There are some shortcomings of animal experiments applied in chemical toxicity testing, such as long period, large cost, species differences and dose differences, which limit the use of animal experiments' results in predicting human toxicity.Accordingly, we established a toxicity testing alternative screening system in line with the toxicity endpoints which required in chemical safety evaluation and risk assessment (genotoxicity, carcinogenicity, reproductive toxicity, acute toxicity and general toxicity) based on "3R" principles (replacement, reduction, refinement) for animal experiments.This system covers most of the endpoints of toxicity assessment, and molecular biology technology was also applied to integrate the toxicity test, as well as some operation was optimized in order to shorten the experimental period, reduce experimental costs, improve animal welfare.Furthermore, the results from the screening system have higher clinical relevance because it is based on the toxicity mechanisms.

目的 采用1H NMR的代谢组学技术揭示大黄素的肾毒性机制,寻找肾脏损害的早期生物标志物.方法 雄性SD大鼠20只,随机分为溶剂对照,大黄素170、500、1 500 mg/(kg·d)3个剂量组,连续给药16d,给药结束后收集24 h尿液,血浆及肾组织,测定1H NMR谱,并进行血浆生化指标测定和肝脏组织病理学检查.结果 1 500 mg/(kg·d)大黄素服用16d可引起大鼠血肌酐下降,大黄素可导致肾细胞胞浆中出现明显的空泡化改变.代谢成分的改变主要表现为血液中乳酸、糖、氨基酸和脂肪酸成分下降;尿液中乳酸、糖和氨基酸成分增加;肾脏组织中醋酸盐和肌酐/肌酸明显升高,乳酸和胆碱/磷酸卵磷脂水平下降,饱和与不饱和脂肪酸及磷脂的成分比例明显改变.结论 代谢组学分析在识别药物诱导代谢成分改变方面较传统技术更灵敏;脂肪和能量代谢紊乱参与了大黄素的肾毒性,尿液中氨基酸、葡萄糖氧化三甲胺(TMAO)及肌酐可作为大黄素诱导肾组织损害的潜在生物标志物.

Mitochondrial dysfunction and oxidative damage may be the initial events of copper nanoparticle (CuNP)-induced hepato and nephrotoxicity.

药物性肝损伤是导致药物临床实验失败和上市后撤回的主要因素,而导致肝毒性药物进入临床实验的一个重要原因,是临床前动物模型与人类的种属差异.因此,开发新的临床前药物安全评价模型显得至关重要,临床前药物安全性评价模型中以人类肝细胞的体外培养为最可靠、快速、经济的模型.本文综述了在新药研发早期安全性评价中,不同体外模型的优点和不足之处,以期为药物安全性评价模型的进一步发展提供参考.

目的 探讨亚硫酸氢钠穿心莲内酯(ASB)对人肾小管上皮细胞HK-2的细胞毒性作用.方法 HK-2细胞在DMEM/F12培养液培养24 h后,在经ASB作用2h前,分别加入N-乙酰-L-半胱氨酸(NAC)10 mmol· L-1、2-氨基-4(丁基磺酸亚氨)-丁酸(BSO)2 mmol· L-1及30 min前分别加入过氧化氢酶(CAT)1000 U· L-1、环孢素A (CsA)50 μmol·L-1,再经ASB作用24 h,MTT法检测细胞存活率;用酶标仪检测乳酸脱氢酶(LDH)和丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)、总ATP酶(T-ATP)活性;流式细胞仪检测细胞周期、细胞凋亡、线粒体膜电位(MMP)和活性氧(ROS);Western蛋白质印迹法检测胱天蛋白酶酶原3、Bcl-2、Bax和细胞色素c(Cyt c)蛋白的表达.结果 ASB呈浓度依赖性地抑制HK-2细胞增殖,作用24和48 h后IC5o值分别为19.1±3.6和(11.6±3.9)g·L-1.ASB 20 9·L-1处理HK-2细胞24 h后,G2/M期细胞由对照组的(23.3±1.0)％增至(37.0±0.8)％(P＜0.01).与正常对照组相比,ASB组LDH漏出率和ROS水平明显增加,MMP呈下降趋势;与ASB 20 9·L-1组相比,加入还原剂NAC后,细胞存活率和MMP水平明显增加(P＜0.05),LDH和ROS水平明显降低(P＜0.05);加入CsA后,细胞存活率显著降低(P＜0.01),LDH和细胞内ROS水平明显增加(P＜0.05).随着ASB浓度增加,HK-2细胞上清液中的MDA水平明显增加,GSH,SOD和T-ATP酶活性逐渐减弱;同时细胞内胱天蛋白酶原3、Bax和Cyt c蛋白表达升高.结论 ASB对HK-2细胞的毒性作用可能是通过干扰HK-2细胞线粒体能量代谢,改变细胞内氧化还原状态,耗竭细胞内GSH和SOD,导致细胞内ROS大量堆积,引发脂质过氧化作用,破坏细胞膜的通透性,导致Cyt c等凋亡因子从线粒体释放,进而引发细胞凋亡或坏死.

