Effects of treatment with azathioprine and cyclosporin A on interferon-gamma production by peripheral blood leukocytes of renal allograft recipients

We have studied the concanavalin A (ConA)-induced interferon gamma (IFN-gamma) production of peripheral blood mononuclear cells (PBMC) of renal allograft recipients. Both under immunosuppressive treatment with azathioprine and with cyclosporin A (CsA) the PBMC of these patients proved deficient for IFN-gamma production when compared to those of healthy controls. After conversion from conventional azathioprine to CsA medication the ConA-induced IFN-gamma production increased.     

高效液相色谱法测定人全血中环孢素A浓度

目的:建立正相高效液相色谱法(NP-HPLC)测定肾移植患者全血中环孢素A(CsA)的浓度.方法:患者全血经乙醚单步萃取后,以正己烷-无水乙醇-乙腈(88∶10∶2)为流动相,采用CN色谱柱分析,柱温为50℃,于210 nm波长处检测.结果:本方法在50～1 000μg·L-1范围内具有良好线性关系,平均回收率为101.06%,日内和日间RSD均小于5.2%.结论:本法简便、准确,适用于肾移植患者术后CsA的常规治疗药物监测.     

人全血中环孢素A浓度的HPLC测定及药物动力学研究

建立了测定人全血中环孢素A的HPLC法.采用液-液萃取法处理血样,以环孢素D为内标,色谱柱Diamonsil Ci8柱,流动相为乙腈-甲醇-水(73:5:22),流速1.5ml/min,柱温60℃,检测波长210nm.环孢素A在50～1200ng/ml的浓度范围内线性关系良好,方法回收率为93.6%～108.0%,批内、批间RSD均小于10%.5名健康志愿者单剂量口服环孢素A 500mg,Cmax Tmax、T1/2β和AUC0～24分别为(2090.40±150.52)ng/ml、(1.30±0.27)h、(6.84±0.61)h和(9977.19±1260.63)h·ng·ml-1.     

应用高效液相色谱法中的高速扫描技术测定全血中环孢素A浓度

目的建立一种运用高速扫描技术测定血中环孢素A浓度的HPLC法.方法取全血2.0 ml,加盐酸溶液2 ml后,以3 ml乙醚两次提取;分离乙醚液后用氢氧化钠液洗涤后蒸干;以乙腈/水溶液60 μl重组,再用正己烷洗涤后进样20 μl.色谱条件Elite 220 mm×4.6 mm C18柱,柱温65℃,以甲醇-水(88∶12)为流动相,流速1.0 ml·min-1,扫描波长205～250 nm(间隔5 nm),积分波长210 nm.结果CsA浓度在50～1000 ng·     

测定全血中环孢素A样品处理方法的优化

目的：探讨提高全血中环孢素Ａ（ＣｓＡ）萃取率的可行性，方法：采用正交设计法优化全血中的ＣｓＡ的萃取条件，并加以考察验证，结果：影响萃取率的主要因素为：盐酸，甲醇，氢氧化钠量及乙醚萃取次数，改变萃取条件可使全血中ＣｓＡ萃取率由７８％提高至９０．１４％，日内日间变异均〈９．０％，线性范围为２５～１６００ｎｇ／ｍｌ，最低检测浓度为１０ｎｇ／ｍｌ。结论：本法能提高全血中ＣｓＡ的萃取率，可广泛用于临床血药浓     

环孢素A植入微球的制备

目的制备长效环孢素A微球。方法以聚乳酸为基质,采用乳化-挥发法制备,正交设计优化制备工艺。结果优选制备工艺的重复性良好,环孢素A微球平均粒径为(84.6±3.4)μm,跨距为(0.68±0.13)μm,载药量为20.1%±1.2%,包封率为92.6%±2.7%。结论该处方和制备工艺合理可行。     

环孢素A气雾吸入对哮喘大鼠气道炎症的影响

目的研究环孢素A(CsA)气雾吸入对抗原诱导的大鼠过敏性气道炎症的作用。方法用卵白蛋白(OA)致敏大鼠,2周后气雾吸入CsA(5，10，20 g·L^-1),每天1次，连续7 d。大鼠致敏后d 20和d 21用OA(10 g·L^-1，每天1次)攻击，观察第2次OA攻击24 h后支气管肺泡灌洗液及外周血中嗜酸性粒细胞的数量和支气管肺组织病理学改变情况，测定支气管肺泡灌洗液中TNF-α含量。结果CsA气雾吸入能明显降低支气管肺泡灌洗液及外周血中嗜酸性粒细胞的数量，减轻肺组织中炎症细胞特别是嗜酸性粒细胞的浸润,减轻组织水肿及上皮损伤等气道炎症状况,降低支气管肺泡灌洗液中TNF-α含量。结论CsA气雾吸入对大鼠过敏性气道炎症具有抑制作用，其作用机制与细胞因子TNF-α释放减少有关。     

