Microwave-assisted Micellar Extraction and Determination of Glycyrrhizic Acid and Liquiritin in Licorice Root by HPLC

The feasibility of emploving non-ionic surfactant （Triton X-100） as an alternative and effective solventfor the microwave-assisted extraction of glycyrrhizic acid （GA） and liquiritin （LQ） from licorice root was studied.The optimal extraction parameters based on the microwave-assisted micellar extraction technique were determined.Under the optimal conditions, i.e. 5% （by volume） Triton X-100, microwave-assisted extraction for 3--5min at 373K, the percentage extraction of active ingredients reached the highest value. The preconcentration tactor for GA and L＇Q （about 13.5） and the extraction efficiency for these two ingredients approached 100% showed the coupling of microwave-assisted extraction and cloud-point extraction could be employed as a new and. effective techniquefor the rapid extraction and preconcentration of pharrnacologically active ingredients from medicinal plants SUCh aslicorice root without disturbing chromatographic analysis.

Protective effect of liquiritin on oxidative stress injury of SH-EP1 cell lines caused by hydrogen peroxide

Objective:To study the protective effect of liquiritin on the oxidative stress injury of SH-EP1 cell lines caused by hydrogen peroxide.Methods: SH-EP1 cell lines were cultured and randomly divided into control group, H2O2 group and liquiritin group that were treated with the culture medium without serum, 200 μmol/L H2O2 as well as 100 μmol/L, 200 μmol/L and 400 μmol/L liquiritin combined with 200 μmol/L H2O2 respectively. After treatment, cell viability values as well as the content of mitochondrial apoptosis molecules and antioxidant molecules in cells were determined.Results:After 12 h, 24 h, 36 h and 48 h of treatment, the cell viability values of H2O2 group were significantly lower than those of control group, the cell viability values of 100 μmol/L, 200 μmol/L and 400 μmol/L liquiritin group were significantly higher than those of H2O2 group and the larger the liquiritin dosage, the higher the cell viability value;after 24 h of treatment, Bax, Caspase-3, Nrf2 and ARE content of H2O2 group were significantly higher than those of control group while Bcl-2, XIAP, SOD, GHS-Px and HO-1 content were significantly lower than those of control group;Bax and Caspase-3 content of 100 μmol/L, 200 μmol/L and 400 μmol/L liquiritin group were significantly lower than those of H2O2 group while Bcl-2, XIAP, Nrf2, ARE, SOD, GHS-Px and HO-1 content were significantly higher than those of H2O2group, and the larger the liquiritin dosage, the lower the Bax and Caspase-3 content while the higher the Bcl-2, XIAP, Nrf2, ARE, SOD, GHS-Px and HO-1 content.Conclusions:Liquiritin can inhibit the mitochondrial apoptosis and enhance the antioxidant system function to relieve the oxidative stress injury of SH-EP1 cell lines caused by hydrogen peroxide.

Intestinal permeability of liquiritin and isoliquiritin in the Caco-2 cell monolayer model

The intestinal permeability of two flavonoid compounds liquiritin （LQ） and isoliquiritin （ILQ） was investigated using the Caco-2 cell monolayer model. In order to evaluate the permeability and predict the absorption mechanism of the two compounds, the study on bidirectional permeability from the apical （AP） side to the basolateral （BL） side as well as from the BL side to the AP side was carried out. The determination was performed by HPLC-UV method. And the permeability parameters, especially the apparent permeability coefficients （Papp）, were then calculated. The Papp values of LQ and ILQ are （5.40±0.16）× 10^-7 and （8.69±0.15）× 10^-7 cm/s, respectively. The results of time- and concentration-dependent transport experiments indicate that both LQ and ILQ are poor absorbed mainly through passive diffusion.

