塞来昔布对人乳腺癌SKBR-3细胞生长的影响

目的探讨选择性环氧化酶-2(COX-2)抑制剂塞来昔布对乳腺癌SKBR-3细胞生长的影响及机制。方法用不同浓度的塞来昔布处理SKBR-3细胞后,采用CCK-8法检测塞来昔布对SKBR-3细胞增殖活性的影响;流式细胞仪检测细胞周期;酶联免疫吸附试验(ELISA)检测前列腺素E2(PGE2)的释放水平;Western Blot法测定各浓度塞来昔布刺激SKBR-3细胞后Caspase-3被酶解激活情况。结果塞来昔布对SKBR-3细胞的增殖抑制作用呈剂量-时间依赖性;随着塞来昔布浓度的增加,G0/G1期细胞阻滞,S期细胞比例明显减少;塞来昔布明显减少PGE2的释放水平;Caspase-3在细胞凋亡早期被激活,在凋亡晚期则无表达。结论塞来昔布能有效抑制乳腺癌SKBR-3细胞的增殖,诱导其凋亡;其作用机制可能与COX-2表达下调、抑制PGE2水平和促进Caspase-3的活化有关。     

COX-2抑制剂塞来昔布临床用药对胃肠道安全性观察

目的使用COX-2抑制剂塞来昔布和传统NSAIDS双氯芬酸;观察对患者止痛效果和胃肠道安全的影响。方法选择骨关节炎（OA）患者120例,分为两组。一组使用传统NSAIDS双氯芬酸,一组使用COX-2抑制剂塞来昔布胶囊,两组用药分阶段至半年评估对胃肠道耐受性影响。结果NSAIDS组胃肠道不耐受性半年停药80％,塞来昔布组胃肠道不耐受性半年停药只占38％。两组止痛效果相当。结论COX-2抑制剂塞来昔布用于骨关节炎治疗止痛效果满意,胃肠道安全性好。     

白三烯B4在塞来昔布介导的抗癌效应中的作用

目的 研究白三烯B4(LTB4)在环氧合酶2(COX-2)抑制剂塞来昔布介导的抗癌效应中的作用.方法 MTT法检测塞来昔布、LTB4和去甲二氢愈创木酸(NDGA)对人结肠癌HT-29细胞和人前列腺癌PC-3细胞生存的影响以及LTB4对塞来昔布抗癌效应的影响.ELISA法检测塞来昔布对癌细胞中前列腺素E2(PGE2)和LTB4表达的影响.结果 塞来昔布抑制HT-29和PC-3细胞的生存及LTB4表达(P<0.05或P<0.01),但仅下调HT-29细胞的PGE2表达(P<0.01).LTB4能拮抗塞来昔布对HT-29细胞生长的抑制作用(P<0.01),NDGA明显抑制HT-29细胞的生存(P<0.01),而对PC-3细胞则没有这些作用.结论 塞来昔布对HT-29和PC-3细胞的抑制效应是COX-2非依赖性的,且只有在HT-29细胞中,塞来昔布的抗癌作用主要是通过下调LTB4实现的.     

ELISA法检测外周血环氧合酶-2活性在亚临床冠状动脉粥样硬化中的应用探讨

目的:探讨酶联免疫法(ELISA)检测外周血环氧合酶-2(COX-2)活性在亚临床冠状动脉粥样硬化(AS)中的应用价值。方法:应用选择性COX-2抑制剂(塞来昔布)对亚临床冠状动脉粥样硬化进行短期干预,ELISA法检测各分组血浆COX-2水平,并结合临床资料进行分析。结果:亚临床AS组COX-2水平[(10.84±3.11)U/L],明显高于正常对照组[(6.97±1.25)U/L](P<0.05);塞来昔布组在接受塞来昔布后COX-2水平显著降低(P<0.05),接近正常对照组水平(P>0.05),与安慰剂组有明显差异(P<0.05)。结论:亚临床AS血浆COX-2水平明显升高,ELISA法测定血浆COX-2活性可作为亚临床AS的诊断、预后的参考指标。     

