Nimesulide联合奥沙利铂对人肝癌细胞增殖与凋亡的影响

目的:探讨选择性环氧合酶-2(cyclooxy-genase-2,COX-2)抑制剂Nimesulide联合奥沙利铂(Oxaliplatin,L-OHP)对人肝癌细胞株SMMC-7721增殖与凋亡的影响,为肝癌的药物治疗提供依据.方法:预实验筛选Nimesulide有效终浓度与不同浓度L-OHP(0.5、1.0、2.0、5.0mg/L)联合处理肝癌细胞48h;另外选取L-OHP有效终浓度与Nimesulide筛选的浓度单独或联合处理肝癌细胞24h、48h、72h;采用倒置显微镜观察细胞形态学改变,四氮唑盐比色法(MTT法)观察细胞增殖活力的改变,流式细胞仪检测细胞凋亡率.结果:不同浓度Nimesulide及L-OHP对SMMC-7721表现出一定程度的生长抑制作用,并呈剂量依赖性.Nimesulide(50mmol/L)或L-OHP(1mg/L)单用时可显著抑制SMMC-7721细胞的增殖,选用Nimesulide 50mmol/L与L-OHP(0.5、1.0、2.0、5.0mg/L)联合使用时,联合抑制作用呈现协同作用(Q>1.15).L-OHP 1mg/L、Nimesulide 50mmol/L及两药联合处理SMMC-7721细胞24h、48h、72h,各处理组各时间点对肝癌细胞的抑制作用随着时间的延长而增大,并且两药联合作用有协同作用(Q>1.15).流式细胞仪检测分析发现两种药物均可有效诱导SMMC-7721细胞的凋亡,且在上述浓度联合应用时具协同效应.结论:Nimesulide与L-OHP对人肝癌细胞株SMMC-7721均有抑制增殖和促进凋亡作用,而且两者联合应用有协同作用.

COX-2抑制剂Nimesulide抗肺癌作用及其机制研究

目的:流行病学资料显示,非甾体类抗炎药物阿司匹林能降低肺癌的发病率。我们的目的是探讨Nimesulide-一种选择性COX-2抑制剂对肺癌A549细胞增殖的影响及其机制。方法:采用MTT比色法观察Nimesulide对A549细胞生长的影响,采用放射免疫法(ELISA)测定Nimesulide对肺癌A549细胞上清液中前列腺E2(PGE2)释放的影响,免疫细胞化学法测定肺癌A549细胞中COX-2、PCNA、bcl-2基因蛋白表达水平。结果:Nimesulide呈时间、剂量依赖性抑制A549细胞增殖,抑制率最低22.73%,最高72.04%;以剂量依赖性方式抑制A549细胞PGE2释放;Nimesulide可抑制A549细胞PCNA、bcl-2的蛋白表达,而对COX-2蛋白表达无影响。结论:Nimesulide以时间、剂量依赖性方式抑制肺癌A549细胞增殖,其机制可能与抑制肺癌A549细胞PGE2释放,并下调A549细胞PCNA、bcl-2的蛋白表达有关,而对COX-2的蛋白表达无影响。选择性COX-2抑制剂Nimesulide有可能在肺癌防治中发挥重要作用。

5-aminosalicylic acid in combination with nimesulide inhibits proliferation of colon carcinoma cells in vitro

AIM: To investigate the effects of 5-aminosalicylic acid (5-ASA) in combination with nimesulide on the proliferation of HT-29 colon carcinoma cells and its potential mechanisms. METHODS: Inhibitory effects of drugs (5-ASA,nimesulide and their combination) on HT-29 colon carcinoma cells were investigated by thiazolyl blue tetrazolium bromide (MTT) assay. Cellular apoptosis and proliferation were detected by TUNEL assay and immunocytochemical staining,respectively. RESULTS: Pretreatment with 5-ASA or nimesulide at the concentration of 10-1000 μmol/L inhibited proliferation of HT-29 colon carcinoma cells in a dose-dependent manner in vitro (t = 5.122,P < 0.05; t = 3.086,P < 0.05,respectively). The inhibition rate of HT-29 colon carcinoma cell proliferation was also increased when pretreated with 5-ASA (100 μmol/L) or nimesulide (100 μmol/L) for 12-96 h,which showed an obvious time-effect relationship (t = 6.149,P < 0.05; t = 4.159,P < 0.05,respectively). At the concentration of 10-500 μmol/L,the apoptotic rate of HT-29 colon carcinoma cells significantly increased (t = 18.156,P < 0.001; t = 19.983,P < 0.001,respectively),while expression of proliferating cell nuclear antigen (PCNA) was remarkably decreased (t = 6.828,P < 0.05; t = 14.024,P < 0.05,respectively). 5-ASA in combination with nimesulide suppressed the proliferation of HT-29 colon carcinoma cells more than either of these agents in a dose-dependent and time-dependent manner (t = 5.448,P < 0.05; t = 4.428,P < 0.05,respectively). CONCLUSION: 5-ASA and nimesulide may inhibit the proliferation of HT-29 colon carcinoma cells and coadministration of these agents may have additional chemopreventive potential.

