人参皂苷Rg1对心肌细胞氧化应激损伤的抑制作用

目的 通过构建H₂O₂诱导的H9c2细胞氧化应激模型,观察人参皂苷Rg₁对氧化应激的抑制作用,并探讨mi R-499c是否参与人参皂苷Rg₁抑制氧化应激的作用机制。方法 采用600μmol/L的H₂O₂诱导H9c2细胞氧化应激模型,给予人参皂苷Rg₁预处理24 h,检测乳酸脱氢酶(lactate dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD)活性及丙二醛(malondialdehyde,MDA)、活性氧(reactive oxygen species,ROS)水平和线粒体膜电位(mitochondrial membrane potential,MMP)变化;采用Western blotting检测凋亡相关蛋白表达。采用Lipofiter 3.0将miR-499c转染至H9c2细胞再制备H₂O₂诱导的氧化应激模型,给予人参皂苷Rg     

人参皂苷Rg1通过激活PINK1/parkin增强线粒体自噬保护Aβ损伤的PC12细胞

阿尔茨海默病(Alzheimer′s disease,AD)患者脑中的β-淀粉样蛋白(amyloidβ-protein,Aβ)沉积是造成神经元线粒体损伤的直接原因。线粒体自噬是清除受损线粒体,保护神经细胞的重要方式。人参皂苷Rg₁具有神经保护作用,在防治AD方面有广阔的应用前景,但人参皂苷Rg₁减轻Aβ造成神经损伤的机制尚未阐明。为探讨人参皂苷Rg₁通过影响自噬保护神经细胞的作用机制,该文利用Aβ_25-35诱导PC12细胞损伤模型,观察了人参皂苷Rg₁的保护作用及对细胞自噬的影响,运用自噬诱导剂雷帕霉素和自噬抑制剂氯喹验证Rg₁的神经保护作用与自噬的相关性。发现人参皂苷Rg₁能够增强Aβ损伤PC12细胞的活力,并提高线粒体膜电位,减轻Aβ引起的线粒体损伤,这种保护效果能被自噬抑制剂氯喹阻断;人参皂苷Rg₁还能增加LC3Ⅱ/Ⅰ蛋白比例并促进p62蛋白消耗,同时增加PINK1及parkin蛋白,减少线粒体自噬接头蛋白OPTN的数量,表现出增强PC12细胞的自噬水平的作用。在运用PINK1 shRNA减低线粒体自噬调控位点PINK1的表达后,人参皂苷Rg₁不能再增加PINK1及其下游parkin的表达,并且不能显著影响线粒体自噬接头蛋白NDP52的数量,但其增强自噬的作用没有受到明显影响,仍能够显著增加LC3Ⅱ/Ⅰ比例,并且促进OPTN的消耗。该文研究表明人参皂苷Rg₁能促进Aβ损伤的PC12细胞的自噬水平,此外还可能通过促进PINK1介导的线粒体自噬,减轻线粒体受到的损伤,这可能是其改善Aβ造成的细胞损伤的作用机制之一。     

母鸡油制三七的质量评价

目的 对母鸡油制三七的质量进行评价。方法 采用LC-MS、HPLC法对皂苷类成分进行定性定量分析,GC-MS法鉴定脂肪酸类成分,并与生三七进行比较。结果 共鉴定出15种皂苷类成分,母鸡油制三七比生三七多出人参皂苷Rg₆、Rg₄、Rk₃、Rh₄、Rk₁、Rg₅,并且三七皂苷R₁及人参皂苷Rg₁、Re、Rb₁含量更低,人参皂苷Rh₁含量更高,其中人参皂苷Rk₃、Rg₅含量分别为2.464 1、20.387 1 mg/g;共鉴定22种脂肪酸类成分,其中母鸡油制三七19种,生三七8种,前者比后者多出棕榈油酸、肉豆蔻酸、花生四烯酸、11Z-十四碳烯酸等脂肪酸。结论 母鸡油制三七比生三七多出人参皂苷Rg₆、Rg₄等成分,可为该炮制品今后开发利用及相关新药研制奠定基础。     

