40种香豆素类化合物对人胃癌细胞株BGC和人肝癌细胞株BEL-7402细胞生长抑制活性的筛选

目的:寻找中药中抗肿瘤活性化合物和先导化合物,为开发新药奠定基础。方法:采用人胃癌细胞株BGC和人肝癌细胞株BEL-7402体外培养法,鉴定受试的40种香豆素类化合物对其生长的抑制作用。结果:伞形花内酯刘寄奴酸酯、木桔素、环氧酸橙皮油素、酸橙皮油素、前胡酮、芸香苦素、牛防风素和异香柑内酯对人胃癌细胞株BGC细胞的生长有一定程度的抑制作用,且呈浓度-效应关系;伞形花内酯刘寄奴酸酯、异虎耳草素、前胡素和二氢山芹醇当归酸酯对人肝癌细胞株BEL-7402细胞的生长有一定程度的抑制作用,且呈浓度-效应关系。结论:香豆素类化合物是潜在的抗肿瘤药物。     

Quantitative and qualitative analysis of common peaks in chemical fingerprint of Yuanhu Zhitong tablet by HPLC-DAD–MS/MS

A quality control (QC) strategy for quantitative and qualitative analysis of “common peaks” in chemical fingerprint was proposed to analyze Yuanhu Zhitong tablet (YZT), using high performance liquid chromatography with diode array detector and tandem mass spectrometry (HPLC-DAD–MS/MS). The chromatographic separation was achieved on an Agilent Eclipse plus C₁₈ column with a gradient elution using a mixture of 0.4‰ ammonium acetate aqueous (pH 6.0 adjusted with glacial acetic acid) and acetonitrile. In chemical fingerprint, 40 peaks were assigned as the “common peaks”. For quantification of “common peaks”, the detection wavelength was set at 254 nm, 270 nm, 280 nm and 345 nm, respectively. The method was validated and good results were obtained to simultaneously determine 10 analytes (protopine, jatrorrhizine, coptisine, palmatine, berberine, xanthotoxin, bergapten, tetrahydropalmatine, imperatorin and isoimperatorin). For qualification of “common peaks”, 33 compounds including 10 quantitative analytes were identified or tentatively characterized using LC–MS/MS. These results demonstrated that the present approach may be a powerful and useful tool to tackle the complex quality issue of YZT.     

福参化学成分研究

目的：从福参Angelica morri Hayata饮片提取分离活性成分。方法：95％乙醇提取、 石油醚萃取、硅胶柱层析、重结晶等。结果：分离到7个化合物，其中5个为香豆素，分别欧前胡素（imperatorin Ⅲ）、异欧前胡素（isoimperatorin Ⅳ、珊瑚菜素（phellopterin V)、别异欧前胡素（alloimperatorin Ⅵ）和紫花前胡内酯（nodakenetin Ⅶ），另两个分别为β－谷甾醇（β－sitosterol I和二十二烷酸（docosanoic acid Ⅱ）结论：上述化合物均为首次从该植物中分得。     

地骨皮化学成分的研究

目的：对地骨皮（枸杞Lycium chinese Mill.根皮）的化学成分进行研究。方法：利用各种色谱技术进行分离纯化，根据化合物的理化性质和光谱数据进行结构鉴定。结果：分离到多个化合物，主要为莨菪亭（scopoletin,I)、大黄素甲醚（physcion,Ⅱ）、大黄素（emodin,Ⅲ）、东莨菪甙（scopolin(Ⅳ)、Fabiatrin(Ⅴ)等五个化合物。结论：除莨菪亭外，其余四个化合物均为从地骨皮中首次分得。     

花椒毒酚的药效学研究 Ⅰ抗心律失常作用

花椒毒酚（ｘａｎｔｈｏｔｏｘｏｌ，ＸＴ）是从蛇床子乙醇提取物中分离出的一种香豆素。ＸＴ对氯仿诱发的小鼠室颤、氯化钙诱发的大鼠室颤均有明显的预防作用，对乌头碱诱发的大鼠心律失常有显著的治疗效果，且能明显提高兔心室电致颤阈。此外，ＸＴ还能降低蟾蜍离体坐骨神经干的动作电位振幅。这些均表明ＸＴ有抗心律失常的作用。ＸＴ的小鼠ｉｖＬＤ＿（５０）为４７．０ｍｇ·ｋｇ￣（－１）。     

杀鼠药中毒所致凝血功能障碍20例临床分析

目的 总结杀鼠药中毒导致的凝血功能障碍的临床诊断及治疗。 方法 对20例抗凝血杀鼠药中毒致凝血功能障碍患者予以维生素K₁治疗,比较患者治疗前后的凝血功能改变:凝血酶原时间(PT)、国际正常化比率( INR ),活化部分凝血活酶时间(APTT)、凝血因子Ⅱ、Ⅶ、Ⅸ、Ⅹ活性。 结果 20例患者全部治愈,治疗前后凝血功能明显改变,PT治疗前后分别为:(103.2±20.3)s,(12.0±0.5)s。 INR治疗前后分别为:4.2±2.4,1.4±0.2。APTT 治疗前后分别为:(93.2±25.4)s,(30.2±3.4)s。凝血因子Ⅱ:活性成分治疗前后分别为:(6.2±2.5)%,(92.3±4.5)%。凝血因子Ⅶ:活性成分治疗前后分别为:(10.8±4.9)%,(86.2±3.4)%。凝血因子Ⅸ:活性成分治疗前后分别为:(22.5±6.8)%,(96.4±7.8)%。凝血因子X:活性成分治疗前后分别为:(14.2±3.2)%,(83.2±3.4)%,差异有统计学意义(P<0.001)。 结论 杀鼠药中毒致凝血功能障碍,容易误诊,维生素K₁是特效解毒药。     