Fe_3O_4 Magnetic Nanoparticles Modified Electrode as a Sensor for Determination of Nimesulide

A novel type of Fe3O4 nanoparticles modified glass carbon electrode（Fe3O4/GCE） was constructed and the electrochemical properties of N-（4-nitro-2-phenoxyphenyl）methanesulfonamide（nimesulide） were studied on the Fe3O4/GCE.In 0.4mol/L HAc-NaAc buffer solution（pH=5.0）,the electrode process of nimesulide was irreversible at bare GCE and Fe3O4/GCE.The Fe3O4/GCE exhibited a remarkable catalytic and enhancement effect on the reduction of nimesulide.The reduction peak potential of nimesulide shifted positively from-0.683 V at bare GCE to-0.625 V at Fe3O4/GCE,and the sensitivity was increased by ca.3 times.Some experimental conditions were optimized.The linear range between the peak current and the concentration of nimesulide was 2.6×10-6 ＂1.0×10-4mol/L（R=0.993） with a detection limit of 1.3×10-7mol/L.This method has been used to determine the content of nimesulide in medical tablets.The recovery was determined to be 96.9% ＂101.9% by means of standard addition method.The method is comparable to UV-Vis spectrometry.

Nimesulide抗肺癌作用的体内及体外研究

目的：探讨选择性CoX-2抑制剂Nimesulide对肺腺癌体内、体外的影响及其机制。方法：（1）采用MrITr比色法观察Nimesulide对肺腺癌A549细胞生长的影响，免疫细胞化学法测定肺癌A549细胞中PCNA、bcl-2基因蛋白表达水平；（2）建立肺腺癌裸鼠移植瘤模型，绘制肿瘤生长曲线并计算肿瘤抑制率。结果：（1）Nimesulide呈时间、剂量依赖性抑制A549细胞增殖，抑制率最低22．73％，最高72．04％；Nimesulide可抑制A549细胞PCNA、bcl-2的蛋白表达；（2）尼关舒利处理组的裸鼠移植瘤瘤体的平均重量为0．76±0．4g，显著低于对照组的1．56±1．02g（P〈0．05）；实验结束时尼美舒利对裸鼠移植瘤的体积抑制率为41．59％，重量抑制率为51．28％。结论：Nimesulide以时间、剂量依赖性方式抑制肺癌A549细胞增殖，其机制可能与下调A549细胞Pc—NA、bcl-2的蛋白表达有关。选择性COX-2抑制剂Nimesulide有可能在肺癌防治中发挥重要作用。

Combined anti-tumor effects of mDRA-6 and nimesulide on human hepatocellular cancer cell line SMMC-7721

Objective: The aim of this work was to evaluate anti-tumor effects of mDRA-6 plus nimesulide on a human hepatocellular cancer cell line, SMMC-7721, and study the main mechanisms. Methods: The DR5 receptor of SMMC-7721 cells was detected by flow cytometry (FCM). For further experimental application, SMMC-7721 cells were treated with proper dose of mDRA-6, nimesulide, or mDRA-6 plus 200 μmol/L nimesulide; untreated SMMC-7721 cells were comparably set as control. Cytotoxicity was tested by MTT assay; cell morphology was examined using Hoechst 33258 staining; and apoptosis was determined by FCM. Results: The positive rate of DR5 on SMMC-7721 was 95.0%. Either mDRA-6 or nimesulide alone induces SMMC-7721 cell death in a dose-dependent manner. Treatment of 1,600 ng/mL mDRA-6 for 12h led to a cell-death rate of 35.0%, while an increased cell-death rate (91.1%) was found under the same condition of mDRA-6 treatment supple- mented with 200 μmol/L nimesulide. Hoechst 33258 and Annexin V/PI staining confirmed apoptosis as the main cause of this anti-tumor response. Conclusion: Both mDRA-6 and nimesulide can induce apoptosis of SMMC-7721 cells, and they have synergistic anti-tumor activities against SMMC-7721.

