老年类风湿关节炎患者应用MTX或LEF联合糖皮质激素治疗的效果观察

目的:探讨甲氨蝶呤(MTX)或来氟米特(LEF)联合泼尼松(PDN)对老年类风湿关节炎的疗效。方法:选取112例老年RA患者,按随机数表法将患者分为LEF组(n=28)、MTX组(n=28)、LET+PDN组(n=28)、MTX+PDN组(n=28),治疗3个月后统计四组关节肿胀数、关节压痛数、DAS28评分、VAS评分、RF、晨僵时间,并记录不良反应事件。结果:MTX组和LEF组治疗后各项指标均低于治疗前(P0.05);MTX+PDN组与LEF+PDN组治疗后各项指标均明显低于治疗前(P0.001);MTX+PDN组治疗效果明显优于MTX组(P0.001);LEF+PDN组治疗效果明显优于LEF组(P0.001);MTX+PDN组治疗总有效率为53.57%,高于LEF+PDN组的42.86%,但差异无统计学意义(x~2=2.426,P=0.119)。结论:四组对RA均有治疗效果,联合使用效果更佳,且服药后无严重不良反应,安全性较高。     

不同剂量甲氨蝶呤联合不同剂量叶酸治疗活动期类风湿关节炎的临床研究

摘要 目的：探讨不同剂量甲氨蝶呤（MTX）联合不同剂量的叶酸治疗活动期类风湿关节炎（RA）的疗效与安全性。方法：选取100例符合纳入、排除标准的RA患者,按照28关节疾病活动指数（DAS28）分为高疾病活动度组（使用MTX 15 mg 每周1次）50例和低疾病活动度组（使用MTX 10 mg 每周1次）50例。高疾病活动度组按叶酸使用10 mg 每周1次 或5 mg 每周1次随机分为两组。低疾病活动度组按叶酸使用5 mg 每周1次或不使用随机分为两组,对比治疗6个月后的临床疗效和安全性。结果：治疗后,高疾病活动度叶酸5 mg组DAS28评分、视觉模拟评分量表（VAS）评分、血沉（ESR）、超敏C反应蛋白（hs-CRP）以及总有效率均优于叶酸10 mg组（P<0.05）,但是两组健康评定问卷（HAQ）评分、不良反应发生率和MTX浓度比较无明显差异（P>0.05）。低疾病活动度叶酸5 mg 组与无叶酸组患者治疗后hs-CRP、ESR、MTX浓度比较差异有统计学意义（P<0.05）,但是两组总有效率和不良反应发生率比较无明显差异（P>0.05）。结论：RA高疾病活动时,使用MTX 15 mg 每周1次联合叶酸5 mg 每周1次疗效更优,且不良反应发生率及MTX浓度变化不明显。RA低疾病活动时,MTX 10 mg 每周1次,与是否使用叶酸在疗效和安全性上无显著差异,但未使用叶酸患者的MTX浓度更高。     

甲氨蝶呤联合羟基氯喹对类风湿关节炎合并糖尿病患者糖化血红蛋白水平的影响

目的:观察类风湿关节炎(rheumatoid arthritis,RA)合并糖尿病(diabetes mellitus,DM)患者使用甲氨蝶呤(methotrexate,MTX)联合羟基氯喹(hydroxychloroquine,HCQ)治疗前后糖化血红蛋白(glycosylated hemoglobin,HbA1C)水平的变化。方法:通过回顾性分析,选取联合使用MTX+HCQ或单独使用MTX治疗的RA合并DM患者,且在治疗前及开始治疗12个月内分别至少有1次HbA1C值,记录其年龄、性别、诊断、体重指数、使用糖皮质激素情况。计算用药前到用药后12个月内HbA1C最低值的变化。结果:40例使用HCQ+MTX和45例使用MTX患者符合入选标准。两组间年龄、性别、体重指数、用药前HbA1C水平相似,MTX+HCQ组糖皮质激素使用比例(40.00%)比HCQ组更多(26.67%)(P=0.25)。MTX+HCQ组治疗后HbA1C有明显下降(0.42%,P=0.00),且MTX+HCQ组HbA1C降低幅度高于MTX组(0.42%,0.12%,P=0.02)。结论:与单独使用MTX相比,RA合并DM患者联合使用HCQ+MTX可明显降低HbA1C水平。     