De novo biosynthesis of liquiritin in Saccharomyces cerevisiae

Liquiritigenin(LG),isoliquiritigenin(Iso-LG),together with their respective glycoside derivatives liquiritin(LN)and isoliquiritin(Iso-LN),are the main active flavonoids of Glycyrrhiza uralensis,which is arguably the most widely used medicinal plant with enormous demand on the market,including Chinese medicine prescriptions,preparations,health care products and even food.Pharmacological studies have shown that these ingredients have broad medicinal value,including anti-cancer and antiinflammatory effects.Although the biosynthetic pathway of glycyrrhizin,a triterpenoid component from G.uralensis,has been fully analyzed,little attention has been paid to the biosynthesis of the flavonoids of this plant.To obtain the enzyme-coding genes responsible for the biosynthesis of LN,analysis and screening were carried out by combining genome and comparative transcriptome database searches of G.uralensis and homologous genes of known flavonoid biosynthesis pathways.The catalytic functions of candidate genes were determined by in vitro or in vivo characterization.This work characterized the complete biosynthetic pathway of LN and achieved the de novo biosynthesis of liquiritin in Saccharomyces cerevisiae using endogenous yeast metabolites as precursors and cofactors for the first time,which provides a possibility for the economical and sustainable production and application of G.uralensis flavonoids through synthetic biology.

甘草苷通过TLR4/IL-6信号通路改善癫痫幼鼠认知功能及海马神经元损伤

目的探讨甘草苷改善癫痫幼鼠认知功能、海马神经元损伤及对Toll样受体4(TLR4)/白细胞介素-6(IL-6)信号通路的调节作用。方法将60只幼鼠随机分为正常组、模型组、甘草苷组、激动剂组和激动剂+甘草苷组,每组12只。除正常组外,其余组幼鼠腹腔注射氯化锂-匹罗卡品建立癫痫模型。采用水迷宫实验检测幼鼠认知功能,酶联免疫吸附法检测血清IL-1β、肿瘤坏死因子α(TNF-α)和高迁移率族蛋白B1(HMGB1)水平,苏木精-伊红染色法观察海马神经元组织病理形态,末端脱氧核苷酸转移酶介导的dUTP标记测定法检测海马神经元凋亡率,蛋白印迹法检测海马B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、TLR4、IL-6、IL-1β、TNF-α和HMGB1蛋白表达水平。结果模型组和激动剂组幼鼠海马神经元形态不规则,排列紊乱,细胞核固缩、碎裂,细胞数减少,甘草苷组和激动剂+甘草苷组幼鼠海马神经元损伤程度明显改善。与模型组比较,甘草苷组幼鼠逃避潜伏期缩短,穿越平台次数增加,血清IL-1β、TNF-α和HMGB1表达水平,海马神经元凋亡率以及Bax、TLR4、IL-6、IL-1β、TNF-α和HMGB1蛋白表达水平均降低,Bcl-2蛋白表达水平升高(P<0.05);与激动剂组比较,激动剂+甘草苷组幼鼠逃避潜伏期缩短,穿越平台次数增加,血清IL-1β、TNF-α和HMGB1表达水平,海马神经元凋亡率以及Bax、TLR4、IL-6、IL-1β、TNF-α和HMGB1蛋白表达水平均降低,Bcl-2蛋白表达水平升高(P<0.05)。结论甘草苷可减轻癫痫幼鼠炎症反应,降低神经元损伤程度,改善认知功能和空间记忆能力。甘草苷可能是通过抑制TLR4/IL-6信号通路发挥作用。