目的 探讨大黄素对大鼠肝脏细胞色素P450酶(CYP450)及其主要亚型的影响.方法 20只雄性SD大鼠,随机分成4组,每组5只,分别为溶剂对照组,170、500和1 500mg/kg大黄素染毒组,大黄素蒸馏水混悬后连续经口给药16d,结束后次日取大鼠肝脏组织制作微粒体,分别采用CO还原差示光谱法、分光光度法及化学发光法检测大鼠肝脏微粒体总CYP450水平,红霉素脱甲基酶(CYP3A)、氨基比啉-N-脱甲基酶,CYP1A、CYP2B和CYP2E1酶活性变化.结果 大黄素连续经口给药16d,能够引起大鼠肝脏微粒体总CYP450显著升高、可轻度诱导CYP3A、CYP1A、CYP2E1和CYP2B酶,500 mg/kg剂量组最明显.结论 大黄素对大鼠肝脏中CYP3A、CYP1A、CYP2B和CYP2E1酶均有诱导作用.

Objective: Apoptosis plays a dominant role in both spontaneous spermatogenesis and germ cell death. This study was aimed to investigate the functions of related genes in testicular germ cell death induced by Hydroxyurea (HU).Method: Wild-type (WT) and FasL transgenic (TG) DBA/C57BL mice were intraperitoneal injected with 400mg/kg HU. Twelve hours later, testes were collected. Histomorphology of testis was observed by staining with Periodic Acid Schiff (PAS). Apoptosis was assessed by TUNEL assay. mRNA and protein levels of related genes were evaluated by quantitative RT-PCR and Western blot, respectively.Results: The 2x2 factorial design comparative experiments between the WT and TG mice showed that the TG mice exhibited a higher basal apoptotic index. The basal mRNA levels of Fas and FasL and protein levels of Fas, FasL, Caspase-3, Caspase-8 and Caspase-9 in the TG mice were also higher than that in the WT mice. Twelve hours after injection of HU, the testicular tubules exhibited no significantly morphological changes but apoptosis index remarkably increased in both the WT and TG mice, with the latter having the higher amplitude. Although, HU up-regulated the mRNA of apoptosis-related genes, such as Fas and FasL, in both the TG and WT mice, the increased amplitude was more obvious in the TG mice. By Western blot analysis, apoptosis-related proteins Fas, FasL Caspase-3, Caspase-8 and Caspase-9 were significantly increased in both the WT and TG mice, with the TG mice exhibiting a greater up-regulation.Conclusion: Germ cell apoptosis induced by the HU treatment may be related to the FasL-mediated signal transduction pathway.

LPS-induced inflammatory response could be used to establish screening models for ILT and provides a new way to reduce liver toxicity of Z24.