高效液相色谱法测定羟甲烟胺对环孢菌素A药物动力学参数的影响

目的：建立简单、快速、准确的高效液相色谱法（HPLC）测定环孢菌素A（cyclosporin A,CsA)血药浓度的方法，研究羟甲烟胺对CsA药物动力学参数的影响。方法：采用HPLC法测定18名健康志愿单服CsA及同服羟甲烟胺后CsA的血药浓度，用3p97分析处理数据，比较两组药动学参数的变化。结果：单服CsA及同服羟甲烟胺后CsA的Cmax和AUC0－∞有极显性差异（P＜0．01），而t1/2(β）、Tmax无显性差异（P＞0．05）。结论：本实验建立的HPLC法准确可靠，操作简便，适用于临床药动学研究及常规血药浓度监测。羟甲烟胺可促进CsA的吸收，提高CsA的峰浓度及药时曲线下面积，但不影响CsA的代谢。     

环孢素A和维拉帕米对动物实验性银屑病的影响

目的 研究环孢素Ａ（ＣＳＡ）、维拉帕米（Ｖｅｒ）对银屑病的作用。方法 用鼠尾鳞片表皮模型和小鼠阴道上皮模型，观察ＣＳＡ和Ｖｅｒ对鼠尾表皮鳞片中颗粒层形成数及小鼠阴道上皮细胞有丝分裂指数的影响。结果ＣＳＡ既能增加鼠尾鳞片表皮颗粒层的形成数（Ｐ＜０．０５）；又能减少小鼠阴道上皮细胞有丝分裂指数，抑制细胞增殖（Ｐ＜０．０１）。而Ｖｅｒ只能减少小鼠阴道上皮细胞有丝分裂数，抑制细胞增殖（Ｐ＜０.０５）。结论 ＣＳＡ对两个银屑病模型均有效，能控制表皮过度增生和使表皮角化不全转向正常角化，其治疗作用好，而Ｖｅｒ只对阴道上皮模型有效，表明其对银屑病有治疗作用，也有毒副作用。     

Effects of sulindac on in vitro immunologic function and on prostaglandin production by human peripheral blood mononuclear leukocytes

Prostaglandins (PG) appear to regulate immune-mediated inflammation, but the mechanisms involved remain unclear. Several families of nonsteroidal antiinflammatory drugs (NSAID) have been observed to inhibit PG synthesis. Among these drugs, sulindac sulfide is a potent inhibitor of PG production while its parent pro-drug, sulindac sulfoxide (sulindac), lacks PG synthesis inhibitory activity in cell-free systems. We have studied the effects of sulindac sulfoxide on the blastogenic response of human peripheral blood mononuclear cells (PBMC) stimulated by exposure to alloantigens and mitogens in vitro. Sulindac inhibited proliferation of activated PBMC in a dose-dependent manner but had little effect on the proliferation of unstimulated cells. The inhibition of mitogen-induced blastogenesis correlated with both the uptake of radiolabeled drug and the inhibition of in vitro production of PG (PGE and PGF) by mitogen-activated PBMC. These data indicate a functional relationship between PG synthesis and immune cell activation which may also apply to PBMC activated in vivo by autoimmune disease. Metabolism of sulindac sulfoxide by PBMC in vitro produced too little sulindac sulfide to adequately explain the inhibition of PG production. These data suggest that immunomodulation by sulindac may be due to a direct inhibition of cellular activation. Thus, it is proposed that decreased PG production may be a result rather than the cause of the hypoproliferative response.     

Effect of interferon alpha,interferon beta,and interferon gamma on the in vitro growth of human renal adenocarcinoma cells

Interferon-, interferon-, and interferon- differ in their antiproliferative effects for several cell lines. Interferons were thus assessed for their activity in inhibiting proliferation of three renal cell carcinoma cell lines. The malignant epithelial phenotype of each of these cell lines was confirmed by electron microscopy, histology, karyotype and tumorigenicity. When compared on an anti-viral unit basis, naturally produced interferon- was more effective than natural interferon- for all cell lines and clones. Proliferation of each of the cell lines was inhibited by interferon-. In all cases, removal of interferons from culture media resulted in resumption of the rate of cell growth after a variable delay of 6–10 days. If the antiproliferative effects of interferons predominate in mediating tumor regression, clinical response may depend upon the type of interferon to which the tumor is exposed.     

