机体保护多肽对大鼠重复给药毒性研究
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摘要
目的:检测机体保护多肽(HuBPP)对大鼠产生的毒性反应,为临床提供无毒性反应剂量,从而为临床更安全地用药以及不良反应的监控提供参考信息。方法:80只SD大鼠单次静脉和肌内注射给药以确认给药剂量。选择0.2 mg/kg、1 mg/kg和4 mg/kg为本次实验的低、中、高剂量,并设置溶媒对照组,每组30只大鼠,雌雄各半。连续肌内注射给药一个月,实验期间观察大鼠质量、摄食量等指标,给药末期和恢复期采血检测各临床指标,病理学检测各主要脏器毒性变化。结果:急性毒性实验表明,20 mg/kg给药剂量下,实验组与对照组相比各观察指标未发现明显改变。长期毒性实验结果表明,与对照组相比,各剂量组大鼠质量、摄食量、主要脏器指标均未发现明显毒性反应(P>0.05)。长期毒性实验给药结束后,高剂量组雌鼠MCHC高于溶媒对照组,MPV低于溶媒对照组(P<0.01和P<0.05);中剂量对照组雌鼠TG高于溶媒对照组,Na~+浓度低于溶媒对照组(P<0.05);雄鼠高、中剂量组活化部分凝血活酶时间显著低于溶媒对照组(P<0.05),高剂量组Cl~-浓度显著高于溶媒对照组(P<0.05)。恢复期结束后高剂量组雄鼠红细胞体积分布宽度值低于溶媒对照组(P<0.05),低剂量组雌鼠丙氨酸氨基转移酶、血糖、三酰甘油显著高于溶媒组(P<0.05～P<0.01),但均处于文献报道正常变化范围内;其他指标差异无统计学意义。结论:在本实验条件下,HuBPP对大鼠的相对安全剂量为4 mg/kg及其以下剂量。
Objective:To evaluate the toxic reaction of human body protection peptide(HuBPP) in rat,and determine non-toxic reaction dose for clinical application and providing the reference information in clinical safe medication and monitoring of adverse reactions.Methods:The dosage of administration in 80 SD rats were confirmed by single intravenous and intramuscular injection.The dose of 0 mg/kg,0.2 mg/kg,1 mg/kg and 4 mg/kg were intramuscularly injected into the control group,low,medium high dose groups for 1 month,respectively(30 rats each group).The body weight and food intake in four groups were recorded during the test,the biochemical index at the end of drug administration and recovery period were detected,and the toxicity change of main organs was detected using histopathology.Results:The acute toxicity test showed that there was no significant change in the observed indicators between control group and experiment group.The long-term toxicity test showed that there was no statistical significance in toxicity reaction of body weight,food intake and main organ indicators between control group and experiment group(P>0.05).After the long-term poison test,the levels of MCHC and MPV in male rats of high-dose group were higher and lower than that in control group,respectively(P<0.01 and P<0.05);the levels of TG and Na~+ in female rats of middle-dose group were higher and lower than that in control group,respectively(P<0.05);the levels of APTT in male rats of high-and middle-dose group were significantly lower than that in control group(P<0.05);the Cl~- concentration in high-dose group was significantly higher than that in control group(P<0.05).At the end of the recovery period,the values of RDW in male rats of high-dose group were lower than those in control group(P<0.05),the levels of ALT,GLU,and TG in female rats of low-dose group were significantly higher than those in control group(P<0.05 to P<0.01).However,the parameters were within the normal range as reported in literature,and the significant difference of which was not found.Conclusions:Under the condition of the test,the 4 mg/kg or less than 4 mg/kg of HuBPP is safe dose in rats.
