Objective: To reveal the pharmacodynamic material basis and mechanism of Pogostemon cablin in the intervention of viral pneumonia based on serum pharmacochemistry and network pharmacology. Methods: The components in P. cablin decoction, rat blank serum and drug-containing serum were analyzed by ultra-performance liquid chromatography-quadrupole/orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS）, and the absorbed components of P. cablin in serum were identified by Compound Discoverer 3.1 in combination with MS/MS profiles and existing literature. The targets of the absorbed components of P. cablin were retrieved from SwissTargetPrediction, and the target genes of viral pneumonia were searched from GeneCards, DisGeNET and Online Mendelian Inheritance in Man（OMIM） by input of the keyword viral pneumonia. Cytoscape 3.9.1 was employed to construct the protein-protein interaction（PPI） network and the key targets were screened, followed by Gene Ontology（GO） function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis of the intersection targets by Database for Annotation, Visualization and Integrated Discovery（DAVID）. Results: Twelve compounds were identified from rat serum,including 7 prototype components（luteolin-7-glucuronide, apigenin-7-O-glucuronide, eriodictyol-7, 3 ′-dimethyl ether,genkwanin, pogostone, 4 ′,7-di-O-methylnaringenin, dibutyl phthalate） and 5 metabolites. Serine/threonine kinase 1（Akt1）,tumor necrosis factor（TNF）, epidermal growth factor receptor（EGFR） and Caspase-3（CASP3） were the hub genes in PPI network. Enrichment analysis revealed that the absorbed components of P. cablin exerted the antiviral and anti-inflammatory effects via regulating phosphoinositide 3-kinase（PI3K）-Akt, Kaposi sarcoma-associated herpesvirus infection, human immunodeficiency virus 1 infection, TNF, mitogen-activated protein kinase（MAPK）, Janus kinase（JAK）-signal transducer and activator of transcription（STAT） and interleukin-17（IL-17） signaling pathways. Conclusion: The absorbed components of P. cablin may play a role in the treatment of viral pneumonia by interfering with proteins and pathways related to antivirus, anti-inflammation, cell proliferation and apoptosis.

目的 观察电针预处理"足三里"穴和"尺泽"穴对脓毒症急性肺损伤(ALI)大鼠肺组织中NEK7-NLRP3炎症小体激活的影响,探讨电针预处理在脓毒症ALI中发挥的保护效应及可能机制.方法 将SD大鼠随机分为对照组、电针预处理+对照组、模型组、电针预处理+模型组,每组10只.各模型组采用腹腔注射脂多糖(LPS)的方法建立脓毒症ALI大鼠模型.各电针预处理组于LPS造模前1周进行连续7 d电针预处理,疏密波,频率4 Hz/20 Hz,强度1～2 mA,持续30 min.检测各组大鼠肺功能;HE染色法观察大鼠肺组织病理学变化;测定大鼠肺组织湿/干质量比值(W/D);ELISA法检测大鼠血浆及肺组织中炎症因子IL-1β、IL-18含量;免疫荧光观察大鼠肺组织中ASC蛋白阳性表达;Western blot法检测大鼠肺组织中NEK7、NLRP3、Caspase-1及IL-1β蛋白的表达.结果 与对照组相比,模型组大鼠用力肺活量(FVC)、第0.1秒用力呼气量(FEV0.1)、第0.3秒用力呼气量(FEV0.3)、FEV0.1/FVC、FEV0.3/FVC均显著降低(P<0.001);肺泡结构紊乱,肺组织内炎性细胞浸润明显,肺组织充血水肿;W/D比值显著升高(P<0.001);血浆及肺组织内IL-1β、IL-18含量明显升高(P<0.001);ASC蛋白阳性表达明显增多(P<0.001);肺组织中炎症小体相关蛋白NEK7、NLRP3、Caspase-1及IL-1β的表达水平均显著升高(P<0.001).与模型组相比,电针预处理+模型组大鼠FVC、FEV0.1、FEV0.3、FEV0.1/FVC、FEV0.3/FVC均明显升高(P<0.05,P<0.001);肺组织内炎症细胞浸润及充血水肿有明显改善;W/D比值显著降低(P<0.01);血浆及肺组织内IL-1β、IL-18含量显著降低(P<0.01,P<0.001);ASC蛋白阳性表达降低(P<0.001);肺组织中炎症小体相关蛋白NEK7、NLRP3、Caspase-1、IL-1β表达水平均显著降低(P<0.05,P<0.01,P<0.001).结论 电针预处理可以减轻肺部炎性反应,改善肺功能,其机制与电针抑制脓毒症ALI大鼠肺组织中NEK7-NLRP3炎症小体激活相关.

