Objective:To explore the effect of Oxymatrine(OMT)on hepatocellular carcinoma cells and its possible action pathways.Methods:The core targets of Oxymatrine action in hepatocellular carcinoma were identified using netw...Objective:To explore the effect of Oxymatrine(OMT)on hepatocellular carcinoma cells and its possible action pathways.Methods:The core targets of Oxymatrine action in hepatocellular carcinoma were identified using network pharmacology and validated using in vitro experiments.CCK8 assay and scratch assay were used to detect the effects of Oxymatrine on the proliferation and migration of MHCC97H(TP53 mutant)cells,and Western blot assay and qPcr assay were used to find out the expression of the core targets after Oxymatrine acted on MHCC97H cells at the protein level and mRNA level,respectively.Results:Network pharmacology showed that the core target was TP53 protein;CCK8 assay showed that the IC50 at 24 h and 48 h was 9.64 mg/ml and 7.47 mg/ml,respectively;Scratch assay proved that Oxymatrine markedly inhibited the migratory ability of MHCC97H cells;Western blot assay and qPcr assay found that TP53 gene expression was down-regulated.The western blot experiment and qPcr experiment found that TP53 gene expression was down-regulated.Conclusions:Oxymatrine significantly inhibited the proliferation and migration of hepatocellular carcinoma MHCC97H cells,and its mechanism of action may be related to TP53.展开更多
探讨氧化苦参碱(OMT)乳膏对小鼠皮肤屏障功能的修复作用。将70只雄性ICR小鼠随机分为7组,除正常组小鼠外,采用胶带粘贴法模拟机械损伤,复制皮肤屏障功能障碍小鼠模型。造模后经皮连续给药5天,拍摄照片并测定造模区域的经表皮失水量(TEWL...探讨氧化苦参碱(OMT)乳膏对小鼠皮肤屏障功能的修复作用。将70只雄性ICR小鼠随机分为7组,除正常组小鼠外,采用胶带粘贴法模拟机械损伤,复制皮肤屏障功能障碍小鼠模型。造模后经皮连续给药5天,拍摄照片并测定造模区域的经表皮失水量(TEWL)。实验结束后,HE染色观察皮肤组织形态学变化,酶联免疫吸附法(ELISA)检测皮肤组织中肿瘤坏死因子α(TNF-α)、白细胞介素-8(IL-8)的含量,通过实时定量PCR(RT-qPCR)检测紧密连接蛋白(ZO-1)、丝聚蛋白(FLG)、水通道蛋白3(AQP3)mRNA的表达量,通过Western blot检测屏障相关蛋白如封闭蛋白(OCC)、FLG、紧密连接蛋白(Claudin)的表达量。结果显示,与模型组相比,OMT各剂量组均能降低皮肤TEWL,减轻炎症浸润,改善皮肤病理状态,同时高剂组皮肤组织中TNF-α等炎症因子的含量显著降低(P<0.05),ZO-1、FLG m RNA及OCC、FLG、Claudin蛋白的表达显著升高(P<0.05)。因此,OMT具有修复皮肤屏障功能的作用。展开更多
建立基于前处理技术结合液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)对鲤鱼基质中苦参碱和氧化苦参碱残留量测定的分析方法.样品采用乙腈均质提取的萃取方式,用硫酸钠作为除水剂,样品经过C18色谱柱...建立基于前处理技术结合液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)对鲤鱼基质中苦参碱和氧化苦参碱残留量测定的分析方法.样品采用乙腈均质提取的萃取方式,用硫酸钠作为除水剂,样品经过C18色谱柱分离,以水-乙腈为流动相,基质匹配标准曲线外标法定量.苦参碱和氧化苦参碱在质量浓度范围1—50 ng·mL^(−1)内,具有良好的线性关系,相关系数R^(2)均大于0.99.苦参碱方法检出限为0.21 ng·mL^(−1),定量限为0.69 ng·mL^(−1);氧化苦参碱方法检出限为0.17 ng·mL^(−1),定量限为0.52 ng·mL^(−1).添加浓度水平1.0、2.0、5.0 ng·mL^(−1),平均回收率为72.5%—108.7%,相对标准偏差(relative standard deviation,RSD,n=6)为4.2%—8.2%.苦参碱和氧化苦参碱的基质效应均表现为抑制.该方法前处理方法简单,灵敏度高,稳定性好,适合鲤鱼基质苦参碱和氧化苦参碱残留量的测定,同时为后续同类样品检测提供参考,也为今后相关标准制定提供数据积累.展开更多
