抗结核药物致大鼠肝脏损害的病理研究

目的 研究常用抗结核药物及不同抗结核药物组合对肝脏损害的程度及光、电镜下病理表现,分析抗结核药物对肝细胞超微结构的影响.方法 健康成年SD大鼠152只,体重250～300 g,雌雄各半,随机分为8组.对照组:生理盐水;H(INH)组:异烟肼;R(RFP)组:利福平;Z(PZA)组:吡嗪酰胺;HR组:异烟肼 利福平;HZ组:异烟肼 吡嗪酰胺;RZ组:利福平 吡嗪酰胺;HRZ组:异烟肼 利福平 吡嗪酰胺.大鼠用药的常规剂量:每公斤体重人的常用剂量乘以5.5倍.标本制备:根据各组的不同要求,每天对大鼠进行1次灌胃,10 d后断头取血,分离肝脏,一部分以20%甲醛固定并制备病理切片,另一部分戊二醛固定后做电镜检查.结果 肝组织病理实验组与对照组比较,R组差异显著(P＜0.05);HR组、HZ组、HRZ组、RZ组、Z组差异极显著(P＜0.01);将HRZ组、HR组、HZ组、R组、RZ组比较,HRZ组差异显著(P＜0.05);HR组、Z组、RZ组、HZ组组间差异无显著性(P＞0.05).结论 异烟肼、利福平、吡嗪酰胺均可引起大鼠肝脏的损害,多种药物联用比单药应用损害更明显;随着药物应用种类的增多,肝脏损害逐渐加重;Z组可能比其他单用药组肝脏毒性更大.     

槲皮素对异烟肼肝脏损伤大鼠的保护作用研究

目的：研究槲皮素对异烟肼（INH）致大鼠肝脏损伤的保护作用。方法：给予INH（100 mg/kg）用药14d制备肝脏损伤动物模型,分别给予低和高剂量槲皮素（50、100m g/kg）,14d后测定大鼠血清中天冬氨酸氨基转移酶（A ST）和丙氨酸氨基转移酶（ALT）的活性,计算肝脏脏器系数,并作肝脏组织病理学检测。结果：与INH组比较,槲皮素组可降低肝脏脏器系数（P〈0.05）、血清A ST和ALT的活性（P〈0.05或P〈0.01）;并减轻肝细胞变性、坏死及炎症反应程度,缓解肝脏组织的病理改变。结论：槲皮素对INH所致的大鼠肝脏损伤具有保护作用。     

番茄红素对异烟肼和利福平致大鼠肝损伤的保护作用

目的观察番茄红素对异烟肼(INH)和利福平(RFP)合用致大鼠肝损伤的保护作用并探讨其作用机制。方法给予异烟肼和利福平(各75mg.kg-1)用药4wk制备肝损伤动物模型,分别给予低、中、高剂量番茄红素(10、20、30mg.kg-1),4wk后测定大鼠血清中AST和ALT含量、肝脏指数,以及肝组织匀浆中的MDA、GSH的含量和SOD的活性,并作肝组织病理学检测。结果番茄红素能降低肝脏指数(P<0.05)、降低大鼠血清AST和ALT的含量、降低大鼠肝匀浆中的MDA、增加肝匀浆中GSH、升高SOD(P<0.05或P<0.01),减轻肝组织变性、坏死程度,缓解肝组织的病理改变。结论番茄红素对INH和RFP合用所致的大鼠肝损伤具有保护作用,其作用机制可能与番茄红素的抗脂质过氧化作用有关。     

抗结核药物致大鼠肾脏损害电镜研究

目的观察透射电镜下不同抗结核药物对SD大鼠肾脏超微结构的改变。方法健康成年SD大鼠(清洁级)152只,体重250～300g,雌雄各半,随机分为8组。对照组:0.9%氯化钠溶液;H(INH)组:异烟肼;R(RFP)组:利福平;Z(PZA)组:吡嗪酰胺;HR组:异烟肼+利福平组;HZ组:异烟肼+吡嗪酰胺组;RZ组:利福平+吡嗪酰胺组;HRZ组:异烟肼+利福平+吡嗪酰胺组。大鼠用药的常规剂量:每公斤体重人的常用剂量乘以5.5倍。标本制备:根据各组的不同要求,每天对大鼠进行一次灌胃,10d后断头取血,分离肾脏,戊二醛固定后做电镜检查。结果肾小球细胞部分线粒体溶解、髓样变,粗面内质网脱颗粒,滑面内质网池扩张;肾近曲小管上皮细胞核固缩,细胞器减少。异烟肼、利福平、吡嗪酰胺均可引起大鼠肾脏的损害,多种药物联用比单药应用损害更明显(P<0.01)。结论电镜诊断对抗结核药物所致大鼠肾脏损害有重要价值。     

