3,5-O-二咖啡酰基奎宁酸的制备及其对HeLa细胞的抗增殖活性研究

目的 制备高纯度3，5-O-二咖啡酰基奎宁酸，并评价其对人宫颈癌HeLa细胞的抗增殖活性。方法 采用柱色谱提取法和中压液相色谱法从奇蒿花中分离、纯化得到高纯度的3，5-O-二咖啡酰基奎宁酸。采用MTT法评价该化合物对人宫颈癌HeLa细胞的体外抗增殖活性。结果 柱色谱提取的提取率和中压液相色谱法的回收率分别为99.0%，61.2%，总回收率为54.0%。随着3，5-O-二咖啡酰基奎宁酸浓度升高，HeLa细胞存活率下降，细胞形态损伤增加，受试药物的IC₅₀值为26.5µg·mL^-1。结论 本研究提供了一种简单、高效、节能的3，5-O-二咖啡酰基奎宁酸制备方法。3，5-O-二咖啡酰基奎宁酸对HeLa细胞具有一定的体外抗增殖活性。     

Synthesis,molecular docking,and pharmacological evaluation of N‐(2‐(3,5‐dimethoxyphenyl)benzoxazole‐5‐yl)benzamide derivatives as selective COX‐2 inhibitors and anti‐inflammatory agents

A series of N‐(2‐(3,5‐dimethoxyphenyl)benzoxazole‐5‐yl)benzamide derivatives ( 3am ) was synthesized and evaluated for their in vitro inhibitory activity against COX‐1 and COX‐2. The compounds with considerable in vitro activity (IC₅₀ < 1 μM) were evaluated in vivo for their anti‐inflammatory potential by the carrageenan‐induced rat paw edema method. Out of 13 newly synthesized compounds, 3a , 3b , 3d , 3g , 3j , and 3k were found to be the most potent COX‐2 inhibitors in the in vitro enzymatic assay, with IC₅₀ values in the range of 0.06–0.71 μM. The in vivo anti‐inflammatory activity of these six compounds ( 3a , 3b , 3d , 3g , 3j , and 3k ) was assessed by the carrageenan‐induced rat paw edema method. Compounds 3d (84.09%), 3g (79.54%), and 3a (70.45%) demonstrated significant anti‐inflammatory activity compared to the standard drug ibuprofen (65.90%) and were also found to be safer than ibuprofen, by ulcerogenic studies. A docking study was done using the crystal structure of human COX‐2, to understand the binding mechanism of these inhibitors to the active site of COX‐2.

     

保健食品中新型二硫代卡地那非类似物的分析鉴定

目的 鉴定保健食品中非法添加的一个未知结构的二硫代卡地那非类似物。方法 采用高效液相色谱-二极管阵列检测器联用（HPLC-DAD）技术进行补肾壮阳类保健食品非法添加筛查时发现一个未知结构的二硫代卡地那非类似物,经过正相硅胶薄层色谱分离纯化得到目标化合物后,用超高效液相色谱-二级质谱联用（UPLC-MS/MS）技术获得其准分子离子和二级质谱图,用核磁共振得到碳谱和氢谱数据,结合文献分析,最终鉴定该化合物的结构。结果 在保健食品中发现了一个新型二硫代卡地那非类似物,结构为3,5-二甲基哌嗪基二硫代去甲卡地那非。结论 该化合物不在现有补肾壮阳类中成药检验标准的13种目标化合物范围内,是一种新的非法添加化学物质。     

指纹图谱结合一测多评法评价拟草果质量的研究

目的建立拟草果Amomumparatsao-ko药材的质量评价方法。方法以鼠李柠檬素为参照峰确定共有峰,建立拟草果的HPLC指纹图谱;以鼠李柠檬素为内标,确定鼠李素、3,5-二羟基-7,4’-二甲氧基黄酮的相对校正因子并计算含量,实现一测多评。同时采用外标法测定3种成分的含量,比较计算值和实测值的差异。结果建立了拟草果的HPLC指纹图谱,10批样品相似度均在0.9以上;以鼠李柠檬素为内标建立的鼠李素、3,5-二羟基-7,4’-二甲氧基黄酮的相对校正因子分别为1.300、0.885,重现性良好,10批拟草果样品一测多评法计算值与实测值无显著差异。结论建立的HPLC指纹图谱和一测多评法可用于拟草果的质量评价。     