甘草苷抑制急性心肌梗死大鼠心肌细胞铁死亡的作用研究

目的探讨甘草苷对急性心肌梗死(AMI)大鼠心肌细胞铁死亡的作用及调控机制。方法动物体内实验:将大鼠按照随机数字表法分为假手术组(0.9%氯化钠溶液)、AMI模型组(建模后0.9%氯化钠溶液灌胃)、低剂量组(建模后40 mg/kg甘草苷灌胃)和高剂量组(建模后80 mg/kg甘草苷灌胃)后,3组大鼠均在左心耳下端距离冠状动脉左前降支2 mm处进行结扎,构建AMI大鼠模型。观察并比较4组大鼠给药7 d后心肌缺血面积、组织病理学改变、心肌损伤标志物[血清肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、心肌肌钙蛋白I(cTnI)]水平、铁死亡标志物[活性氧(ROS)、谷胱甘肽(GSH)、Fe^(2+)、丙二醛(MDA)]水平及沉默调节蛋白3(SIRT3)、谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLC7A11)表达水平。体外细胞实验:将培养的H9c2细胞分别设空白组、模型组、低剂量组(40μmol/L甘草苷)、高剂量组(80μmol/L甘草苷)、高剂量+Erastin组(80μmol/L甘草苷+5μmol/L Erastin)、高剂量+si-NC组(80μmol/L甘草苷+0.8μg si-NC)和高剂量+si-SIRT3组(80μmol/L甘草苷+0.8μg si-SIRT3)予相应药物预处理;除空白组外,其余5组均进行低氧处理(1%O_(2)、5%CO_(2)、94%N2)建立缺氧细胞模型。观察并比较6组细胞活性、细胞凋亡率、铁死亡标志物及SIRT3、GPX4、SLC7A11蛋白表达水平。结果动物体内实验结果显示,与AMI模型组比较,低剂量组和高剂量组心肌组织损伤减轻且梗死面积较小,低剂量组和高剂量组CK、CK-MB、cTnI、MDA、ROS和Fe^(2+)水平均下降,GSH、SIRT3、GPX4和SLC7A11表达水平均升高(均P<0.05)。体外细胞实验结果显示,与模型组比较,低剂量组和高剂量组细胞活性、GSH、SIRT3、GPX4和SLC7A11表达水平均升高(均P<0.05),细胞凋亡率、MDA、ROS和Fe^(2+)水平均降低(均P<0.05)。与高剂量组比较,高剂量+Erastin组和高剂量+si-SIRT3组细胞活性、GSH、SIRT3、GPX4和SLC7A11表达水平均降低(均P<0.05),细胞凋亡率、MDA、ROS和Fe^(2+)水平均升高(均P<0.05)。结论在AMI大鼠的心肌细胞中甘草苷通过调控SIRT3减轻铁死亡,保护心肌细胞免受损伤。

甘草苷调节丙酮酸激酶M2改善过氧化氢诱导的人脐静脉内皮细胞功能障碍

目的:探讨甘草苷(LIQ)调节丙酮酸激酶M2(PKM2)对过氧化氢(H_(2)O_(2))诱导的人脐静脉内皮细胞(HUVECs)功能障碍的影响。方法:使用H_(2)O_(2)诱导HUVECs损伤模型,将细胞分为对照组(常规培养的HUVECs)、H_(2)O_(2)组(400μmol/L H_(2)O_(2)诱导的HUVECs模型)、H_(2)O_(2)+LIQ组(400μmol/L H_(2)O_(2)+1μmol/L LIQ)、H_(2)O_(2)+LIQ+紫草素(PKM2抑制剂)组(400μmol/L H_(2)O_(2)+1μmol/L LIQ+1μmol/L紫草素)。使用qRT-PCR检测PKM2 mRNA水平,荧光探针检测活性氧(ROS)水平,Western Blotting检测Nrf2、HO-1、PKM2、p-PKM2、Bcl-2、Bax及TNF-α、IL-1β蛋白表达变化。结果:与H_(2)O_(2)组相比,LIQ干预显著上调PKM2 mRNA水平、PKM2及p-PKM2的蛋白水平,抑制了ROS水平及TNF-α、IL-1β蛋白表达水平,Bcl-2/Bax比例显著增加。与H_(2)O_(2)+LIQ组相比,使用紫草素显著逆转了上述指标变化趋势(P<0.01)。结论:LIQ可上调PKM2表达,改善H_(2)O_(2)诱导的HUVECs细胞功能障碍。