地塞米松的诱导效应对异烟肼大鼠肝毒性的影响

目的观察大鼠孕烷X受体(pregnane X receptor,PXR)激活剂地塞米松(dexamethasone,DEX)对异烟肼(isoniazid,INH)肝毒性的影响,探讨其发生机制,为临床上抗结核药物所致肝损伤的防治策略提供理论依据。方法 SD♂大鼠随机分成4组(n=6):对照组、INH给药组、DEX给药组和INH-DEX合并给药组,分别给予含DEX 20 mg.kg-1的饲料和含1 000 mg.L-1INH的水喂养28 d后,测定肝脏指数(liver index))和药物代谢酶CYP3A活性,并制备肝组织切片观察各组大鼠肝毒性的程度,同时分析血清中酶学指标ALT、AST、ALP和血脂水平。结果使用DEX组的大鼠肝脏指数与对照组相比明显上升(P<0.01),INH合用DEX后肝细胞CYP3A活性与对照组相比增加了近3.0倍(P<0.01),其肝组织损伤程度加剧,肝细胞脂肪变性并伴有大片梗死,血清中ALT、AST、ALP、血脂水平均较单用INH组明显升高。结论大鼠PXR激活剂DEX能增强INH导致的大鼠肝毒性,其机制可能与DEX上调CYP3A的活性有关。     

抗结核药物致肝损害108例诊治体会

多数抗结核药物对肝脏有不同程度的损害,尤以含利福平(RFP),吡嗪酰胺(PZA),异烟肼(INH)方案为甚,这3种药物是目前倡导的短程化疗方案中必不可少的药物.现将我们从2000-2007年临床资料完整,应用抗结核药物所致的肝功能损害108例肺结核患者的临床资料报告如下.     

甘草酸单铵对异烟肼致大鼠肝损伤的保护作用及其机制

目的:考察甘草酸单铵(MAG)对异烟肼(INH)诱导的大鼠肝损伤的保护作用及其对肝脏转运体Na⁺-牛磺胆酸共转运多肽(Na⁺-taurocholate cotransporting polypeptide,NTCP)及多药耐药相关蛋白2(multidrug resistance protein 2,Mrp2)的调节作用。方法:Wistar雄性大鼠45只,随机分为9组,即对照组,异烟肼肝损伤组(INH组),甘草酸单铵治疗组(MAG组),以上每组又各分为干预7,14,21 d组。INH组大鼠灌胃INH 60 mg·kg^-1·d^-1;MAG组大鼠灌胃MAG 45 mg·kg^-1·d^-1 3 h后灌胃INH。干预结束后,处死大鼠,通过血清生化指标、肝脏病理及Ntcp、Mrp2蛋白表达对各组大鼠进行评价对比。结果:血清生化指标与肝脏病理结果显示,INH诱导的大鼠肝损伤具有时间依赖性,而MAG对INH诱导的大鼠肝损伤具有保护作用,且存在时间依赖性;与INH组相比,MAG干预21 d对肝脏转运体Ntcp及Mrp2表达具有明显下调作用(P<0.05)。结论:MAG对INH诱导的肝损伤的保护作用可能与下调肝细胞膜转运体Ntcp与Mrp2的表达有关。     

山茱萸提取物对大鼠肝脏缺血再灌注损伤的保护作用及对SIRT1-p53通路的影响

目的探讨山茱萸提取物(FCE)对大鼠肝脏缺血再灌注损伤(HIRI)的保护作用及可能的分子机制。方法建立大鼠HIRI模型,并灌胃给予FCE预处理,使用试剂盒检测各组大鼠血清谷丙转氨酶(ALT)和谷草转氨酶(AST)水平,取肝脏组织做HE染色观察肝脏病理改变,采用Western blot检测肝脏组织中SIRT1、Ac-p53、Bax、Bcl-2和Cleaved-caspase-3等蛋白表达变化。结果与假手术组相比,模型组血清ALT和AST水平显著增加;病理切片显示肝脏组织表现出炎性细胞浸润、肝细胞坏死等现象;SIRT1表达水平减少,Ac-p53表达水平增高,导致Bax和Cleaved-caspase-3表达增加,Bcl-2表达减少(P<0.05)。与模型组相比,FCE组大鼠血清ALT和AST水平降低,肝组织炎性反应、肝细胞坏死等病理改变有明显改善,SIRT1的表达水平增加,Ac-p53的表达减少,引起Bax和Cleaved-caspase-3的表达减少,Bcl-2的表达增加。结论 FCE可能通过调控SIRT1-p53信号通路对大鼠HIRI起到保护作用。     