目的:探究何首乌中成分含量随生长年限和采收季节的变化规律.方法:建立超高效液相色谱-三重四极杆质谱法(UPLC-QQQ-MS/MS),测定德庆地区何首乌药材中 14 个成分反式-2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷(trans-THSG)、顺式-THSG(cis-THSG)、2,3,5,4'-四羟基二苯乙烯-2-O-(2"-O-阿魏酰)-β-D-吡喃葡萄糖苷、虎杖苷、白藜芦醇、儿茶素、表儿茶素、金丝桃苷、芦丁、没食子酸、对羟基苯甲酸、对羟基苯甲醛、对香豆酸和决明酮-8-O-β-D-葡萄糖苷含量.结果:建立了准确可靠的UPLC-QQQ-MS/MS测定何首乌中 14 个成分含量的方法;随生长年限的增加,何首乌中二苯乙烯苷类、黄酮类、酚类化合物含量多降低;随采收季节的变化,二苯乙烯苷类、黄酮类、决明酮-8-O-β-D-葡萄糖苷成分多春季含量最高、秋季含量最低,酚类化合物多夏季含量最高、春秋含量低.结论:成分的含量与何首乌药材生长年限及采收季节密切相关,其中采收季节变化较为明显;以trans-THSG、决明酮-8-O-β-D-葡萄糖苷作为潜在肝毒性物质,cis-THSG作为有效成分,秋季为何首乌的适宜采收期,与《中华人民共和国药典》2020 年版规定相符.

Objective: To revise and improve quality standards of Pogostemonis Herba medicinal materials and decoction pieces, further provide scientific basis for rational control of the quality. Methods: A total of 23 batches of Pogostemonis Herba and 32 batches of representative decoction pieces were collected. The leaf powder was microscopically identified with reference to the Chinese Pharmacopoeia（2020 edition）. The existing pre-treatment method for thin-layer chromatographic（TLC） identification was simplified to establish a new TLC identification using Pogostemonis Herba reference material and patchouli alcohol reference substance as the controls. Moreover, the sample preparation method for content determination was improved, and the limits of index components in medicinal materials were revised. The inspection on Pogostemonis Herba decoction pieces standard, leachate and content determination were added. Results: It was suggested that the microscopic identification of glandular squamous head cells should be described as having a fissured cell wall with multiple ruptures. Ultrasonic extraction with petroleum ether and re-dissolution with ethyl acetate were adopted instead of original extraction of volatile oil as a pretreatment in TLC. With 5% anisaldehyde concentrated sulfuric acid solution was used as the chromogenic agent, all samples, patchouli reference medicinal material and patchouli alcohol reference substance all showed the spots in the same color at the same position. The pretreatment was simplified after improvements, the development efficacy was good, and the spots were distinct. It was suggested that the limit of alcohol-soluble extracts of medicinal materials should be appropriately increased to no less than 4.0% when ethanol was used as the solvent. Anhydrous ethanol was used to replace the original toxic chloroform as the extraction solvent for content determination. The methodological investigation met with the requirements. The mass fractions and average value of patchouli alcohol in the 23batches of Pogostemonis Herba medicinal materials were 0.34%-0.92% and 0.58%, respectively. It was recommended that the limit of patchouli alcohol content in the medicinal materials should be increased to 0.22%. Moreover, new items for inspection were added for decoction pieces, including moisture content no higher than 14.0%, total ash content no higher than 11.0%, and acid insoluble ash content no higher than 4.0%. The item for leachate from decoction pieces was added and the limit was the same to that of the medicinal material standards. The item for content determination of decoction pieces was added, the mass fraction and average of patchouli alcohol in the 32 batches of decoction pieces were 0.12%-0.46% and 0.29%, respectively, and it was recommended that the content of patchouli alcohol should be no less than 0.20%. Conclusion:The method is simple, feasible, accurate, reliable, and highly reproducible. The new items and the standard limits are reasonable. They can be used for the quality control of Pogostemonis Herba medicinal materials and decoction pieces.