Objective:To evaluate the toxic reaction of human body protection peptide(HuBPP) in rat,and determine non-toxic reaction dose for clinical application and providing the reference information in clinical safe medication and monitoring of adverse reactions.Methods:The dosage of administration in 80 SD rats were confirmed by single intravenous and intramuscular injection.The dose of 0 mg/kg,0.2 mg/kg,1 mg/kg and 4 mg/kg were intramuscularly injected into the control group,low,medium high dose groups for 1 month,respectively(30 rats each group).The body weight and food intake in four groups were recorded during the test,the biochemical index at the end of drug administration and recovery period were detected,and the toxicity change of main organs was detected using histopathology.Results:The acute toxicity test showed that there was no significant change in the observed indicators between control group and experiment group.The long-term toxicity test showed that there was no statistical significance in toxicity reaction of body weight,food intake and main organ indicators between control group and experiment group(P>0.05).After the long-term poison test,the levels of MCHC and MPV in male rats of high-dose group were higher and lower than that in control group,respectively(P<0.01 and P<0.05);the levels of TG and Na~+ in female rats of middle-dose group were higher and lower than that in control group,respectively(P<0.05);the levels of APTT in male rats of high-and middle-dose group were significantly lower than that in control group(P<0.05);the Cl~- concentration in high-dose group was significantly higher than that in control group(P<0.05).At the end of the recovery period,the values of RDW in male rats of high-dose group were lower than those in control group(P<0.05),the levels of ALT,GLU,and TG in female rats of low-dose group were significantly higher than those in control group(P<0.05 to P<0.01).However,the parameters were within the normal range as reported in literature,and the significant difference of which was not found.Conclusions:Under the condition of the test,the 4 mg/kg or less than 4 mg/kg of HuBPP is safe dose in rats.
引文
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[15] DU QS,XIE NZ,HUANG RB.Recent development of peptide drugs and advance on theory and methodology of peptide inhibitor design[J].Med Chem,2015,11 (3):235.
[2] 袁伯俊,廖明阳,李波.药物毒理学实验方法与技术[M].北京:化学工业出版社,2007:212.
[3] 国家食品药品监督管理局.化学药物长期毒性研究技术指导原则[Z].2005:3.
[4] 国家食品药品监督管理局.药物非临床研究质量管理规范[J].中国医药导刊,2003,5(5):364.
[5] KARL-HEINZ DIEHL,ROBIN HULL,DAVID MORTON,et al.A good practice guide to the administration of substances and removal of blood,including routes and volumes[J].J Appl Toxicol,2001,21:15.
[6] FISH R,DANNEMAN PJ,BROWN M,et al.Anesthesia and analgesia in laboratory animals [M].2nd ed.San Diego:Academic Press,2008.
[7] National Research Council.Guide for care and use of laboratory animals[S].8th ed.Washington DC:National Academy Press,2011.
[8] 美国《实验动物饲养管理和使用指南》修订委员会,美国国家学术研究委员会.实验动物饲养管理和使用指南[M].王建飞,周艳,刘吉宏,等译.8 版.上海:上海科学技术出版社,2012.
[9] WALLACE HAYES A,KRUGER CL.Hayes′ principles and methods of toxicology[M].6th ed.Boca Raton:CRC Press.2014:572.
[10] HARDISTY JF,EUSTIS SL.Toxicologic pathology:a critical stage in study interpretation,in progress in predictive toxicology[M].Amsterdam:Elsevier,1990:41.
[11] GILL SS,KABANAGH M,CHERRY W,et al.A 28-day gavage toxicity study in male Fischer 344 rats with 2-methylfuran[J].Toxicol Pathol,2014,2(42):352.
[12] MARK S.The study pathologist′s role in GLP studies:a regulator′s perspective[J].Toxicol Pathol,2014,42(1):285.
[13] REICHL FX,SCHWENK M.Regulatory Toxicology[M]//KAUFMANN W,JACOBSEN M.Examination of Organ Toxicity.Berlin Heidelberg:Springer,2014:89.
[14] 陈力田,张葆樽,蒋国彦,等.内科学原理[M].北京:人民卫生出版社,2005:116.
[15] DU QS,XIE NZ,HUANG RB.Recent development of peptide drugs and advance on theory and methodology of peptide inhibitor design[J].Med Chem,2015,11 (3):235.