Background:Reliable animal models are crucial to drug development for focal segmental glomerulosclerosis(FSGS),a rare kidney disease.Variability in success rates in literature and significant ethical concerns with ani...Background:Reliable animal models are crucial to drug development for focal segmental glomerulosclerosis(FSGS),a rare kidney disease.Variability in success rates in literature and significant ethical concerns with animal welfare necessitate further optimization of adriamycin(ADR)-induced FSGS model developed on BALB/c mice.Methods:High-performance liquid chromatography(HPLC)was used to assess ADR stability in water and upon light exposure.To identify the optimal ADR level,single intravenous ADR injections with dosing levels from 10 to 17 mg/kg body weight were administered to BALB/c mice to induce FSGS-like pathology.Body weight and proteinuria of FSGS mice were monitored and analyzed for FSGS model-associated morbidity.Animals were euthanized for hematological and kidney histological assessments 8 weeks post induction.To identify the suitable experiment time frame of the ADR-induced FSGS mouse model,a longitudinal study was performed,with an 11-week continuous monitoring of the symptoms.Results:ADR was found to be unstable in aqueous media and light sensitive.A dosing level of 10.5 mg/kg of ADR was optimal for consistent FSGS mouse model induction on BALB/c strain,characterized by minimal mortality and sustained FSGS-like symptoms.Findings from the longitudinal study suggest that 6 weeks post ADR induction may represent the peak of FSGS pathology severity in this mouse model.This time frame may be used for FSGS drug development projects.Conclusion:Based on the outcome from this study,we identified the optimal ADR dosing level and model testing duration.A standard operating procedure(SOP)for the ADR-induced FSGS mouse model was established to facilitate FSGS basic research and drug development.展开更多
在诊疗行为中,用药错误、药物相互作用、药品质量问题、药物滥用和依赖、静脉注射感染、药物耐药性、药品说明书错误等均属于药品不良反应(Adverse Drug Reaction,简称“ADR”)。对于医疗机构来说,开展药品不良反应监测(以下简称“ADR...在诊疗行为中,用药错误、药物相互作用、药品质量问题、药物滥用和依赖、静脉注射感染、药物耐药性、药品说明书错误等均属于药品不良反应(Adverse Drug Reaction,简称“ADR”)。对于医疗机构来说,开展药品不良反应监测(以下简称“ADR监测”)是药品安全体系的基石,通过科学的方法收集和分析数据,实现药品不良反应从“被动收集”向“主动防控”的转变,具有重要的医学、社会和经济意义。展开更多
本研究探讨中药大黄素(emodin)对人白血病耐药细胞株HL-60/ADR的增殖、凋亡影响及其相应的可能作用机制。采用MTT法绘制细胞生长曲线;集落培养法观察大黄素对HL-60/ADR克隆形成的影响;细胞周期分析、线粒体跨膜电位检测、Caspase-3酶活...本研究探讨中药大黄素(emodin)对人白血病耐药细胞株HL-60/ADR的增殖、凋亡影响及其相应的可能作用机制。采用MTT法绘制细胞生长曲线;集落培养法观察大黄素对HL-60/ADR克隆形成的影响;细胞周期分析、线粒体跨膜电位检测、Caspase-3酶活性检测、Annexin V FITC/PI法、TdT酶介导的原位缺口末端标记法(TUNEL)分析凋亡细胞;RT-PCR及Western blot法检测大黄素作用后不同时间段bcl-2、c-myc mRNA及Bcl-2、c-Myc、Caspase-3前体蛋白表达水平的变化。结果表明,大黄素对HL-60/ADR细胞增殖具有明显的抑制作用,呈时间和浓度依赖性。较低浓度大黄素即可抑制HL-60/ADR细胞集落形成,IC50值为5.79μmol/L。细胞周期分析显示,与对照组比较,40、80μmol/L浓度组细胞被阻滞于G0/G1期比率明显增高(p<0.01),G2/M期细胞比率降低(p<0.01),而20μmol/L浓度组细胞周期改变差异不具有显著性(p>0.05),各浓度组均检测到典型的亚二倍体峰(凋亡峰)。大黄素作用12小时HL-60/ADR细胞线粒体跨膜电位降低,Annexin V FITC/PI法检测到早期凋亡细胞,作用24小时caspase-3酶活性显著升高,作用48小时TUNEL法检测到晚期凋亡细胞,细胞凋亡率呈药物浓度依赖性。大黄素作用HL-60/ADR细胞不同时间段,bcl-2、c-myc mRNA和Bcl-2、c-Myc、Caspase-3前体蛋白表达水平均有不同程度下调,并呈时间依赖性。结论:大黄素能够有效抑制HL-60/ADR细胞增殖,并诱导其凋亡,bcl-2、c-myc表达水平下调,线粒体跨膜电位水平降低和caspase-3激活可能参与了该过程。展开更多