环氧化酶-2抑制剂减轻重症急性胰腺炎合并肺损伤的研究

目的研究环氧化酶-2(COX-2)在重症急性胰腺炎(SAP)中的表达,通过应用COX-2抑制剂检验对SAP并发肺损伤的作用,并研究SAP时COX-2抑制剂对细胞因子肿瘤坏死因子(TNF)、白细胞介素(IL)-6的影响及与肺损伤的关系,探讨其可能的机制。方法 SD大鼠54只,随机分为3组:对照组、SAP组、用药组,用药用在造模前30 min经股静脉注射COX-2抑制剂塞来昔布,SAP组和对照组注射等剂量生理盐水。用药组和SAP组将3.5%牛磺胆酸钠溶液用微量注射泵逆行注入胰胆管制造胰腺炎模型。各组分别于手术后30 min、6h、24 h取6只动物抽血测定脂肪酶、TNF、IL-6;胰腺、肺行病理检查,比色法测定肺的髓过氧化物酶(MPO)的活性显示中性粒细胞浸润;放射免疫法测定胰腺前列腺素E2(PGE2)的水平;用实时定量-聚合酶链反应(RT-PCR)测定胰腺COX-2mRNA的表达。结果 COX-2mRNA在SAP表达增强,相对应胰腺的PGE2水平升高,应用COX-2抑制剂塞来昔布后各时间点COX-2mRNA表达明显抑制,PFE2下降;6、24 h胰腺、肺的病理损害减轻,肺的MPO减少,肺的病理损害及肺的MPO与COX的表达有相关性;用塞来昔布后TNF、IL-6在SAP早期无抑制,而24 h明显减低,两者与肺的MPO有相关性。结论 COX-2在SAP病程进展中起重要作用;COX-2抑制剂可以减轻SAP时肺的病理损害,可能与减轻中性粒细胞在肺部的浸润有关;COX-2抑制剂在SAP时对TNF、IL-6的抑制不是直接的,有可能是通过减少中性粒细胞等炎性细胞在肺等器官的聚积后抑制其释放的。     

塞来昔布对SLE患者关节炎症及血浆内皮素1、降钙素基因相关肽水平的影响

目的 探讨特异性环氧合酶2(COX-2)抑制剂塞来昔布对系统性红斑狼疮(SLE)患者关节炎症及血浆内皮素1(ET-1)、降钙素基因相关肽(CGRP)水平的影响.方法 采用随机抽样将100例SLE患者分为治疗组50例和对照组50例,观察治疗前后关节疼痛数的变化并用放免法测定各组治疗前后血浆ET-1、CGRP浓度.结果 塞来昔布治疗组较对照组关节痛数减少,血浆ET-1水平下降, CGRP水平升高.结论 塞来昔布可改善SLE患者关节疼痛,降低血浆ET-1水平,升高血浆CGRP水平.     

选择性环氧合酶-2抑制剂体外筛选模型的建立

目的:建立选择性环氧合酶-2(COX-2)抑制剂筛选模型。方法:以脂多糖(LPS)为刺激剂刺激大鼠腹腔巨噬细胞产生前列腺素E2(PGE2),采用放免法确定最佳刺激浓度和时间,以选择性COX-2抑制剂戊地昔布和达布非隆(darbufelone)为阳性对照药验证实验模型。结果:达布非隆和戊地昔布对COX-2和COX-1的半数抑制浓度(IC50)的比值分别为3．175×10-4和3．576×10-3。结论:本实验建立的COX-2抑制剂筛选模型比较灵敏,可靠,可用于选择性COX-2抑制剂的筛选。     