不同干燥方式对人参果浆中稀有皂苷生成的影响

目的 研究采不同干燥方法对人参Panax ginseng果浆中原型皂苷降解和稀有皂苷生成的影响。方法 采用HPLC建立了同步检测11种人参皂苷含量的分析方法，对采用冷冻、电热、汽热3种方法干燥的人参果浆样品中原型皂苷和稀有皂苷的含量进行了测定。结果 与未干燥处理的人参果浆进行对比，冷冻干燥样品中11种人参皂苷含量变化较小；人参果浆电热干燥和汽热干燥样品中原型皂苷发生降解，稀有皂苷生成出现不同程度增加。结论 人参果浆经电热干燥和汽热干燥后原型人参皂苷Re、Rg₁、Rb₁、Rc、Rb₂、Rb₃发生降解，稀有人参皂苷F₁、Rh₁、Rg₃、Rk₁、Rh₂含量增加。     

人参皂苷治疗纤维化疾病的分子作用机制研究进展

纤维化是由于炎症导致器官实质细胞发生坏死,组织内细胞外基质异常增多和过度沉积引起的一种病理改变,以器官组织内纤维结缔组织增多、实质细胞减少为主要特征。纤维化的持续发展可致器官结构破坏和功能减退,乃至衰竭,严重威胁人类健康。人参皂苷包括Rg₁、Rg₃、Rd、Rb₁、Rh₁和AD₂等天然单体成分,其多种有益的生物学效应有助于治疗纤维化疾病,可通过调控转化生长因子-β(transforming growth factor-β,TGF-β)/Smad(small mothers against decapentaplegic)信号通路、核因子-κB(nuclear factor-κB,NF-κB)信号通路、p62/Kelch样环氧氯丙烷相关蛋白1(Kelch-like epichlorohydrinassociated protein 1,Keap1)/核因子红系2相关因子2(nuclear factor erythroid 2 related factor 2,Nrf2)通路、Jan...     

三七化学成分和药理作用研究概况及质量标志物的预测

三七植物含三萜皂苷、黄酮、氨基酸、多糖、挥发油等活性成分,具有活血、止血、散瘀等功效。该文对三七的本草考证、化学成分和主要药理活性进行总结,并基于中药质量标志物(Q-marker)的理论,从植物亲缘关系、药效、药性、化学成分可测性等方面对三七的质量标志物进行预测分析,推断三七中特定比例的人参皂苷Rg₁、Re、Rb₁,以及人参皂苷Rb₂、Rb₃、Rc、Rd、Rh₂、Rg₃和三七皂苷R₁、三七素、槲皮素可作为三七潜在的质量标志物,为反映三七功效的质量标准制定提供了参考。     

人参皂苷Rg1通过调控AMPK/NLRP3通路介导的细胞焦亡抑制大鼠肺纤维化研究

目的 探究人参皂苷Rg₁对大鼠肺纤维化(pulmonary fibrosis,PF)的影响及作用机制。方法 50只雄性SD大鼠随机分为对照组、模型组、人参皂苷Rg₁(72 mg/kg)组、腺苷酸活化蛋白激酶(adenosine monophosphate activated protein kinase,AMPK)激动剂(200 mg/kg)组、人参皂苷Rg₁(72 mg/kg)+AMPK抑制剂(20 mg/kg)组,每组10只。除对照组外,其余各组大鼠气管内注射博来霉素(5 mg/kg)构建大鼠PF模型。造模成功后2 d开始给药,连续给药28 d后检测大鼠肺功能指标;采用苏木素-伊红(HE)、Masson染色观察肺组织病理变化;采用免疫组化检测肺组织I型胶原(collagen I)和α-肌动球蛋白(α-smooth muscle actin,α-SMA)表达;采用试剂盒测定肺组织羟脯氨酸(hydroxyproline,Hyp)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-...     

基于人参皂苷和多糖含量的蒸制软支和硬支西洋参质量评价研究

目的 建立蒸西洋参的8个人参皂苷类成分以及多糖的含量测定方法。方法 采用高效液相色谱法(HPLC)测定自制蒸西洋参中8个人参皂苷类成分(人参皂苷Rg₁, Re, Rb₁, Rc, Rg₂, Rb₂, Rb₃, Rd)的含量,利用紫外-可见分光光度法(UV-Vis)对其总多糖的含量进行测定,并结合化学计量学(聚类分析、主成分分析、正交偏最小二乘判别分析等)的方法对所得结果进行分析。结果 来源于不同产地蒸西洋参的人参皂苷和多糖的含量有一定的差异,主要表现为软支西洋参蒸制后的各人参皂苷以及多糖的含量高于硬支,主成分分析(PCA)和偏最小二乘判别分析(OPLS-DA)结果也印证了这一结论。由以上研究可知,采用的蒸西洋参的方法较为稳定,所建立的基于8个人参皂苷类成分和多糖得率的含量测定方法可用于蒸西洋参的质量综合评价。结论 本研究为制定科学合理的蒸西洋参的加工方法和质量评价方法提供了依据。     

