去甲斑蝥素对喜树碱引起的白细胞减少症的缓解作用
|
摘要
目的:观察去甲斑蝥素(norcantharidin,NCTD)能否缓解喜树碱(camptothecin,CPT)用药引起的骨髓抑制。方法:取Balb/c小鼠,采用概率单位法计算CPT半数致死量(median lethal dose,LD50)。以1 mg·kg-1作为CPT给药剂量,使用全血分析仪检测24 h内不同时间点白细胞浓度变化,为后续联合用药提供检测时间点参考。随后取Balb/c小鼠随机分为6组,分别为空白组,CPT(1 mg·kg-1)组,NCTD低、高剂量(10,20 mg·kg-1)组,CPT(1 mg·kg-1)+NCTD(10 mg·kg-1)组和CPT(1 mg·kg-1)+NCTD(20 mg·kg-1)组。连续灌胃2周,采用眼球采血法,应用全血分析仪检测进行相关指标的测定。流式细胞仪检测各组小鼠骨髓中性粒细胞的差异。病理组织切片评估肠道的毒性情况。结果:CPT LD50为1 mg·kg-1。CPT单独给药后6 h小鼠白细胞数目降至最少。24 h内与CPT单独用药比较,CPT+NCTD联合给药组白细胞、中性粒细胞数目明显上升(P<0.05),嗜酸性粒细胞、嗜碱性粒细胞等无明显变化。连续灌胃2周,比较CPT单独用药,CPT+NCTD联合给药组白细胞数目明显上升(P<0.05),其中淋巴细胞升高显著(P<0.01),其他细胞无明显变化。流式细胞仪检测各组小鼠骨髓中性粒细胞的无明显差异,苏木素-伊红(HE)染色显示CPT主要毒靶器官小肠的病理变化短期、长期均不明显。结论:NCTD可减轻CPT用药引起的白细胞减少症,并没有增加其肠道毒性,为临床CPT应用,并减轻其毒性提供一定理论参考。
Objective: To explore whether norcantharidin(NCTD) could reduce camptothecin(CPT)-caused myelosuppression. Method: Balb/c mice were collected. Probit analysis was made to calculate CPT median lethal dose(LD50). With 1 mg·kg-1 dose as CPT,whole blood analyzer was used to detect white blood cell(WBC) concentration at different time points within 24 h,so as to provide a reference for the subsequent detection time point in the subsequent combined administration. Subsequently,Balb/c mice were randomly divided into six groups: control group,CPT(1 mg·kg-1) group,low-dose NCTD(10 mg·kg-1) group,high-dose NCTD(20 mg·kg-1) group,CPT(1 mg·kg-1) + low-dose NCTD(10 mg·kg-1) group and CPT(1 mg·kg-1) +high-dose NCTD(20 mg·kg-1) group. After continuous gavage for 2 weeks,eyeball sampling method and whole blood analyzer were used to measure relevant indicators. Flow cytometry was used to detect the differences of bone marrow neutrophils in each group. Histopathological sections evaluated the intestinal toxicity. Result: The CPT LD50 is 1 mg·kg-1. The number of white blood cells in mice was reduced to a minimum at 6 h after the single administration with CPT. Within 24 hours after the single treatment with CPT,the numbers of leukocytes and neutrophils in CPT and NCTD combined groups increased significantly(P < 0. 05),with no significant change in the number of eosinophils and basophils. After continuous gavage for 2 weeks,compared with the single treatment with CPT,the number of white blood cells in CPT and NCTD combined groups also increased significantly(P <0. 05),and lymphocyte was significantly increased(P < 0. 01). No significant changes were observed in other cells. No significant difference in the mouse bone marrow neutrophil was detected by flow cytometry,hematoxylineosin(HE) staining showed no significant short-term or long-term pathological changes in intestine,the main target organ of toxicity of camptothecin. Conclusion: Norcantharidin can reduce the incidence of leucopenia caused by camptothecin,without increasing the intestinal toxicity. It provides a theoretical reference for the clinical application of camptothecin and its toxicity reduction.
Objective: To explore whether norcantharidin(NCTD) could reduce camptothecin(CPT)-caused myelosuppression. Method: Balb/c mice were collected. Probit analysis was made to calculate CPT median lethal dose(LD50). With 1 mg·kg-1 dose as CPT,whole blood analyzer was used to detect white blood cell(WBC) concentration at different time points within 24 h,so as to provide a reference for the subsequent detection time point in the subsequent combined administration. Subsequently,Balb/c mice were randomly divided into six groups: control group,CPT(1 mg·kg-1) group,low-dose NCTD(10 mg·kg-1) group,high-dose NCTD(20 mg·kg-1) group,CPT(1 mg·kg-1) + low-dose NCTD(10 mg·kg-1) group and CPT(1 mg·kg-1) +high-dose NCTD(20 mg·kg-1) group. After continuous gavage for 2 weeks,eyeball sampling method and whole blood analyzer were used to measure relevant indicators. Flow cytometry was used to detect the differences of bone marrow neutrophils in each group. Histopathological sections evaluated the intestinal toxicity. Result: The CPT LD50 is 1 mg·kg-1. The number of white blood cells in mice was reduced to a minimum at 6 h after the single administration with CPT. Within 24 hours after the single treatment with CPT,the numbers of leukocytes and neutrophils in CPT and NCTD combined groups increased significantly(P < 0. 05),with no significant change in the number of eosinophils and basophils. After continuous gavage for 2 weeks,compared with the single treatment with CPT,the number of white blood cells in CPT and NCTD combined groups also increased significantly(P <0. 05),and lymphocyte was significantly increased(P < 0. 01). No significant changes were observed in other cells. No significant difference in the mouse bone marrow neutrophil was detected by flow cytometry,hematoxylineosin(HE) staining showed no significant short-term or long-term pathological changes in intestine,the main target organ of toxicity of camptothecin. Conclusion: Norcantharidin can reduce the incidence of leucopenia caused by camptothecin,without increasing the intestinal toxicity. It provides a theoretical reference for the clinical application of camptothecin and its toxicity reduction.