塞来昔布安全性与耐受性评价

目的:评价COX-2抑制药塞来昔布的安全性和耐受性.方法:采用随机双盲阳性对照方法,将430例RA和OA病人分成3组,塞来昔布100 mg bid组、塞来昔布200mg bid组和双氯芬酸钠75 mg bid组,比较3组在安全性和耐受性的差异.结果:内窥镜下塞来昔布两组病人胃十二指肠溃疡的发生率比双氯芬酸钠组低4倍,具有统计学差异(P＜0.01).塞来昔布对血小板聚集和出血时间无影响,而双氯芬酸钠可减少血小板聚集,延长出血时间.结论:塞来昔布通过选择性抑制COX-2有效减少了胃肠道的毒性作用,避免了与COX-1抑制相关的生理效应,具有良好的安全性和耐受性.     

The Effect of Celecoxib on Memory Function and Expression of Caspase-3 Protein following Traumatic Brain Injury in Rats

目的:探讨选择性环氧合酶-2(COX-2)抑制剂塞来昔布对重型颅脑创伤大鼠脑组织中COX-2和半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达及学习记忆功能的影响.方法:将成年Wistar大鼠48只随机分为脑创伤组、塞来昔布治疗组、假手术组和正常对照组,每组12只.建立大鼠重型闭合性颅脑创伤模型,各组均在相应的时间点处死取材,应用免疫组化法和Western blot法检测COX-2及Caspase-3蛋白表达.再取24只大鼠随机分为脑创伤组、塞来昔布治疗组、假手术组及正常对照组,进行水迷宫测试.结果:脑创伤组COX-2及Caspase-3的水平高于正常对照组及假手术组,塞来昔布治疗组的COX-2及Caspase-3水平低于脑创伤组,差异有统计学意义(P＜0.05).脑创伤组大鼠脑创伤后7～10d搜索平台所需的时间均大于其他3组,差异有统计学意义(P＜0.05),其他3组间比较差异无统计学意义(p＞0.05).结论:塞来昔布对大鼠重型颅脑创伤有脑保护作用,其可降低COX-2、Caspase-3的表达,抑制脑损伤后的炎症反应和细胞凋亡,能够改善脑创伤后的学习记忆障碍.     

塞来昔布抗肿瘤作用的研究进展

到目前为止,人们对塞来昔布（celecoxib）的抗肿瘤机制还不是完全清楚,但是越来越多的研究集中于抑制环氧化酶-2（cyclooxygenase-2,COX-2）活性、降低肿瘤细胞内前列腺素E2（PGE2）水平方面。COX-2是一种诱导型表达的蛋白酶,     

硝普钠对门脉高压兔肝血流阻断后肝内NO,NOS,MDA及SOD的影响

目的 :观察实验性门脉高压家兔肝血流阻断前预注硝普钠 (SNP)对肝组织一氧化氮 (NO)及其合成酶 (NOS)的影响。方法 :门脉高压模型家兔2 4只 ,随机平均分为 3组。肝门阻断前 10min按组分别静脉注射氯化钠注射液 3mL ,硝普钠 (SNP)5.0 μg·kg- 1·min- 1和左旋硝基精氨酸甲酯 (L NAME) 10mg·kg- 1,计 75min ,当肝血流恢复 6 0min后测定肝组织NO ,NOS ,丙二醛 (MDA)及超氧化物歧化酶 (SOD)活性。结果 :SNP组 ,L NAME组与对照组NO分别为 (2 .2± 0 .5) ,(1.15± 0 .2 7)和 (1.6 4± 0 .2 8) μmol·g- 1Pro(P <0 .0 5和P <0 .0 1)。MDA分别为 (1.14± 0 .12 ) ,(1.70± 0 .2 9)和(1.4 3± 0 .2 2 )nmol·mg- 1Pro(P <0 .0 1)。结论 :硝普钠是一有效的外源性NO供体     