Determination of Camylofin Dihydrochloride and Nimesulide in Pharmaceutical Preparation by Gas Chromatography

This research paper describes simple analytical method for determination of Camylofin dihydrochloride and Nimesulide in tablet formulation by Gas chromatography method. Benzoic acid was used as internal standard. Validation was carried out in compliance with the International Conference on Harmonization guidelines. The method utilized GC (Agilent Technologies 6890 N Network GC system with FID detector), and RTX-5 capillary column (5% diphenyl-95% dimethyl polysiloxane), 30 m × 0.53 mm, 1.5 μm as stationary phase. Helium was used as the carrier gas at a flow rate of 1.5 mL?min–1. The proposed method was validated for linearity, LOD, LOQ, accuracy, precision, ruggedness and solution stability. It can be conveniently adopted for routine quality control analysis.

选择性COX-2抑制剂Nimesulide对肝肿瘤细胞HepG2增殖与凋亡的影响

目的 观察环氧合酶 (COX) 2抑制剂尼美舒利 (Nimesulide)对肝肿瘤细胞HepG2增殖和凋亡的影响。方法 用不同浓度 (10 0、2 0 0、3 0 0 μmol/L)的Nimesulide处理HepG2细胞 ,采用四甲基偶氮唑蓝法检测Nimesulide对HepG2细胞增殖的影响 ;Western印迹法检测HepG2细胞半胱氨酸 天冬氨酸特异性蛋白酶 (Caspase) 3表达 ,流式细胞术检测细胞凋亡。用放射免疫测定法检测Nime sulide对HepG2细胞PGE2释放水平 ,反映Nimesulide对COX 2活性的影响。结果 10 0、2 0 0、3 0 0μmol/L的Nimesulide作用于HepG2细胞 ,其A值分别为 1.69± 0 .16,1.0 6± 0 .0 9,0 .62± 0 .0 6,与对照组 (3 .0 9± 0 .2 8)相比 ,差异有非常显著性 (P <0 .0 1)。Nimesulide诱导HepG2细胞的凋亡率分别为 (4 .5 3± 2 .15 )、(6.0 2± 3 .5 0 )、(16.2 4± 7.92 ) ,与对照组比较 (2 .84± 1.88,P <0 .0 1)。Nimesulide干预的HepG2细胞的Caspase 3的蛋白表达均明显升高 ,A值分别为 (4 6.96± 2 .86)、(60 .2 8± 3 .0 5 )、(76.5 9± 4.2 6) ,与对照组相比 (3 2 .5 7± 1.83 )差异有非常显著性 (P <0 .0 1)。Nimesulide可明显诱导HepG2细胞Caspase 3蛋白活性的升高 ,A值分别为 (0 .46± 0 .0 3 )、(0 .73± 0 .0 6)、(0 .91± 0 .12 ) ,?

A simple and robust HPLC-MS method for the quantitative determination of nimesulide in human plasma and its application to bioequivalence study in Chinese volunteers

A rapid and sensitive high performance liquid chromatography-mass spectrometry （HPLC-MS） method for the quantification of nimesulide in human plasma was developed and validated. Sample aliquots of 100μL were extracted by one-step liquid-liquid extraction after addition of hydrochlorothiazide as the internal standard （IS）. Analytes were separated on a reverse phase C18 column using methanol-water （84：16, v/v） as the mobile phase and detected by a single quadrupole mass spectrometer in selected ion monitoring （SIM） negative mode. Monitored m/z values for nimesulide and IS were 307.00 and 295.90, respectively. The overall run time was 4.2 min. Validation experiments demonstrated good precision and accuracy over a wide concentration range of 20.0-7000 ng/mL with a lower limit of quantification （LLOQ） at 20.0 ng/mL. No interference by endogenous substances or matrix effect was observed. Average extraction recoveries for nimesulide and IS were all greater than 84.4%. The assay was successfully applied to a bioequivalence study of nimesulide dispersible tablets in Chinese male volunteers after oral administration.