甘草苷抑制破骨细胞分化缓解骨丢失

背景:破骨细胞的骨吸收功能相对或绝对活跃是骨质疏松症的诱因之一,因此,如何抑制破骨细胞形成、降低骨吸收活性是预防和治疗骨质疏松症的重点内容。甘草苷来源于传统中药甘草,对临床骨相关疾病具有治疗作用,但目前关于甘草苷在骨质疏松症方面的研究较少且机制不明。目的:通过体内外实验共同验证甘草苷抑制破骨细胞分化和缓解骨丢失的作用。方法:CCK-8实验检测甘草苷对小鼠来源骨髓巨噬单核细胞是否具有毒性或增殖作用,抗酒石酸酸性磷酸酶染色观察甘草苷抑制破骨细胞分化情况;通过网络药理学验证甘草苷与破骨细胞分化相关蛋白结合的亲和力大小;RT-PCR和Western blot实验检测甘草苷对破骨细胞相关特异性蛋白和基因表达以及相关信号通路的抑制作用;最后通过C57BL/6J小鼠骨质疏松模型验证甘草苷缓解骨丢失的作用。结果与结论:甘草苷在浓度20μmol/L及以下时能够起到抑制破骨细胞形成和分化的作用,同时与破骨细胞相关特异性蛋白如活化T细胞核因子c1、组织蛋白酶K、c-Fos、基质金属蛋白酶9具有较高的亲和力,能够降低上述基因和蛋白的相对表达水平,甘草苷也能够在干预第15,30,45,60分钟时有效降低MAPK信号通路中JNK的磷酸化表达水平,同时在第60分钟时能够挽救核因子κB信号通路中IκB-α的降解现象,体内实验证明甘草苷能够缓解由破骨细胞引起的骨质丢失现象,改善骨小梁相关参数。结果表明:甘草苷能抑制破骨细胞分化和缓解骨丢失,从而起到抗骨质疏松症的作用。

HPLC法测定参苓白术散中三种成分的含量

目的:建立参苓白术散中苍术酮、白术内酯Ⅰ和甘草苷的定量检测方法。方法:采用HPLC法,以C_(18)色谱柱(5μm,4.6 mm×250 mm),乙腈作为流动相A,测定了参苓白术散中苍术酮、白术内酯Ⅰ、甘草苷的含量。结果:苍术酮在0.0082~0.1308 mg/mL浓度范围内线性关系良好,平均加样回收率97.8%(RSD=0.7%,n=6);白术内酯Ⅰ在0.0016~0.0256 mg/mL浓度范围内线性关系良好,平均加样回收率103.4%(RSD=1.8%,n=6);甘草苷在0.0153~0.2444 mg/mL浓度范围内线性关系良好,平均加样回收率104.2%(RSD=0.5%,n=6)。结论:本研究成功建立了参苓白术散中苍术酮、白术内酯I和甘草苷的检测方法,且方法专属性强,精密度、重复性良好。

甘草颗粒特征图谱及其质量相关性研究

建立甘草颗粒中甘草苷、甘草酸铵含量检测方法并进行指纹图谱研究。采用高效液相色谱法,色谱柱为Eclipse Plus C_(18)柱(4.6 mm×250.0 mm,5μm),检测波长237 nm,以乙腈(A)-0.05%磷酸(B)为流动相,梯度洗脱,流速为1.0 mL/min,柱温为30℃,进样量10μL。结果显示,10批样品有12个共有峰,相似度大于0.901。经对照品比对,指认甘草苷和甘草酸铵两个指标性成分并对其进行定量分析,线性关系良好(R^(2)>0.9998),平均加样回收率为99.67%~102.42%。该方法操作简单、快捷,结果准确,可以甘草颗粒中甘草苷、甘草酸铵含量的测定对甘草颗粒进行质量评价。

