摘要
目的研究赤芍801（propyl gallate，PrG）对环氧酶（cyclooxygenase，COX）活性、mRNA及蛋白表达的影响。方法采用基于小鼠腹腔巨噬细胞的COX-1和COX-2体外筛选模型，用钙离子导入剂（calcium ionophore A23187）短时刺激小鼠腹腔巨噬细胞，测定培养上清液中的6-酮-前列腺素F1α（6-keto-PGF1α）的量反映COX-1活性；用脂多糖（lipopolysaccharide，LPS）长时间刺激细胞，测定培养上清液中的前列腺素E，2（PGE2）的量反映COX-2活性。半定量RT-PCR法检测PIG对LPS刺激的Raw小鼠巨噬细胞COX-1、COX-2 mRNA表达的影响。Western blot法检测PrG对LPS刺激的Raw小鼠巨噬细胞COX-1、COX-2蛋白表达的影响。结果PrG体外10，5μmol·L^-1浓度下不影响6-keto-PGF1α生成（P〉0．05），而在1，0．5，0．1，0．05μmol·L^-1浓度下则诱导6-keto-PGF1α生成（P〈0．01），并呈较好的剂量依赖性。PrG体外1，10μmol·L^-1可抑制PGE2生成（P〈0．05）。PrG不同浓度对LPS刺激的Raw小鼠巨噬细胞COX-1、COX-2 mR-NA表达无明显影响。PrG（100，10，1μmol·L^-1）浓度下可抑制COX-2蛋白表达，但对COX-1蛋白表达无影响。结论在0．05～10μmol·L^-1。浓度范围内，PrG低浓度时促进6-keto-PGF1α生成，高浓度时抑制PGE2生成，提示其高浓度时抑制COX-2，低浓度时激活COX-1；不同浓度PrG对COX-1、COX-2 mRNA表达均无明显影响。PrG（1～100μmol·L^-1）浓度下可抑制COX-2蛋白表达，但对COX-1蛋白表达无影响。
Aim This study aimed to investigate effects of propyl gallate （PrG） on cyclooxygenase-1 and-2 （COX-1/-2）activities, mRNA and protein expression in murine macrophages. Methods A screening model for COX inhibitors in vitro based on murine peritoneal maerophages was used. COX-1 aetivity was determined by the level of 6-keto-PGF1α in supernatants of euhured maerophages whieh were stimulated with calcium ionophore A23187 for 1 h, while COX-2 activity was determined by the level of PGE2 in supernatants of cultured macrophages which were stimulated with lipopolysaccharide （LPS） for 9 hours. Semi-quantative RT-PCR was used to determine the effect of PrG on mRNA expression of COX-1 and COX-2 in RAW murine macrophage cell line stimulated by LPS. Western blotting was used to determine the effect of PrG on protein expression of COX-1 and COX-2 in RAW cells stimulated by LPS. Results PrG did not affect 6-keto-PGF1α synthesis at concentrations of 5 and 10 μmol · L^-1 （ p 〉0.05 ） , but enhanced 6-keto-PGF1α synthesis at concentrations of 1, 0. 5,0. 1 and 0.01μmol · L^- 1 （ P 〈 0. 01 ） in vitro. It inhibited PGE2 synthesis at concentrations of 10 and 1 μmol · L^-1（P〈0.05）. PrG at tested concentrations did not affect mRNA expression of COX-1 and COX-2. PrG at concentrations of 100,10 and 1 μmol · L^-1 inhibited COX-2 protein expression, while showed no effect on COX-1 protein expression. Conclusion Within the range of 0. 05 10 μmol · L^-1, PrG activated COX-1 at lower concentrations and inhibited COX-2 at higher concentrations in murine peritoneal macrophages. PrG had no effect on COX-1 and COX-2 mRNA expression, while PrG could inhibit COX-2 protein expression.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2006年第10期1188-1194,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No90409021)
关键词
赤芍801
COX-1
COX-2
炎症
propyl gallate
cyclooxygenase-1
cyclooxygenase-2
inflammation
作者简介
蒋跃绒（1976-），女，博士，助理研究员，研究方向：心血管病防治，Tel：010-2874093，E-mail：jiang_yr@hotmail．com
殷惠军（1971-），男，副教授，研究方向：心血管病学，通讯作者，Tel：010-62875599-6408，E-mall：huijunyin@yahoo．com.cn
