Paeoniflorin protects HUVECs from AGE-BSA-induced injury via an autophagic pathway by acting on the RAGE

The aim of our study was to investigate the protective effects of Paeoniflorin (PF) against injury induced by AGE-modified bovine serum albumin (AGE-BSA) in human umbilical vein endothelial cells (HUVECs), and to examine the underlying mechanisms of these effects. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to determine cell viability. Protein expression levels were determined by western blotting. For function-blocking experiments, we used small interfering RNA molecules (siRNA) for function-blocking experiments. At 6 h, we found that 100 μg/mL AGE-BSA reduced the viability of HUVECs. However, pretreatment with PF restored cell viability in a dose-dependent manner. AGE-BSA increased the levels of microtubule-associated protein light chain 3-II (LC3-II) and the receptor for advanced glycation end products (RAGE). Expression of p62 protein was also increased, but not at a statistically significant level. Pretreatment with PF further increased levels of LC3-II and RAGE, but reduced the expression of p62. In cells transfected with Atg5 and RAGE siRNA, cell viability and expression of LC3-II decreased in both the AGE-BSA and PF + AGE-BSA treatments. PF can protect HUVECs from AGE-BSA-induced injury by upregulating autophagy and promoting the completion of autophagy flux. RAGE plays an important role in this autophagic protection effect.     

养阴退热颗粒提取物中用高效液相色谱法测芍药苷含量

目的:建立一个采用高效液相色谱法测定养阴退热颗粒提取物中芍药苷含量的方法。方法:Odshypersil色谱柱(4.6 mm×250 mm,5μm),乙腈-0.1%磷酸溶液洗脱,柱温度32℃,流量0.8 m L/min,检测波长230 nm。结果:芍药苷在在5~200μg/m L范围内线性关系良好,平均回收率为99.83%,RSD为0.76%,A类不确定度Ua=0.003417.结论:3批样品的抽检结果显示,该方法准确、方便、重复性好,可用于测定芍药苷在养阴退热颗粒提取物中的含量。     

Antidepressant-like effects of paeoniflorin on post-stroke depression in a rat model

ABSTRACT

Background: Post-stroke depression (PSD) is one of the most prevalent emotional disorders after stroke and often results in poor outcomes. However, the underlying physiopathologic mechanism and effective treatment of PSD remain poorly elucidated.

Objective: To investigate whether paeoniflorin has antidepressant-like activity in a rat model of PSD.

Methods: Rats were randomly divided into four groups: sham-operated control (Sham), PSD, paeoniflorin (with PSD) and fluoxetine group(with PSD). PSD was developed by the right middle cerebral artery occlusion followed 21 days chronic unpredictable mild stress combined (CUMS) with raised alone. Tests of sucrose preference and open field were used to assess the depression-like behavior. Neurological function was evaluated by neurological deficit score and beam balance test. Expression of phosphorylated CREB (p-CREB) and brain-derived neurotrophic factor (BDNF) in the CA1 region of the hippocampal complex was evaluated by western blot and immunofluorescence.

Results: Te depressive-like behaviors markedly improved after paeoniflorin and fluoxetine treatment. Furthermore, paeoniflorin treatment significantly increased BDNF and p-CREB expression in the CA1 region.

Conclusions: Observed results suggested that paeoniflorin could ameliorate the symptoms and improve the functional capability of PSD rats, similar to the effect of fluoxetine.

Abbreviations: PSD: post-stroke depression; CUMS: chronic unpredictable mild stress stimulation; MCAO: middle cerebral artery occlusion; OFT: open field test; SPT: sucrose preference test, NDS: neurological deficit score, BBT: beam balance test; BDNF: brain-derived neurotrophic factor protein; p-CREB: phosphorylated Cyclic-AMP responsive element binding protein     

不同生产厂家赤芍配方颗粒中单萜苷类化合物含量比较

目的比较不同厂家生产的赤芍配方颗粒中单萜苷类化合物(吡啶芍药苷、牡丹皮苷F、氧化白芍苷、氧化芍药苷、10-羟基芍药苷、白芍苷、芍药苷、oxypaeonidanin、4-甲氧基-氧化芍药苷、没食子酰基芍药苷、4-甲氧基芍药苷、白芍苷R_1、paeonidanin、苯甲酰氧化芍药苷、苯甲酰芍药苷)的含量差异,为制定统一的质量标准奠定基础。方法采用Zorbax SB-Aq C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈和磷酸二氢钾缓冲盐(pH 2.8)溶液,梯度洗脱,体积流量1.0 mL/min,柱温30℃,检测波长260 nm。结果芍药苷、白芍苷和氧化芍药苷是赤芍配方颗粒中含量最高的3种单萜苷类化合物,且不同厂家生产的赤芍配方颗粒中主要单萜苷类化合物的含量差异较大。样品CSPFKL-KRT中芍药苷和氧化芍药苷的含量最高,样品中质量分数分别为73.214mg/g和16.935mg/g,白芍苷的质量分数最低,为2.343mg/g。而样品CSPFKL-XLS中芍药苷和氧化芍药苷的质量分数最低,样品中分别为26.327 mg/g和4.165 mg/g,白芍苷的质量分数最高18.893 mg/g。结论不同厂家生产的赤芍配方颗粒中主要单萜苷类化合物的含量差异较大,可能会对临床应用产生影响。建立统一的质量标准对于赤芍配方颗粒的质量控制具有重要作用。     