液相色谱-质谱联用同时测定芪参益气滴丸中黄芪甲苷、丹参素、原儿茶醛、人参皂苷Rg1和Rb1含量

目的 建立同时测定芪参益气滴丸中黄芪甲苷、丹参素、原儿茶醛、人参皂苷Rg₁和Rb₁含量的液相色谱-质谱联用方法。方法 采用Wonda Sil C₁₈色谱柱 (4.6 mm×150 mm, 5 μm ),流动相A为10 mmol·L甲酸铵,0.2%甲酸水溶液,B为0.2%甲酸乙腈溶液,采用线性梯度洗脱;流速为1 mL·min,柱温30 ℃。质谱条件采用电喷雾离子化（ESI）方式,负离子模式,以选择性离子监测（SIM）模式对黄芪甲苷、丹参素、原儿茶醛、人参皂苷Rg₁和Rb₁进行含量测定。结果 黄芪甲苷、丹参素、原儿茶醛、人参皂苷Rg₁和Rb₁分别在10.72~536.0,89.10~4 455,13.37~668.7,22.60~1 130,23.55~1 177 ng·mL内与峰面积呈良好线性关系。芪参益气滴丸中黄芪甲苷、丹参素、原儿茶醛、人参皂苷Rg₁和Rb₁的平均加样回收率均介于98%~102%之间。结论 本法简单、快速、灵敏、准确,可用于芪参益气滴丸中黄芪甲苷、丹参素、原儿茶醛、人参皂苷Rg₁和Rb₁含量测定,为该药的质量控制提供依据。     

人参皂苷Rg3和大黄酸纳米乳的制备及其联合aPD-L1治疗三阴性乳腺癌的药效学研究

目的 根据中医治疗肿瘤时提倡的“扶正祛邪”治则，选用中药人参和大黄的主要成分人参皂苷Rg₃(ginsenoside Rg₃,G-Rg₃)和大黄酸(rhein,Rhe)进行组合，制备G-Rg₃/Rhe纳米乳(G-Rg₃/Rhe NE)，同时提高G-Rg₃和Rhe溶解度以利于注射给药，并利用该纳米乳联合程序性死亡受体配体1单抗(anti-programmed cell death ligand 1,a PD-L1)治疗三阴性乳腺癌(triple negative breast cancer,TNBC)。方法 采用紫外分光光度法分别测定G-Rg₃和Rhe在不同辅料中的溶解能力，筛选出合适的乳化剂、助乳化剂和油相。利用伪三元相图考察乳化剂与助乳化剂质量比(Km值)对纳米乳体系的影响，根据形成纳米乳区域面积大小选择适宜的Km值，优选G-Rg₃/RheNE的处方，并对其进行表征和稳定性评价。选用4T1Fluc原位乳腺肿瘤小鼠...     

���ں����������������ɷֺ���������ɷ��о�

??OBJECTIVE To establish an HPLC-UV method for the quantitative determination of ginsenosides Rg₁, Re, Rf, Rb₁, Rc, Rb₂ and the qualitative determination of ginsenosides Rb₃ and Rd in Red Ginseng. This method is used to make different between the imported Red Ginseng and China Red Ginseng.METHODS The analysis were performed on a YMC-Pack ODS-A column(4.6 mm??100 mm,3 ??m), the mobile phase was acetonitrile -0.1% phosphoric acid at the flow rate of 0.6 mL??min^-1, the detection wavelength was set at 203 nm,the column temperature was maintained at 30 ??.RESULTS The method herein is effective. The data of samples was subjected to t test and principal component analysis(PCA)in order to find the marker constituents. According to the information of t test and PCA,ginsenoside Rg₁ and ginsenoside Rd were the main factor to classify Korean Red Ginseng and Chinese Red Ginseng.CONCLUSION The peak area ratio of ginsenoside Rg₁ to ginsenoside Rd is used as the quality control parameters. This method is suitable to classify Korean Red Ginseng and Chinese Red Ginseng.     