赛来昔布和帕瑞昔布钠用于围术期镇痛的效果观察

目的观察赛来昔布和帕瑞昔布钠用于围术期镇痛的效果。方法将髋关节置换、膝关节置换和脊柱手术患者72例随机分为2组,试验组42例采用赛来昔布和帕瑞昔布钠进行围术期镇痛,在术前2h给予赛来昔布400mg、术后即刻给予帕瑞昔布钠40mg肌内注射、以后每12小时1次,共6次,进行围术期镇痛;对照组30例使用静脉镇痛泵镇痛,药液为芬太尼混合液。2组均在术后24、48、72h用VAS评分评价疼痛程度,在术前、术后即刻和48h测定血清中COX-2和人前列腺素E（PGE2）。结果 2组的VAS评分在术后24h和48h具有统计学差异（P〈0.05）。术前2组血清COX-2和PGE2差异无统计学意义（P〉0.05）;试验组术后即刻血清COX-2和PGE2均下降,但与术前对比无统计学意义（P〉0.05）;术后48h血清COX-2和PGE2均下降,且与术后即刻对比较差异有统计学意义（P〈0.05或P〈0.01）;对照组术后即刻血清COX-2和PGE2均升高,而且PGE2浓度术后即刻与术前对比差异有统计学意义（P〈0.05）;术后48h血清COX-2和PGE2均较术后即刻下降,但术后48h与术后即刻血清COX-2比较差异无统计学意义（P〉0.05）;而血清PGE2浓度差异有统计学意义（P〈0.01）;术后即刻2组血清PGE2浓度差异有统计学意义（P〈0.05）,术后48hCOX-2和PGE2差异均有统计学意义（P〈0.05）。结论围术期使用赛来昔布和帕瑞昔布钠具有较强的镇痛作用。     

两种一氧化氮合酶抑制剂的急性毒性及对海洛因依赖性小鼠戒断综合征的作用

目的··:研究一氧化氮合酶抑制剂NAME和NABE的急性毒性对海洛因依赖小鼠的戒断症状的治疗作用并测定体内一氧化氮 (NO )的含量。方法·· :用NAME和NABE1000mg.kg-1 经灌胃及37.5mg.kg-1 静脉注射给药进行急性毒性实验 ;采用海洛因递增法形成小鼠依赖模型 (海洛因剂量从10mg.kg-1增至50mg.kg-1) ,皮下注射给药4.5d ,采用上述小鼠模型 ,观察NAME及NABE对海洛因依赖小鼠戒断综合征的治疗效果。结果··:NAME和NABE在尾静脉注射37.5mg.kg-1 及灌胃1000mg.kg-1 剂量下未见明显急性毒性 ;NAME16mg.kg-1 、8mg.kg-1 剂量组可明显延长戒断小鼠跳跃潜伏期 ,减少小鼠的戒断跳跃反应数 ,且抑制其腹泻症状 ,而NAME4mg.kg-1剂量组及NABE8mg.kg-1剂量组除对小鼠戒断反应的个别指标有影响外 ,其余作用均不明显 ;NAME对胸腺有一定的保护作用。NAME及NABE减轻海洛因依赖小鼠血液NO水平明显降低。结论··:NAME及NABE毒性均较小 ,NAME减轻海洛因依赖小鼠戒断反应的作用较为明显     

一氧化氮与索曼中毒

目的：探索索曼中毒机制是否有ＮＯ参与，方法：预先侧脑室注射Ａｒｇ、ＮＡＭＥ，观察索曼中毒小鼠惊厥潜伏期、死亡率及脑中ＮＯＳ活性变化。结果：Ａｒｇ预处理时，潜伏期从５．２ｍｉｎ（对照组）缩短到４．３ｍｉｎ（Ａｒｇ１６０ｎｍｏｌ），死亡率由５０％（对照组）增加至８１％（Ａｒｇ１６０ｎｍｏｌ）。ＮＡＭＥ预处理时，潜伏期从４．０ｍｉｎ延长到１４．５ｍｉｎ（ＮＡＭＥ２．２０μｍｏｌ），死亡率由８７％（对照组     