目的 探究《中华人民共和国药典》(2020 年版)收载的黑豆汁拌蒸炮制法,在不同炮制时间(0～48 h)下对何首乌中主要化学成分含量的影响,为何首乌用药安全提供参考.方法 制备药典黑豆汁拌蒸样品;建立超高效液相色谱-三重四极杆质谱分析方法(UPLC-QQQ-MS/MS),测定何首乌药材中24种成分含量.结果 建立UPLC-QQQ-MS/MS方法,测定何首乌中二苯乙烯苷、蒽醌、黄酮、酚类化合物含量;随着蒸制时间的延长,24种成分的含量发生了明显变化;多数二苯乙烯苷类、蒽醌类、黄酮类经长时间炮制含量降低,酚类成分随炮制时间延长含量增加.结论 本方法操作简单、准确度高、重复性好,可以用于炮制前后何首乌中 24 种化学成分的定量检测分析.制何首乌中 24种成分的含量与蒸制时间密切相关,蒽醌类、二苯乙烯苷类、黄酮类、酚类发生规律性变化,对何首乌毒副作用和补益功效产生重要影响.

In order to study the mechanism of tonifying kidney and strengthening Yang by Rougui（Cinnamomi Cortex）,the expression of hypothalamic-pituitary-target gland related mRNA and histopathological changes of kidney-Yang deficiency rats were investigated. The rats were divided into normal group（CON）,kidney-Yang deficiency model group（MOD）,low-dose Rougui（Cinnamomi Cortex） group（RGL） and high-dose Rougui（Cinnamomi Cortex） group（RGH）. Rats in RGH group and RGL group were given different doses of Rougui（Cinnamomi Cortex） extract(8.1 g·kg -1 ·d -1 and 4.05 g·kg -1 ·d -1 ) for 4 weeks, respectively. Rats in CON group and MOD group were given the same volume of normal saline. At the end of the experiment, the levels of serum biochemistry, hypothalamic-pituitary-target gland related organ index, plasma hormone, mRNA expression and the pathological changes of related tissues were measured. The results showed that the hypothalamus index, adrenal index, thymus index, thyroid index and testis index in MOD group were significantly lower than those in CON group（P＜0.01）. The levels of adrenocorticotropic hormone（ACTH）,corticosterone（CORT）, triiodothyronine（T3）, tetraiodothyronine（T4）, testosterone（T） and hypothalamic thyrotropin releasing hormone（TRH） mRNA, gonadotropin releasing hormone（GnRH） were significantly decreased（P＜0.05）. The level of plasma estradiol（E 2 ） and hypothalamic corticotropin-releasing hormone（CRH） mRNA expressions were significantly increased（P＜0.05）. The pathological changes of hypothalamus, adrenal gland, thymus and testis were also observed. Compared with the MOD group, RGL group and RGH group showed significant regression（P＞0.05）. The pathological damage has also abated. In this experiment, the regulation of Rougui（Cinnamomi Cortex） on kidney-Yang deficiency syndrome is related to the regulation of hypothalamus-pituitary-target gland axis. This study may reveal the mechanism of Rougui（Cinnamomi Cortex） tonifying kidney and lay a foundation for clinical prevention and treatment of kidney-yang deficiency syndrome.