现代科技的发展与人类文明的进步,对新药研发带来了前所未有的机遇与挑战。无论是人口老龄化所带来的人类疾病谱转变和多药治疗的日益普及,还是尚未全面了解之药物新靶点的逐次发现与应用;无论是难以临床前预测的特异质毒性,还是公众对药物安全性愈来愈高的要求和预期;都不同程度地影响到新药发现与开发完整链条的成功率,并导致诸多药物因安全性原因导致终止研发、黑框标识、撤出市场之类的研发失败,并对制药企业、患者和药政管理部门带来一系列的经济、健康和形象等方面的问题或危机。近10年来,随着IOM"Innovation/Stagnation:Challenge and Opportunity on the Critical Path to New Medical Products"报告的发布,国外药物毒理学研究模式正在酝酿巨大的变革。国家层面,FDA与EMEA分别制订了提高新药安全性的"关键路线计划CPI"和"创新药物计划IMI";美国NRC于2007年正式发表"21世纪的毒性试验:愿景与策略"报告,呼吁实施基于"化学物鉴定、毒性通路、靶向测试以及剂量反应与外推建模"的新毒性试验范式变革。新分支学科建设层面,则注重开展创新药物的转化毒理学、循证毒理学、本体论毒理学、系统毒理学、进化毒理学等理论体系的建立与发展;研究策略方面,则注重以体外系统、高内涵筛选、药理学作用谱分析为基础的早期毒理学系统的建立、验证与运用,强化对新药毒性相关的线粒体毒性、人类特异性毒性代谢物、特异质肝毒性、心脏毒性的体内外模型的研究与探索,积极引进人源化小鼠、计算机虚拟筛选、模式生物、干细胞、体内成像、3D打印等新技术与新模型体系,进而逐渐形成以基于毒性作用机制的毒性评价研究的模式体系。在管控新药安全风险方面,FDA与EMEA分别制订了"风险管理计划(RMP)"和"风险评估与减轻策略REMS"指导原则。国内的药物毒理学研究方面应当以创新药物研发为主导,综合跨领域、多学科研究方法为手段,建立和完善与国际先进水平同步的药物毒性机制研究体系与安全性评价平台,为国内新药研发的大发展提供强有力的支持和保证。

目的建立体外肾毒性优化筛选模型,降低药物先期研发费用、加快研究开发进程。方法永生化的人肾小管上皮细胞(HK-2)进行体外药物肾毒性评价的可能性进行评估,选择临床上常用的具有明显肾毒性的药物(顺铂、环孢素A、两性霉素B等),以及临床已经证实没有肾毒性的几种药物(阿司匹林、阿莫西林等),检测上述药物对HK-2细胞的细胞活性以及增值等作用,然后利用MTT和LDH漏出检测三代铂类抗肿瘤药物的细胞毒性,流式细胞仪PI染色检测细胞周期以及An-nexin-Ⅴ/PI双染色检测细胞凋亡,Western蛋白印迹检测热休克蛋白(HSP70)和免疫荧光法检测金属硫蛋白(MT)的表达,从细胞和分子水平对顺铂、卡铂和双环铂,三代铂类抗肿瘤药的HK-2细胞毒性进行比较研究,并结合体内外实验来验证该细胞用于体外肾毒性评价的特异性和敏感性。结果研究结果显示HK-2细胞对药物的肾毒性比较敏感,能够在体外反映药物肾毒性的大小,例如顺铂引起HK-2细胞周期以及LDH漏出改变的剂量是在1～5μmo·L-1,环孢素A和两性霉素B发生相同效应是25～50μmo·L-1,即使1000μmo·L-1的阿司匹林、阿莫西林和环磷酰胺也没能够引起HK-2细胞毒性的显著性改变。三代铂类抗肿瘤药的评价结果显示:顺铂在1μmo·L-1就能够引起HK-2细胞发生周期抑制,在5μmo·L-1就能够诱导HK-2细胞凋亡,但卡铂、双环铂对HK-2细胞产生的毒性作用的剂量约是顺铂剂量的50～100倍。三代铂类抗肿瘤药的SD大鼠体内肾毒性结果发现顺铂高剂量组(0.9 mg·kg-1)血BUN和Crea与空白对照组相比均呈显著升高,至恢复期此种改变仍存在。病理检测现广泛的小管退变、小管浓缩与扩张。卡铂、双环铂上述指标在给药结束时和恢复期与相应空白对照组相比均无显著差异,可见卡铂、双环铂对SD大鼠的肾毒性明显低于顺铂,这与体外实验结果是吻合的。结论 HK-2细胞对肾毒性药物比较敏感,能够在体外反映药物肾毒性的大小,利用该细胞建立模型来筛选药物体外肾毒性是可行的。