多基原甘草质量标准探讨

目的分析多基原甘草质量现状及存在的问题,探索制约因素并提出解决方案。方法通过和企业座谈、实地调研、文献查询及甘草质量标准研究工作,分析当前甘草产业现状、药用标准问题及发展契机。结果以《中华人民共和国药典》(2020年版)含量测定标准,我国胀果甘草、光果甘草中甘草苷含量不合格率偏高,但3种基原的甘草存在其他具有药理作用的黄酮类化学成分,仅以甘草苷作为质量控制指标评价不够全面。结论建议进一步提高和完善《中华人民共和国药典》中胀果甘草和光果甘草质量标准,采用多种黄酮类成分作为甘草质量评价指标,进而推进我国甘草种植业发展。

LC–MS/MS Method for Quantification of Liquiritigenin in Rat Plasma: Application to Pharmacokinetic Study of Liquiritin

Objective A simple, sensitive, and rapid LC-MS/MS method has been established and validated for the determination of liquiritigenin （LG） in rat plasma. Methods Naringenin was chosen as internal standard （IS）. LG and IS were separated on a Diamonsil C18 analytical column with a mobile phase of methanol-10% methanol in water containing 0.5 mmol/L ammonium formate and 0.2% formic acid （55：45） at the isocratic flow rate of 0.6 mL/min for 10 min. The multiple reaction monitoring （MRM） was performed on a mass spectrometer in the negative ion mode with electro-spray ionization （ESI） source and the transition from precursor ion to product ion was m/z255.0~119.0 for LG and m/z 271.04151.0 for IS, respectively. Results The linearity was acceptable in the range of 5-5000 ng/mL （r= 0.9973）. The inter-day and intra-day accuracies were in the ranges of -0.09%-3.25% and -5.02%-9.21%, respectively. The precision was in the ranges of 3.60%-12.4% and 0.909%-6.89%, respectively. LG was stable in the course of anarysis and storage. Conclusion The LC-MS/MS method was successfully applied to the pharmacokinetic study for the first time in rats after ig and iv administration of liquiritin （LQ）, a glycoside of LG, at pharmacologically effective levels.

甘草防治阿尔茨海默病的作用机制研究进展

阿尔茨海默病(Alzheimer's disease,AD)是一种中枢神经系统退行性疾病,临床表现为学习记忆障碍以及日常生活能力减退等,对老年人的健康造成巨大威胁。近些年,对中药治疗AD的研究逐渐兴盛起来,其具有多组分、多途径、多靶点整体调节以及安全性高,毒副作用小等特点,符合疾病的复杂性需求。甘草作为我国传统药食同源补益类中草药具有抗病毒、抗氧化、抗炎等多种药理活性。研究表明,甘草对AD具有潜在的治疗作用,但其作用机制尚不清晰。研究发现,甘草提取物可以通过抗炎、抗氧化应激,抑制Tau蛋白磷酸化的作用,进而发挥抗AD的作用,甘草单体活性成分及其复方不仅可以通过上述机制达到抗AD的作用,而且能通过抑制β-淀粉样蛋白沉积,保护胆碱能神经元,抑制神经细胞凋亡等达到防治AD的目的。本文就近年来甘草提取物、甘草单体活性成分及甘草复方防治AD的作用机制进行总结,以期为甘草防治AD提供理论依据。

