Pepper Leaf Polyphenol Biological Activity Test

Objective:To investigate the biological activities of polyphenol extracts of Capsicum anmum leaves both crude and purified. These were detected by anti-inflammatory test, bacteriostatic test, and autitumor experiment in vitro and in vivo, which provided a theoretical basis for the further research of comprehensive development of Capsicum an.Methods:1. The effects of polyphenol extracts of Capsicum anmum leaves on the proliferation of mice macrophage RAW264.7cell was evaluated by CCK-8assay; the effects of polyphenol extracts on the cycle of RAW264.7cell by Flow Cytometry Method; the effects of polyphenol extracts on inflammatory factors such as NO-PGE2、TNF-α、 IL-6released by RAW264.7cell stimulated by LPS and intracellular iNOS vigour.2. By detecting the inhibition ratio of auricle swelling restrained by polyphenol extracts of Capsicum annuum leaves both crude and purified with mouse auricle swelling model which was induced by xylene, the inhibition of acute inflammation treated with polyphenol extracts were valued.3. Detect the inhibition of common contaminated strains, for instance, Staphylococcus aureus, Bacillus cereus, Eschehchia coli, and Bacillus subtilis, with polyphenol extracts of Capsicum annuum leaves both crude and purified by micro-broth dilution method.4. The cytotoxicity and antitumor activity of purified polyphenol extracts of Capsicum annuum leaves were valued by CCK-8assay, including its proliferous effects on mouse fibroblast cell line L929, human glioma U251cells, human neuroblastoma SH-SY5Y cells, human fibrosarcoma HT-1080cells, and human liver HepG2cells.Results:1. The result of the CCK-8test indicates that after24h of<400μg/mL purified polyphenol extracts of Capsicum anmum leaves treatment, the proliferation of RAW264.7cells have not been inhibited; the cell cycle detection shows that polyphenol extracts of Capsicum annuum leaves could induce cell cycle rest on G2/M phase, which promotes the proliferation of RAW264.7cells; the results of anti-inflammation test demonstrates that after the cells were stimulated by LPS, the concentrations of inflammatory factors such as NO、PGE2、TNF-α、IL-6in the supernatant of the RAW264.7cells cultured with polyphenol extracts of Capsicum anmum leaves were significantly lower than that of control, as well as iNOS activities. It shows that there was a dose-effect relationship between them.2. The result of anti-acute inflammation in vivo test indicated that mouse auricle swelling which was induced by xylenecould be remarkably inhibited not only by purified polyphenol extracts of Capsicum anmum leaves but also by crude extracts. The swelling inhibition ratios were59.84%and62.69%(p<0.01), respectively. Furthermore, the content of PGE2in mice edema ear was respectively decreased from32.41A/g to19.27,16.51A/g(p<0.05); the acute toxicity assay showed that the extract were non-toxic which was proved by the normal increasing of mouse weight, and both alanine transaminase (ALT) and aspartate transanimase(AST) were in a normal range.3. Crude extracts were the best choice for inhibition of bacillus cereus, which was proved by bacteriostasis experiment, and Staphylococcus aureus and Bacillus subtilis were in the second place. Only in high concentration condition, Escherichia coli would perform its inhibition; the purified extracts played a positive role in promoting the proliferation of Bacillus cereus, Staphylococcus aureus, Escherichia coli, and Bacillus subtilis, which may be related to synergistic effect between chemical substances. It needs further study.4. Polyphenol extracts of Capsicum anmnim leaves had no effect of inhibition on proliferation of L929cells, which meant this dose of polyphenol extracts of Capsicum anmum leaves had nontoxicity on normal cells; on one hand the polyphenol extracts could strongly inhibit the proliferation of U251cells, on the other hand the extracts performed certain inhibitory effect on SH-SY5Y, but there were no inhibitory effects on HepG-2cells and HT-1080cells, which explained that the polyphenol extracts had targeted anti-tumor effects.Conclusions: This research showed that polyphenols extracts of Capsicum annuum leaves had anti-inflammatory activity, and its mechanism might be achieved by inhibiting NO、PGE2、 TNF-α、IL-6released from cells and reducing iNOS vigour; the animal experiment further proved that polyphenols extracts had the effect of anti-acute inflammation and were nontoxicity. Bacteriostasis experiment showed that crude polyphenols extracts had biacteriostatic activity but the purified had not, which may be related to synergism; cells experiment suggested that the polyphenols extracts had no any toxic effects on normal cells, but had inhibition effects on U251cells and SH-SY5Y cells, which explained that the polyphenol extracts had targeted anti-tumor effects.