核苷(酸)类似物(NUC)是治疗乙型病毒性肝炎的一线药物。目前,应用较广的有阿德福韦酯、恩替卡韦、替诺福韦酯、克拉夫定等,但这些药物都或多或少存在,比如耐药性,肾脏性或生物利用度低等缺点。M478新酯为阿米福韦酯类似物,相对于其他化合物,其主要特点是抗毒活性大,不形成毒性代谢中间产物,药物的稳定性和水溶性更好些。本研究在采用体内外方法对M478新酯,M478和替诺福韦(PMPA)毒性比较研究。方法 Derek软件进行计算毒理学预测分析,体外细胞毒性采用MTT比较方法,AMES波动比较遗传毒性,体内采用经口单次和经口重复给药14 d毒性比较研究。结果采用Derek软件进行计算毒理学预测分析,结果致癌性、急性中毒性肾病和染色体损伤毒性可能出现于代谢中间体;M478,M478新酯在大鼠、犬、人类中都可能表现一定的肝肾毒性。MTT试验结果表明对于HepG2细胞,毒性大小顺序为M478新酯>M478>单酯>PMPA。Ames波动试验结果 M478及M478新酯均对TA100菌株无致突变性。上下法进行体内急性毒性研究,结果阿德福韦酯经口染毒大鼠LD50=1296 mg·kg-1(♀),95%CI(760～2000 mg·kg-1);M478新酯经口染毒大鼠LD50>2000 mg·kg-1(♀);大鼠灌胃M478新酯14 d重复染毒毒性试验结果表明阿德福韦阳性对照组组动物示肾近曲小管上皮细胞有轻度损伤。M478新酯高剂量组动物肝细胞肿胀等轻度损伤。结论 M478和M478新酯在有无S9活化条件下均无致突变性;对SD大鼠重复经口给予均无明显肝肾毒性,结合药效研究综合判断适合后续研发。

毒性病理学是药物安全性评价研究中的关键学科之一,可分为解剖病理学与临床病理学两个亚学科。药物急性毒性试验、长期毒性试验、局部刺激毒性试验、致癌性试验及生殖毒性试验等安全性评价研究的结论均有赖毒性病理学的检查结果。因此,毒性病理工作的质量直接影响药物安评工作的质量和水平。由于我国药物临床前安全性评价(Good Laboratory Pratice,GLP)和毒性病理学的规范性实施起步较晚,与美国、欧盟及日本等发达国家和地区相比存在较大差距,这在相当程度上制约了我国新药研发的质量与进步。本中心在国家重大新药创制重大专项的资助下,立足国内GLP行业毒性病理学现状,以OECD,FDA及STP等有关毒性病理学的指导原则与最佳实践指南及其它相关学术成果为依据,建立了一套基于中国国情,基本符合国际标准的药物安全性评价研究毒性病理学实践规范。解剖病理学方面,调研并购置了蜡块与玻片打号机、自动脱水机、石蜡包埋机、自动切片机、染色封片机与显微成像系统,并建立了基于数字切片扫描系统的远程病理会诊技术平台。以上硬件设备均按照标准操作规程进行规范化的管理、认证、使用及维护,从而为毒性病理学规范化建设提供了必要的硬件保障。系统地规范了动物解剖、脏器称量、标本留样修切、组织固定、脱水、包埋、切片、染色封片、切片镜检、同行评议及报告撰写等毒性病理学操作各流程的操作方法、操作细节及操作要点。建立了毒性病理学规范的诊断术语和诊断标准,毒性病理学常用实验动物脏器重量、脏器系数、自发疾病的背景数据库及常见自发肿瘤病理组织图像数据库。这些术语及标准对规范化毒性病理学的诊断具有重要意义。此外,对药物安全性评价研究中毒性病理学不同岗位从业人员的教育背景及资质认证等提出了具体要求和实施办法,建立了人员培训及考核上岗制度体系。临床病理学方面,购买了生化分析仪、免疫分析仪、流式细胞仪、PCR等一批先进高端仪器,实现了常规检测仪器有备份、生物技术药物安全性评价有仪器保彰的建设目标。对上述仪器设备,从购置、安装、使用、人员培训、维护、维修、SOP制定等环节制定符合GLP要求的管理制度,制定了仪器设备的计量检定管理制度和期间核查制度,建立了完善的大型仪器的安装验证、操作验证和性能验证制度。以上措施保证了仪器的良好运行,检测结果的准确可靠。建立了临床病理学组织管理体系、人员分工和培训体系、质量控制体系,并建立了分析前、分析中和分析后的全程质量控制模式。建立了SD大鼠、Beagle犬、恒河猴等安全性评价研究中常用实验动物的临床病理学背景数据。以上毒性病理学实践规范从"物"(硬件建设)、"法"(软件建设)、"人"(人员建设)三个方面初步尝试了毒性病理学规范化建设的方法和内容,多层次立体地构建了GLP条件下规范化的毒性病理学技术平台,对我国毒性病理学的规范化建设具有一定的指导意义。