HPLC-VWD-ELSD法同时测定消炎宁胶囊中7个成分的含量

目的 采用高效液相色谱-紫外检测器-蒸发光散射检测器(HPLC-VWD-ELSD)法同时测定消炎宁胶囊中苦玄参苷IA、异嗪皮啶、迷迭香酸、金丝桃苷、甘草苷、冬青素A和毛冬青皂苷A1的含量。方法 采用Waters Symmetry C_(18)色谱柱(250 mm×4.6 mm, 5μm),流动相为乙腈(A)-0.15%醋酸水溶液(B),梯度洗脱,流速为1.0 mL·min^(-1)。苦玄参苷IA、异嗪皮啶、迷迭香酸、金丝桃苷、甘草苷采用VWD进行检测,切换波长分别为270 nm及342 nm;冬青素A和毛冬青皂苷A1采用ELSD进行检测,漂移管温度为85℃,载气流速3.5 L·min^(-1)。结果 苦玄参苷IA、异嗪皮啶、迷迭香酸、金丝桃苷、甘草苷、冬青素A和毛冬青皂苷A1进样量分别在1.764~35.28μg、0.6903~13.81μg、0.9524~19.05μg、0.4067~8.134μg、0.2136~4.272μg、0.9182~18.36μg和0.7665~15.33μg范围内与峰面积呈良好的线性关系(r≥0.9991),平均回收率及RSD分别为100.01%(0.81%)、99.14%(0.84%)、99.40%(0.92%)、99.34%(1.02%)、99.20%(1.04%)、98.83%(1.09%)、98.99%(1.08%)。4批消炎宁胶囊中上述7个成分的含量测定结果分别为6.812~6.901 mg·g^(-1)、1.726~1.789 mg·g^(-1)、2.391~2.422 mg·g^(-1)、1.134~1.155 mg·g^(-1)、0.6087~0.6105 mg·g^(-1)、2.306~2.324 mg·g^(-1)和1.995~2.016 mg·g^(-1)。结论 所建立的方法简捷,重复性好,可用于消炎宁胶囊中7个成分的含量测定,为消炎宁胶囊全面的质量控制提供科学依据。

甘草苷对胃癌荷瘤小鼠免疫功能的调节作用及机制研究

目的基于Janus激酶2(JAK2)/信号转导及转录激活蛋白3(STAT3)信号通路研究甘草苷(LIQ)对胃癌荷瘤小鼠免疫功能的调节作用及机制。方法皮下注射胃癌细胞MFC建立小鼠胃癌荷瘤模型,将造模成功的小鼠分为模型组(Model组)、低剂量LIQ组(LIQ-L组,20 mg/kg)、高剂量LIQ组(LIQ-H组,40 mg/kg)、高剂量LIQ+JAK2激活剂香豆霉素A1组(LIQ-H+香豆霉素A1组,40 mg/kg LIQ+1 mg/kg香豆霉素A1),每组12只;另取12只不造模小鼠设为正常组(Normal组)。各组小鼠灌胃/腹腔注射相应药物或生理盐水,每日给药1次,连续14 d。末次给药后,测定小鼠胃癌肿瘤的体积、质量以及脏器指数;检测外周血中CD4+、CD8+T淋巴细胞百分率;观察胃癌肿瘤组织病理形态学变化;检测胃癌肿瘤组织中JAK2/STAT3信号通路相关蛋白和白细胞介素6(IL-6)蛋白表达水平。结果与Normal组相比,Model组小鼠胸腺指数、脾脏指数、外周血中CD8+T淋巴细胞百分率均显著升高(P<0.05),外周血中CD4+T淋巴细胞百分率显著降低(P<0.05)。与Model组相比,LIQ-L组、LIQ-H组小鼠上述指标均显著逆转(P<0.05),胃癌肿瘤的体积、质量以及肿瘤组织中JAK2、STAT3蛋白的磷酸化水平和IL-6蛋白的表达水平均显著降低(P<0.05),且LIQ的作用具有剂量依赖性(P<0.05);肿瘤组织细胞出现不同程度的排列疏松、空泡化、分布不均匀等情况。加入JAK2激活剂香豆霉素A1减弱了LIQ对胃癌荷瘤小鼠免疫功能的改善作用和对胃癌肿瘤的抑制作用(P<0.05)。结论LIQ可通过抑制JAK2/STAT3信号通路,改善胃癌荷瘤小鼠的免疫功能,从而发挥抗肿瘤作用。

