罗非昔布对足肿胀小鼠花生四烯酸代谢产物影响的研究

发布时间：2018-02-23 04:49

本文关键词： 罗非昔布花生四烯酸环氧酶抑制剂前列腺素 HETE　出处：《大连医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：罗非昔布为特异性环氧化酶-2抑制药，有抗炎、镇痛作用，持续服用罗非昔布的患者中发生确定的心血管事件（如高血压、脑卒中等）的相对危险性显著增加。罗非昔布以及同类药物（如：塞来昔布）致心血管风险机制的研究日益引起重视。本研究旨在分析罗非昔布对花生四烯酸代谢产物血浆含量的影响，从而探讨罗非昔布导致心血管风险的可能机制，为其他同类药物的临床安全使用提供参考。方法：将雄性C57小鼠随机分组为对照组、足肿胀组和罗非昔布组。在所有处置之前测小鼠足部厚度。足肿胀组和罗非昔布组于足部皮下注射1%角叉菜胶；罗非昔布组于注射角叉菜胶前1小时腹腔注射罗非昔布20mg/kg；对照组于相应时间注射生理盐水。处置完成2小时后测小鼠足部厚度、取血及足部组织。将足组织做病理切片、染色、显微镜下观察病理切片形态学变化。采用BCA蛋白浓度测定试剂盒及小鼠肿瘤坏死因子(tumornecrosisfactor-α，TNF-α)ELISA试剂盒测定小鼠足组织和血浆中的TNF-α含量，采用超高压液相色谱-串联质谱（ultrahighpressureliquidchromatography-tandemmassspectrometry，UPLC-MS/MS）测定血浆中花生四烯酸代谢产物6-酮-前列腺素F1α(6-keto-prostaglandinF1α，，6-keto-PGF1α)、前列腺素E2(prostaglandinE2，PGE2)、白三烯B4（leukotrieneB4，LTB4)、8-羟-二十烷四烯酸（8-hydroxy-eicosatetraenoicacid，8-HETE）、11-羟-二十烷四烯酸（11-hydroxy-eicosatetraenoicacid，11-HETE）、12-羟-二十烷四烯酸（12-hydroxy-eicosatetraenoicacid，12-HETE）以及8,9-环氧二十碳三烯酸(8,9-epoxyeicosatrienoicacid，8,9-EET)的含量。采用统计学软件进行数据分析。结果：足肿胀组小鼠足部组织病理切片显微镜下示：大量炎性细胞浸润，组织明显水肿，提示角叉菜胶制造模型成功；罗非昔布组小鼠足组织肿胀显著改善。与足肿胀组小鼠相比，罗非昔布组血浆6-keto-PGF1α、PGE2、11-HETE含量显著降低（p0.01、p0.05、p0.05）；与对照组相比，罗非昔布组血浆LTB4、8-HETE、12-HETE、8,9-EET含量亦显著降低（p0.01、p0.01、p0.05）。结论：罗非昔布可显著降低足肿胀小鼠花生四烯酸的代谢产物PGI2、PGE2、8-HETE、11-HETE、12-HETE以及8,9-EET的血浆含量；且此作用可能与其心血管风险发生的机制有关。
[Abstract]:Objective: rofecoxib is a specific cyclooxygenase-2 inhibitor with anti-inflammatory and analgesic effects, and certain cardiovascular events (such as hypertension) occur in patients who continue to take rofecoxib. The relative risk of stroke was significantly increased. The mechanism of cardiovascular risk caused by rofecoxib and similar drugs (such as celecoxib) has attracted increasing attention. The purpose of this study was to analyze the effects of rofecoxib on arachidonic acid. The effect of the content of Xie product on plasma, To explore the possible mechanism of cardiovascular risk caused by rofecoxib, and to provide reference for the safe use of other similar drugs. Methods: male C57 mice were randomly divided into control group, foot swelling group and rofecoxib group. The foot thickness was measured before all the treatments. 1% carrageenin was injected subcutaneously into the foot in the foot swelling group and rofecoxib group. Rofecoxib group was injected rofecoxib 20 mg / kg one hour before carrageenin injection, and control group was injected with normal saline at the corresponding time. Foot thickness, blood and foot tissue were measured 2 hours after disposal. BCA protein concentration assay kit and tumor necrosis factor- 伪 (TNF- 伪) Elisa kit were used to detect the content of TNF- 伪 in mouse foot tissue and plasma. Determination of arachidonic acid metabolites 6-keto-prostaglandin F1 伪 6-keto-prostaglandin F1 伪 6-keto-PGF 1 伪, prostaglandin F 1 伪 6-keto-PGF 1 伪, prostaglandin E 2 prostaglandin PGE 2 伪, leukotrienol B4 leukotrieneB4LTB4- 8-hydroxy-hydroxy-eicosatetraenoacididine 8-HETEX 11-hydroxy-#number1# keto-prostaglandin F _ 1 伪, prostaglandin E _ 2 (PGE _ 2), leukotrienotrieneB _ 4 (LTB4) -8-hydroxy-hydroxy-eosatetraenoacididine (8-hydroxy-hydroxy-#number1# enoacidate) in plasma, prostaglandin F _ (1 伪) 6-keto-PGF _ (1 伪), prostaglandin E _ 2 (PGE _ (2)), leukotriene B _ (4) (LTB _ 4) -8-hydroxy-eosoic acid traenoacididine (8-hydroxy--#number1# enoic acid). The contents of enoic acid (12-hydroxy-eicosatetraenoic acididine) and 89-epoxyeicosatrienoic acididine (89-EET-89-EET-89-epoxyeicosatrienoic acididine) were analyzed by statistical software. Results: in the foot swelling group, the pathological sections of the feet showed that a large number of inflammatory cells infiltrated and the tissues were obviously edema, which suggested that carrageenin was a successful model. Rofecoxib group significantly improved the swelling of podocyte tissue. Compared with the paw swelling group, the plasma 6-keto-PGF1 伪 PGE2F1-HETE content in rofecoxib group was significantly lower than that in rofecoxib group, and the content of plasma LTB48-HETE12-HETE89-EET in rofecoxib group was also significantly lower than that in rofecoxib group. Conclusion: rofecoxib can significantly reduce the plasma levels of PGI2PGE28-HETETE-11-HETETE-12-HETE and 89-EET in paw swelling mice, and this effect may be related to the mechanism of its cardiovascular risk.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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