液相色谱-串联质谱法测定肾移植患者全血中环孢素A及其2种代谢物浓度

目的:建立液相色谱-串联质谱法(LC-MS/MS)测定肾移植患者体内环孢素A及其两个代谢物浓度。方法:全血样品用乙腈沉淀蛋白后直接进样分析,流动相为乙腈-10mmol.L-1甲酸铵溶液-甲酸(85∶15∶0.5,v/v/v),流速0.5mL.min-1,采用电喷雾离子化源,检测方式为正离子多离子反应监测(MRM)。血药浓度数据用Topfit2.0软件,按非室模型计算药动学参数。结果:对所建分析方法进行了较全面的验证,考察了环孢素A及其代谢物的药动学特征。环孢素A的主要药动学参数谷浓度C0为(74.0±25.9)μg.L-1,峰浓度Cmax为(566.2±170.3)μg.L-1,达峰时间tmax为(1.8±0.4)h,消除半衰期t1/2为(4.6±0.9)h,0~12h药时曲线下面积AUC0-12为(2 173.5±580.1)μg.h.L-1。结论:所建方法灵敏,重现性好,选择性强,样品处理简单,可用于环孢素A及其代谢物浓度的监测和药动学研究。     

高效液相色谱法及荧光偏振免疫法测定环孢素A血药浓度的比较

本文建立了特异性测定全血中环孢素Ａ（ＣｓＡ）的高效液相法（ＨＰＬＣ），并与荧光偏振免疫法（ＦＰＩＡ）比较，结果提示两法相关性好，回归方程ＦＰＩＡ值＝７５．６８＋２．６８ＸＨＰＬＣ值（Ｎ＝９４），ｒ＝０．９０５（Ｐ＜０．００１），ＨＰＬＣ法测定的是ＣｓＡ主药和代谢物，ＦＰＩＡ／ＨＰＬＣ值＞３．提示患者体内ＣｓＡ代谢物积蓄，肝功能异常。     

荧光偏振免疫法测定环孢素A血样的保存与稀释条件

目的：考察FPIA测定全血环孢素A浓度时样品的保存条件和保存时间，以及不同稀释方法对测定结果的影响。方法：取临床肾移植术后患者的全血标本分别置室温和冰箱冷藏1周或2周，重复测定标本，考察其稳定性；采用不同溶剂稀释后测定标本与不经稀释测定对比，考察稀释溶剂。结果：室温下或冷藏的标本测定结果均与保存前无统计学差异。但冷藏者变异较小；用空白全血稀释后的样品测定结果与未稀释者无统计学差异，但其他两种稀释方法则与未稀释者有统计学差异。结论：全血标本在室温下至少可稳定保存2周，但冷藏仍是最佳保存方法；高浓度标本应该用空白全血稀释，而不能用注射用水和缓冲液稀释。     

反相高效液相色谱法测定全血中环孢素A的浓度

目的：建立检测患者全血中环孢素A（CsA）浓度的高效液相色谱法。方法：血样经乙醚-石油醚（70：30）萃取后,以C18硅胶键合相为固定相,乙腈-甲醇（80：20）为流动相,70℃柱温下进行色谱分析,紫外检测波长为210nm。结果：CsA全血浓度在50-1600ng／mL范围内呈良好的线性关系,r=0．9998,方法的平均回收率为97．6％,日内和日间精密度RSD均小于6％（n=5）,最低检测限为10ng／mL。结论：本法简便、快速,易于开展,用于肾移植术后患者CsA的血药浓度监测,效果良好。     

Hepatotoxic effect of cyclosporin A in the mitochondrial respiratory chain

Cyclosporin A (CyA), a potent immunosuppressant, was used to determine the hepatotoxic effect in long-term treatments. Male Wistar rats were used in these experiments. They were given CyA chronically at doses used in patients for 120 days, and at doses of 5, 10, 15 and 20 mg kg(-1) day(-1). These doses amount to CyA values in blood of 200 +/- 24, 314 +/- 40, 445 +/- 33 and 598 +/- 53 ng ml(-1), respectively. A significant increase in glutamate dehydrogenase (GLDH) was found in the groups treated with 15 and 20 mg kg(-1) day(-1), which would point to mitochondria as the potential target of the toxic action of CyA. The mitochondrial respiratory chain of rat livers was studied in enzyme complexes I and II. Enzyme complex I was determined by spectrophotometry at 340 nm using NADH oxidase with the respirable substrate 10 mm NADH; enzyme complex II was determined by monitoring succinate dehydrogenase by oxymetry using the respirable substrate 10 mm succinate. The results show the inhibition of NADH oxidase in the groups treated with 10, 15 and 20 mg kg(-1) day(-1), an effect dependent both on time and on CyA concentration. Enzyme complex II showed a decrease in oxygen consumption. These findings were confirmed by histological studies (hematoxylin-eosin technique). CONCLUSIONS: Long-term treatment with CyA at doses of 15 and 20 mg kg(-1) day(-1), amounting to concentrations in blood of 445 +/- 33 and 598 +/- 53 ng ml(-1), causes alterations in the mitochondria, revealed by the increase in serum GLDH and by the functional alteration of enzyme complexes I and II of the mitochondrial respiratory chain.     

