| Catharanthus roseus is among the most important medicinal plants, mainly due to its anticancer alkaloids. In this paper, a novel and green enzyme-assisted negative pressure cavitation extraction (E-NPCE) of five main indole alkaloids vindoline (VDL), catharanthine (CTR), vincristine (VCR), vinblastine (VLB) and anhydravinblastine (AVLB) from C. roseus leaves was investigated and finished the middle experiment. The content of these five indole alkaloids in the root, stem and leaf of C. roseus at various developmental stages were determined, and the biomass allocation was also determined to find the best harvest time. To investigate the regulating effect of five indole alkaloids and C. roseus leaves extracts on lipopolysaccharide (LPS)-induced inflammatory factor release from murine RAW264.7cells and C. roseus leaves extracts antioxidant potential.1. In this paper, a novel and green enzyme-assisted negative pressure cavitation extraction (E-NPCE) of five indole alkaloids VDL, CTR, VCR, VLB and AVLB from C. roseus leaves was investigated.The incubation conditions were studied using Box-Behnken design (BBD) with response surface methodology (RSM), which revealed these as follows:enzyme amount0.61%(CB&P, w/w of anhydration material), pH4.73, incubation temperature35.87℃and time8.62h. Under the optimal conditions, the extraction yields of VDL, CTR, VCR, VLB and AVLB reached4.940±0.215,6.283±0.307,0.049±0.002,0.066±0.003and0.038±0.001mg·g-1which represented an increase of38.8%,51.5%,48.4%,37.5%and58.3%, compared to standard NPCE, respectively.2. The content of VDL, CTR, VCR, VLB and AVLB in the root, stem and leaf of C. roseus at various developmental stages were determined, and the biomass allocation was also determined to find the best harvest time. The content of VDL, CTR, VCR, VLB and AVLB in the root, stem and leaf of C. roseus were determined by HPLC.The content of these alkaloids were influenced by season and it varied in the different tissues of the plant. The content of VDL and CTR in the leaves were the highest, and there was VBL in the root is higher, after leaf; the content of VDL and CTR reached the maximum at August, and the content of VLB reached the highest at September. The biomass was the highest in the initial stage of August. Conclusion is the best harvest time was in the initial stage of August.3. Objective to study the effect of investigate the antioxidant activity of C. roseus extracts in vitro. The radical scavenging, ABTS and1,1-diphenyl-2-picrylhydrazyl radical (DPPH) radical by C. roseus extracts were detected and analyzed by using spectrophotometry and multiple linear regression analysis. The correlation between contents of C. roseus extracts and their antioxidant activity in vitro was surveyed.Result of he radical scavenging capacity of C. roseus extracts could be observed in ABTS and DPPH radical showed performance. IC50of root, stem and leaf is153μg·mL-1,476μg·mL-1and447μg·mL-1, respectively. DPPH radical root extracts>leaf extracts>steam extracts. Conclusion of C. roseus extracts showed strong antioxidant activities. The scavenging activities of all the three extracts on ABTS radical were more than80%, and the highest scavenging capacity reached96.11%.4. Objective To study the effect of C. roseus extracts on IL-6and TNF-a expression in lipopolysaccharide (LPS) induced RAW264.7cells. Methods The contents of IL-6and TNF-a were detected by ELISA methods.In the presence or absence of10μg-mL"1LPS or C. roseus extracts (0.012,0.06,0.3mg·mL-1). Results Compared with control group, the contents of IL-6and TNF-a were significantly induced by LPS, C. roseus extracts inhibited the expression of IL-6and TNF-a significantly elicited by LPS in RAW264.7cells, in dose dependent manners. Conclusion C. roseus extracts has anti-inflammatory effect, which might be mediated by down-regulating inflammatory factors IL-6. |
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