雷公藤多甙片与甲氨喋呤联合治疗类风湿关节炎的临床疗效及对炎症因子的影响

目的:研究雷公藤多苷片联合甲氨蝶呤治疗类风湿关节炎(RA)的临床疗效及对血清炎症因子的影响。方法:选取2014年1月-2015年8月我院收治的RA患者60例,按治疗方法的不同分为观察组和对照组各30例,观察组应用雷公藤多苷片联合甲氨蝶呤治疗,对照组单纯应用甲氨蝶呤治疗,对比两组的临床疗效及治疗前后临床症状、红细胞沉降率(ESR)、C反应蛋白(CRP)及类风湿因子(RF)水平。结果:观察组的总有效率为93.33%,明显高于对照组的73.33%(P0.05);治疗后观察组的临床症状(晨僵时间、关节压痛及肿胀数、关节疼痛度及肿胀指数)均较对照组明显改善(P0.05);治疗后两组血清ESR、CRP、RF水平均降低,且观察组明显低于对照组(P0.05);观察组不良反应率为13.33%,低于对照组的10.00%,差异无统计学意义(P0.05)。结论:雷公藤多苷片联合甲氨蝶呤治疗RA能够有效控制炎症反应,改善临床症状,临床疗效显著,且不增加副反应,是一种安全可靠的联合治疗方案,值得推广应用。     

戈利木单抗联合雷公藤多甙对甲氨蝶呤治疗反应不佳活动性类风湿关节炎的效果及安全性分析

目的:分析戈利木单抗联合雷公藤多苷对甲氨蝶呤(MTX)治疗反应不佳活动性类风湿关节炎(RA)的临床效果及安全性。方法:选择MTX治疗反应不佳(MTX治疗超3个月,但应答不足)的活动性RA患者66例,按照随机数字表法分为对照组与试验组各33例,对照组单用雷公藤多苷治疗,试验组加用戈利木单抗治疗,测定两组治疗前后血沉(ESR)、C反应蛋白(CRP)、类风湿因子(RF)水平的变化,记录晨僵时间、关节肿胀数目、关节压痛数目,评定治疗效果,采用视觉模拟评分法(VAS)评定患者关节疼痛程度、患者对疾病总体症状的耐受情况,监测两组不良事件的发生情况。结果:(1)治疗后,两组ESR、CRP、RF均降低,与同组治疗前对比差异有统计学意义(P0.05),试验组ESR、CRP降低幅度高于对照组(P0.05);(2)治疗后,两组晨僵时间减少、关节肿胀及压痛数目均减少,与治疗前对比差异有统计学意义(P0.05),试验组各指标改善幅度均高于对照组(P0.05);(3)治疗后,两组VAS评分均降低,与治疗前对比差异有统计学意义(P0.05),试验组降低幅度高于对照组(P0.05);(4)试验组ACR50、ACR70所占比例均高于对照组(P0.05);(5)试验组、对照组不良反应发生率对比差异无统计学意义(P0.05)。结论:戈利木单抗联合雷公藤多苷治疗MTX反应不佳活动性RA疗效肯定,可下调患者ESR、CRP水平,缩短晨僵时间,显著改善患者症状,减轻关节疼痛程度,且安全性高。     

超声引导下局部注射甲氨蝶呤治疗异位妊娠的临床应用

目的:探讨超声引导下局部注射甲氨蝶呤(MTX)治疗异位妊娠的疗效。方法:对30例异位妊娠患者在超声引导下行MTX病灶局部注射,术后给予米非司酮口服(25mg,2次/天,共3天)。治疗前及治疗后每周一次抽血查β-hCG直至其降至正常范围内。结果:30例患者中,29例(96.7%)通过治疗后β-hCG降至正常,1例(3.3%)治疗失败后行外科手术。结论:超声引导下MTX局部注射治疗异位妊娠技术操作简单,疗效肯定,对适应范围内的病人是值得考虑的治疗方法。     