Objective: To investigate the differences in metabolic kinetics between monomeric magnoflorine and magnoflorine in Coptidis Rhizoma water extract. Methods: The linkage rat model of "intestinal penetrating fluid-liver microsome incubation" in vitro was adopted to study the metabolic kinetic characteristics of magnoflorine in vitro liver microsome and to identify the metabolites by high performance liquid chromatography-linear ion trap/electrostatic field orbitrap high resolution mass spectrometry（HPLC-LTQ/Orbitrap MS）. The metabolic pathways of magnoflorine were analyzed, and the interaction between magnoflorine and coexisting components in Coptidis Rhizoma was investigated.Results: The Michaelis constant [K m ,（0.53±0.16） μmol·L –1 ] of magnoflorine in Coptidis Rhizoma water extract was lower than that [（0.85±0.12） μmol·L –1 ] of magnoflorine monomer（P＜0.05）, which indicated that the affinity of magnoflorine in Coptidis Rhizoma water extract in the rat liver microsome was higher than that of magnoflorine monomer. The maximum reaction velocity(V max ), hepatic intrinsic clearance(CL int ), and hepatic clearance（CL） of magnoflorine in Coptidis Rhizoma water extract were lower than those of magnoflorine monomer（P＜0.05）, and the half-time(T 1/2 ) of magnoflorine in Coptidis Rhizoma water extract（44.22 min） was higher than that（37.35 min） of magnoflorine monomer（P＜0.01）, suggesting that the metabolism and elimination rates of magnoflorine in Coptidis Rhizoma water extract in the rat liver microsome were slower,and the half-life period was longer. The results indicated that monomeric magnoflorine and magnoflorine in Coptidis Rhizoma water extract had different metabolites. Five metabolites were found in monomeric magnoflorine, while eight metabolites were found in magnoflorine in Coptidis Rhizoma water extract. The pathways to these metabolites were hydroxylation, demethylation, dehydrogenation, ketonization, glucosylation, and glucuronidation. Mono-hydroxylation and mono-demethylation metabolites of magnoflorine in Coptidis Rhizoma water extract in the intestinal penetrating fluid could be further metabolized in the rat liver microsome. Conclusion: Magnoflorine can metabolize in both intestinal tract and liver microsomes. Coexisting components in Coptidis Rhizoma reduce the metabolic rate of magnoflorine in the rat liver microsome and increase the number of its metabolites.

目的 评价紫苏籽油与亚麻籽油合用的安全性及降血脂作用.方法 通过急性经口毒性试验、Ames试验、骨髓细胞微核试验、小鼠精子畸形试验和大鼠30天喂养试验对紫苏籽油与亚麻籽油合用的安全性进行考察;通过建立大鼠混合型高脂血症模型,经口灌胃不同剂量(250、750、1 500 mg/kg.BW)的紫苏籽油与亚麻籽油混合物连续30天,测定血脂指标,对紫苏籽油与亚麻籽油合用的降血脂作用进行考察.结果 安全性试验结果显示,紫苏籽油与亚麻籽油合用对大、小鼠急性经口 MTD值均＞15.0 g/kg.BW,属无毒级;Ames试验、骨髓细胞微核试验、小鼠精子畸形试验结果均为阴性;30天喂养试验未引起动物中毒性损伤改变.降血脂试验结果显示,与模型对照组比较,紫苏籽油与亚麻籽油合用的三个剂量组的总胆固醇(TC)、甘油三酯(TG)显著降低(F = 4.405,P = 0.010;F=4.601,P = 0.008),降血脂作用明显.结论 紫苏籽油与亚麻籽油合用的安全性高,无致突变性及亚急性毒性,并且具有较好的降血脂作用.