药物毒理学是现代毒理学中研究药物的毒性作用机制,评价新药安全性的药理学分支学科;其主要目的在于指导临床合理用药,降低药物的副作用,减少因药物毒性导致的新药开发失败。药物毒理学科的两大任务是:结合药品注册为药物开发提供技术支撑,即实施GLP,规范常规的药物毒性试验;同时,为学科发展开展基础或应用基础研究,即开展毒性机理和新技术/新方法应用的研究。经过多年的发展,无论在规范性还是技术上,我国GLP都获得的巨大的发展,多数GLP中心都有能力承接国际项目,已有5家安评机构通过OECD成员国的GLP检查。作为创新药物研发体系的重要组成部分,药物毒理学的学科建设也同样取得了快速进展。研究对象已从群体转向个体;研究模式已转变为早期发现毒理学、转化毒理学、全程毒理学相结合的新范式等等;同时,国外药物毒理学研究范式也在发生着巨大的变革,这些变革也预示药物毒理学未来的发展展望。变革的起因为2004年FDA下属的医学研究所发布的1份题目为"Innovation/Stagnation:Challenge and Opportunity on the Critical Path to New Medical Products"的报告。基于这一报告,FDA当年即制订"关键路线计划(CriticalPath Initiative,CPI)"。2007年,欧盟也制订了类似的"创新药物计划(Innovation Medicines Initiative,IMI)",计划设立"欧洲药物安全研究中心",试图从整个欧盟层面解决因安全性原因所导致的新药研发失败。美国EPA下属的"环境化学物毒性试验与评价委员会"正式发表"21世纪的毒性试验:愿景与策略"报告,呼吁实施以"化学物鉴定、毒性途径、靶向测试以及剂量反应与外推建模"为模块、新的毒性试验范式的变革。为了应对这一变革进程的需要,制药企业、学术界和药政监管部门各司其责,共同推进药物毒理学学科的专业发展,走出了一条"加强桥接,通过转化毒理学认证毒性生物标志物和新的检测方法;提早决策,缩短新药开发时间和降低成本;抓住重点,关注最可能导致新药开发失败的线粒体毒性和人体特有的代谢物毒性问题;兼容并蓄,积极从现代生物医学领域引进并应用新技术和新方法和制药企业、学术界和药政监管部门三方协作,以联合体(Consortium)形式实施药物毒理学重大课题的合作攻关"的发展药物毒理学的新思路。

Objective: To establish the Beagle dog model with implanted telemetry transmitter,and analyze the parameters of cardiovascular and respiratory systems in the conscious Beagle dogs.Methods: We implanted the telemetry TL11M3-D70-PCTP transmitters to the dogs.The 24-h parameters of cardiovascular and respiratory systems in the conscious telemetered dogs were recorded and analyzed using Dataquest ART 4.8 system.Terfenadine 30 mg·kg-1 was used as a positive control.Results: Telemetry transmitters were successfully implanted.A circadian rhythm was obvious in the parameters from the conscious dogs;at 09:00AM,most parameters reached their peaks.Terfenadine prolonged QTc intervals in Beagle dogs.Conclusion: The conscious and telemetered Beagle dogs can be used for simultaneously assessing cardiovascular and respiratory changes for a longer term in the studies of safety pharmacology.

