基于血清药理学的肝龙胶囊含药血清制备方法的初步探究

R363.2%R446.11%R285.5; 目的:利用血清药理学,探究用于肝细胞脂肪变性药效观察的肝龙胶囊(GLC)含药血清的最佳取血时间和含药血清体积分数.方法:C57BL/6J小鼠60只,灌胃GLC(90 mg/kg),每日两次,连续7 d,分别在末次给药0.5、1、2和4 h心脏采血,收集血清.对照组小鼠灌胃同体积的生理盐水.LC-MS/MS技术检测空白血清及不同采血时间含药血清肌苷含量,确定最佳采血时间.应用0%、1%、3%和10%GLC血清处理棕榈酸(PA)刺激的小鼠肝细胞NCTC1469,尼罗红染色检测细胞脂滴生成,RT-qPCR检测细胞内脂肪酸生成基因Fabp1和Scd1,及脂...

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Published in中国病理生理杂志 Vol. 40; no. 5; pp. 956 - 960
Main Authors 张珂珂, 白豪, 吕金宇, 金悦, 罗芮
Format Journal Article
LanguageChinese
Published 暨南大学基础医学与公共卫生学院病理生理学系,国家中医药管理局病理生理实验室,广东 广州 510632%暨南大学药学院实验教学中心,广东 广州 510632 01.05.2024
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ISSN1000-4718
DOI10.3969/j.issn.1000-4718.2024.05.023

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Abstract R363.2%R446.11%R285.5; 目的:利用血清药理学,探究用于肝细胞脂肪变性药效观察的肝龙胶囊(GLC)含药血清的最佳取血时间和含药血清体积分数.方法:C57BL/6J小鼠60只,灌胃GLC(90 mg/kg),每日两次,连续7 d,分别在末次给药0.5、1、2和4 h心脏采血,收集血清.对照组小鼠灌胃同体积的生理盐水.LC-MS/MS技术检测空白血清及不同采血时间含药血清肌苷含量,确定最佳采血时间.应用0%、1%、3%和10%GLC血清处理棕榈酸(PA)刺激的小鼠肝细胞NCTC1469,尼罗红染色检测细胞脂滴生成,RT-qPCR检测细胞内脂肪酸生成基因Fabp1和Scd1,及脂肪酸β-氧化基因Pparα的表达.结果:(1)LC-MS/MS检测发现在末次给药2 h后血清中肌苷浓度最高.(2)尼罗红染色显示GLC血清对PA诱导的肝细胞脂质积累具有剂量依赖性的减少趋势.(3)1%和3%和10%GLC血清处理均能显著下调PA刺激的肝细胞内Fabp1和Scd1的表达,且具有剂量依赖性的减少趋势;1%、3%和10%GLC血清处理均能显著上调PA刺激的肝细胞内Pparα的表达,且具有剂量依赖性的增加趋势.结论:小鼠给药后2 h的10%GLC血清是用于肝细胞脂肪变性药效观察的最佳血清干预浓度.
AbstractList R363.2%R446.11%R285.5; 目的:利用血清药理学,探究用于肝细胞脂肪变性药效观察的肝龙胶囊(GLC)含药血清的最佳取血时间和含药血清体积分数.方法:C57BL/6J小鼠60只,灌胃GLC(90 mg/kg),每日两次,连续7 d,分别在末次给药0.5、1、2和4 h心脏采血,收集血清.对照组小鼠灌胃同体积的生理盐水.LC-MS/MS技术检测空白血清及不同采血时间含药血清肌苷含量,确定最佳采血时间.应用0%、1%、3%和10%GLC血清处理棕榈酸(PA)刺激的小鼠肝细胞NCTC1469,尼罗红染色检测细胞脂滴生成,RT-qPCR检测细胞内脂肪酸生成基因Fabp1和Scd1,及脂肪酸β-氧化基因Pparα的表达.结果:(1)LC-MS/MS检测发现在末次给药2 h后血清中肌苷浓度最高.(2)尼罗红染色显示GLC血清对PA诱导的肝细胞脂质积累具有剂量依赖性的减少趋势.(3)1%和3%和10%GLC血清处理均能显著下调PA刺激的肝细胞内Fabp1和Scd1的表达,且具有剂量依赖性的减少趋势;1%、3%和10%GLC血清处理均能显著上调PA刺激的肝细胞内Pparα的表达,且具有剂量依赖性的增加趋势.结论:小鼠给药后2 h的10%GLC血清是用于肝细胞脂肪变性药效观察的最佳血清干预浓度.
Abstract_FL AIM:To investigate the optimal time of blood collection and the best concentration of Ganlong capsule(GLC)-containing serum for steatohepatitis.METHODS:Sixty C57BL/6J mice were gavaged with GLC at a dose of 90 mg/kg body weight,twice a day for 7 days.Control mice were administered the same volume of 0.9%saline.After 0.5,1,2 and 4 h from the last gavage,blood was collected to prepare GLC-containing serum.The LC-MS/MS was used to detect the inosine content of blank and drug-containing serum at different blood collection times to determine the optimal time.Subsequently,NCTC1469 cells stimulated with palmitic acid(PA)were treated with different concentrations of the GLC-containing serum.Nile Red staining was used to detect lipid droplet formation,and RT-qPCR was employed to as-sess the expression of intracellular fatty acid synthesis(Fabp1,Scd1)and fatty acid β-oxidation gene(Pparα).RE-SULTS:(1)LC-MS/MS detection revealed that the inosine concentration peaked 2 h after the last administration.(2)Nile red staining demonstrated that the GLC-containing serum exerted a dose-dependent decreasing trend of lipid droplets in hepatocytes induced by PA.(3)GLC-containing serum significantly downregulated the mRNA expression levels of lipo-genesis-genes(Fabp1 and Scd1)with a dose-dependent decreasing trend in the PA-induced NCTC1469 cells.The mRNA expression of genes associated with fatty acid β-oxidation(Pparα)were upregulated with a dose-dependent increasing trend.CONCLUSION:The 10%GLC-containing serum collected 2 hours after the last administration in mice has the best therapeutic effect for steatohepatitis.
Author 罗芮
张珂珂
白豪
吕金宇
金悦
AuthorAffiliation 暨南大学基础医学与公共卫生学院病理生理学系,国家中医药管理局病理生理实验室,广东 广州 510632%暨南大学药学院实验教学中心,广东 广州 510632
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Author_FL Lü Jinyu
JIN Yue
LUO Rui
ZHANG Keke
BAI Hao
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DocumentTitle_FL Preliminary serum pharmacological exploration of method to preparate Ganlong capsule-containing serum
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Keywords 取血时间
volume fraction
blood collection time
体积分数
Ganlong capsule
drug-containing serum
含药血清
肝龙胶囊
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PublicationTitle 中国病理生理杂志
PublicationTitle_FL Chinese Journal of Pathophysiology
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Publisher 暨南大学基础医学与公共卫生学院病理生理学系,国家中医药管理局病理生理实验室,广东 广州 510632%暨南大学药学院实验教学中心,广东 广州 510632
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