RESUMO
BACKGROUND: Apocynum venetum leaves are used as a kind of phytomedicine and the main ingredient in some traditional Chinese medicine products for the relief of colitis. To understand the bioactive constituents of A. venetum L., we did a phytochemistry study and investigated anti-Inflammatory effects of compounds and explored the underlying mechanisms. METHODS: We isolated compounds from ethanol extract of A. venetum L. leaf and detected the most effective compound by NO inhibition assay. We investigated anti-Inflammatory effects on dextran sulfate sodium (DSS)-induced colitis mice and lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The disease activity index was determined by scores of body weight loss, diarrhea and rectal bleeding; histological damage was analyzed by H&E staining; macrophages change in the colon were analyzed by immunohistochemistry (IHC); myeloperoxidase activity was measured by myeloperoxidase assay kits; levels of proinflammatory cytokines were determined by qPCR and ELISA; protein production such as COX-2, iNOS, STAT3 and ERK1/2 were determined by western blotting. RESULTS: We isolated uvaol from ethanol extract of A. venetum L. leaf and found uvaol has excellent potential of inhibiting NO production. We further found uvaol could attenuate disease activity index (DAI), colon shortening, colon injury, and colonic myeloperoxidase activity in DSS-induced colitis mice. Moreover, uvaol significantly reduces mRNA expression and production of pro-inflammatory cytokines (TNF-α, IL-6, IL-1ß, and MCP-1) and infiltration of macrophages in colonic tissues of colitis mice. Studies on LPS challenged murine macrophage RAW246.7 cells also revealed that uvaol reduces mRNA expression and production of pro-inflammatory cytokines and mediators. Mechanically, uvaol inhibits the pro-inflammatory ERK/STAT3 axis in both inflamed colonic tissues and macrophages. CONCLUSIONS: A. venetum leaf contains uvaol and uvaol has potent anti-inflammatory effects on DSS-induced experimental colitis and LPS-stimulated RAW264.7 macrophage cells. These results suggest uvaol is a prospective anti-inflammatory agent for colonic inflammation.
RESUMO
By cross breeding, the maize pepc gene in the pepc transgenic rice was successfully incorporated into the parents of two-, three-line hybrid rice, including sterile lines (Peiai64S, 2302S, 2304S, 2306S and Shuangjiu A) and restorer lines (5129, 02428 and Wanjing97) to breed the high-photosynthetic efficiency parents of hybrid rice and utilize heterosis between C4 and C4/C3 rice. Some lines of pepc transgenic rice (LPTR) have been developed. The study on the generations of LPTR suggests the following: (1) The segregation observed in F2 and BC1 progenies demonstrated that pepc transgene inherited as a single dominant gene in the progenies of LPTR. (2) The maize pepc gene is actively expressed at high level in LPTR, and changes of pepc gene expression in the progenies of LPTR may be related to position effect, difference of gene copy number and environmental factors. (3) Through the selection method of soaking seeds into hygromycin solution to germinate, tracing the pepc gene by PCR analysis, evaluating the performance of the rice plants in the field and examining PEPC activities, the segregation of the pepc transgene in LPTR was controlled effectively. Based on the above strategy three pepc transgene lines, H1596, H1597 and Y1470, have been selected. The result suggests that it is possible to breed practical, stable and high-expression pepc transgenic rice by conventional crossing.