Response Of Taxus Chinensis And Ginkgo Biloba Leaves Under UV Radiation And Proteomics Analysis Of Rheumatoid Arthritis In Rats Treated By Achyranthes Aspera

Proteomics technology is a scientific method for gloable research of disease pathogenesis, cell model, and functional connection at protein level. China has abundant resources of medicinal plants, however, the research on stress response and treated target of medicinal plants using morden molecular biology is rarely reported. In this paper, proteomics, metabolomics, molecular biology, and plant physiology were combined to reveal the defense mechanisms of response to UV radiation and regular network on physiological pathway in Taxus chinensis and Ginkgo biloba leaves at first time. Several key enzymes were identified and confirmed related to secondary metabolism. That provided a theoretical basis for quality improment and value in use of medicinal plants. Achyranthes aspera is commonly used for treatment of rheumatoid arthritis as ethnodrug. We first reported biomarkers relevant to rheumatoid arthritis pathogenesis and targets of Achyranthes aspera treatment using Proteomics and other modern medical methods. It is benefit for searching efficient and low toxicity candidate medicines. The main contents and results of this paper are as following:(1) Response and Defense Mechanisms of Taxus chinensis leaves under UV-A Radiation are Revealed Using Comparative Proteomics and Metabolomics AnalysesTaxus chinensis var. mairei is an endemic species to southeastern China and one of the natural sources for the anticancer medicine paclitaxel. To investigate the molecular response and defense mechanisms of T. chinensis leaves to enhanced ultraviolet-A (UV-A) radiation, gel-free/label-free and gel-based proteomics and GC-MS analyses were performed. The transmission electron microscopy results indicated damage on chloroplast under UV-A radiation. Proteomics analyses in leaves and chloroplast showed that photosynthesis-, glycolysis-, secondary metabolism-, stress-, and protein synthesis, degradation, and activation-related systems were mainly changed under UV-A radiation. Forty-seven PS II proteins and six PS I proteins were identified changed in leaves and chloroplast under UV-A treatment. It indicated that PS Ⅱ was more sensitive to UV-A than PS Ⅰ as UV-A light target. Enhanced glycolysis, with four glycolysis-related key enzymes increased, provided precursors for secondary metabolism. The 1-deoxy-D-xylulose-5-phosphate reductoisomerase and 4-hydroxy-3-methylbut-2-enyl diphosphate reductase were identified significantly increased during UV-A radiation, which resulted in paclitaxel enhancement. Additionally, mRNA expression levels of genes involved in the paclitaxel biosynthetic pathway indicated a down-regulation under UV-A irradiation and up-regulation in dark incubation. These results reveal that a short-term high dose of UV-A radiation could stimulate the plant stress defense system and paclitaxel production.(2) Improved Metabolites of Pharmaceutical Ingredient Grade Ginkgo biloba and the Correlated Proteomics AnalysisGinkgo biloba is an attractive and traditional medicinal plant, and has been widely used as a phytomedicine in the prevention and treatment of cardiovascular and cerebrovascular diseases. Flavonoids and terpene lactones are the major bioactive components of Ginkgo, whereas the ginkgolic acids (GAs), with strong allergenic properties, are strictly controlled. In this study, we tested the content of flavonoids and GAs under ultraviolet-B (UV-B) treatment and performed comparative proteomic analyses to determine the differential proteins that occur upon UV-B radiation. That might play a crucial role in producing flavonoids and GAs. Our phytochemical analyses demonstrated that UV-B irradiation significantly increased the content of active flavonoids, and decreased the content of toxic GAs. We conducted comparative proteomic analysis of both whole leaf and chloroplasts proteins. In total,27 differential proteins in the whole leaf and 43 differential proteins in the chloroplast were positively identified and functionally annotated. The proteomic data suggested that enhanced UV-B radiation exposure activated antioxidants and stress-responsive proteins as well as reduced the rate of photosynthesis. We demonstrate that UV-B irradiation pharmaceutically improved the metabolic ingredients of Ginkgo, particularly in terms of reducing GAs. With high UV-absorption properties, and antioxidant activities, the flavonoids were likely highly induced as protective molecules following UV-B irradiation.(3) Identification of Candidate Synovial Membrane Biomarkers after Achyranthes aspera Treatment for Rheumatoid ArthritisRheumatoid arthritis (RA) is a systemic autoimmune disease whose main symptom is a heightened inflammatory response in synovial tissues. Despite its common occurrence, the pathogenic mechanism of RA is still relatively unknown. Historically, Achyranthes aspera L. has been used in the clinical treatment of rheumatoid arthritis in the She ethnic minority group of China. To verify the anti-arthritic activities of A. aspera and its possible therapy-related factors on the pathogenesis of RA, the main compounds in A. aspera root were isolated and identified by nuclear magnetic resonance (NMR). Using the collagen-induced arthritis (CIA) model, Wistar rats were then immunized with type II collagen and the effects of Achyranthes saponins were evaluated by rats’paw swelling, radiological, and histopathological examinations. Two-dimensional DIGE (2D-DIGE) was then performed to determine the differentially expressed proteins in either inflamed or drug-treated synovium of CIA rats. Treatment with Achyranthes saponins resulted in dramatically decreased paw swelling, proliferation of inflammatory cells, and bone degradation. Fibrinogen, procollagen, protein disulfide-isomerase A3, and apolipoprotein A-I were all increased in inflamed synovial tissues and were found to decrease when administered drug therapy. Furthermore, Alpha-1-antiproteinase and manganese superoxide dismutase were both increased in drug-treated synovial tissues. The inhibition of RA progression shows that A. aspera is a promising candidate for future treatment of human arthritis. Importantly, that the total saponins found within A. aspera are the active component. Finally, that autoantigens such as fibrinogen and collagen could act as inducers of RA due to their aggravation of inflammation. Given this, it is possible that the vimentin and PDIA3 could be the candidate biomarkers specific to Achyranthes saponins therapy for rheumatoid arthritis in synovial membrane.