引文
[1]Torre L A,Bray F,Siegel R L,et al.Global cancer statistics,2012[J].Ca Cancer J Clin,2011,65(2):87-108.
[2]黄庆英,肖静.肝癌病人介入治疗术的护理[J].家庭护士,2006,4(2):27-28.
[3]Pommier Y,Leo E,ZHANG H,et al.DNA topoisomerases and their poisoningby anticancer and antibacterial drugs[J].Chem Biol,2010,17(5):421-433.
[4]宋云龙,张万年,季海涛,等.DNA拓扑异构酶Ⅰ结构、功能及喜树碱类抗癌药物研究进展[J].中国药学杂志,2002,37(9):6-10.
[5]Oguma T.Antitumor drugs possessingtopoisomerase I inhibition:applicable separation methods[J].J Chromatogr B Biomed Sci Appl,2002,764(1/2):49-58.
[6]杨玉焕,邵长敏,骆沙曼,等.10-羟基喜树碱缓释微胶囊的组织分布学研究[J].中成药,2013,35(8):1644-1648.
[7]Ikegami T,HA L,Arimori K,et al.Intestinal alkalization as a possible preventive mechanism in irinotecan(CPT-11)-induced diarrhea[J].Cancer Res,2002,62(1):179-187.
[8]文柳静.10-羟基喜树碱经2种途径给药后在大鼠动、静脉血浆中的分布[J].中国药房,2013,24(13):1174-1176.
[9]陈军,方芸,张海霞,等.雾化吸入羟基喜树碱在兔体内的组织分布[J].药学学报,2004,39(9):747-751.
[10]王蓉,胡巍,吴松,等.静脉注射不同内酯型比例的羟基喜树碱后原形药物经小鼠尿液和粪便排泄的研究[J].中国医院药学杂志,2015,35(7):593-596.
[11]梁斌.羟喜树碱抑制肝癌介入栓塞后缺氧诱导因子1α表达及血管生成的实验研究[D].武汉:华中科技大学,2010.
[12]LIAN Q,XU J,YAN S,et al.Chemotherapy-induced intestinal inflammatory responses are mediated by exosome secretion of double-strand DNA via AIM2inflammasome activation[J].Cell Res,2017,27(6):784-800.
[13]于睿鹏,范潇婷,童宏选,等.喜树碱对小鼠骨髓中性粒细胞XIAP、TLR4、CXCR2表达及细胞凋亡影响的研究[J].江苏中医药,2017,49(6):80-82.
[14]刘亚楠.中药斑蝥研究进展[J].中药与临床,2013,4(4):50-52.
[15]李晓飞,陈祥盛,王雪梅.贵州含斑蝥素昆虫资源调查及斑蝥素含量的测定[J].湖北农业科学,2007,46(2):300-302.
[16]周长华,张青,欧瑞明,等.急性髓系白血病应用GCSF后对中性粒细胞碱性磷酸酶活性的影响[J].现代医院,2013,13(3):13-14.
[17]王绮如,汪保和,夏晖.粒-巨噬细胞集落刺激因子对小鼠骨髓内皮细胞增殖的刺激作用[J].中华血液学杂志,1998,19(6):41-42.
[18]李先茜,李晓丽,吴嘉熙,等.去甲斑蝥素诱导肿瘤细胞凋亡的研究进展[J].检验医学与临床,2013,10(9):1145-1146.
[19]顾兵,张政,李玉萍,等.半数致死量及其计算方法概述[J].中国职业医学,2009,36(6):507-508,511.
[20]李翠萍,吴民耀,王宏元.3种半数致死浓度计算方法之比较[J].动物医学进展,2012,33(9):89-92.
[21]范潇婷,于睿鹏,董瑞娟,等.斑蝥素及去甲斑蝥素对小鼠毒性靶器官的影响[J].中国实验方剂学杂志,2017,23(15):118-123.
[22]Fischer M A,Davies M L,Reider I E,et al.CD11b(+),Ly6G(+)cells produce type I interferon and exhibit tissue protective properties followingperipheral virus infection[J].PLo S Pathog,2011,7(11):e1002374.
