矮紫堇生物碱成分

目的:研究藏药矮紫堇生物碱部位的化学成分。方法:通过反复硅胶柱色谱,羟丙基葡聚糖凝胶(Sephadex LH-20),制备薄层色谱和半制备高效液相色谱等技术进行分离纯化,通过理化性质,运用NMR,MS等波谱方法以及结合文献数据对分离得到的化合物进行结构鉴定。结果:从矮紫堇乙醇提取物的总生物碱部位中分离鉴定了11个化合物,分别为9-methyldecumbenine C(1),原阿片碱(2),别隐品碱(3),四氢黄连碱(4),四氢小檗碱(5),四氢巴马亭(6),tetrahydrocolumbamine(7),奥柯紫堇明碱(8),fumariline(9),二氢血根碱(10),小檗碱(11)。结论:化合物3和5~11为首次从该植物中分离得到。     

延胡索中生物碱类成分的质量控制

目的:建立RP-HPLC同步测定延胡索中11种生物碱成分的方法,为该药材的质量控制提供参考。方法:Kromasil Eternity C18色谱柱(4.6 mm×250 mm,5μm),流动相乙腈(A)-0.1 mol·L-1乙酸铵溶液(每1 L溶液中加冰乙酸0.5m L)(B)梯度洗脱(0~50 min,20%~55%A;50~55 min,55%~65%A;55~60 min,65%A),流速1.0 m L·min-1,柱温25℃,检测波长280 nm,进样量10μL。结果:11种待测成分的线性关系良好,相关系数均0.999;精密度试验RSD 0.1%~2.0%,稳定性试验RSD 0.2%~2.1%,重复性试验RSD 1.7%~4.1%,平均加样回收率98.2%~100.8%(RSD 1.6%~3.5%),均符合中药质量分析要求。结论:建立的RP-HPLC灵敏、简便、可靠,可用于延胡索药材中多种生物碱成分的同步测定。     

延胡索中1个新生物碱

目的 研究延胡索Corydalis yanhusuo块茎的化学成分。方法 运用常压硅胶柱色谱分离等现代分离手段,对延胡索块茎生物碱成分进行分离,结合理化性质和核磁共振波谱数据鉴定化合物结构。结果 从延胡索块茎乙醇提取物中分得3个化合物,分别鉴定为1-[2-(N-甲基铵乙基)]-3,4,6,7-四甲氧基菲(1)、1-乙胺甲基-3,4,6,7-四甲氧基菲(2)、氧代海罂粟达林碱(3)。结论 化合物1为新天然产物,化合物2为新化合物,命名为延胡索菲碱A;化合物3为首次从延胡索中分离得到。     

基于一测多评法对延胡索中生物碱类成分的质量控制研究

目的建立同时测定延胡索中原阿片碱、黄连碱、黄藤素、去氢延胡索甲素、D-四氢药根碱、延胡索乙素和延胡索甲素7种生物碱类成分的HPLC方法,在此基础上,建立一测多评法的方法学考察方式,验证该方法在延胡索质量控制中应用的可行性和技术适用性。方法以延胡索所含的7种成分为指标成分,采用2种校正方法分别建立各成分与延胡索乙素的相对校正因子(fk/s),计算各成分的量,实现一测多评;同时采用外标法测定延胡索中该7种成分的量,并比较2种一测多评法所得计算值与实测值的差异,以验证一测多评法的准确性和可行性。结果建立了延胡索中7种生物碱类成分一测多评法方法学的考察模式;不同的色谱柱和仪器一测多评法的计算值与外标法实测值之间没有显著性差异。结论同时测定延胡索中原阿片碱、黄连碱、黄藤素、去氢延胡索甲素、D-四氢药根碱、延胡索乙素和延胡索甲素7种生物碱的一测多评法可靠,结果准确,可用于延胡索药材的质量控制。     

岩黄连生物碱自微乳的制备及体外评价

 目的以岩黄连生物碱自乳化药物为模型,研究自微乳释药系统的制备及体外评价方法。方法制备岩黄连生物碱自微乳,以自微乳时间、所成微乳的形态、粒径分布、Zeta电位、含量、稳定性以及体外模拟人体释放等指标为考察项目,进行体外评价。结果所制备的自微乳载药量2.29 g·L^-1,3 min内已基本乳化完全,乳化后粒子粒径大多数在20 nm左右,呈高斯分布,包封率大于98.7%,自微乳效率高。结论优选出的自乳化处方所制备的产品外观澄清透明,稳定性好,质量稳定,检测方法可靠,重现性好,展现了自微乳药物传统系统良好的释放特性。     

