目的探讨车前方联合缬沙坦对糖尿病肾病小鼠干预的协同增效作用。方法(1)60只小鼠分为空白对照组(Ctrl,n=10)和造模组(n=50)。Ctrl予普通饲料,造模组小鼠选用高糖高脂饮食(HFD)、单侧肾切除联合腹腔注射链脲佐菌素(STZ)的方法诱导糖尿...目的探讨车前方联合缬沙坦对糖尿病肾病小鼠干预的协同增效作用。方法(1)60只小鼠分为空白对照组(Ctrl,n=10)和造模组(n=50)。Ctrl予普通饲料,造模组小鼠选用高糖高脂饮食(HFD)、单侧肾切除联合腹腔注射链脲佐菌素(STZ)的方法诱导糖尿病肾病模型。空腹血糖≥16.7 mmol·L^(-1)者认定为造模成功,小鼠随机分为模型组(DKD)、缬沙坦组(Val,12 mg·kg^(-1))、车前方组(CQF,2.1 g·kg^(-1)生药)和车前方联合缬沙坦组(CV,Val 12 mg·kg^(-1)+CQF 2.1 g·kg^(-1)生药),每组10只,连续给药干预12周。定期测量各组体质量、日饮水量、尿量;采用胰岛素耐量实验(ITT)以及空腹血糖(FBG)评估糖代谢功能;采用血尿素氮(BUN)、内生肌酐清除率(CCr)、24 h尿蛋白(UP)、24 h尿肌酐(Cr)、24 h尿中肾损伤分子-1(KIM-1)与中性粒细胞明胶酶相关脂质运载蛋白(NGAL)排泄量等指标评估肾功能;采用苏木精-伊红(HE)、马松(Masson)、过典酸雪夫氏(PAS)染色观察肾脏组织病理变化;分析肾小球超微结构变化。(2)基于超高效液相色谱-质谱联用(UPLC-MS/MS)方法测定正常小鼠单次灌胃缬沙坦(12 mg·kg^(-1))、车前方(生药2.1 g·kg^(-1))与联合给药的药代动力学指标,选择非房室模型方法计算药代动力学参数。结果(1)与Ctrl比较,DKD小鼠体质量明显下降,饮水量、尿量、FBG、ITT-葡萄糖曲线下面积(ITT-AUCG)、CCr、BUN、24 h UP、24 h Cr、24 h KIM-1、24 h NGAL水平异常升高;与DKD比较,各给药干预组饮水量、尿量、糖代谢及肾功能部分指标明显下降(P<0.05)。与Ctrl比较,DKD肾小球肥大、系膜区增宽,肾小管出现上皮细胞肿胀、小空泡样变性(P<0.05),肾小球窗孔消失,足突融合增宽,基底膜均质性增厚;与DKD比较,给药干预组肾脏形态与肾小球超微结构明显逆转(P<0.05)。各给药干预组组间比较,与Val相较,CV的CCr、24 h KIM-1、24 h NGAL排泄率下降更为显著(P<0.05);与CQF比较,CV的FBG、UACR值改善更为显著(P<0.05)。(2)与单药(缬沙坦或车前方)干预比较,联合给药(车前方联合缬沙坦)可增加缬沙坦血药浓度(提高1.59倍,P<0.05);促进车前方中主要活性成分小檗碱(BBR)、黄连碱(COP)、木兰花碱(MAG)和京尼平苷酸(GPA)的吸收(血药浓度时间曲线下面积0-t分别提高37.54%、36.05%、85.80%、26.11%,P<0.05)。结论车前方联合缬沙坦可提高活性成分的生物利用度,从而有效改善糖尿病肾病的糖代谢紊乱和肾功能损伤,发挥协同增效作用。展开更多
以中药为代表的天然药物具有悠久的药用历史,其功效物质基础解析和质量评价对于促进中医药的传承和创新发展具有重要意义。由于中药化学组成复杂,准确分析和鉴定与临床药效密切相关的指标成分较为困难,亟需开发和运用新型分离表征手段...以中药为代表的天然药物具有悠久的药用历史,其功效物质基础解析和质量评价对于促进中医药的传承和创新发展具有重要意义。由于中药化学组成复杂,准确分析和鉴定与临床药效密切相关的指标成分较为困难,亟需开发和运用新型分离表征手段。实时直接分析质谱(direct analysis in real-time mass spectrometry,DART-MS)是近年来新兴的常压敞开式离子化质谱技术,具有分析速度快、原位、样品前处理简单等特点。自2005年被报道以来,DART-MS在生物医学、环境监测、公共安全、药物分析等领域应用广泛。该技术为解析中药复杂化学成分和质量控制提供了新方式,具有较好的应用前景。本文概述了DART-MS技术的原理、特点、影响因素及技术进展,总结其在中药分析中的应用情况,并展望其发展前景。展开更多
Bile acids(BAs)are synthesized by the liver from cholesterol through several complementary pathways and aberrant cholesterol metabolism plays pivotal roles in the pathogeneses of cholesterol gallbladder polyps(CGP)and...Bile acids(BAs)are synthesized by the liver from cholesterol through several complementary pathways and aberrant cholesterol metabolism plays pivotal roles in the pathogeneses of cholesterol gallbladder