富含anthocyanin的水果提取物对血管内皮细胞产生PGE2的影响研究

目的研究富含anthocyanin的水果提取物对血管内皮细胞产生PGE2的影响。方法将人类正常的血管内皮细胞CRL-2606在Kaighn’s F12K培养基中（补加有10％FBS，0．1mg／ml肝素，0．03mg／ml的ECGS，50μg／ml链霉素和500U／ml青霉素）37℃、5％CO2的条件下培养至接近融合，然后在含有从水果中提取出的anthocyanin的F12K培养基中添加或不添加100ng／mlLPS，继续培养18小时，测定细胞生存率，并将培养液离心，收集上清液，用STAT-Prostaglandin E2 EIA Kit试剂盒测定培养基上清液中的PGE2浓度。结果当CRL-2606细胞暴露于300μg／ml或以上的Chokeberry提取物中时，细胞的生存率低于60％，且表现出一定的剂量效应关系。而50～400μg／ml的蓝莓提取物和100～700μg／ml的黑醋栗提取物未显示细胞毒性。LPS刺激可使细胞PEG2释放量增加一倍以上，蓝莓提取物可明显抑制由于LPS刺激引起的PGE2释放增加，100μg／ml的蓝莓提取物可完全抑制此作用而使其含量维持于正常水平。700μg／ml的黑醋栗提取物对处于LPS刺激下的CRL-2606细胞具有一定的抑制PGE2释放作用。而对于处于LPS刺激下的细胞，500tμg／ml以上的Chokeberry提取物能抑制PGE2的释放。结论富含anthocyanin的黑醋栗、蓝莓以及Chokeberry提取物可以抑制由LPS刺激引起的血管内皮细胞释放PEG2增加的作用，具有一定的抗炎、抗氧化作用。

Effect of anthocyanin rich fruit extract on PGE2 produced by endothelia cells

OBJECTIVE: To study the effect of anthocyanin rich fruit extract on PGE2 produced by Endothelia Cells. METHODS: Normal human endothelia cells, CRL-2606, were cultured in F12K medium (complemented with 10% FBS, 0.1 mg/ml heparin, 0.03 mg/ml ECGS, 50 microg/ml streptomycin and 500U/ml penicillin) in 5% CO2 with 95% air at 37 degrees C. When the cells grow close to confluence, anthocyanin rich fruit extract with/without 100 ng/ml of LPS were added to the medium. After 18 hours of incubation, cells were harvested and the supernatant were collected. Cell viability was assayed. After centrifugation, PGE2 concentration in the supernatant was measured with the STAT-Prostaglandin E2 EIA Kit. RESULTS: 300 microg/ml or higher Chokeberry extract showed cytotoxicity effect on CRL-2606 cells, the viability was lower than 60% and showed a dose-response manner. Under using dosage, Blackcurrant extract (100 - 700 microg/ml) and Blueberry extract (50 - 400 microg/ml) did not show any cytotoxicity. When stimulated by LPS, the production of PGE2 by endothelial cells were increased two fold. Blueberry extract inhibit such action. 100 microg/ml of Blueberry extract keeps the production of PGE2 in normal level. 700 microg/mL of Blackcurrant extract and 500 microg/ml Chokeberry extract also inhibit the releasing of PGE2. CONCLUSION: Anthocyanin rich fruit extract from Blueberry, Blackcurrant, and Chokeberry inhibit PGE2 produced by endothelia cells, there exist antiinflammation and antioxidation.