The comparison of the effect of fibrinolytic activity of recombinant human prourokinase (rhpro-UK) and urokinase(UK) with rabbits indicated rhpro-UK and UK all decreased resolution time of globular proteins, and the affect of rhpro-UK on parameter of fibrinolysis of rabbits was less than UK. The solution of thrombin of rhpro-UK in vitro was increased with dosage of rhpro-UK, and when rhpro-UK was compared with isodose UK, the difference is significant. Both rhpro-UK and UK all was marked solution of thrombin from Chinese piglets, and it were not evident effect on fibrinogen, plasminogen, and a2-antiplasmin of the piglets. The affect of rhpro-UK on bleeding time, clotting time, bleeding amount of unit time was less than UK. It indicated that the side effect of rhpro-UK was distinguished less than UK. The investigation of pharmacology of ileums of mice, rats, dogs, and guinea pigs indicated that rhpro-UK was not marked affect on behavior, state, central nervous system, cardiovascular system, respiratory system, and digestive system of the tested animals. But tested dogs were bleeding at location of operation, and the blood of the animal was heparinized. The studis of toxicology of rhpro-UK indicated that rhpro-UK was median lethal dose of 97.5 mg/kg in mice. When The Beagle dogs was administed rhpro-UK of 2, 8, 28 mg/kg, the toxic reaction was not observed, except for temporal hyperemia of gum, extend of clotting time, increasing of total cholerythrin, total proteins, albumin of the dogs, the direct damage of organs for the medicine was not observed in examination of pathology in the dosages of 8 and 28 mg/kg, any toxic reaction was not observed in dosage of 2 mg/kg. The special toxic experiments proved that rhpro-UK was not of mutagenesis and teratogenesis.

<正>啮齿类致癌性试验是创新药物安全性评价和上市风险控制内容的重要组成部分。新药上市申请时通常需提供必要的整体动物致癌性试验结果。对于明显存在可能引起致癌风险的情况,药政管理部门均要求在早期提交该项试验结果以支持长期用药的临床试验。而愈来愈多的证据表明,利用2种啮齿类动物评价药物致癌风险的经典范例存在诸多的局限性

Objective:To study the hypersusceptibility of two Lianbizhi injections with various purity by passive cutaneous anaphylaxis test(PCA) and all system anaphylaxis test(ASA).Method:The PCA and ASA tests were carried out according to the Chinese traditional medicine,natural medicine immunity toxicity study technical directed principle.Result:Anaphylaxis responses were exhibited in the two kinds of Lianbizhi injections in the PCA tests.In the ASA test,two guinea pigs showed weak-positive reaction with high dose of Lianbizhi injection A.One guinea pig showed weak-positive reaction,five showed positive reaction and two showed strong-positive reaction with high dose of Lianbizhi injection B.Conclusion:Anaphylaxis response induced by Lianbizhi injections may be related to the impurity.

军事医学科学院的药物毒理学研究源远流长、与时俱进,经过近30年的积累与发展,已建立一整套服务于新药发现、临床前开发、临床实验及上市后监督再评价等完整研发链条的药物毒理学研究体系和学科群,涵盖新药早期发现毒理学、非临床安全性评价以及药物毒性作用机制研究等内容,通过与新药研发体系中其他学科互动与协作,为军事医学科学院乃至全国的新药研发提供了良好的非临床安全性评价的技术平台和保障.