Optimization and characterization of nimesulide bilayer tablets by response surface methodology

The objectives of this present investigation were to develop and formulate nimesulide bilayer tablets by using different polymer combinations and fillers, to optimize the formulations for different drug release variables by orthogonal design and central composite design-surface methodology and to evaluate drug release pattern of the optimized product. The bilayer tablet containing a fast release layer（FRL） and a sustained release layer（SRL） provided an initial burst release of nimesulide, followed by the sustained release for a period of time. The optimal formulation obtained was as follows：（I） the formulation of FRL： nimesulide, 50 mg; lactose, 92 mg; starch, 22 mg; CCMC-Na, 14 mg; PVP K30, 1 mg; micronized silica gel, 1 mg; magnesium stearate, 0.9 mg; and iron oxide red, 0.1 mg; and（II） the formulation of SRL： nimesulide, 150 mg; HPMC K100LV, 26 mg; HPMC K4M, 33 mg; lactose, 54 mg; PVP K30, 1 mg; micronized silica gel, 1 mg; and magnesium stearate, 0.9 mg. According to the optimal formulation, the biphasic type of release was identified. The in vitro drug dissolution from the bilayer tablets was sustained for about 16 h after releasing 15% of drug in the first 10 min. The developed nimesulide bilayer tablets with improved efficacy can perform therapeutically better than the conventional tablets.

环氧化酶-2抑制剂nimesulide对乳腺癌细胞株增生、凋亡的影响

目的探讨环氧化酶-2抑制剂nimesulide(NIM)对雌激素受体(ER)阴性(MDA-MB-231)和阳性(MCF-7)人乳腺癌细胞株增生、凋亡的影响。方法应用MTT比色法分析细胞生长抑制作用,流式细胞技术测定细胞周期分布和凋亡率,透射电镜观察细胞形态与超微结构,AnnexinV法检测细胞的凋亡。结果NIM以时间、剂量依赖性方式抑制MDA-MB-231(COX-2阳性)、MCF-7(COX-2阴性)细胞生长,阻滞细胞周期于G0/G1期,诱导细胞的凋亡,MDA-MB-231细胞对NIM的作用更为敏感。NIM对COX-2表达阴性的MCF-7细胞同样具有抑制增生、诱导凋亡的作用。结论NIM对ER阳性和ER阴性乳腺癌细胞均有抑制增生、诱导凋亡的作用。NIM的抗肿瘤作用存在环氧化酶-2依赖性与非依赖性两种途径。

Nimesulide诱导胃癌细胞凋亡的作用机制研究

目的 非甾体抗炎药诱导胃癌细胞凋亡的机制不清楚。通过研究选择性COX 2抑制剂Nimesulide对人胃癌细胞系SGC 790 1的COX 2mRNA表达和c myc、Bcl 2、caspase 3凋亡基因蛋白表达的影响 ,探索其诱导胃癌细胞凋亡的机制。方法 胃癌细胞的凋亡用电子显微镜、AnnexinV FITC染色、流式细胞仪技术测定。COX 2基因表达用RT PCR法测定。c myc、Bcl 2和caspase 3蛋白表达用免疫细胞化学技术测定。结果 Nimesulide在浓度为 50 μmol/L时作用 48、72h和浓度为 1 0 0和2 0 0 μmol/L 时作用 2 4、48、72h均可诱导胃癌细胞凋亡 ,凋亡率分别为 7.51 % ,9.86 % ;1 1 .58% ,1 2 .45 % ,1 6 .66 %和 1 2 .2 1 % ,1 5 .38% ,2 0 .2 8% ,呈浓度和时间依赖性。Nimesulide可降低COX 2mRNA表达和Bcl 2蛋白表达 ,增加c myc和caspase 3蛋白表达。 2 0 0 μmol/LNimesulide作用 72h ,Bcl 2 ,c myc和caspase 3蛋白表达阳性率分别为 (2 0 .2± 7.6) % ,(49.2± 1 5 .1 ) %和 (34 .6± 1 2 .9) % ,对照组为(44.6± 1 2 .1 ) % ,(2 4 .7± 9.5) %和 (1 4 .8± 6 .4) % ,三者对照比较 ,差异有显著性 (P <0 .0 1。)结论 Nimesulide可诱导胃癌细胞SGC 790 1凋亡 ,其机制涉及Bcl 2表达下调、c myc表达上调及caspase 3激活。

In vitro-in vivo correlation study on nimesulide loaded hydroxypropylmethylcellulose microparticles

This study involves mathematical simulation model such as in vitro-in vivo correlation(IVIVC) development for various extended release formulations of nimesulide loaded hydroxypropylmethylcellulose(HPMC) microparticles(M1,M2 and M3 containing 1,2,and 3 g HPMC,respectively and 1 g drug in each) having variable release characteristics.In vitro dissolution data of these formulations were correlated to their relevant in vivo absorption profiles followed by predictability worth analysis of these Level A IVIVC.Nimaran was used as control formulation to validate developed formulations and their respective models.The regression coefficients of IVIVC plots for M1,M2,M3 and Nimaran were 0.834 9,0.831 2,0.927 2 and 0.898 1,respectively.The internal prediction error for all formulations was within limits,i.e.,<10%.A good IVIVC was found for controlled release nimesulide loaded HPMC floating M3 microparticles.In other words,this mathematical simulation model is best fit for biowaiver studies which involves study parameters as those adopted for M3 because the value of its IVIVC regression coefficient is the closest to 1 as compared to M1 and M2.