不同免疫分析仪在环孢素A血药浓度监测中的差异

目的:考察不同分析方法对环孢素A(CsA)血药浓度监测中的影响.方法:71例肝肾移植术后CsA样品分别用AxSYM和TDx测定.结果:TDx监测结果大于AxSYM监测结果.结论:AxSYM和TDx在环孢素A血药浓度监测中存在差异.     

环孢酶素A的临床应用机制研究进展

免疫抑制剂环孢酶素A广泛应用于临床器官移植后排斥反应和自身免疫性疾病的防治。此外，环孢酶素A在防治心肌肥大、心肌缺血再灌注损伤、脑损伤等疾病都显示了极好的疗效。其临床应用机制主要包括干扰T细胞胞内信号转导通路，抑制致心肌细胞肥大基因的表达，保护线粒体功能、抑制细胞凋亡等。     

环孢素A血药浓度对肾移植术后患者血总胆固醇水平的影响

目的:研究接受肾移植手术后常规服用环孢素A(CsA)的患者其全血谷浓度C0及服药后2h的血药浓度C2对其总胆固醇水平是否有影响。方法:选择我院接受肾移植术病人其中28例,采用荧光偏振免疫分析法(FPIA)测定C0、C2,并按C0分为Ⅰ组:150~250μg.L-1,Ⅱ组:250~400μg.L-1;按C2进行分组,Ⅰ组:<1 000μg.L-1,Ⅱ组:1 000~1 300μg.L-1,Ⅲ组:>1 300μg.L-1。检测其术前及术后的血总胆固醇水平。结果:按C0,Ⅰ组C0(215.2±23.1)μg.L-1,总胆固醇升高(2.1±1.0)mmol.L-1,Ⅱ组C0(302.4±54.1)μg.L-1,总胆固醇升高(2.2±1.1)mmol.L-1,两组手术前后血总胆固醇水平均显著升高(P<0.01)。C0与血总胆固醇水平相关性分析,r=0.200(P>0.05);按C2,Ⅰ组C2(748.5±155.6)μg.L-1,总胆固醇升高(2.3±1.0)mmol.L-1,Ⅱ组C2(1131.6±71.3)μg.L-1,总胆固醇升高(2.0±1.2)mmol.L-1,Ⅲ组C2(1 578.2±376.0)μg.L-1,总胆固醇升高(2.3±0.5)mmol.L-1,3组手术前后血总胆固醇水平均显著升高(P<0.01)。C2与血总胆固醇水平相关性分析,r=0.237(P>0.05)。,结论:术后一个月,各组患者的总胆固醇均有明显增加(P<0.01),但这种改变用C0或C2分析均无明显相关性。     

环孢素对硫唑嘌呤药代动力学影晌

目的　研究合用环孢素（Ｃｙｓ）前后硫唑嘌呤（ＡＺＰ）血药浓度及药物动力学变化。方法　９只家兔灌胃法给硫唑嘌呤（２０ｍｇ·ｋｇ－１）后,高效液相色谱法（ＨＰＬＣ）测定ＡＺＰ和其主要代谢物６ 巯基嘌呤（６ ＭＰ）的血药浓度,１ｗｋ后,灌胃给Ｃｙｓ（１５ｍｇ·ｋｇ－１,ｑｄ）共４ｄ,并于ｄ４灌胃给Ｃｙｓ后ｌｈ再以同样方式给ＡＺＰ,同法测定ＡＺＰ和６ ＭＰ血药浓度,以３Ｐ８７药物动力学程序拟合药物动力学参数。结果　正常家兔合用环孢素后,６ ＭＰ的Ｖｃ升高,由合用前（１５９－１±２４－２）升至合用后（２６９－９±１１８－０）Ｌ·ｋｇ－１（Ｐ＜０－０５）,ＣＬ升高,由（１－６±０－４）升至（３－２±２－０）Ｌ·ｍｉｎ－１·ｋｇ－１（Ｐ＜０－０５）,ＡＵＣ降低,由（７７－９７±２６－０２）降至（４７－０１±２２－７０）ｍｉｎ·μｍｏｌ·Ｌ－１（Ｐ＜０－０５）。其他药动学参数变化差异无显著性。结论　合并用药可引起Ｖｃ、ＣＬ升高,ＡＵＣ降低。