�˲�����Rg1�ԡ�Ѫ΢͸��̽��������������о�

??OBJECTIVE To establish a measuring method for microdialysis probe recovery of ginsenoside Rg₁ and investigate the effects of flow rate, concentration and using times of probe on the recovery in vivo and in vitro. METHODS Dialysis method and retrodialysis method were used for the study. The concentration of ginsenoside Rg₁ in brain and blood dialysate was determined by LC-MS/MS and the probe recovery was calculated. RESULTS The recoveries of brain and blood microdialysis probes showed good stability within 10 h, with average values of 17.0% and 34.4% respectively for ginsenoside Rg₁ at 1.5 ??L??min^-1. Concentrations (50,200,500,1 000 ng??mL^-1) had no obvious effect on recovery. At the same concentration, the recovery of brain and blood probes for ginsenoside Rg₁ decreased with the increase of flow rate (0.5, 1.0, 1.5, 2.0, 3.0 ??L??min^-1) in vitro and in vivo. The dialysis recoveries of brain and blood probes in vitro were (40.6??4.3)%, (23.5??2.3)%, (17.7??0.8)%, (12.2??1.1)%, (8.8??0.6)% and (70.6??3.6)%, (46.0??2.1)%, (32.9??1.6)%, (25.6??0.7)%, (18.2??1.3)%, respectively. The recoveries of dialysis and retrodialysis in vitro were approximately equal, and the recovery detected by retrodialysis in vivo was similar with the in vitro results. Probe used for no more than 3 times still kept high transmittance by flushing with 2% heparin sodium and ultrapure water successively. CONCLUSION Retrodialysis method can be used to study brain and blood probe recovery in vivo, and microdialysis can be used for simutaneous pharmacokinetic studies of ginsenoside Rg₁ in intercelluar fluid and blood.     

HPLC-ELSD�ⶨ���������ĵ��������˯����Ч������5��������ɷֵĺ���

??OBJECTIVE To establish a method for content determination of ginsenoside Rg₁, ginsenoside Rb₁, saikosaponin a, saikosaponin d, and saikosaponin B₂ in 70% ethanol elution effective fraction of Chaihu plus Longgu Muli decoction by HPLC-ELSD. METHODS A Diamonsil C₁₈ column(4.6 mm??250 mm, 5 ??m) was used as the stationary phase and the mobile phase consisted of acetonitrile and water. Gradient elution was carried out at the flow rate of 1.0 mL??min^-1. The column temperature was maitained at 30 ??. The ELSD detector was operated at 105 ?? with nebulizing gas at the optimum flow rate of 2.5 L??min^-1. RESULTS The average contents of ginsenoside Rg₁, ginsenoside Rb₁, saikosaponin a, saikosaponin d, and saikosaponin B₂ were 0.474%, 1.372%, 1.554%, 0.883%, and 2.073%, respectively. The calibration curves were linear in the ranges of 1.820-9.10, 1.810-9.050, 1.130-10.170, 0.420-2.100, and 3.125-15.625 ??g, respectively. The RSDs of precision, reproducibility and recovery were all less than 3.0%. CONCLUSION The method is rapid, simple, reliable and accurate, and has been successfully used to the quantification of five components in 70% ethanol elution effective fraction of Chaihu plus Longgu Muli decoction, which can provide a basis for the quality evaluation of Chaihu plus Longgu Muli decoction.     

��ЧҺ��ɫ�׷�ͬʱ�ⶨ�˲ν�Ƣ�����˲�����Rg1��Re��Rb1������������A��B�ĺ���

??OBJECTIVE To establish an HPLC method for simultaneous determination of ginsenoside Rg₁, Re, Rb₁ and jujuboside A, B in Renshen Jianpi Pellets (RSJPW). METHODS The determination was conducted on an Eclipse XDB C₁₈ column (4.6 mm??250 mm, 5 ??m) with the mobile phase of acetonitrile (A)-water (B) gradiently eluted at the following flow rates:0-90 min, 1.0 mL??min^-1; 90-130 min, 1.0-0.5 mL??min^-1; 130-140 min, 0.5 mL??min^-1. And the column temperature was maintained at 35 ??; the detection wavelength was set at 203 nm. RESULTS The calibration curves of ginsenoside Rg₁, Re, Rb₁ and jujuboside A, B were in good linearity over 0.32-2.24 ??g (r=0.998 2), 0.16-1.12 ??g (r=0.995 3), 0.32-2.24 ??g (r=0.999 6), 0.16-1.12 ??g (r=0.991 5), 0.08-0.56 ??g (r=0.999 6). The corresponding average recovery rates were 97.18%, 97.62%, 98.79%, 98.48%, 94.51%; the standard deviations were 1.10%, 0.98%, 0.34%, 1.09%, 1.88%, respectively. CONCLUSION The established HPLC method is accurate, reliable and reproducible, which can be used as a reference for the quality control of RSJPW.     

