism of the active constituents in both NGF-differentiated PC12 cells and human skin cells. As well as to estimate the feasibility of plant tissues cultures applying in the development of H. pogonocalyx.