大蒜油对利福平和异烟肼联合所致大鼠肝损伤的保护作用

目的探讨大蒜油对利福平(rifampicin,RIF)和异烟肼(isoniazid,INH)所致大鼠肝损伤的保护作用。方法将60只雄性Wistar大鼠随机分为5组:大蒜油低、中、高剂量处理组、RIF+INH模型组和正常对照组,每组12只。大蒜油低、中、高处理组分别给予相应浓度的大蒜油20、40和80 mg/kg,正常对照组给予等体积的玉米油;2 h后RIF+INH模型组和大蒜油各处理组均给予RIF(75 mg/kg)+INH(75 mg/kg),正常对照组给予等体积的5%淀粉溶液,所有处理每天1次,连续灌胃28 d。观察大鼠体重变化,28 d后测定大鼠血清中丙氨酸转氨酶(alanine transaminases,ALT)、天冬氨酸转氨酶(aspartate transaminases,AST)、总胆红素(total bilirubin,TB)含量、肝重及肝脏系数,同时对肝脏进行病理组织学检查。结果与正常对照组比较,RIF+INH模型组体重无统计学意义,肝重和肝脏系数明显增大(P<0.01),血清中ALT、AST及TB的含量明显升高(P<0.01);与RIF+INH模型组比较,大蒜油低、中和高剂量处理组大鼠体重差异无统计学意义,肝重分别减小3.79%、3.84%和7.73%(P<0.05),脏系数分别减小0.32%、3.25%和3.57%(P<0.05),血清中ALT、AST及TB的含量明显降低(P<0.01或P<0.05)。肝脏病理结果显示,模型组有不同程度细胞浊肿、气球样变,大蒜油处理组没有明显的病理结构改变。结论大蒜油可对利福平和异烟肼联合诱导的肝损伤有保护作用。     

抗结核药在结核病合并乙肝病毒感染中的合理应用

结核病合并乙肝病毒感染者并非少见，如何选用抗结核药，减少或避免肝毒性作用成为最棘手的问题。临床医生应熟悉和掌握各类抗结核药性能、剂量，给药途径及其对肝脏的毒性作用，以制定出一个有效的、快速的最佳治疗方案，并应定期复查肝功能及血清标志物，给以相应保肝治疗。本文就以下几个问题进行阐述。1抗结核药的肝毒性作用抗结核药中除链霉素（SM）、乙胺丁醇（EMB）、卡那霉素（KM）PF均有肝毒性，由异烟胁（INH）、毗喀酬胺（PZA）和利福平（RFP）类引起的肝脏损害发生率分别为10％～20％、2％～3％及l％，而联合用药后毒性…     

常见抗痨药物的肝损害分析

随着临床用药种类不断增多,药物性肝炎的发生率增加。目的：对药物性肝炎患者资料进行分析,提高对该病的认识,及早做出正确诊断和治疗,分析异烟肼（INH）、利福平（RFP）、丙硫异烟胺（Th1321）、乙硫异烟胺（Th1314）、对氨水杨酸钠（PAS）以及吡嗪酰胺（PZA）和氨硫脲（TB）等药物对于不同年龄段、不同异常指标的反映。方法：对我院2011年1月～2012年12月收治典型的96例确诊为药物性肝炎的病历进行分析总结。根据患者以往的药物史、临床试验、血常规、病史以及肝功能测试、B超等综合结果进行比较,最终做出判断。结果：使用含异烟肼（INH）、利福平（RFP）、丙硫异烟胺（Th1321）、乙硫异烟胺（Th1314）、对氨水杨酸钠（PAS）以及吡嗪酰胺（PZA）和氨硫脲（TB）等抗痨药物的化疗方案时,药物性肝炎的发生率较高。结论：抗结核药物对结核病患者而言是必须采用的,但是药物引起的对肝脏的影响也是不能忽视的。对肝脏的影响大小由于个体差异引起的毒性代谢产物积累量也是不同的。但是抗结核药物引起的肝损伤问题要早期发现,及时处理,绝大部分患者是可以恢复的。     

