从环氧化酶和5-脂氧化酶途径探讨虎杖痛风颗粒的抗炎机制

Inhibiting cyclooxygenase and 5-1ipoxygenase activities is an anti-inflammatory mechanism of Huzhang Gout Granule

目的:观察虎杖痛风颗粒对环氧化酶(cyclooxygenase,COX)和5-脂氧化酶(5-lipoxygenase,5-LOX)活性的影响,探讨虎杖痛风颗粒抗炎的可能机制。方法:健康志愿者全血标本加入钙离子载体 A23187予以刺激,以阿司匹林为对照,通过酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测血栓素 B_2(thromboxane B_2,TXB_2)含量考察不同浓度的虎杖痛风颗粒溶液对 COX-1活性的影响;健康志愿者全血经阿司匹林灭活 COX-1后,予脂多糖(lipopolysaccharide,LPS)刺激后,以塞来昔布为对照,通过 ELISA 检测前列环素 I_2(prostaglandin I_2,PGI_2)含量考察不同浓度虎杖痛风颗粒溶液对 COX-2活性的影响;大鼠皮下植入含0.5%花生四烯酸溶液的棉球,收集渗液中的多形核白细胞(polymorphonuclear leukocyte,PMN),采用反相高效液相色谱法测定白细胞三烯 B_4(leukotriene B_4,LTB_4)含量考察不同剂量虎杖痛风颗粒对5-LOX 活性的影响。结果:低浓度虎杖痛风颗粒组 TXB_2浓度较高浓度虎杖痛风颗粒组和阿司匹林组升高(P<0.05);高、低浓度虎杖痛风颗粒组 PGI_2浓度较塞来昔布组明显升高(P<0.05),高、低浓度虎杖痛风颗粒组之间比较,差异无统计学意义(P>0.05);空白对照组 LTB_4含量较高、低剂量虎杖痛风颗粒组和地塞米松组高(P<0.05)。结论:虎杖痛风颗粒能通过抑制 COX 和5-LOX 的活性减少 TXB_2、PGI_2和 LTB_4等炎症介质的释放。

Objective： To observe the effects of Huzhang Gout Granule （HZGG）, a compound traditional Chinese herbal medicine, on cyclooxygenase （COX） and 5-lipoxygenase （5-LOX） activities, the two important oxidases in the course of inflammation, so as to investigate the possible anti-inflammatory mechanism of HZGG. Methods： After stimulating the blood sample of healthy volunteer with calcium ionophore A23187, concentration of thromboxane B2 （TXB2） in the healthy volunteer’s blood was detected by enzyme-linked immunosorbent assay （ELISA） to observe the effects of HZGG at low- and high-dose on the activity of COX-1, with aspirin as control drug. The concentration of prostaglandin I2 （PGI2） in the healthy volunteer’s blood sample, in which aspirin was added to destroy activity of COX-1 beforehand and which was stimulated with lipopolysaccharide, was detected by ELISA method to observe the effects of HZGG on the activity of COX-2, with celecoxib as control drug. In the animal experiment, 40 rats were implanted with sponges soaking in 0.5% arachidonic acid solution in the back to induce inflammatory effusion. Content of leukotriene B4 （LTB4） in the polymorphonuclear leukocytes （PMNs） from the inflammatory effusions was detected with reversed-phase high-performance liquid chromatography （RP-HPLC） to observe the impacts of different doses of HZGG on the activity of 5-LOX, with dexamethasone as control drug. Results： The concentration of TXB2 in the low-dose HZGG group was higher than those in the high-dose HZGG group and the aspirin group （P〈0.05）. The concentrations of PGI2 in the low- and high-dose HZGG groups were higher than that in the celecoxib group （P〈0.05）, but there was no significant difference between the low-dose HZGG group and the high-dose HZGG group （P〉0.05）. The content of LTB4 in the blank control group was higher than those in the low-dose HZGG group, the high-dose HZGG group or the dexamethasone group （P〈0.05）. Conclusion： HZGG can reduce the releasing of inflammatory mediators, such as TXB2, PGI2 and LTB4, by inhibiting the activities of COX and 5-LOX.