阿达木单抗联合甲氨蝶呤治疗类风湿关节炎的疗效研究

目的:评估阿达木单抗(ADA)联合甲氨蝶呤(MTX)治疗类风湿关节炎(RA)的疗效与安全性。方法:选取在我院风湿科治疗的203例RA患者,将患者随机分为MTX单药治疗组(MTX组,n=101)与ADA联合MTX(ADA-MTX组,n=102)治疗组。记录患者的临床资料,检测患者实验室指标,比较患者的临床特征与不良反应发生率。结果:ADA-MTX组患者肿胀减轻、VAS评分降低、压痛减轻的患者比率与总有效率显著高于MTX组,差异有统计学意义(P0.05)。与MTX组比较,ADA-MTX组第12周、24周临床特征(晨僵时间、关节压痛数、关节肿胀数与DAS28评分)显著下降,差异有统计学意义(P0.05)。与MTX组第24周值比较,ADA-MTX组患者的生活质量评分显著提高,差异有统计学意义(P0.05)。ADA-MTX组与MTX组不良反应发生率比较无统计学差异(12.7%vs 14.9%,P=0.651)。结论:阿达木单抗联合甲氨蝶呤可明显改善类风湿关节炎患者实验室指标与临床体征、症状,从而减少患者功能障碍,提高患者生活质量,且具有良好的耐受性与安全性。     

甲氨蝶呤联合艾得辛可降低RA患者血清中IL-37及PD-1的水平

分析甲氨蝶呤联合艾得辛治疗对类风湿关节炎患者血清IL-37及PD-1水平的影响。选择2015年4月至2017年3月本院接诊的100例类风湿关节炎患者作为研究对象,按随机数字表法分组,每组50例,观察组患者采用甲氨蝶呤与艾得辛联合治疗,对照组患者单独采用甲氨蝶呤治疗,比较两组患者血清中细胞因子IL-6、IL-18和IL-18BP的表达,探讨IL-37、PD-1的临床意义。观察组患者的血清IL-18(63.23±7.89) pg/mL、IL-6 (3.25±0.98) pg/mL、IL-18BP (210.56±24.51) pg/mL表达水平显著低于对照组患者的血清IL-18 (140.67±18.65) pg/m L、IL-6 (8.27±1.23) pg/mL、IL-18BP (308.89±30.67) pg/mL表达水平;观察组患者的血清IL-37 (23.65±4.28) pg/m L及PD-1 (19.58±3.78) ng/mL表达水平显著低于对照组患者的血清IL-37 (58.98±5.29) pg/mL及PD-1 (36.72±4.62) ng/mL表达水平,差异具有统计学意义(p0.05),相关性分析发现,血清IL-37与细胞因子IL-18、IL-6和IL-18BP均呈正相关;但是PD-1仅与细胞因子IL-6呈正相关,按照RA评分标准,患者血清中IL-37与诊断评分呈正相关,PD-1与诊断评分呈正相关。甲氨蝶呤联合艾得辛治疗类风湿关节炎具有较为明确的疗效,能够短期内控制RA患者疾病的发展,临床的表现稳定,有效降低患者的血清IL-18、IL-6、IL-18BP以及IL-37、PD-1的表达水平,对患者的积极意义较大,可在临床的治疗中推广应用。     