polyps(CGP)and cholesterol gallstones(CGS).To date,there is neither systematic study on BAs profile of CGP or CGS,nor the relationship between them.To explore the metabolomics profile of plasma BAs in healthy volunteers,CGP and CGS patients,an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was developed and validated for simultaneous determination of 42 free and conjugated BAs in human plasma.The developed method was sensitive and reproducible to be applied for the quantification of BAs in the investigation of plasma samples.The results show that,compared to healthy volunteers,CGP and CGS were both characterized by the significant decrease in plasma BAs pool size,furthermore CGP and CGS shared aberrant BAs metabolic characteristics.Chenodeoxycholic acid,glycochenodeoxycholic acid,l-muricholic acid,deoxycholic acid,and 7-ketolithocholic acid were shared potential markers of these two cholesterol gallbladder diseases.Subsequent analysis showed that clinical characteristics including cysteine,ornithine and body mass index might be closely related to metabolisms of certain BA modules.This work provides metabolomic information for the study of gallbladder diseases and analytical methodologies for clinical target analysis and efficacy evaluation related to BAs in medical institutions.展开更多
文摘目的探讨车前方联合缬沙坦对糖尿病肾病小鼠干预的协同增效作用。方法(1)60只小鼠分为空白对照组(Ctrl,n=10)和造模组(n=50)。Ctrl予普通饲料,造模组小鼠选用高糖高脂饮食(HFD)、单侧肾切除联合腹腔注射链脲佐菌素(STZ)的方法诱导糖尿病肾病模型。空腹血糖≥16.7 mmol·L^(-1)者认定为造模成功,小鼠随机分为模型组(DKD)、缬沙坦组(Val,12 mg·kg^(-1))、车前方组(CQF,2.1 g·kg^(-1)生药)和车前方联合缬沙坦组(CV,Val 12 mg·kg^(-1)+CQF 2.1 g·kg^(-1)生药),每组10只,连续给药干预12周。定期测量各组体质量、日饮水量、尿量;采用胰岛素耐量实验(ITT)以及空腹血糖(FBG)评估糖代谢功能;采用血尿素氮(BUN)、内生肌酐清除率(CCr)、24 h尿蛋白(UP)、24 h尿肌酐(Cr)、24 h尿中肾损伤分子-1(KIM-1)与中性粒细胞明胶酶相关脂质运载蛋白(NGAL)排泄量等指标评估肾功能;采用苏木精-伊红(HE)、马松(Masson)、过典酸雪夫氏(PAS)染色观察肾脏组织病理变化;分析肾小球超微结构变化。(2)基于超高效液相色谱-质谱联用(UPLC-MS/MS)方法测定正常小鼠单次灌胃缬沙坦(12 mg·kg^(-1))、车前方(生药2.1 g·kg^(-1))与联合给药的药代动力学指标,选择非房室模型方法计算药代动力学参数。结果(1)与Ctrl比较,DKD小鼠体质量明显下降,饮水量、尿量、FBG、ITT-葡萄糖曲线下面积(ITT-AUCG)、CCr、BUN、24 h UP、24 h Cr、24 h KIM-1、24 h NGAL水平异常升高;与DKD比较,各给药干预组饮水量、尿量、糖代谢及肾功能部分指标明显下降(P<0.05)。与Ctrl比较,DKD肾小球肥大、系膜区增宽,肾小管出现上皮细胞肿胀、小空泡样变性(P<0.05),肾小球窗孔消失,足突融合增宽,基底膜均质性增厚;与DKD比较,给药干预组肾脏形态与肾小球超微结构明显逆转(P<0.05)。各给药干预组组间比较,与Val相较,CV的CCr、24 h KIM-1、24 h NGAL排泄率下降更为显著(P<0.05);与CQF比较,CV的FBG、UACR值改善更为显著(P<0.05)。(2)与单药(缬沙坦或车前方)干预比较,联合给药(车前方联合缬沙坦)可增加缬沙坦血药浓度(提高1.59倍,P<0.05);促进车前方中主要活性成分小檗碱(BBR)、黄连碱(COP)、木兰花碱(MAG)和京尼平苷酸(GPA)的吸收(血药浓度时间曲线下面积0-t分别提高37.54%、36.05%、85.80%、26.11%,P<0.05)。结论车前方联合缬沙坦可提高活性成分的生物利用度,从而有效改善糖尿病肾病的糖代谢紊乱和肾功能损伤,发挥协同增效作用。