Objective:To observe the effects of a single injection of Lianbizhi on the endogenous metabolite in SD rat urine using metabonomics technique in order to explore the mechanisms of Lianbizhi-induced renal damage. Methods:Two kinds of Lianbizhi injections were used in this experiment as follow:Lianbizhi A containing andrographolide sodium bisulfis 98.7% and other related substance 1.3%,Lianbizhi B containing andrographolide sodium bisulfis 49.1% and other related substance 50.9%. Twenty five SPF male SD rats were randomly divided into five groups:the high-dose(1 600 mg/kg)and low-dose (400 mg/kg) Lianbizhi A groups,the high-dose(400 mg/kg)and low-dose (100 mg/kg) Lianbizhi B groups,and the control group (normal saline). Each group comprised 5 rats. All rats received respectively a single intravenous injection of Lianbizhi 10 mL/kg via caudal vein. The rat urine samples were collected over a 12 hours period before dosing,as well as within 0-6,7-12,13-24 and 25-48 hours after dosing respectively,and 1H nuclear magnetic resonance (1H NMR) spectra of urine samples were measured. The blood biochemical indices were measured 48 hours after dosing. Results:The results of blood biochemical indices testing showed that the ALP level in the high-dose Lianbizhi A group and high-dose Lianbizhi B group were respectively (306±13) and (294±45) U/L ,which were higher than that [(207±47) U/L] in the control group. The ALB levels in the high-dose Lianbizhi A and Lianbizhi B groups were respectively (28.26±1.07) and (27.34±1.01) U/L,which were higher than that [(25.70±0.37) U/L] in the control group. The TP levels in the high-dose Lianbizhi A and Lianbizhi B groups were respectively (51.32±3.36) and (50.10±2.36) g/L,which were higher than that [(45.76±1.73 )g/L] in the control group,all differences were statistically significant (all P<0.05). In addition,the BUN levels in the high-and low-dose Lianbizhi A groups as well as high-dose Lianbizhi B group were respectively (6.94±0.49),(6.69±0.31),and (6.81±0.38) mmol/L,which were higher than that [(5.90±0.45 ) mmol/L] in the control group,the differences were statistically significant ( P<0.05). The TG levels in the high-and low-dose Lianbizhi B groups were respectively (1.13±0.36) and (0.84±0.18)mmol/L,which were higher than that [(0.57±0.10) mmol/L] in the control group,the differences were statistically significant ( P<0.05). No marked changes were found in other blood biochemical indices. The time-course trajectories diagram of the urinary 1H NMR revealed the urinary metabolic profiles in all rats after dosing initially deviated from and subsequently returned to the levels of the control group. The principle components analysis (PCA) of 1H NMR at different time periods showed there was no significant difference in the rat urinary metabolic profiles among the 5 groups before dosing. The metabolic profiles in the drug-exposed groups could be distinguished from those in the control group,and the metabolic profile in the high-dose Lianbizhi B group deviated farthest from those in the control group 0-6 hours after dosing. Meanwhile,the trimethylamine(δ2.90)and dimethylglycine(δ2.94)levels increased and the citric acid (δ2.54,δ2.66)and α-ketoglutarate(δ2.46,δ3.02)levels decreased in the drug-exposed groups compared with the control group. The metabolic profiles in the drug-exposed groups could be distinguished from those in the control group 7-12 hours after dosing. Compared with the control group,citric acid and α-ketoglutarate in the drug-exposed groups decreased and the TMA and DMG levels in the high-and low-dose Lianbizhi A groups and high-dose Lianbizhi B group increased. The metabolic profiles in the drug-exposed groups could not distinguished from those in the control group within 13-48 hours after dosing and the metabolic profiles in the drug-exposed groups gradually returned to the levels of the control group. Conclusion:There is a correlation between the contents of other related substance in the Lianbizhi injection and the changes in the levels of rat urine endogenous metabolites. And the decrease in the levels of citric acid and α-ketoglutarate as well as the increase in the levels of trimethylamine and dimethylglycine suggest that the mechanism of renal damage caused by Lianbizhi injection may be related to its effects affecting the energy metabolism of mitochondria and osmotic pressure in renal medulla.