通过探讨并梳理苦参碱和氧化苦参碱的发展概况,为预测其发展趋势与挖掘研究热点提供参考.以中国知网和Web of Science为数据源,用VOSviewer、CiteSpace和Excel软件分别对中文和英文文献的发文量、作者、发文机构和关键词等进行分析.共纳...通过探讨并梳理苦参碱和氧化苦参碱的发展概况,为预测其发展趋势与挖掘研究热点提供参考.以中国知网和Web of Science为数据源,用VOSviewer、CiteSpace和Excel软件分别对中文和英文文献的发文量、作者、发文机构和关键词等进行分析.共纳入431篇中文文献和223篇英文文献,研究机构主要以贵州医科大学和郑州大学为代表.结果表明,对苦参碱和氧化苦参碱的研究可以致力于探究苦参碱和氧化苦参碱治疗炎症性疾病、肿瘤,以及抗氧化应激和抗细胞凋亡等的作用靶点和分子机制方面;探究其诱导肝毒性、生殖毒性和发育毒性等不良反应的作用机制,明确作用靶点和通路;扩大其在畜牧业和林业等领域的应用范围.苦参碱和氧化苦参碱的研究处于稳步发展阶段,该领域最新的研究主要集中在苦参碱和氧化苦参碱的转录组学测序分析、相关中药膳食风险的研究和相关制剂的临床应用,以及其治疗炎症性疾病、肿瘤和对氧化应激、细胞凋亡的影响及其作用机制和信号通路方面.展开更多
使用超高效液相色谱⁃串联质谱仪,采用同位素内标法定量,建立枸杞中苦参碱和氧化苦参碱的测定方法。样品经1.0%磷酸水溶液超声提取,强阳离子固相萃取柱净化,10 mL 5%氨水甲醇溶液洗脱,采用0.1%甲酸水溶液和甲醇为流动相,梯度洗脱,通过T3...使用超高效液相色谱⁃串联质谱仪,采用同位素内标法定量,建立枸杞中苦参碱和氧化苦参碱的测定方法。样品经1.0%磷酸水溶液超声提取,强阳离子固相萃取柱净化,10 mL 5%氨水甲醇溶液洗脱,采用0.1%甲酸水溶液和甲醇为流动相,梯度洗脱,通过T3色谱柱分离,电喷雾离子源正离子扫描模式、多反应监测模式检测。通过考察不同种类提取溶剂、超声时间、固相萃取条件下目标化合物峰面积,确定最优前处理方式。结果表明:苦参碱和氧化苦参碱在0.5~50.0μg/kg范围内呈现良好线性关系,相关系数(R2)均大于0.999。方法检出限0.05μg/kg,定量限0.2μg/kg。低、中、高3个浓度加标回收试验,苦参碱回收率范围在76.3%~90.7%,相对标准偏差(relative standard deviation,RSD)在2.2%~7.4%,氧化苦参碱回收率范围在79.2%~89.0%,RSD在3.1%~6.6%。该方法适用于枸杞中苦参碱和氧化苦参碱检测。展开更多
AIM:To determine the antiviral mechanism or target of oxymatrine against hepatitis B virus(HBV).METHODS:HepG2.2.15 cells were incubated with culture medium containing 500 μg/mL of oxymatrine for 2 and 5 d.The surface...AIM:To determine the antiviral mechanism or target of oxymatrine against hepatitis B virus(HBV).METHODS:HepG2.2.15 cells were incubated with culture medium containing 500 μg/mL of oxymatrine for 2 and 5 d.The surface antigen of HBV(HBsAg) and e antigen of HBV(HBeAg) in supernatant were determined by ELISA.HBV DNA in supernatant,and intracellular covalently closed circular DNA(cccDNA),relaxed circular DNA(rcDNA) and pregenomic RNA(pgRNA) were quantif ied by specif ic real-time polymerase chain reaction(PCR) or reverse transcription(RT)-PCR.RESULTS:Treatment with oxymatrine for 2 d and 5 d reduced the production of HBV by the cell line,as indicated by the decline of HBsAg(22.67%,t = 5.439,P = 0.0322 and 22.39%,t = 5.376,P = 0.0329,respectively),HBeAg(55.34%,t = 9.859,P = 0.0101 and 43.97%,t = 14.080,P = 0.0050) and HBV DNA(40.75%,t = 4.570,P = 0.0447 and 75.32%,t = 14.460,P = 0.0047) in the supernatant.Intracellular cccDNA was also markedly reduced by 63.98%(t = 6.152,P = 0.0254) and 80.83%(t = 10.270,P = 0.0093),and intracellular rcDNA by 34.35%(t = 4.776,P = 0.0413) and 39.24%(t = 10.050,P = 0.0097).In contrast,intracellular pgRNA increased by 6.90-fold(t = 8.941,P = 0.0123) and 3.18-fold(t = 7.432,P = 0.0176) after 500 μg/mL of oxymatrine treatment for 2 d and 5 d,respectively.CONCLUSION:Oxymatrine may inhibit the replication of HBV by interfering with the process of packaging pgRNA into the nucleocapsid,or inhibiting the activity of the viral DNA polymerase.展开更多
文摘Objective:To explore the effect of Oxymatrine(OMT)on hepatocellular carcinoma cells and its possible action pathways.Methods:The core targets of Oxymatrine action in hepatocellular carcinoma were identified using network pharmacology and validated using in vitro experiments.CCK8 assay and scratch assay were used to detect the effects of Oxymatrine on the proliferation and migration of MHCC97H(TP53 mutant)cells,and Western blot assay and qPcr assay were used to find out the expression of the core targets after Oxymatrine acted on MHCC97H cells at the protein level and mRNA level,respectively.Results:Network pharmacology showed that the core target was TP53 protein;CCK8 assay showed that the IC50 at 24 h and 48 h was 9.64 mg/ml and 7.47 mg/ml,respectively;Scratch assay proved that Oxymatrine markedly inhibited the migratory ability of MHCC97H cells;Western blot assay and qPcr assay found that TP53 gene expression was down-regulated.The western blot experiment and qPcr experiment found that TP53 gene expression was down-regulated.Conclusions:Oxymatrine significantly inhibited the proliferation and migration of hepatocellular carcinoma MHCC97H cells,and its mechanism of action may be related to TP53.
文摘探讨氧化苦参碱(OMT)乳膏对小鼠皮肤屏障功能的修复作用。将70只雄性ICR小鼠随机分为7组,除正常组小鼠外,采用胶带粘贴法模拟机械损伤,复制皮肤屏障功能障碍小鼠模型。造模后经皮连续给药5天,拍摄照片并测定造模区域的经表皮失水量(TEWL)。实验结束后,HE染色观察皮肤组织形态学变化,酶联免疫吸附法(ELISA)检测皮肤组织中肿瘤坏死因子α(TNF-α)、白细胞介素-8(IL-8)的含量,通过实时定量PCR(RT-qPCR)检测紧密连接蛋白(ZO-1)、丝聚蛋白(FLG)、水通道蛋白3(AQP3)mRNA的表达量,通过Western blot检测屏障相关蛋白如封闭蛋白(OCC)、FLG、紧密连接蛋白(Claudin)的表达量。结果显示,与模型组相比,OMT各剂量组均能降低皮肤TEWL,减轻炎症浸润,改善皮肤病理状态,同时高剂组皮肤组织中TNF-α等炎症因子的含量显著降低(P<0.05),ZO-1、FLG m RNA及OCC、FLG、Claudin蛋白的表达显著升高(P<0.05)。因此,OMT具有修复皮肤屏障功能的作用。
文摘通过探讨并梳理苦参碱和氧化苦参碱的发展概况,为预测其发展趋势与挖掘研究热点提供参考.以中国知网和Web of Science为数据源,用VOSviewer、CiteSpace和Excel软件分别对中文和英文文献的发文量、作者、发文机构和关键词等进行分析.共纳入431篇中文文献和223篇英文文献,研究机构主要以贵州医科大学和郑州大学为代表.结果表明,对苦参碱和氧化苦参碱的研究可以致力于探究苦参碱和氧化苦参碱治疗炎症性疾病、肿瘤,以及抗氧化应激和抗细胞凋亡等的作用靶点和分子机制方面;探究其诱导肝毒性、生殖毒性和发育毒性等不良反应的作用机制,明确作用靶点和通路;扩大其在畜牧业和林业等领域的应用范围.苦参碱和氧化苦参碱的研究处于稳步发展阶段,该领域最新的研究主要集中在苦参碱和氧化苦参碱的转录组学测序分析、相关中药膳食风险的研究和相关制剂的临床应用,以及其治疗炎症性疾病、肿瘤和对氧化应激、细胞凋亡的影响及其作用机制和信号通路方面.