OBJECTIVE:To observe the effect of electroacupuncture (EA) at "Zusanli" (ST36) and "Feishu" (BL13) on the activation and secretion of calcitonin gene-related peptide (CGRP) and 5-hydroxytryptamine (5-HT) of pulmonary neuroendocrine cells (PNECs) and inflammatory response in rats with chronic obstructive pulmonary disease (COPD), so as to explore its underlying mechanisms in treating COPD.METHODS:Male SD rats were randomly divided into normal control, COPD model and EA groups, with 7 rats in each group. The COPD model was established by forced inhale of cigarette smoke for 1 h in a self-made box (1 m×1 m×1 m in volume), twice daily for 12 weeks. EA (4 Hz/20 Hz, 1-3 mA) was applied at bilateral ST36 and BL13 acupoints for 30 min, once a day for 14 consecutive days. The pulmonary function including the forced vital capacity (FVC), forced expiratory volume at 0.1 second (FEV0.1), FEV0.3, FEV0.1/FVC and FEV0.3/FVC was detected using a lung function analyzer for small animals. The lung tissue was sampled for observing histopathological changes by using H.E. staining, for observing expression and distribution of PNECs by Grimelius silver staining, and for detecting the immunoactivity (integrated optical density) of CGRP and 5-HT by using immunohistochemistry. The contents of CGRP, 5-HT, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and transforming growth factor-β1 (TGF-β1) in the bronchoalveolar lavage fluid (BALF) and lung tissue were detected by ELISA, and the correlations between TNF-α and CGRP, IL-1β and CGRP, TNF-α and 5-HT, and IL-1β and 5-HT levels were analyzed. The mRNA and protein expression levels of nerve fiber markers of CGRP and purinergic receptor P2X ligand gated ion channel 3 (P2X3) which dominate PNECs in the lung tissue were detected by real-time fluorescence quantitative PCR and Western blot, respectively.RESULTS:Compared with the normal control group, the levels of FVC, FEV0.1, FEV0.3, and the ratios of FEV0.1/FVC and FEV0.3/FVC were significantly decreased (P<0.05, P<0.01), while the immunoactivity of PNECs, CGRP and 5-HT, the contents of CGRP, 5-HT, TNF-α, IL-1β and TGF-β1 in the BALF and lung tissue, and the expression levels of CGRP and P2X3 mRNAs and proteins in the lung tissue significantly increased in the COPD model group (P<0.01, P<0.05). Following EA intervention, both the increased and decreased levels of all the indexes mentioned above were reversed (P<0.05, P<0.01) except FEV0.3. H.E. staining showed severe deformed bronchial lumen with thickened wall and alveolar septum, and obvious inflammatory cell infiltration and reduced number of alveolar lumen fusion in the COPD model group, which was mild in the EA group. A positive correlation was found between TNF-α and CGRP, IL-1β and CGRP, TNF-α and 5-HT,IL-1β and 5-HT levels in both BALF and lung tissues (P<0.01).CONCLUSION:EA at ST36 and BL13 can improve lung function and reduce inflammatory response in COPD rats, which may be related to its function in inhibiting the activation of PNECs and release of neuroactive substances.

目的 分析探讨何首乌肝毒性作用机制及肝毒性物质基础.方法 通过查阅何首乌文献对肝毒性机制和争议成分相关报道进行整理、分析、归纳.结果 何首乌的肝毒性具体表现在影响肝脏代谢酶的功能与表达、使胆汁代谢循环及转运蛋白的功能受阻、以及免疫应激与线粒体功能紊乱等,文献报道其可能的毒性成分为脂溶性蒽醌类、特征性二苯乙烯苷类、鞣质类和二蒽酮类化合物,推测上述物质对机体的单独或协同作用可能与肝毒性发生有关.结论 何首乌肝毒性机制可能众多,相关研究可为何首乌现代研发和安全运用奠定理论基础.

目的 采用多指标结合化学计量学的方法综合分析龙葵Solanum nigrum果质量及关键指标,为其质量评价提供科学依据.方法 采用HPLC法建立龙葵果特征指纹图谱及澳洲茄碱、澳洲茄边碱含量测定方法,并测定浸出物含量,使用SAS14.0、SIMCA14.1 软件对上述指标进行聚类分析(hierarchical clustering analysis,HCA)、主成分分析(principal component analysis,PCA)及偏最小二乘判别分析(discriminant analysis of partial least squares analysis,PLS-DA),通过对吉林产地 25 批不同产区、不同采收期药材的分析,综合评价龙葵果质量及标志性成分.结果 建立的特征指纹图谱及澳洲茄碱、澳洲茄边碱含量测定方法稳定性较好.HCA分析可将25批样品聚为2大类,分类主要与产区有关,Ⅰ类药材为铁东区及梨树县的孤家子、榆树台产区;Ⅱ类药材为梨树县的杏山、北夏家、泉眼沟、獾子洞产区.用3个主成分对龙葵果药材进行综合评价,结果表明Ⅰ类药材综合得分及排名较高,质量较优.根据PLS-DA分析,造成两类药材质量差异的指标为醇溶性浸出物含量及指纹图谱相似度.色谱峰2、3(澳洲茄碱)、4(澳洲茄边碱)、14、15(薯蓣皂苷元)、16所代表的成分是2类龙葵果药材质量差异的标志性物质.结论 建立的龙葵果质量综合评价方法操作简便、重复性好、稳定可靠,可为龙葵果质量评价提供基础.