Analysis of the mechanism for the anti-inflammatory effect of the anti-rheumatic drug auranofin

Effects of auranofin, an orally active chrysotherapeutic agent, were examined on the production of prostaglandin E2 (PGE2) and nitric oxide (NO) in rat peritoneal macrophages and in RAW 264.7 cells, a murine macrophage-like cell line. Auranofin (1-10 microM) inhibited PGE2 production in rat peritoneal macrophages stimulated with 12-O-tetra-decanoylphorbol 13-acetate (TPA, 16.2 nM) at 8-20 h, but did not affect PGE2 production at 4 h. However, in non-stimulated rat peritoneal macrophages, auranofin increased PGE2 production at 4 h and had no effect on PGE2 production at 8-20 h. It was proved that auranofin (1-10 microM) increased COX (cyclooxygenase)-1-dependent PGE2 production and inhibited COX-2-dependent PGE2 production in rat peritoneal macrophages. Auranofin showed no effect on the enzyme activities of the purified COX-1 and COX-2 proteins. Furthermore, auranofin did not affect the COX-1 protein level, but inhibited the TPA-induced expression of COX-2 protein. Therefore, it was suggested that auranofin inhibited PGE2 production by inhibiting the COX-2 protein induction in TPA-stimulated macrophages. In RAW 264.7 cells, auranofin (0.3-3 microM) inhibited lipopolysaccharide-induced NO synthesis by inhibiting the induction of NO synthase (NOS) protein expression. Auranofin did not affect the enzyme activity of iNOS (inducible NOS). Finally, using rat peritoneal macrophages, the effects of auranofin on PGE2 production and NO production were determined. Auranofin (10 microM) strongly inhibited the production of PGE2 and NO, and the induction of COX-2 protein and NOS protein by TPA. Indomethacin, a COX inhibitor, partially inhibited NO production at the concentration at which PGE2 production was completely inhibited. On the other hand, L-NG-monomethyl-L-arginine acetate (L-NMMA), a NOS inhibitor, partially inhibited PGE2 production. NO production was completely inhibited at the same concentration as shown above. These findings suggest that PGE2 production and NO production partially affect each other. Therefore, the inhibition of PGE2 production by auranofin might be partly due to the inhibition of NO production, and the inhibition of NO production by auranofin be partly due to the inhibition of PGE2 production. In conclusion, auranofin inhibits both PGE2 production and NO production by inhibiting the upregulation of mRNA levels of COX-2 and NOS.     

骨性关节炎患者血清及膝关节液白细胞介素-1、肿瘤坏死因子-α的水平变化及意义

目的 观察骨性关节炎（OA）患者膝关节液及血清中白细胞介素-1（IL-1）及肿瘤坏死因子-α（TNF-α）水平与病变程度的关系. 方法 50例单侧膝关节OA患者中早期、中期和晚期分别为18例、15例和17例,检测患侧膝关节液及血清中IL-1和TNF-α水平,以健康志愿者15例为对照组. 结果 OA患者膝关节液及血清中IL-1及TNF-α水平高于对照组（P＜0.05）,随着疾病程度加重,膝关节液及血清中的IL-1及TNF-α水平显著增加,IL-1和TNF-α水平存在线性相关（均P＜0.01）. 结论 IL-1和TNF-α与OA患者免疫炎症有关,其水平高低可反映病情活动变化情况,与病变程度密切相关,联合检测这两种指标有助于病情判断.     