利福平致药物热1例报告

目的 认识利福平导致的药物热的临床特征,提高对该不良反应的认识.方法 回顾分析1例服用利福平导致药物热病人的临床资料及药物治疗情况.结果 1例57岁女性病人因结核性胸腔积液给予异烟肼0.3 g,1 d 1次,口服,利福平0.45 g,1 d 1次,口服,乙胺丁醇0.75 g,1 d 1次,口服,吡嗪酰胺0.5 g,1 d 3次,口服,四联抗结核方案.用药15 d后,病人无明显诱因下出现畏寒发热,体温39~40.3 ℃,伴腹泻4次/天,恶心、呕吐、头晕.停用利福平、乙胺丁醇、吡嗪酰胺1 d后,体温恢复至正常,体温37.4 ℃.再次加用利福平、乙胺丁醇、吡嗪酰胺,病人再次发热,体温39 ℃.考虑抗结核药导致的药物热,遂停用所有抗感染药物,病人体温平.第2次停药5 d后,根据药物热发生率从小到大依次加用阿米卡星、乙胺丁醇、异烟肼、吡嗪酰胺、左氧氟沙星,在加药的过程中和加药后,病人体温维持在36.8~37.4 ℃.结论 病人的药物热很可能与利福平的使用相关.     

The inhibition of hepatic bile acids transporters Ntcp and Bsep is involved in the pathogenesis of isoniazid/rifampicin-induced hepatotoxicity

Abstract

Co-treatment of isoniazid (INH) and rifampicin (RFP) is well known for clinically apparent liver injury. However, the mechanism of INH/RFP-induced liver injury is controversial. Emerging evidence shows links between inhibition of bile acids transporters and drug-induced liver injury (DILI). The present study investigates whether sodium taurocholate cotransporting polypeptide (NTCP/Ntcp; SLC10A1) and bile salt export pump (BSEP/Bsep; ABCB11) are involved in the anti-tuberculosis medicines induced liver injury. ICR female mice were intragastrically treated with INH (50 or 100?mg/kg), RFP (100 or 200?mg/kg), or the combination of INH/RFP (50?+?100?mg/kg or 100?+?200?mg/kg) for 14 consecutive days. Liver histopathological examination, serum biochemical and liver malondialdehyde tests were evaluated. Apparent histopathological alterations and hepatic oxidative stress showed in INH (100?mg/kg), RFP (200?mg/kg) and their combination group. The hepatoxic effect was also indicated by increased serum biomarkers, such as aspartate transaminase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), direct bilirubin (DBil), total bilirubin (TBil) and total bile acids (TBA). Both doses of INH/RFP administration significantly down-regulated the expression of Ntcp and Bsep in liver. Furthermore, the combination of INH and RFP displayed stronger effect on the expression of Ntcp compared with the corresponding dose of INH or RFP alone. In conclusion, down-regulated expression of hepatic Ntcp and Bsep might play an important role in the development of INH and RFP induced liver injury.     

Serum biomarkers of isoniazid-induced liver injury: Aminotransferases are insufficient,and OPN,L-FABP and HMGB1 can be promising novel biomarkers

Isoniazid (INH)-induced liver injury is a great challenge for tuberculosis treatment. Existing biomarkers cannot accurately determine the occurrence of this injury in the early stage. Therefore, developing early specific sensitive biomarkers of INH-induced liver injury is urgent. A rat model of liver injury was established with gastric infusion of INH or INH plus rifampicin (RFP). We examined seven potential novel serum biomarkers, namely, glutamate dehydrogenase (GLDH), liver-fatty acid-binding protein (L-FABP), high-mobility group box-1 (HMGB1), macrophage colony-stimulating factor receptor (MCSF1R), osteopontin (OPN), total cytokeratin 18 (K18), and caspase-cleaved cytokeratin-18 (ccK18), to evaluate their sensitivity and specificity on INH-induced liver injury. With the increase of drug dosage, combining with RFP and prolonging duration of administration, the liver injury was aggravated, showing as decreased weight of the rats, upgraded liver index and oxidative stress level, and histopathological changes of liver becoming marked. But the activity of serum aminotransferases decreased significantly. The area under the curve (AUC) of receiver-operating characteristic (ROC) curve of OPN, L-FABP, HMGB1, MCSF1R, and GLDH was 0.88, 0.87, 0.85, 0.71, and 0.70 (≥0.7), respectively, and 95% confidence interval of them did not include 0.5, with statistical significance, indicating their potential abilities to become biomarkers of INH-induced liver injury. In conclusion, we found traditional biomarkers ALT and AST were insufficient to discover the INH-induced liver injury accurately and OPN, L-FABP, and HMGB1 can be promising novel biomarkers.     