一测多评法同时测定蒲元和胃胶囊中7种成分含量

目的建立同时测定蒲元和胃胶囊中延胡索乙素、盐酸巴马汀、α-香附酮、咖啡酸、木犀草素、甘草苷、甘草酸铵的含量的一测多评法。方法采用高效液相色谱法测定,色谱柱为Diamonsil C_(18)柱(250 mm×4.6 mm,5µm),流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为1.0 mL/min,柱温为30℃,检测波长为237 nm,进样量为10µL。以木犀草素为内参物,建立与延胡索乙素、盐酸巴马汀、α-香附酮、咖啡酸、甘草苷、甘草酸铵的相对校正因子;采用一测多评法和外标法检测7种成分的含量,比较计算值与实测值的差异。结果延胡索乙素、盐酸巴马汀、α-香附酮、咖啡酸、木犀草素、甘草苷、甘草酸铵的质量浓度分别在2.67~133.41µg/mL、0.89~44.60µg/mL、1.36~67.85µg/mL、1.53~76.65µg/mL、2.00~100.12µg/mL、3.00~150.20µg/mL、8.51~425.61µg/mL范围内与峰面积线性关系良好(r≥0.9995,n=6);精密度、稳定性、重复性试验结果的RSD均小于2.0%(n=6);平均加样回收率分别为99.09%,97.34%,97.42%,96.85%,98.60%,99.20%,99.09%,RSD分别为0.72%,1.68%,1.75%,1.29%,0.69%,0.79%,0.97%(n=9)。延胡索乙素、盐酸巴马汀、α-香附酮、咖啡酸、木犀草素、甘草苷、甘草酸铵的相对校正因子分别为1.3159,0.7059,1.1805,1.2241,1.2385,6.7178;在不同色谱柱、色谱仪、流速、柱温下的重复性均良好,结果的RSD均低于1.50%。一测多评法与外标法的含量计算结果相对误差的绝对值均小于2.0%。结论该方法重复性好、结果准确可靠,可用于蒲元和胃胶囊的质量控制。

UHPLC-MS/MS法同时测定苏桔口服液中4种成分的含量

目的建立超高效液相色谱-三重四级杆串联质谱(UHPLC-MS/MS)同时快速测定苏桔口服液中甘草苷、迷迭香酸、桔梗皂苷D、甘草酸4种成分质量浓度的分析方法,以期更全面控制苏桔口服液的质量。方法样品采用Agilent Extend C18(2.1 mm×100 mm,1.7μm)色谱柱分离,柱温为35℃,以0.1%甲酸水(A)-乙腈(B)为流动相进行梯度洗脱,流速为0.3 mL/min;质谱采用电喷雾离子源,多重反应监测模式下负离子同时测定,检测离子对为甘草苷417.1/255.1(135)、迷迭香酸359.0/197.1(160.9)、桔梗皂苷D 1223.6/681.0、甘草酸821.4/350.9。结果4种成分在一定质量浓度范围内呈良好线性关系(r>0.999),平均加样回收率为98.80%~100.84%,RSD为1.82%~2.53%;10批苏桔口服液样品中,甘草苷、迷迭香酸、桔梗皂苷D、甘草酸的质量浓度分别为0.1408~0.1592、0.0824~0.1421、0.0359~0.0698、0.6659~0.7238 mg/mL。结论建立的多成分质量浓度检测方法快速、准确、重现性高,可为有效评价苏桔口服液的质量提供依据。

复方美登木片中甘草苷含量测定研究

目的:建立复方美登木片中甘草苷含量的高效液相色谱测定方法。方法:采用高效液相色谱法,Agilent C_(18)色谱柱(4.6 mm×150 mm,5μm),以乙腈-0.05%磷酸水溶液为流动相梯度洗脱,流速为1.0 mL/min,检测波长为237 nm,柱温30℃。结果:甘草苷线性范围为0.052~0.312μg(r^(2)=0.9997),平均加样回收率为101.04%,RSD=1.06%(n=6)。结论:该方法简便,准确可靠,可用于复方美登木片的质量控制。