Effects of an endogenous nitric oxide synthase inhibitor on phorbol myristate acetate-induced acute lung injury in rats

1. In the present study, we determined whether the endogenous nitric oxide (NO) synthase (NOS) inhibitor Nomega-nitro-l-arginine methyl ester (l-NAME) could ameliorate the acute lung injury (ALI) induced by phorbol myristate acetate (PMA) in rat isolated lung. 2. Typical ALI was induced successfully by PMA during 60 min of observation. At 2 micro g/kg, PMA elicited a significant increase in microvascular permeability (measured using the capillary filtration coefficient Kfc), lung weight gain, lung weight/bodyweight ratio, pulmonary arterial pressure (PAP) and protein concentration of bronchoalveolar lavage fluid. 3. Pretreatment with the NOS inhibitor l-NAME (5 mmol/L) significantly attenuated ALI. None of the parameters reflective of lung injury showed significant increase, except for PAP (P < 0.001). The addition of l-arginine (4 mmol/L) blocked the protective effective of l-NAME. Pretreatment with l-arginine exacerbated PMA-induced lung injury. 4. These data suggest that l-NAME significantly ameliorates ALI induced by PMA in rats, indicating that endogenous NO plays a key role in the development of lung oedema in PMA-induced lung injury.     

Hippo-YAP信号通路在人膝骨性关节炎中表达的意义

目的探讨Hippo-YAP信号通路在膝骨关节炎中的作用及其意义。方法研究对象包括膝骨关节炎患者71例,正常人28例,应用ELISA测定YAP和BLC-2在关节滑液中的表达程度。结果实验组与对照组的滑液中YAP与BCL-2的表达水平作单因素方差分析,结果见表1。YAP的实验组与对照组比较差异有统计学意义P〈0.01,KOA患者的滑液中YAP的水平显著高于对照组。BCL-2的实验组与对照组比较差异有统计学意义P〈0.01,KOA患者的滑液中BCL-2的水平显著低于对照组。实验组中YAP与BCL-2水平作Spearman相关分析,相关系数为r=0.921（P〈0.01）,两者呈正相关。结论 Hippo-YAP信号通路可能是软骨细胞的抗凋亡过程,深入探讨Hippo-YAP信号通路在骨关节炎中的各种促进机制将有可能建立骨关节炎新的治疗靶点。     

Central ghrelin gastroprotection involves nitric oxide/prostaglandin cross-talk

BACKGROUND AND PURPOSE: Ghrelin, a gut-brain peptide, is considered a gastroprotective factor in gastric mucosa. We investigated the role of prostaglandins (PG) and the possible interplay between PGs and nitric oxide (NO) in ghrelin gastroprotection against ethanol (EtOH)-induced gastric lesions. EXPERIMENTAL APPROACH: We examined the effects of (1) central ghrelin (4 mug per rat) injection on PGE(2) accumulation in normal or EtOH-lesioned gastric mucosa, (2) pretreatment with indomethacin (10 mg kg(-1), p.o.), a non-selective cyclooxygenase (COX) inhibitor, and with a selective COX-1, SC560 (5 mg kg(-1), p.o.) or COX-2 inhibitor, celecoxib (3.5 mg kg(-1), p.o.) on ghrelin gastroprotection against 50% EtOH (1 mL per rat)-induced gastric lesions, (3) the NO synthase inhibitor, L-NAME (70 mg kg(-1), s.c), on gastric PGE(2) content in ghrelin-treated rats and (4) central ghrelin on the expression of constitutive and inducible NOS and COX mRNA and on the localization of the immunoreactivity for COX-2 in the gastric mucosa exposed to EtOH. KEY RESULTS: Ghrelin increased PGE(2) in normal mucosa, whereas, it reversed the EtOH-induced PGE(2) surge. Ghrelin had no effect on mucosal COX-1 expression but reduced the EtOH-induced increase in COX-2 expression and immunoreactivity. Indomethacin and SC560, but not celecoxib, removed ghrelin gastroprotection. L-NAME prevented the PGE(2) surge induced by ghrelin and, like indomethacin, reduced EtOH-induced PGE(2) increase. Ghrelin enhanced eNOS expression and reduced iNOS mRNA. CONCLUSIONS AND IMPLICATIONS: This study shows that COX-1-derived PGs are mainly involved in ghrelin gastroprotection and that the constitutive-derived NO together with PGE(2) are involved in ghrelin gastroprotective activity.