目的 探究不同采收季节和生长年限对广东省肇庆市德庆地区何首乌蒽醌类化合物含量的影响.方法 采用超高效液相色谱-三重四极杆质谱(UPLC-MS/MS),测定何首乌中大黄素-1-O-葡萄糖苷、大黄素-8-O-β-D-葡萄糖苷、大黄素-8-O-(6'-甲基丙二酰)吡喃葡萄糖苷、大黄素-6-O-葡萄糖苷、大黄素甲醚-8-O-β-D-葡萄糖苷、大黄素甲醚-8-O-(6'-甲基丙二酰)吡喃葡萄糖苷、大黄素-1-甲醚、大黄素、大黄素甲醚共9种蒽醌类成分的含量,归纳总结其随采收季节和种植年限的变化趋势.结果 秋季采收样品9种蒽醌总量介于1465.82～1707.76μg·g-1之间,远低于春季(2826.56～3100.61μg·g-1)和夏季(3616.38～5777.23μg·g-1)采收样品;除大黄素甲醚-8-O-β-D-葡萄糖苷和大黄素甲醚-8-O-(6'-甲基丙二酰)吡喃葡萄糖苷的含量随季节变化呈现逐渐下降趋势外,其余7种成分春夏升高、入秋后急剧降低;种植3～4年后,何首乌中大黄素、大黄素-8-O-β-D-葡萄糖苷等成分出现含量谷值;秋季作为传统采收季节,其4年生样品中大黄素-8-O-β-D-葡萄糖苷和大黄素甲醚-8-O-β-D-葡萄糖苷在4年生何首乌中含量最高.结论 采收季节对蒽醌类成分含量的影响大于生长年限;以蒽醌类成分作为肝毒性指标,秋季采收样品含量低于春夏采收样品,为适宜采收期,与药典规定相符;3～4年生何首乌蒽醌类化合物含量较低,其含量变化规律可指导药材的种植采收.

目的 考察连续42 d灌胃何首乌水提物(PMW)后大鼠肝、肾、血浆和胆汁中的成分累积特征,为何首乌肝毒性研究提供数据支撑.方法 采用UPLC-LTQ-Orbitrap MS技术,通过与对照品比对及通过scitific library模块分析,对大鼠不同组织中何首乌原型成分及代谢产物进行定性鉴别和推测.结果 大鼠经过长期灌胃(PMW)诱导后,体内可检测到24个何首乌相关化合物,其中7个为原型成分,5个为Ⅰ相代谢产物,12个为Ⅱ相代谢产物.主要成分为二苯乙烯苷和蒽醌类原型及其代谢物.各类成分具有选择蓄积性,肾脏中蓄积成分最多,肝脏和胆汁中相当,血浆中最少.二苯乙烯苷以硫酸化和葡萄糖醛酸化反应为主,大黄素多发生氧化反应和葡萄糖醛酸化反应.结论 原型成分大黄素、芦荟大黄素、大黄素-8-O-葡萄糖苷、二苯乙烯苷、大黄酸、没食子酸很可能与何首乌引发的不良反应相关,应引起关注;大黄素-葡萄糖醛酸化物和二苯乙烯苷-葡萄糖醛酸化物-硫酸化物仅在胆汁中检出,没食子酸及原儿茶酸以及大量大黄素Ⅱ相代谢物仅在肾脏中检出,这类成分的潜在毒性风险应进一步研究.

目的:旨在通过考察何首乌70％乙醇提物(PME)和水提物(PMW)在大鼠体内不同组织的成分蓄积差异,推测何首乌引起不良反应的原因.方法:建立大鼠不同组织中二苯乙烯苷、大黄素和大黄素-8-O-β-D-葡萄糖苷的高效液相色谱串联质谱法(UPLC-MS/MS)定量分析方法;并定量分析连续42 d灌胃PME和PMW后大鼠肝、肾、血浆和胆汁中3个成分的蓄积情况.结果:PME给药组大鼠体内不同组织中相同成分蓄积量明显高于PMW给药组.2个给药组中3个成分选择性蓄积的靶器官基本相同,二苯乙烯苷主要蓄积在肝脏中,大黄素主要蓄积在肾脏中,大黄素-8-O-β-D-葡萄糖苷主要蓄积在胆汁中,而血浆中3个成分的含量均较低.结论:何首乌中二苯乙烯苷、大黄素和大黄素-8-O-β-D-葡萄糖苷在大鼠体内的蓄积情况具有明显的组织选择性.与PMW给药组相比,PME组大鼠给药后体内蓄积成分含量高,可能是其导致临床不良反应的主要原因.其中,二苯乙烯苷大量蓄积于大鼠肝脏中,成为何首乌引发肝损伤的重要潜在毒性物质;大黄素-8-O-β-D-葡萄糖苷主要蓄积在胆汁中,有引发胆汁淤积的风险;大黄素主要蓄积部位为肾脏,其与肝毒性是否直接相关仍需进一步探讨.