Metabolomics investigation on antiobesity effects of Corydalis bungeana on high-fat high-sugar diet-induced obese rats

Objective: Corydalis bungeana (CB) is a well-used medicinal herb in Mongolian folk medicine and has been traditionally applied?as?an antiobesity agent. However, the evidence-based pharmacological effects of CB and its specific metabolic alterations?in the obese model are not entirely understood. This study aimed to utilize untargeted metabolomic techniques to identify biomarkers and gain mechanistic insight into the serum metabolite alterations associated with weight loss and lipid metabolism in obese rats. Methods: A high-fat high-sugar (HFHS) diet was used to induce obese models in rats. CB extract was orally gavaged at 0.18, 0.9 and 1.8 g/kg doses for six weeks, and feed intake, body weight, fat pad weight, and blood indexes were measured. Blood serum metabolites were evaluated by gas chromatography/quadrupole time-of-flight tandem mass spectrometry (GC-TOF/MS). Results: The results showed that compared with the obese group, the administration of CB extract caused significant decreases in body weight (P < 0.05), feed intake, Lee’s index, and perirenal, mesenteric, epididymal fat weight. CB extract also reduced blood triglyceride and total cholesterol levels (P < 0.05) of obese rats. Metabolomic findings showed that nine differential metabolites, including pyruvic acid, D-glucuronic acid, malic acid, dimethylglycine, oxoglutaric acid, pantothenic acid, sorbitol acid, fumaric acid and glucose 6-phosphate were identified under CB treatment and altered metabolic pathways such as TCA cycle, pantothenate and CoA biosynthesis, and glycolysis/gluconeogenesis. Conclusion: This study demonstrated weight loss and lipid lowering effects of CB on HFHS diet-induced obese rats and identified nine metabolites as potential biomarkers for evaluating the favorable therapeutic mechanism of CB via regulation of lipid and glucose metabolism.     

苦地丁化学成分和体内代谢产物的LC-MS鉴定

目的 研究苦地丁Corydalis bungeana提取物中的化学成分和体内代谢产物,探究其代表性成分质谱裂解规律。方法采用高效液相色谱-电喷雾质谱联用技术（HPLC-Q-TOF/MS）,正离子模式扫描,对苦地丁提取物和大鼠口服苦地丁提取物后不同时间段内的血浆、胆汁、尿液和粪便分别进行检测分析,通过与对照品和文献比对保留时间和质谱碎片信息,确定化学成分归属及其裂解方式,预测代谢产物的动态变化。结果 在苦地丁提取物中共鉴定得31个化合物,其中共有21个化合物以原型进入体内;并以紫堇灵为研究对象,在大鼠血浆、胆汁、尿液和粪便中分别发现3、3、4、4个代谢产物。结论 借助HPLC-Q-TOF/MS方法鉴定了苦地丁体外化学成分和体内以原型存在的化学成分,阐明了代表性成分紫堇灵的代谢产物及代谢规律,推测苦地丁在体内发生了I相和II相代谢,其中以II相代谢方式为主（硫酸化、糖基化、甲基化和去甲基化）,为阐明苦地丁的药效物质基础提供依据。     

Discovery of N-methyltetrahydroprotoberberines with κ-opioid receptor agonists-opioid receptor agonist activities from corydalis yanhusuo W. T. Wang by using two-dimensional liquid chromatography

Ethnopharmacological relevance

The need for an efficacious analgesic without unwanted side effects is urgent. κ-opioid receptor agonists are known to exhibit potent analgesic effects and elicited fewer side effects than other opioid agonists. Thus in this study we chose the κ-opioid receptor as the target to identify the active components from traditional Chinese medicines (TCMs).