[23]Othmani S,M'Saddek F,Zidi B,et al.Bicytopenia(leukopenia and thrombopenia)induced by cimetidine.Apropos of a new case[J].Therapie,1992,47(1):81-82.
[24]李静,任小沧.香菇多糖联合化疗治疗肺癌的疗效及免疫功能影响[J].中华肿瘤防治杂志,2016,23(S1):98-99.
[25]戴书静,王继峰,莫日根,等.去甲斑蝥素增强Bel7402中IκBα表达和抑制细胞增殖的研究[J].解剖学报,2001,32(2):155-158.
[2]黄庆英,肖静.肝癌病人介入治疗术的护理[J].家庭护士,2006,4(2):27-28.
[3]Pommier Y,Leo E,ZHANG H,et al.DNA topoisomerases and their poisoningby anticancer and antibacterial drugs[J].Chem Biol,2010,17(5):421-433.
[4]宋云龙,张万年,季海涛,等.DNA拓扑异构酶Ⅰ结构、功能及喜树碱类抗癌药物研究进展[J].中国药学杂志,2002,37(9):6-10.
[5]Oguma T.Antitumor drugs possessingtopoisomerase I inhibition:applicable separation methods[J].J Chromatogr B Biomed Sci Appl,2002,764(1/2):49-58.
[6]杨玉焕,邵长敏,骆沙曼,等.10-羟基喜树碱缓释微胶囊的组织分布学研究[J].中成药,2013,35(8):1644-1648.
[7]Ikegami T,HA L,Arimori K,et al.Intestinal alkalization as a possible preventive mechanism in irinotecan(CPT-11)-induced diarrhea[J].Cancer Res,2002,62(1):179-187.
[8]文柳静.10-羟基喜树碱经2种途径给药后在大鼠动、静脉血浆中的分布[J].中国药房,2013,24(13):1174-1176.
[9]陈军,方芸,张海霞,等.雾化吸入羟基喜树碱在兔体内的组织分布[J].药学学报,2004,39(9):747-751.
[10]王蓉,胡巍,吴松,等.静脉注射不同内酯型比例的羟基喜树碱后原形药物经小鼠尿液和粪便排泄的研究[J].中国医院药学杂志,2015,35(7):593-596.
[11]梁斌.羟喜树碱抑制肝癌介入栓塞后缺氧诱导因子1α表达及血管生成的实验研究[D].武汉:华中科技大学,2010.
[12]LIAN Q,XU J,YAN S,et al.Chemotherapy-induced intestinal inflammatory responses are mediated by exosome secretion of double-strand DNA via AIM2inflammasome activation[J].Cell Res,2017,27(6):784-800.
[13]于睿鹏,范潇婷,童宏选,等.喜树碱对小鼠骨髓中性粒细胞XIAP、TLR4、CXCR2表达及细胞凋亡影响的研究[J].江苏中医药,2017,49(6):80-82.
[14]刘亚楠.中药斑蝥研究进展[J].中药与临床,2013,4(4):50-52.
[15]李晓飞,陈祥盛,王雪梅.贵州含斑蝥素昆虫资源调查及斑蝥素含量的测定[J].湖北农业科学,2007,46(2):300-302.
[16]周长华,张青,欧瑞明,等.急性髓系白血病应用GCSF后对中性粒细胞碱性磷酸酶活性的影响[J].现代医院,2013,13(3):13-14.
[17]王绮如,汪保和,夏晖.粒-巨噬细胞集落刺激因子对小鼠骨髓内皮细胞增殖的刺激作用[J].中华血液学杂志,1998,19(6):41-42.
[18]李先茜,李晓丽,吴嘉熙,等.去甲斑蝥素诱导肿瘤细胞凋亡的研究进展[J].检验医学与临床,2013,10(9):1145-1146.
[19]顾兵,张政,李玉萍,等.半数致死量及其计算方法概述[J].中国职业医学,2009,36(6):507-508,511.
[20]李翠萍,吴民耀,王宏元.3种半数致死浓度计算方法之比较[J].动物医学进展,2012,33(9):89-92.
[21]范潇婷,于睿鹏,董瑞娟,等.斑蝥素及去甲斑蝥素对小鼠毒性靶器官的影响[J].中国实验方剂学杂志,2017,23(15):118-123.
[22]Fischer M A,Davies M L,Reider I E,et al.CD11b(+),Ly6G(+)cells produce type I interferon and exhibit tissue protective properties followingperipheral virus infection[J].PLo S Pathog,2011,7(11):e1002374.
[23]Othmani S,M'Saddek F,Zidi B,et al.Bicytopenia(leukopenia and thrombopenia)induced by cimetidine.Apropos of a new case[J].Therapie,1992,47(1):81-82.
[24]李静,任小沧.香菇多糖联合化疗治疗肺癌的疗效及免疫功能影响[J].中华肿瘤防治杂志,2016,23(S1):98-99.
[25]戴书静,王继峰,莫日根,等.去甲斑蝥素增强Bel7402中IκBα表达和抑制细胞增殖的研究[J].解剖学报,2001,32(2):155-158.