Liquiritin ameliorates metabolic and endocrine alterations in a mouse model of polycystic ovary syndrome

Objective::Altered bile acid transformation induces low-grade chronic inflammation and may play an important role in the pathophysiology of polycystic ovary syndrome (PCOS). Liquiritincan regulate bile acid metabolism and anti-inflammatory properties;however, limited information is available regarding its therapeutic potential in PCOS.Methods::Female C57BL/6 mice were randomly assigned into four groups ( n = 6 mice/group): the control, letrozole or dehydroepiandrosterone-induced PCOS groups, PCOS + 20 mg/kg liquiritin group, and control + liquiritin groups. After 21 days of treatment, the mice were euthanized, and the associated metabolism indications were investigated. Ovarian histological examinations were performed, and serum hormone concentration was measured. The expression of key genes involved in steroid hormone synthesis, ovarian follicle development, and ovulation was assessed. Results::Liquiritin reduced fasting blood glucose levels and increased insulin sensitivity compared to the PCOS group. Liquiritin also significantly decreased serum levels of total testosterone ( P < 0.001) and dehydroepiandrosterone sulfate ( P < 0.05) in the PCOS group. Histomorphological inspection of ovaries from the liquiritin group revealed fewer cystic dilated follicles than in the PCOS group. Moreover, liquiritinsignificantly ( P < 0.01) decreased Cyp17a1, Cyp19a1, Fshr, Hsd3b2, Runx2, and Ccn2 mRNA expression compared to letrozole-induced PCOS. Conclusion::Liquiritin may be safe and helpful in ameliorating PCOS-associated hyperandrogenemia and hyperglycemia. However, clinical trials investigating different liquiritin dosages are needed to confirm these findings.

甘草苷纳米胶束的制备与评价

目的 制备甘草苷纳米胶束(Lip-NMs)以提高药物的渗透性,并通过衰老模型大鼠评估其抗衰老作用。方法 以聚氧乙烯聚氧丙烯醚嵌段共聚物(Pluronic?F127)和聚乙烯己内酰胺-聚醋酸乙烯酯-聚乙二醇接枝共聚物(Soluplus?)作为载体材料,采用溶剂蒸发-薄膜水化分散法将甘草苷制备成纳米胶束,并以临界胶束浓度、稀释稳定性、粒径分布、多聚分散系数(PDI)、Zeta电位和包封率作为评价指标,通过单因素实验确定Lip-NMs处方中Pluronic?F127和Soluplus?用量的配比;考察甘草苷混悬液和Lip-NMs在不同pH介质溶液中的药物释放情况;评估Lip-NMs对Caco-2细胞的毒性,并进行细胞跨膜转运研究;比较甘草苷混悬液和Lip-NMs对D-半乳糖所致衰老模型大鼠的抗衰老作用。结果 通过综合评估确定Lip-NMs的最优处方:Pluronic?F127和Soluplus?的质量比为25∶75;在透射电镜下可观察到Lip-NMs呈圆整球形;Lip-NMs在不同pH介质溶液中均表现为缓慢释药特性;Lip-NMs能够有效提高药物的渗透性;与对照组相比,Lip-NMs可显著提高衰老模型大鼠脑组织中超氧化物歧化酶(superoxide dismutase, SOD)、谷胱甘肽过氧化酶(glutathione peroxidase, GSH-Px)的含量,降低丙二醛(malondialdehyde, MDA)的含量,有效提高脑的抗氧化能力。结论 将Pluronic?F127和Soluplus?作为载体材料,将甘草苷制备成纳米胶束,有望促进药物吸收,达到更好的抗衰老作用。