Materials and methods

The κ-opioid receptor was expressed in human embryonic kidney-293 T cells (HEK293T). Fluorometric Imaging Plate Reader (FLIPR) assay was used for the determination of Ca²⁺ response when κ-opioid receptor was activated. A novel 2D separation system employing C18HCE as the first dimension and a strong cation exchange column (SCX) as the second dimension was conducted for the purification of the active principles.

Results

With the aid of HPLC-based activity profiling, activities could be linked to two peaks from Corydalis yanhusuo W. T. Wang (C. yanhusuo) extract. Two N-methyltetrahydroprotoberberines with κ-opioid receptor agonist activities were isolated for the first time from C. yanhusuo by using 2D-LC.

Conclusions

Our study suggests that N-methyltetrahydroprotoberberines may serve as a new scaffold for κ-opioid receptor ligands. The strategy that we adopted can be applied to other naturally-occurring active alkaloids acting at different receptors.     

延胡索对缺血再灌注损伤大鼠局灶性脑梗死的作用

目的：探讨延胡索对局灶性脑梗死的作用。方法：总共有30只Sprague-Dawley(SD)大鼠被用来研究。局灶性脑梗死动物模型的建立是将两侧的颈总动脉和右侧的中大脑动脉的血流阻断90min后,再经24h的灌注,然后评估它们的神经状态,大鼠被牺牲取脑,并作成切片用2,3,5-triphenyl-tetrazolium chloride染色。以神经的状态和脑梗死面积的变化作为指标来评估延胡索对脑梗死的作用。另外,24只SD大鼠腹腔注射延胡索,经24h后从心脏采血测量血液的变化。结果：延胡索100mg/kg腹腔注射能够改善大鼠神经缺损的症状和减少脑梗死的面积以及能增加全血中红血球的数目和提高血比容（hematocrit)。结论：延胡索在缺血再灌流损伤大鼠能改善神经状态和减少脑梗死面积。     

基于中医传承辅助平台探索含延胡索-川楝子药对方剂的组方规律

目的通过对《中华人民共和国卫生部药品标准——中药成方制剂》(《中药部颁标准》)含延胡索-川楝子药对(简称"延胡索-川楝子")方剂组方规律的研究,以期为临床应用及方剂的后续产品开发提供理论依据。方法收集整理《中华人民共和国卫生部药品标准——中药成方制剂》中包含延胡索-川楝子的方剂,录入中医传承辅助平台软件(V.2.5)建立数据库,统计主治病症频次、配伍药材频次、组方归经频次;采用关联规则Apriori算法对含延胡索-川楝子方剂的药材核心组合进行统计分析(设置支持度分别为10%、20%、30%,置信度为0.90);选择较高频次归经进行组方规律分析。结果共筛选出51首含延胡索-川楝子方剂,涉及主治疾病8种(频次≥2),包括胃痛、胁痛、带下病、痛经、盆腔炎、月经失调等;涉及主治证候9种(频次≥2),包括肝胃不和证、气滞血瘀证、肝郁气滞证、肝郁血虚证、胞宫血热证等;常配伍药材17味(频次≥5),包括白芍、木香、甘草、香附等;常用药材组合20种(频次≥6),其中核心药材组合包括延胡索-白芍-川楝子、延胡索-甘草-川楝子、延胡索-木香-川楝子、延胡索-香附-川楝子;组方归肝脾胃经频次较高;常与延胡索-川楝子配伍归肝经的组方核心药材有5种,包括木香、白芍、陈皮、砂仁、沉香,其中核心药材组合(频次≥7)包括延胡索-木香-川楝子、延胡索-白芍-川楝子、延胡索-陈皮-川楝子、延胡索-陈皮-白芍-木香-川楝子。常与延胡索-川楝子配伍归脾胃经的组方核心药材有3种,包括沉香、白芍、砂仁,其中核心药材组合(频次≥4)包括延胡索-沉香-川楝子、延胡索-白芍-川楝子、延胡索-砂仁-川楝子。结论《中华人民共和国卫生部药品标准——中药成方制剂》中含延胡索-川楝子方剂所用药物多理气止痛之功效,用药较为集中,组方法度清晰。此外,以3种归经组方规律的对比分析,反映了含延胡索-川楝子方剂不同归经组方的用药规律和配伍特点,为"异病同治"提供理论支持,也为延胡索-川楝子药对更好的应用于临床提供理论支持。