目的 探究《中华人民共和国药典》(2020年版)(简称"《中国药典》")收载的清蒸炮制法,在不同炮制时间(0～48 h)下对何首乌中主要化学成分含量的影响.方法 制备《中国药典》清蒸样品;建立超高效液相色谱-三重四极杆质谱(UPLC-MS/MS)测定何首乌中26种成分的分析方法并进行检测.结果 建立了准确可靠的UPLC-MS/MS测定何首乌中蒽醌、二苯乙烯苷、黄酮、酚类化合物含量的方法;随着蒸制时间的延长,26种成分的含量发生了明显变化:游离蒽醌类成分(大黄素、大黄素甲醚)先降低后升高,结合蒽醌类成分(大黄素-8-O-β-D-葡萄糖苷、大黄素甲醚-8-O-β-D-葡萄糖苷、大黄素-1-O-葡萄糖苷、大黄素-6-O-葡萄糖苷)、大黄酸、大黄素-1-甲醚、cis-THSG、虎杖苷、表儿茶素和芦丁先升高后降低,大黄素二蒽酮、大黄素-8-O-(6'-甲基丙二酰)吡喃葡萄糖苷、大黄素甲醚-8-O-(6'-甲基丙二酰)吡喃葡萄糖苷、trans-THSG、白藜芦醇、2,3,5,4'-四羟基二苯乙烯-2-O-(2"-O-阿魏酰)-β-D-吡喃葡萄糖苷、儿茶素和决明酮-8-O-β-D-葡萄糖苷降低,酚类成分升高.结论 本方法操作简单、准确度高、重复性好,可以用于何首乌中26种化学成分的定量检测分析.制首乌中26种成分的含量与蒸制时间密切相关,蒽醌类、二苯乙烯苷类、黄酮类、酚类发生规律性变化,对何首乌质量产生重要影响.

目的 研究木兰花碱在大鼠体内外的主要代谢产物及代谢途径.方法 SD大鼠ig木兰花碱(50mg/kg),收集0?24h尿液和粪便,0?6h胆汁以及1、2、4、6、8h血浆;体外代谢采用肝微粒体温孵系统和肠菌培养液.利用LC-MS/MS对生物样品中的原型药及代谢产物进行鉴定.采用Agilent TC-C18色谱柱(150mm×4.6mm,5μm),以乙腈-0.1％甲酸水溶液为流动相梯度洗脱,体积流量1.0mL/min,柱温30℃.质谱采用电喷雾电离源(ESI),正离子采集模式;扫描范围m/z 100?1 000.根据药物体内代谢规则,结合木兰花碱的色谱保留时间和多级质谱碎片离子特征,推测其代谢产物的结构.结果 给药后生物样品中共鉴定出12个代谢产物,其中Ⅰ相代谢产物8个,Ⅱ相代谢产物4个.主要的代谢途径为羟基化、去甲基化、脱氢作用、酮基化、葡萄糖化、葡萄糖醛酸化及硫酸酯化.结论 木兰花碱在体内可发生Ⅰ相和Ⅱ相代谢,肠道菌群和肝药酶可催化木兰花碱发生Ⅰ相代谢转化,Ⅱ相代谢存在于肠道以外部位,最有可能的部位是肝脏.

This study is to provide the basis of establishing a quality evaluation system, based on the differences in appearance and internal components of Astragali Radix from different sources. The diameter of 18 batches of Astragali Radix, the content of alcohol(water) extract and 7 kinds of flavonoids were determined. The peak area ratio of flavonoid aglycon to aglycone was calculated. PCA and CA were carried out by synthesizing various indexes. The results of PCA and CA showed that Astragali Radix was obviously clustered into three types. Alcohol extract, formononetin/formosan glycosides,(pilose isoflavones+astragalus flavonoid A)/pilose isoflavone glucoside are the most significant differences in the variable importance projection index(VIP) of Astragali Radix. Combining the diameter, alcohol(water) extract, flavonoid aglycon to aglycone peak area ratio can provide an analysis method for the establishment of the grade evaluation system of Astragali Radix.