文摘以中药为代表的天然药物具有悠久的药用历史,其功效物质基础解析和质量评价对于促进中医药的传承和创新发展具有重要意义。由于中药化学组成复杂,准确分析和鉴定与临床药效密切相关的指标成分较为困难,亟需开发和运用新型分离表征手段。实时直接分析质谱(direct analysis in real-time mass spectrometry,DART-MS)是近年来新兴的常压敞开式离子化质谱技术,具有分析速度快、原位、样品前处理简单等特点。自2005年被报道以来,DART-MS在生物医学、环境监测、公共安全、药物分析等领域应用广泛。该技术为解析中药复杂化学成分和质量控制提供了新方式,具有较好的应用前景。本文概述了DART-MS技术的原理、特点、影响因素及技术进展,总结其在中药分析中的应用情况,并展望其发展前景。
基金supported by the National Natural Science Foundation of China(Grant Nos.:81920108033,and 82274223).
文摘Bile acids(BAs)are synthesized by the liver from cholesterol through several complementary pathways and aberrant cholesterol metabolism plays pivotal roles in the pathogeneses of cholesterol gallbladder polyps(CGP)and cholesterol gallstones(CGS).To date,there is neither systematic study on BAs profile of CGP or CGS,nor the relationship between them.To explore the metabolomics profile of plasma BAs in healthy volunteers,CGP and CGS patients,an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was developed and validated for simultaneous determination of 42 free and conjugated BAs in human plasma.The developed method was sensitive and reproducible to be applied for the quantification of BAs in the investigation of plasma samples.The results show that,compared to healthy volunteers,CGP and CGS were both characterized by the significant decrease in plasma BAs pool size,furthermore CGP and CGS shared aberrant BAs metabolic characteristics.Chenodeoxycholic acid,glycochenodeoxycholic acid,l-muricholic acid,deoxycholic acid,and 7-ketolithocholic acid were shared potential markers of these two cholesterol gallbladder diseases.Subsequent analysis showed that clinical characteristics including cysteine,ornithine and body mass index might be closely related to metabolisms of certain BA modules.This work provides metabolomic information for the study of gallbladder diseases and analytical methodologies for clinical target analysis and efficacy evaluation related to BAs in medical institutions.