雷公藤多苷联合甲氨蝶呤治疗类风湿关节炎活动期患者的疗效及对血清CD62p、CD41的影响

目的:探讨雷公藤多苷联合甲氨蝶呤治疗类风湿关节炎(RA)活动期患者的疗效及对血清释放黏附分子CD62p、CD41的影响。方法:选择2015年6月至2017年6月我院接诊的90例类RA活动期患者作为研究对象,通过随机数表法分为观察组(n=45)和对照组(n=45),对照组口服甲氨蝶呤治疗,观察组联合雷公藤多苷片治疗,均连续用药12周。比较两组临床疗效、治疗前后临床症状评分、实验室指标[红细胞沉降率(ESR)、C反应蛋白(CRP)、血小板计数(PLT)]及血清CD62p、CD41的变化,并比较治疗期间不良反应。结果:观察组临床疗效总有效率为91.11%(41/45),明显高于对照组的66.67%(30/45)(P0.05);治疗后,两组临床症状评分及实验室指标较治疗前均显著降低(P0.05),观察组晨僵时间、关节疼痛、关节肿胀、关节压痛评分及ESR、CRP、PLT均明显比对照组,比较均具有显著差异(P0.05);治疗后,两组血清CD62p、CD41较治疗前均显著降低(P0.05),观察组血清CD62p、CD41均明显低于对照组[(16.58±2.10)%vs(25.46±2.58)%,(67.83±11.03)%vs(76.40±13.45)%](P0.05);两组治疗期间肝肾功能均未发生异常,两组头痛、皮疹、胃肠道反应、感染、脱发发生率均无显著差异(P0.05)。结论:在RA活动期患者中使用雷公藤多苷联合甲氨蝶呤效果显著,可有效改善临床症状及实验室指标,其内在机制可能和降低血清CD62p、CD41的表达相关,且联合用药安全性高,值得应用推广。     

甲氨蝶呤联合来氟米特对类风湿关节炎患者炎症因子和免疫球蛋白的影响

目的:探讨甲氨蝶呤联合来氟米特对类风湿关节炎(RA)患者炎症因子和免疫球蛋白的影响。方法:选取于2016年6月-2017年10月期间我院收治的92例RA患者,根据乱数表法将患者随机分为对照组(n=46)与研究组(n=46)。对照组给予口服甲氨蝶呤片,研究组则在对照组的基础上联合来氟米特片治疗。两组均治疗3个月。比较两组患者临床疗效、临床症状改善情况,检测两组患者治疗前后炎症因子、免疫球蛋白水平,观察两组患者不良反应发生情况。结果:研究组患者治疗后的临床总有效率为95.65%(44/46),高于对照组患者的78.26%(36/46)(P0.05)。两组患者治疗后晨僵时间、压痛关节数、肿胀关节数均较治疗前降低,且研究组低于对照组(P0.05)。两组患者治疗后血细胞沉降率(ESR)、C反应蛋白(CRP)、白介素-8(IL-8)及肿瘤坏死因子(TNF-α)均较治疗前降低,且研究组低于对照组(P0.05)。两组患者治疗后免疫球蛋白G(Ig G)、免疫球蛋白A(Ig A)、免疫球蛋白M(Ig M)均较治疗前降低,且研究组低于对照组(P0.05)。两组患者不良反应发生率比较无差异(P0.05)。结论:甲氨蝶呤联合来氟米特治疗RA患者效果优于单用甲氨蝶呤治疗,可改善患者临床症状,同时降低Ig G、Ig A、Ig M以及炎症因子水平,无严重不良反应发生。     

Methotrexate induced differentiation in colon cancer cells is primarily due to purine deprivation

The folate antagonist methotrexate (MTX) inhibits synthesis of tetrahydrofolate (THF), pyrimidines and purines, and induces differentiation in several cell types. At 1 microM, MTX reduced proliferation and induced differentiation in HT29 colon cancer cells; the latter effect was augmented (P < 0.001) by thymidine (100 microM) but was reversed (P < 0.001) by the purines, hypoxanthine (Hx; 100 microM) and adenosine (100 microM). In contrast 5-fluoro-uracil (5-FU), a specific thymidylate synthase (TS) inhibitor, had no effect on differentiation, suggesting that MTX-induced differentiation is not due to a reduction in thymidine but to the inhibition of purine biosynthesis. Inhibition of cyclic AMP (cAMP) by RpcAMP (25 microM) further enhanced (P < 0.001) MTX induced differentiation, whereas the cAMP activator forskolin (10 microM) reversed (P < 0.001) MTX induced differentiation. These observations implicate a central role of adenosine and cAMP in MTX induced differentiation. By combining Western blot analysis with liquid chromatography-mass spectrometry (LC-MS)and HPLC analyses we also reveal both the expression and activity of key enzymes (i.e. methionine synthase (MS), s-adenosylhomocysteinase, cystathionine beta-synthase and ornithine decarboxylase) regulating methyl cycle, transsulfuration and polyamine pathways in HT29 colon cancer cells. At 1 microM, MTX induced differentiation was associated with a marked reduction in the intracellular concentrations of adenosine and, consequently, S-adenosylmethionine (SAM), S-adenosylhomocysteine, polyamines and glutathione (GSH). Importantly, the marked reduction in methionine that accompanied MS inhibition following MTX treatment was non-limiting with respect to SAM synthesis. Collectively, these findings indicate that the effects of MTX on cellular differentiation and single carbon metabolism are primarily due to the intracellular depletion of purines.     