Acute neck tendonitis with dyspnea:A case report

BACKGROUND Longus colli tendinitis(LCT)with dyspnea is a relatively less-reported condition in the literature,and physicians should be aware of its existence.Misdiagnosis of this condition may cause unnecessary treatment for dyspnea.CASE SUMMARY Herein,we report the case of a 40-year-old man with acute neck tendonitis.The patient presented to the pneumology department clinic with a complaint of acute neck tendonitis with dyspnea.An emergency cervical magnetic resonance examination was performed,and the preliminary diagnosis was“acute longus cervicalis tendinitis.”After aggressive medical treatment,the symptoms obviously improved.CONCLUSION LCT is a self-limiting disease that usually improves after three to seven days of conservative treatment following a definite diagnosis.However,owing to its insidious onset and complex clinical manifestations,most relevant personnel are not fully understood.The definite diagnosis of LCT is based on a comprehensive understanding of the triad,rare symptoms,and the clear identification of cervical 1 and 2 levels calcification and prevertebral edema by medical imaging examination,especially magnetic resonance imaging and computed tomography.

尼美舒利对舌鳞癌Tca8113细胞前列腺素E_2释放和生存素表达的影响

大量研究已表明，环氧化酶-2（cyclooxygenase 2，COX-2）在大多数正常组织中不表达，而在大多数人类肿瘤中过表达，与多种肿瘤的发生发展密切相关。本实验通过研究选择性COX-2抑制剂尼美舒利（Nimesulide，NIM）与人舌鳞癌Tca8113细胞前列腺素E2（PGE2）和生存素（survivin）表达间的关系，探讨NIM对人舌鳞癌细胞生物学行为影响的可能机制，为Nimesulide临床应用提供必要的理论依据。

临床应用尼美舒利治疗关节炎疗效观察

目的:观察临床应用尼美舒利片(NimesulideTablets)治疗关节炎的疗效。方法:将826例关节炎患者随机分组,治疗组421例使用尼美舒利片为主的综合治疗。对照组405例使用传统解热、镇痛、非甾体抗炎药为主的综合治疗。结果:治疗组消炎止痛快、效果好,有效率达86.3%,副作用3.09%;对照组有效率为73.9%,副作用13.08%。结论:两组对比,病例数差异不大,但治疗效果和副作用有明显的差异,治疗组疗效显著,副作用少。具有可比性。

Protective effects of selective and non-selective cyclooxygenase inhibitors in an animal model of chronic stress

Objective Cyclooxygenase isoenzyme is known to be expressed in different regions of brain, and is mainly used for the treatment of pain and inflammation. Recently, it is proposed that cyclooxygenase isoenzyme may also play a key role in the pathophysiology of various brain-related disorders. The present study was aimed to explore the protective effect of cyclooxygenase inhibitors on stress by using an animal model of chronic stress. Methods The animals were forced to swim individually for a period of 6 min every day for 15 d. Then, the behavior （locomotor activity, anxiety and memory） and biochemical （lipid peroxidation, nitrite level, reduced glutathione, and catalase） alterations were assessed. Results Forced swimming for 15 d caused impaired locomotor activity, anxiety-like behavior and decreased percentage of memory retention, as compared to na？ve mice （without chronic fatigue treatment）. Biochemical analysis revealed significant increases in lipid peroxidation and nitrite level, while levels of reduced glutathione and catalase activity were both decreased. Chronic treatment with naproxen （14 mg/kg, i.p.）, rofecoxib （5 mg/kg, i.p.）, meloxicam （5 mg/kg, i.p.）, nimesulide （5 mg/kg, i.p.） and valdecoxib （10 mg/kg, i.p.） significantly attenuated these behavioral and biochemical （oxidative damage） alterations in chronic-stressed mice. Conclusion The cyclooxygenase inhibitors could be used in the management of chronic fatigue-like conditions.