荧光成像技术用于独活根中香豆素类成分积累规律研究

目的：建立独活冰冻切片的新方法,并利用荧光成像技术对不同时期独活根中的香豆素类成分进行组织化学定位、定量研究,明确独活中活性成分香豆素的组织分布及其动态积累规律。方法：以不同时期的独活根为材料,考察不同保护剂、不同切片厚度对独活根最佳制片的影响,以此建立独活根的最佳制片条件。此外,利用显微成像测量技术对不同部位分泌组织进行统计分析,并结合荧光成像对香豆素成分集中分布部位的荧光强度进行测算,以荧光强度对不同时期的独活根中香豆素成分进行相对定量研究,分析不同时期香豆素成分含量的动态变化规律。结果：独活根经15%甘油保护剂的处理,切片厚度为25 μm时切片效果最佳。通过组织化学定位显示,独活香豆素类成分主要分布在韧皮部的分泌腔及分泌道中。不同时期香豆素类成分总体变化趋势呈S形增长,8月、10月含量变化明显,10月香豆素含量达到最大。结论：首次使用了独活根冰冻切片及荧光成像新方法,研究了香豆素成分的产生、分布以及积累规律。发现独活香豆素成分主要分布在韧皮部的分泌腔中;不同时期独活根中香豆素成分动态积累变化趋势呈S形增长,且10月香豆素含量积累达到最大,为拓宽中药分析提供了新思路。     

Protective effects of coumarin and coumarin derivatives against carbon tetrachloride-induced acute hepatotoxicity in rats

The comparison of the antioxidant activity of some coumarins with their molecular structure is well determined. However, the protective function of coumarins with various chemical structures against liver toxicity has not yet been well established. Therefore, the aim of this study was to evaluate the possible cytoprotective properties of coumarin and some coumarin derivatives against CCl₄ (carbon tetrachloride)-induced hepatotoxicity. Coumarin (1,2-benzopyrone) and coumarin derivatives esculetin (6,7-dihydroxycoumarin), scoparone (6,7-dimethoxycoumarin) and 4-methylumbelliferone (7-hyroxy-4-methyl) were examined for their protective effect against CCl₄-induced hepatotoxicity in Male Sprague-Dawley rats. A single toxic dose of CCl₄ (1.25 ml kg⁻¹, orally) produced liver damage in rats, seen histologically as centrilobular necrosis. Administration of CCl₄ increased serum enzyme levels of aspartate transaminase (AST), alanine transaminase (ALT), and alkaline phosphatase (ALP). Pre-treatment of rats with esculetin (31.15 mg kg⁻¹, orally) and scoparone (35 mg kg⁻¹, orally) significantly prevented CCl₄-induced increase in serum enzymes, whereas 4-methylumbelliferone (35 mg kg⁻¹) and coumarin (30 mg kg⁻¹) had no effect against CCl₄-induced rise in serum enzymes. Morphological findings were consistent with the plasma transaminase observations. Among the coumarin analogs, esculetin, which possesses orthodihydroxy coumarins, showed the strongest protective effect against CCl₄-induced liver damage, followed by scoparone, 4-methylumbelliferone and coumarin, respectively. The results of this study indicate that the chemical structures of coumarins play an important role in the prevention of liver toxicity.     

枸橼果实的香豆素和黄酮类成分研究

目的研究枸橼果实的化学成分。方法用硅胶柱色谱法及Sephadex LH-20柱色谱法进行分离纯化,依据理化性质和光谱数据鉴定化合物结构。结果从枸橼果实的95%乙醇提取物中分离鉴定了11个化合物,包括7个香豆素类化合物:7-羟基香豆素(1)、5,7-二羟基香豆素(2)、7-羟基-6-甲氧基香豆素(3)、5,7-二甲氧基香豆素(4)、6,7-二甲氧基香豆素(5)、佛手柑内酯(6)和8-(2',3'-二羟基-3'-甲丁基)-5,7-二甲氧基香豆素(7);4个黄酮类化合物:香叶木素(8)、柚皮素(9)、柚皮苷(10)和橙皮素(11)。结论化合物1,2,5～9为首次从枸橼中分离得到。     

独活中总香豆素组分的含量测定

目的：建立独活中总香豆素含量的测定方法。方法：以蛇床子素为对照品,采用紫外分光光度法测定独活中总香豆素含量。结果：检测波长为322nm,蛇床子素质量浓度在2～12mg/mL范围内与吸光度呈良好线性关系（r=0.9996）,平均回收率99.18%（RSD=1.69%,n=6）。结论：本方法简便易行,结果稳定可靠,可用于独活及其制剂的质量控制。