Methotrexate: pentose cycle and oxidative stress

The effect of methotrexate (MTX) and leucovorin (LCV) on pentose cycle enzymes and the activity of enzymes involved in enzyme defence mechanisms against ROS in HeLa cells, were studied. The effect of MTX was also investigated on the cellular levels of glutathione. MTX inhibited the activity of glucose-6-phosphate and 6-phosphogluconate dehydrogenases. The activities of glutathione reductase and γ-glutamylcysteine synthetase were also inhibited by the drug. No effect was observed on the activities of catalase, superoxide dismutase or transketolase. LCV had no effect on any of the enzymes studied. MTX decreased the cellular levels of glutathione (70 per cent), while the presence of LCV and glutamine did not interfere with the effect of MTX. The net results appear to show that the biological situation resulting from treatment with MTX leads to a reduction of effectiveness of the antioxidant enzyme defence system. Copyright © 1998 John Wiley & Sons, Ltd.     

Proteomic Analysis for the Identification of Proteins Related to Methotrexate Resistance

Methotrexate(MTX) is one of the most important and frequently used drugs in cancer therapy, but the efficacy of this drug is often compromised by the development of resistance in cancer cells. To seek and identify differentially expressed proteins related to MTX resistance and provide clues for the mechanism of MTX resistance, proteins from cell line MTX300 (resistant to 300 μmol/L MTX) and its control cell line 3T3R500 were separated by two-dimensional electrophoresis (2-DE). The colloidal Coomassie brilliant blue-stained 2-DE gels were subjected to image analysis, which revealed several spots with high levels of differential expression between MTX300 and 3T3R500. The protein spot with highest differential expression was submitted for tryptic peptide mass fingerprinting(PMF) for identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). MS analysis and database searches revealed it to be dihydrofolate reductase (DHFR), which was subsequently confirmed by Western blot. The result suggested that DHFR might play an important role in the MTX resistance.     

Methotrexate: studies on cellular metabolism. IV. Effect on the mitochondrial oxidation of cytosolic-reducing equivalents in HeLa cells

The effect of methotrexate (MTX) on the mitochondrial oxidation of cytosolic-reducing equivalents in HeLa cells was studied. MTX inhibited (100 per cent) malate dehydrogenase activity, but no effect was observed on that of GOT. MTX (0.5 mM) inhibited (100 per cent) the activity of reconstituted enzymatic system MDH-GOT, probably as a consequence of inhibition of malate dehydrogenase activity. MTX decreased pyruvate production (54 per cent), demonstrating its inhibitory action on the malate-aspartate shuttle. Blockage of the malate-aspartate shuttle by MTX accounts for the decrease in cellular energetic gain. The results obtained are consistent with the view that in HeLa cells, as well as in other tumour cells, the transport of reducing equivalents from cytoplasmic NADH into the respiratory chain of mitochondria is via the malate-aspartate shuttle.     

