Arachibonate cascade and physiological

(1)测定高脂血症患者尿中前列环素（PGI_2）代谢物、血栓素A_2 （TXA_2）代谢物和8-异前列腺素水平。结合患者的血脂水平，提示患者TXA_2/PGI_2分泌失衡，同时提示二十碳五烯酸对TXA_2、PGI_2和8-异前列腺素分泌有影响。此外，糖尿病患者以及妊娠期糖尿病、宫内发育迟缓等妊娠异常患者体内TXA_2/PGI_2分泌失衡。(2)提示COX-2抑制剂在急性炎症模型中的抗炎作用是以COX-2抑制PGE_2的产生为基础的，而对PLA_2和COX-2基因表达的抑制对地塞米松治疗中血浆渗出的抑制作用并不重要。在血管化模型中，COX-1不是COX-1，而是COX…More -2通过调节PG的产生促进了血管化，也表明诱导的VEGF确实影响了该模型的血管化。我们建立了一个测定活细胞中COX活性的系统，发现COX除了具有COX活性外还具有过氧化物酶活性，通过该系统，我们发现只有COX-2利用内源性花生四烯酸，NO显著抑制COX-2活性。用磷酯诱导人巨核母细胞白血病CMK细胞中COX-1的表达。PGD合成酶的造血类型在血小板中未被发现，在CMK细胞中被发现。COX-2在小鼠成骨细胞MC3T3-E1中被TNFalpha显著诱导，提示NF-kappaB和NFIL-6在诱导过程中发挥转录因子的作用。(3)建立了白三烯（LT） B_4的定量方法；单克隆LTB_4放射免疫分析系统，尿样便捷酶联免疫吸附分析系统，ESI离子源LC/MS微量测定系统。哮喘患者尿中LTE_4和11β - pgf_ <2a>水平明显高于正常对照组，哮喘患者尿中LTE_4水平明显高于正常对照组，肺气肿患者尿中11β - pgf_ <2a>水平明显高于正常对照组。我们发现，新型抗糖尿病药物曲格列酮对大鼠肥大细胞系RBL-2H3具有抑制lt产生的作用。成功克隆的LTB_4受体在CHO细胞中表达时，引起腺苷酸环化酶活性的抑制，细胞内钙浓度的升高，并对细胞活性产生影响。少

(1) We measured the urinary levels of prostacyclin (PGI_2) metabolites, thromboxane A_2 (TXA_2) metabolites, and 8-isoprostane in the patients with hyperlipidemia. Imbalance of the TXA_2/PGI_2 production was suggested in these patients with the progress of the disease, taking their serum lipids levels into consideration, It is also suggested that eicosapentaenoic acid administration influenced their TXA_2, PGI_2, and 8-isoprostane production. Moreover, imbalance of the TXA_2/PGI_2 production was observed in diabetics and in some cases with abnormal pregnancy such as gestational diabetes mellitus or intrauterine growth retardation.(2) It was indicated that the anti-inflammatory action in COX-2 inhibitors in acute inflammatory model was based on the inhibition of the PGE_2 production with COX-2, and that the suppression of the gene expression of PLA_2 and COX-2 were not so important for the suppression of the plasma exudation observed in the treatment with dexamethasone.Not COX-1 but COX … More -2 facilitated the vascularization through the regulation of the PG production in the vascularization model, It was also indicated that the induced VEGF actually influenced the vascularization in this model.We established a system for the measurement of the COX activity in lived cells, noticing that COX possesses the peroxidase activity in addition to the COX activity, With this system, we showed that only COX-2 utilized the endogenous arachidonic acid and that NO significantly suppressed the COX-2 activity. COX-1 was induced by phorbol ester in human megakaryoblast leukemia cells CMK.Hemopoietic type of the PGD synthetase, which was not identified in platelets, was identified in the CMK cells. COX-2 was significantly induced by TNFalpha in murine osteoblast cells MC3T3-E1, It was indicated that NF-kappaB and NFIL-6 played roles as the transcription factors in the course of the induction.(3) We established the quantitative methods of the leukotriene (LT) B_4 ; the radio immunoassay system using monoclonal antibody to LTB_4, the convenient enzyme-linked immunosorbent assay system using urinary samples, and the microdetermination system using LC/MS with ESI ion source.Patients with asthma showed significantly high levels of LTE_4 and 11beta-PGF_<2a> in urine compared with those with the healthy volunteers, Patients with aspirin-induced asthma showed greatly high LTE_4 levels in urine, Significantly high 11beta-PGF_<2a> levels were observed in the urine from the patients with pulmonary emphysema.We revealed that troglitazone, a novel antidiabetics, had LT-production suppressive action in the rat mast cell line RBL-2H3. We also revealed the biochemical aspects of the LTB_4 receptor that we succeeded in the cloning, When the LTB_4 gene expressed in CHO cells, it induced the inhibition of adenylate cyclase activity, increase in the concentration of the intracellular calcium, and it influenced to the wondering cell activity. Less