������Һ��ɫ��-�����ļ���-�������������׷������˲κͺ����������ɷ�

??OBJECTIVE To develop a comprehensive analytical method based on UFLC-QTRAP-MS/MS for simultaneous determination of protopanaxadiol [ginsenoside Rb₁, Rc, Rb₂, Rd, F₂, 20(S)-Rg₃, 20(R)-Rg₃, CK], protopanaxatriol [ginsenoside Re, Rg₁, Rf, 20(S)-Rg₂, 20(S)-Rh₁, 20(R)-Rg₂, 20(R)-Rh₁, F₁] and oleanolic(ginsenoside Ro) in Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra. METHODS Under the optimized chromatographic conditions, good separation for seventeen target compounds was obtained on a Synergi^TM Hydro-RP 100?_ column(2.1 mm??100 mm, 2.5 ??m) at 40 ?? with 0.1% aqueous formic acid (A)/acetonitrile (B) as the mobile phase by gradient elution at a flow rate of 0.4 mL??min^-1. The target compounds were analyzed under multiple reaction monitoring (MRM) mode with an ESI source operated in negative ion mode, and principal component analysis (PCA) and hierarchical cluster analysis (HCA)were used for data processing. RESULTS The calibration curves of the 17 components had good linearity (r>0.999 0). The precision, repeatability and stability were all satisfying.The average recoveries of standard addition for the compounds were between 96.69% and 102.01%,and the relative standard deviations were less than 5%. The results of PCA and HCA showed that Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra were clearly distinguished.The main compositions with significant difference were ginsenoside 20(S)-Rg₃, 20(R)-Rg₃, 20(S)-Rh₁, 20(R)-Rh₁, and 20(R)-Rg₂. CONCLUSION The established method could provide a new technique for the comprehensive evaluation and quality control of Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra, at the same time, it would pave the way for discovering the material basis contributing to the different properties and efficacies of the two medicinal materials.     

芪黄明目胶囊对糖尿病小鼠视网膜保护作用及对VEGF表达的影响

目的:观察芪黄明目胶囊对Ⅱ型糖尿病小鼠视网膜的保护作用及对VEGF表达的影响.方法:40只KK/UpjAy小鼠随机分为模型组、芪黄明目高、中、低剂量组(8.32,4.16,2.08 g·kg-1),另设10只C57BL/6小鼠对照组.灌胃给药3个月,观察一般情况,测定空腹血糖(FBG)及糖化血红蛋白(HbA1c);光镜及电镜观察视网膜形态学变化;Real-time PCR(qPCR)和Western blot法测定视网膜血管内皮生长因子(VEGF)、血管内皮生长因子受体-1(Flt-1)、血管内皮生长因子受体-2(Flk-1)表达.结果:芪黄明目胶囊能不同程度改善模型小鼠症状,降低FBG和HbAlc;改善视网膜病理损伤;降低视网膜VEGF,Flt-1,Flk-1 mRNA和蛋白的表达.结论:芪黄明目胶囊可抑制糖尿病小鼠视网膜VEGF,Flt-1,Flk-1的表达,干预VEGF-VEGFR信号转导通路,起到保护视网膜的作用.     

基于NF-κB/NLRP3信号通路和肺泡巨噬细胞活化研究肺肠合治法抑制炎症反应治疗急性肺损伤的作用机制

中医肺肠合治法治疗急性肺损伤疗效显著,该研究以体现肺肠合治的麻黄汤和大承气汤联合应用,探究肺肠合治法经核转录因子-κB(nuclear factor kappaB,NF-κB)/NOD样受体家族3(nucleotide binding oligomerization domain-like receptors-3,NL...     

Һ�����÷��ⶨ������ͨ������6����Ч�ɷֵĺ���

??OBJECITVE To establish an HPLC-MS/MS method for simultaneous determination of six active components in Dengyinnaotong capsules, ie, scutellarin, bilobalide, ginkgolide A, ginkgolide B, ginsenoside Rg₁ and notoginsenoside R₁. METHODS The chromatographic separation was carried out at 30 ?? on a Phenomenex Luna C₁₈ (4.6 mm??150 mm, 5 ??m) column eluted by gradient program. The flow rate was 0.3 mL??min^-1. The six compounds were separated within 10.0 min. RESULTS The regression curves for the six compounds showed good linearity in wide ranges. The recoveries were around from 94.8% to 108.5%. CONCLUSION The established method is accurate, reliable, specific and reproducible, which can be used for the quality control of Dengyinnaotong Capsules.