阿魏酸钠对异烟肼和利福平肝损害小鼠的保护作用

目的：观察阿魏酸钠对异烟肼(INH)和利福平(RFP)肝毒性的保护作用。方法：分别测定血清谷丙转氨酶(ALT)的活性，肝匀浆中谷胱甘肽(GSH)及脂质过氧化物丙二醛(MDA)的含量，肝微粒体中细胞色素P450及其亚型2E1的活性。结果：阿魏酸钠可对抗INH和RFP合用引起的肝指数、血清ALT水平、肝匀浆中的MDA含量，以及细胞色素P450与亚型P450 2E1活性的升高，增加肝匀浆中GSH含量。病理学检查，阿魏酸钠明显减轻肝细胞的变性和坏死。结论：阿魏酸钠时INH和RFP肝毒性的保护作用与保护肝细胞膜、抑制脂质过氧化反应、清除自由基、降低RFP诱导的细胞色素P450酶系统有关。     

Pharmacokinetic and pharmacodynamic behaviour of antitubercular drugs encapsulated in alginate nanoparticles at two doses

This study was designed to evaluate the pharmacokinetics and tissue distribution of free and alginate-encapsulated antitubercular drugs in mice at different doses. Alginate nanoparticles encapsulating isoniazid (INH), rifampicin (RIF), pyrazinamide (PZA) and ethambutol (EMB) were prepared by controlled cation-induced gelification of alginate. The formulation was orally administered to mice at two dose levels (D1 and D2). A comparison was made in mice receiving free drugs at equivalent doses. Drugs were analysed by high performance liquid chromatography (HPLC). The average size of alginate nanoparticles was found to be 235.5+/-0.0 nm with a polydispersity index of 0.44; drug encapsulation was 70-90% for INH and PZA, 80-90% for RIF and 88-95% for EMB. In the free drug groups, plasma levels of RIF and INH were higher and PZA and EMB levels were lower in the D1 group (per body surface area of mice) compared with the D2 group (recommended human dose). The plasma drug levels of all drugs were higher in the D1 encapsulated group compared with D2, resulting in higher values of area under the plasma drug concentration-time curve (AUC(0-infinity)). The relative bioavailabilities of all drugs encapsulated in alginate nanoparticles were significantly higher compared with free drugs. Drug levels were maintained at or above the minimum inhibitory concentration (MIC(90)) until Day 15 in organs after administration of encapsulated drugs, whilst free drugs stayed at or above the MIC(90) up to Day 1 only irrespective of dose. The levels of drugs in various organs remained above the MIC at both doses for equal periods, demonstrating their equiefficiency. Alginate nanoparticles hold great potential in reducing dosing frequency of antitubercular drugs.     

乌药醚内酯对肝损伤模型大鼠的保护作用

目的 考察不同剂量乌药醚内酯对大鼠肝损伤模型的保护作用。 方法 60只健康雄性SD大鼠随机均分为空白对照组、模型组、乌药醚内酯低、中、高剂量组及阳性药物对照组,除空白对照组外,其余5组采用四氯化碳腹腔注射建立肝损伤大鼠模型,造模期间各组给予相应的药物或等体积的水,造模6w后处死大鼠取血清,测ALT、AST、MDA含量及SOD、GSH-Px的活性,取肝脏组织染色切片观察病理变化。 结果 与模型组对比,乌药醚内酯各剂量组能降低模型大鼠血清ALT、AST且呈剂量依赖性;高剂量组还能显著升高模型大鼠血清SOD、GSH-Px活性,降低MDA含量,显著降低肝脏组织的病理变化评分。 结论 乌药醚内酯可能通过提高抗氧化能力来降低肝损伤模型大鼠的ALT和AST水平及肝脏病理变化。