文摘使用超高效液相色谱⁃串联质谱仪,采用同位素内标法定量,建立枸杞中苦参碱和氧化苦参碱的测定方法。样品经1.0%磷酸水溶液超声提取,强阳离子固相萃取柱净化,10 mL 5%氨水甲醇溶液洗脱,采用0.1%甲酸水溶液和甲醇为流动相,梯度洗脱,通过T3色谱柱分离,电喷雾离子源正离子扫描模式、多反应监测模式检测。通过考察不同种类提取溶剂、超声时间、固相萃取条件下目标化合物峰面积,确定最优前处理方式。结果表明:苦参碱和氧化苦参碱在0.5~50.0μg/kg范围内呈现良好线性关系,相关系数(R2)均大于0.999。方法检出限0.05μg/kg,定量限0.2μg/kg。低、中、高3个浓度加标回收试验,苦参碱回收率范围在76.3%~90.7%,相对标准偏差(relative standard deviation,RSD)在2.2%~7.4%,氧化苦参碱回收率范围在79.2%~89.0%,RSD在3.1%~6.6%。该方法适用于枸杞中苦参碱和氧化苦参碱检测。
基金Supported by The National Natural Scientifi c Foundation of China,No. 30070958The National Key Technologies Research and Development Program of China during the 11th Five-year Plan Period,No. 2008zx1002-006
文摘AIM:To determine the antiviral mechanism or target of oxymatrine against hepatitis B virus(HBV).METHODS:HepG2.2.15 cells were incubated with culture medium containing 500 μg/mL of oxymatrine for 2 and 5 d.The surface antigen of HBV(HBsAg) and e antigen of HBV(HBeAg) in supernatant were determined by ELISA.HBV DNA in supernatant,and intracellular covalently closed circular DNA(cccDNA),relaxed circular DNA(rcDNA) and pregenomic RNA(pgRNA) were quantif ied by specif ic real-time polymerase chain reaction(PCR) or reverse transcription(RT)-PCR.RESULTS:Treatment with oxymatrine for 2 d and 5 d reduced the production of HBV by the cell line,as indicated by the decline of HBsAg(22.67%,t = 5.439,P = 0.0322 and 22.39%,t = 5.376,P = 0.0329,respectively),HBeAg(55.34%,t = 9.859,P = 0.0101 and 43.97%,t = 14.080,P = 0.0050) and HBV DNA(40.75%,t = 4.570,P = 0.0447 and 75.32%,t = 14.460,P = 0.0047) in the supernatant.Intracellular cccDNA was also markedly reduced by 63.98%(t = 6.152,P = 0.0254) and 80.83%(t = 10.270,P = 0.0093),and intracellular rcDNA by 34.35%(t = 4.776,P = 0.0413) and 39.24%(t = 10.050,P = 0.0097).In contrast,intracellular pgRNA increased by 6.90-fold(t = 8.941,P = 0.0123) and 3.18-fold(t = 7.432,P = 0.0176) after 500 μg/mL of oxymatrine treatment for 2 d and 5 d,respectively.CONCLUSION:Oxymatrine may inhibit the replication of HBV by interfering with the process of packaging pgRNA into the nucleocapsid,or inhibiting the activity of the viral DNA polymerase.