基于信息熵理论的正交设计优化消乳增胶囊的提取工艺

目的优选消乳增胶囊中7味药材的提取工艺参数。方法采用HPLC测定芍药苷、阿魏酸、橙皮苷、欧前胡素、藁本内酯的含量,以此5种指标成分的提取率、药材提取的干膏得率和提取物指纹图谱相似度为综合评价指标,通过正交试验考察乙醇浓度、提取溶剂倍量、提取时间和提取次数对提取工艺的影响,采用信息熵赋权法确定各指标的客观权重,优选消乳增胶囊中7味药材的提取工艺参数。结果根据综合评分结果,确定该制剂最佳提取工艺为选用6倍量的50%乙醇,煎煮2次,每次2 h。3批验证综合评分均值为99.72,RSD为0.24%。结论优选后的工艺提取率高、稳定性和重复性好,适用于该制剂的大批量生产。     

HPLC法同时测定乙肝益气解郁颗粒中的11种成分

目的建立HPLC法同时测定乙肝益气解郁颗粒中柴胡皂苷a、柚皮苷、芍药苷、毛蕊异黄酮苷、丹参酮IIA、桂皮醛、五味子醇甲、紫丁香苷、盐酸小檗碱、大黄酚和橙皮苷的含量,并采用主成分分析(PCA)法对其质量进行综合评价。方法采用HPLC法,色谱柱为Caprisil C18-AQ(250 mm×4.6 mm,5.0μm);流动相为0.1%磷酸水溶液-乙腈,梯度洗脱,体积流量0.8mL/min;柱温45℃。最后将定量结果与PCA法相结合对不同批次药物进行科学的质量评价分析。结果乙肝益气解郁颗粒中柴胡皂苷a、柚皮苷、芍药苷、毛蕊异黄酮苷、丹参酮IIA、桂皮醛、五味子醇甲、紫丁香苷、盐酸小檗碱、大黄酚和橙皮苷11种成分分别在1.6～80.0、14～700、10～500、1.6～80.0、1.6～80.0、2.4～120.0、1.2～60.0、1.2～60.0、8.0～400.0、2.0～100.0、2.0～100.0μg/m L线性关系良好;平均加样回收率分别为98.3%、99.2%、98.8%、99.3%、101.9%、97.5%、99.8%、101.7%、101.1%、102.5%、100.9%,RSD均2.0%;16批样品中11种有效成分的质量分数分别为0.233～0.322、3.007～3.142、2.201～2.273、0.320～0.355、0.317～0.399、0.451～0.523、0.265～0.297、0.209～0.226、1.848～1.873、0.380～0.425、0.615～0.647mg/g。结论实验建立的方法简便准确、重复性好,可用于乙肝益气解郁颗粒的质量控制,为乙肝益气解郁颗粒后续质量提高提供参考。     

四逆散干预焦虑症作用机制的网络药理学分析

目的探讨四逆散干预焦虑症的功效作用网络及药效作用机制。方法运用网络药理学方法,多个数据库联合分析四逆散和焦虑症的作用靶点,通过Cytoscape筛选靶点并对其基因功能和信号通路进行分析,运用Bio GPS数据库对所得靶点进行器官定位。结果分析结果表明槲皮素、山柰酚、芍药苷等成分作用于SRC、EGFR、PIK3R1等靶点,通过ERBB2信号通路、EGFRv III信号通路、VEGFA-VEGFR2信号通路等共同发挥作用,器官定位结果表明作用的核心病位为肝。结论四逆散干预焦虑症体现了中药多成分、多靶点、多通路的作用特点。     

经典名方当归四逆汤物质基准量值传递分析

目的 建立经典名方当归四逆汤物质基准的HPLC特征图谱及指标性成分含量测定方法,阐明当归四逆汤物质基准关键质量属性,为后续复方制剂的开发奠定基础。方法 制备18批当归四逆汤物质基准,建立HPLC特征图谱,进行特征峰归属及相似度评价;测定指标性成分的含量,对物质基准进行量值传递分析。结果 18批物质基准的特征图谱的相似度均大于0.90,共确定19个特征峰,分别来自方中当归（峰4、5、19）、桂枝（峰10、11、14）、白芍（峰6、9）、细辛（峰1、3、13、15、18）和炒甘草（峰7、8、12、16、17）;各指标成分从饮片到物质基准的转移率分别为芍药苷78.47%～96.01%、甘草苷43.79%～94.62%、阿魏酸81.20%～131.24%、肉桂酸70.03%～106.75%、桂皮醛3.17%～5.50%、甘草酸31.08%～52.43%、细辛脂素9.63%～13.37%。结论 采用特征图谱结合多指标性成分含量测定对当归四逆汤物质基准进行量值传递研究,该方法科学合理,为当归四逆汤物质基准的质量控制及复方制剂的开发提供科学依据。     

HPLC法测定十味桂芪真武颗粒中芍药苷的含量

目的：建立十味桂芪真武颗粒中芍药苷含量测定方法。方法采用高效液相色谱法对方中芍药苷含量进行测定。结果芍药苷对照品进样量在0．523～8．368μg范围内线性关系良好。方法的平均回收率为97．41％（n=6）,RSD=1．54％。结论该法快速,准确,方法稳定性、重现性好。可以用于控制十味桂芪真武颗粒中芍药苷的含量。     

用高效液相色谱法测定安坤颗粒中栀子苷和芍药苷的含量

目的 建立HPLC法同时测定安坤颗粒中栀子苷和芍药苷的含量.方法 采用安捷伦Eclipse XDB C18色谱柱（250 mm×4.6 mm,5μm）,流动相：乙腈-水（14∶86）,检测波长：230 nm,柱温：35℃.结果 栀子苷在0.071 52 ～0.357 6 μg范围内线性关系良好（r=0.999 8）,平均回收率为98.22％,RSD=1.25％.芍药苷在0.0719 7 ～0.359 85 μg范围内线性关系良好（r=0.9992）,平均回收率为98.40％,RSD=0.68％.结论 该方法操作简便,结果准确,重复性好,可用于安坤颗粒的质量控制.