目的 建立鼻渊净胶囊的高效液相色谱(HPLC)指纹图谱.方法 采用Agilent SB-C18(4.6 mm×250 mm,5μm)色谱柱,乙腈-水为流动相、以1.0 ml/min流速行梯度洗脱,检测波长210 nm,柱温30℃,洗脱时间为80 min.采用中药色谱指纹图谱相似度评价系统(2004A版)对检测出色谱进行指纹图谱相似度评价.结果 建立了鼻渊净胶囊的HPLC指纹图谱,确定了20个共有峰,15个峰归属到各药材,其中5个峰确认了化学成分;10批样品的指纹图谱的整体相似度与对照图谱比较,均在90％以上.结论 所建立的鼻渊净胶囊指纹图谱有助于从整体上控制该制剂的质量.

目的:运用网络药理学方法探讨黄芪抗衰老的作用机制.方法:通过中药系统药理学数据库与分析平台(TCMSP)获得黄芪化学成分,以生物利用度(OB)、类药性(DL)为条件筛选主要活性成分;利用TCMSP和PharmMapper数据库预测黄芪活性成分靶点;在OMIM数据库和GeneCards数据库中输入关键词"aging",搜索与衰老相关的疾病靶点;借助Cytoscape 3.2.1软件构建"药物-疾病-靶点"的可视化网络;通过STRING网站构建蛋白质-蛋白质相互作用(PPI)网络,并筛选核心靶点;通过CB-Dock网站对成分与靶点进行分子对接验证;应用DAVID数据库对靶点进行基因本体(GO)分析和京都基因与基因组百科全书(KEGG)通路分析.结果:筛选得到黄芪24个活性成分,对应385个作用靶点;筛选出衰老相关靶点23997个,取交集得到黄芪与衰老相关靶点377个,其中核心靶点为蛋白激酶B1(Akt1)、丝裂原活化蛋白激酶1(MAPK1)、Jun、白细胞介素-6(IL-6)、MAPK8、表皮生长因子受体(EGFR)、血管内皮生长因子A(VEGFA)、表皮生长因子(EGF)、骨髓细胞瘤病毒癌基因同源物(MYC)和白细胞介素-8(CXCL8);分子对接结果显示,黄芪活性成分与靶蛋白Akt1和MAPK1的结合能力较好;GO功能显著富集在氧化应激反应、活性氧代谢等生物过程;KE GG富集信号通路161条.结论:黄芪中黄酮和皂苷类化合物可能是其抗衰老的物质基础;黄芪可通过Akt1等靶点调节糖基化终末产物及其受体(AGE-RAGE)、缺氧诱导因子-1(HIF-1)等信号通路发挥其抗衰老作用,为其深入研究提供参考.

目的:根据不同提取方法制备的交泰丸治疗大鼠失眠焦虑心肾不交证的量效关系,筛选最佳工艺.方法:采用每天随机束缚3 h且每周分别随机3次禁食禁水(24 h/次),连续21 d,建立大鼠失眠焦虑心肾不交证模型.d22动物分组,灌胃给予梯度剂量8种不同工艺交泰丸(1.86～186.7 mg·kg-1·d-1,k=1.77),连续10周;d77采用戊巴比妥钠睡眠协同实验检测睡眠时间,高架十字迷宫(EPM)和旷场实验(OFT)观察焦虑相关行为学变化;d102处死动物,苏木精-伊红(HE)染色观察海马CA3区病理变化,形态计量神经元细胞个数;ELISA检测血清五羟色胺(5-HT)、去甲肾上腺素(NE)含量;Graphpad Prism 5软件非线型可变斜率回归不同制剂工艺交泰丸的半数有效剂量(ED50).结果:不同工艺制备的交泰丸治疗后,可不同程度延长大鼠睡眠时间,缓解焦虑行为,升高血清5-HT,降低NE水平,减轻海马CA3区病理改变;其量效曲线呈"S"型.优选出交泰丸最佳提取工艺为:黄连、肉桂水提两次(4 h/次),药液合并后减压干燥.结论:本实验通过治疗失眠焦虑心肾不交证量效实验,优选了交泰丸制备工艺,为后续药物研发与临床应用提供了依据.