基金United States Department of Defense Office of the Congressionally Directed Medical Research Programs(CDMRP),Grant/Award Number:W81XWH-22-1-0176。
文摘Background:Reliable animal models are crucial to drug development for focal segmental glomerulosclerosis(FSGS),a rare kidney disease.Variability in success rates in literature and significant ethical concerns with animal welfare necessitate further optimization of adriamycin(ADR)-induced FSGS model developed on BALB/c mice.Methods:High-performance liquid chromatography(HPLC)was used to assess ADR stability in water and upon light exposure.To identify the optimal ADR level,single intravenous ADR injections with dosing levels from 10 to 17 mg/kg body weight were administered to BALB/c mice to induce FSGS-like pathology.Body weight and proteinuria of FSGS mice were monitored and analyzed for FSGS model-associated morbidity.Animals were euthanized for hematological and kidney histological assessments 8 weeks post induction.To identify the suitable experiment time frame of the ADR-induced FSGS mouse model,a longitudinal study was performed,with an 11-week continuous monitoring of the symptoms.Results:ADR was found to be unstable in aqueous media and light sensitive.A dosing level of 10.5 mg/kg of ADR was optimal for consistent FSGS mouse model induction on BALB/c strain,characterized by minimal mortality and sustained FSGS-like symptoms.Findings from the longitudinal study suggest that 6 weeks post ADR induction may represent the peak of FSGS pathology severity in this mouse model.This time frame may be used for FSGS drug development projects.Conclusion:Based on the outcome from this study,we identified the optimal ADR dosing level and model testing duration.A standard operating procedure(SOP)for the ADR-induced FSGS mouse model was established to facilitate FSGS basic research and drug development.
文摘在诊疗行为中,用药错误、药物相互作用、药品质量问题、药物滥用和依赖、静脉注射感染、药物耐药性、药品说明书错误等均属于药品不良反应(Adverse Drug Reaction,简称“ADR”)。对于医疗机构来说,开展药品不良反应监测(以下简称“ADR监测”)是药品安全体系的基石,通过科学的方法收集和分析数据,实现药品不良反应从“被动收集”向“主动防控”的转变,具有重要的医学、社会和经济意义。
文摘本研究探讨中药大黄素(emodin)对人白血病耐药细胞株HL-60/ADR的增殖、凋亡影响及其相应的可能作用机制。采用MTT法绘制细胞生长曲线;集落培养法观察大黄素对HL-60/ADR克隆形成的影响;细胞周期分析、线粒体跨膜电位检测、Caspase-3酶活性检测、Annexin V FITC/PI法、TdT酶介导的原位缺口末端标记法(TUNEL)分析凋亡细胞;RT-PCR及Western blot法检测大黄素作用后不同时间段bcl-2、c-myc mRNA及Bcl-2、c-Myc、Caspase-3前体蛋白表达水平的变化。结果表明,大黄素对HL-60/ADR细胞增殖具有明显的抑制作用,呈时间和浓度依赖性。较低浓度大黄素即可抑制HL-60/ADR细胞集落形成,IC50值为5.79μmol/L。细胞周期分析显示,与对照组比较,40、80μmol/L浓度组细胞被阻滞于G0/G1期比率明显增高(p<0.01),G2/M期细胞比率降低(p<0.01),而20μmol/L浓度组细胞周期改变差异不具有显著性(p>0.05),各浓度组均检测到典型的亚二倍体峰(凋亡峰)。大黄素作用12小时HL-60/ADR细胞线粒体跨膜电位降低,Annexin V FITC/PI法检测到早期凋亡细胞,作用24小时caspase-3酶活性显著升高,作用48小时TUNEL法检测到晚期凋亡细胞,细胞凋亡率呈药物浓度依赖性。大黄素作用HL-60/ADR细胞不同时间段,bcl-2、c-myc mRNA和Bcl-2、c-Myc、Caspase-3前体蛋白表达水平均有不同程度下调,并呈时间依赖性。结论:大黄素能够有效抑制HL-60/ADR细胞增殖,并诱导其凋亡,bcl-2、c-myc表达水平下调,线粒体跨膜电位水平降低和caspase-3激活可能参与了该过程。