运用蛋白质组学技术研究与氨甲蝶呤耐药性相关的蛋白质

氨甲蝶呤（MTX）是一种重要的常用化疗药物,然而由于肿瘤细胞对其耐药性的增强而经常导致其疗效大大降低。为了寻找并鉴定与MTX耐药性相关的蛋白质从而为进一步闸明MTX的耐药机制提供线索,培养来源于小鼠NIH3T3的小鼠胚胎成纤维细胞系3T3R500与其耐300μmol／L MTX的细胞系MTX300,提取上述两种细胞系的总蛋白质,双向凝胶电泳分离蛋白质组成分,扫描并通过软件分析考马斯亮蓝染色的2-DE凝胶,选取表达差异最显著的点,胶内酶切后MALDI-TOF-MS进行肽指纹图谱（PMF）鉴定。图像分析显示,实验组和对照组的蛋白质组图谱之间,一些蛋白质点的表达有明显的变化。通过MALDI-TOF-MS和数据库查询,成功鉴定了耐药后表达变化最显著的蛋白质点为二氢叶酸还原酶（DHFR）,并通过Western blot验证了该结果,提示DHFR在MTX耐药机制中发挥重要作用。     

Methotrexate conjugate with branched polypeptide influences Leishmania donovani infection in vitro and in experimental animals

Methotrexate (MTX) has been coupled to various structurally related, polycationic (poly[Lys(DL-Ala(m))] (AK), poly[Lys(Ser(i)-DL-Ala(m))] (SAK), poly[Lys(DL-Ala(m)-Leu(i))] (ALK)), or amphoteric (poly[Lys(Glu(i)-DL-Ala(m))] (EAK)) synthetic branched polypeptides containing poly[L-Lys] backbone by the aid of BOP reagent. The average degree of MTX incorporation was found to be dependent on the charge properties of the polymer. Under the experimental conditions used, the molar substitution ratio achieved was higher for polycations (25%) than for the amphoteric polypeptide (10%). We have studied the effect of polycationic polypeptides on Leishmania donovani infection. Results demonstrated that MTX conjugates in which the drug is covalently attached to carrier have pronounced leishmanicid activity. In this communication we showed that (a) a branched polypeptide-methotrexate conjugate with a polycationic carrier (ALK) increases the effect of MTX against Leishmania donovani infection in mice; (b) the covalent bond between the carrier and methotrexate is essential for both in vivo and in vitro activity; and (c) the number of Leishmania donovani parasites in infected macrophages are markedly reduced in conjugate treated animals. In vitro observation might also indicate that the MTX conjugate exhibits an effect through an uptake by macrophages which is different from that of the free drug.     

自身代谢缓解甲氨蝶呤对小鼠体内成熟卵母细胞副作用的研究

甲氨蝶呤(methotrexate,MTX)作为抗叶酸代谢类药物,在临床上应用广泛.其副作用之一表现为损伤卵母细胞质量.那么,对于有妊娠需求的女性服药者,停药后多久才适合受孕呢?该文结合双光子荧光成像与三维重构技术研究此问题.通过单次腹腔注射生理盐水及5 mg/kg MTX建立了对照组和MTX组小鼠,随后建立了MTX注...     

Methotrexate: studies on cellular metabolism. III.--Effect on the transplasma-membrane redox activity and on ferricyanide-induced proton extrusion by HeLa cells

The effect of methotrexate (MTX) on transplasma-membrane electron transport and ferricyanide-induced proton extrusion by HeLa cells was studied. Both systems were inhibited by MTX. It is suggested that inhibition of electron transport and proton extrusion caused by MTX could be associated with other metabolic alterations such as response to the increase in NADH levels and decrease in intracellular pH.     

甲氨蝶呤对大鼠尿液蛋白质组的影响

甲氨蝶呤是常见的免疫抑制剂,曾被大剂量用于治疗癌症,近年来其被小剂量用于治疗类风湿性关节炎。文中尝试研究甲氨蝶呤对大鼠尿蛋白质组的影响。大鼠口服甲氨蝶呤构造用药模型,再收集大鼠10 h内的尿液,并且采用液相色谱串联质谱(liquid chromatography tandem mass spectrometry,LC-MS/MS)分析大鼠的尿蛋白。总共鉴定到31个差异蛋白,其中7个蛋白与甲氨蝶呤药物作用和类风湿性关节炎症状有关。部分大鼠的生物学过程反映了甲氨蝶呤对机体谷胱甘肽代谢以及对JAK/STAT信号通路的影响。结果显示,尿蛋白具有反映甲氨蝶呤对大鼠机体影响的能力。对单个大鼠个体差异蛋白的分析体现出不同个体对该药物的反应有较大差异。