海绵共生真菌Chaetomium globosum HXL-1次级代谢产物及其体外抗肿瘤活性研究

目的研究海绵共生真菌Chaetomium globosum HXL-1的次级代谢产物及其抗肿瘤活性。方法利用硅胶柱色谱、ODS柱色谱、高效液相色谱等方法对其次级代谢产物进行分离纯化,运用NMR、MS等多种波谱方法鉴定化合物结构;采用MTT方法评价化合物的体外抗肿瘤活性。结果从该菌株发酵液的醋酸乙酯萃取物中分离得到16个化合物,分别鉴定为4-甲氧基-3,5-二甲基-6-(2-甲基丁酰基)-2氢-2-吡喃酮(1)、吡啶并戊二烯(2)、1-脱乙酰基-吡啶并戊二烯(3)、7-脱乙酰基-吡啶并戊二烯(4)、大黄素-8-甲醚(5)、8-羟基-1,3-二甲氧基-6-甲基蒽醌(6)、cytoglobosins D(7)、isochaetoglobosin D(8)、球毛壳菌素E(9)、球毛壳菌素Vb(10)、monohydroxyisoaflavinine(11)、aflavazole(12)、烟曲酶毒素C(13)、chaetoviridin B(14)、杀锥曲菌素(15)、oxidized-nodulisporic acid B(16)。结论化合物1是吡喃酮类新化合物;化合物7～13为吲哚生物碱类化合物;化合物1、11、12为首次从球毛壳菌中分离得到;化合物2、8、15对选定的人肿瘤细胞具有一定的体外抗增殖活性。     

金银花中抗补体活性酚酸类成分的研究

目的: 研究金银花Lonicera japonica中的抗补体活性酚酸类成分. 方法: 通过溶血试验方法进行抗补体活性成分的导向分离,对所得化合物进行抗补体活性测定,采用现代波谱技术ESI-MS,¹H-NMR,¹³C-NMR进行结构鉴定. 结果: 从金银花乙酸乙酯部位分离得到了14个化合物,其中有8个酚酸类、3个环烯醚萜类和3个黄酮类成分,包括新绿原酸(1)、绿原酸(2)、隐绿原酸 (3)、3,5-O-二咖啡酰奎宁酸(4)、4,5-O-二咖啡酰奎宁酸(5)、3,4-O-二咖啡酰奎宁酸(6)、咖啡酸(7)、咖啡酸甲酯(8)、裂环马钱苷(9)、獐芽菜苷(10)、断氧化马钱子苷(11)、木犀草素(12)、槲皮素(13)、山柰酚(14).抗补体实验表明化合物 1~9, 11~14 对经典的补体激活具有不同程度的抑制作用,以3,5-O-二咖啡酰奎宁酸活性最强. 结论:化合物 14 首次从金银花中分离得到,酚酸类化合物是金银花中的主要抗补体活性成分,3,5-O-二咖啡酰奎宁酸活性最强,值得深入研究.     

HPLC法测定市售杭白菊中2种有机酸和2种黄酮的含量

摘 要 目的： 测定市售杭白菊中有效成分的含量,比较不同产地杭白菊质量。方法: 采用HPLC法测定绿原酸、3,5-O-双咖啡酰基奎宁酸2种有机酸成分以及木犀草苷、木犀草素2种黄酮成分的含量。结果: 收集杭白菊样品各有效成分差异较大,12批市售杭白菊绿原酸含量在0.22%～0.70%之间,木犀草苷含量在0.09%～0.25%之间,3,5-O-双咖啡酰基奎宁酸含量在0.84%～1.67%之间,木犀草素含量在0.008%～0.042%之间。结论: 市售杭白菊质量有较大差异,有必要进一步加强质量控制。     

3,5-Dimethoxy-4-hydroxy myricanol ameliorates photoreceptor cell degeneration in Pde6brd10 mouse model

Introduction: Retinitis pigmentosa (RP) caused by the photoreceptor cell degeneration is currently incurable and leads to partial or complete blindness eventually. 3,5-dimethoxy-4-hydroxy myricanol (DM) is a novel compound isolated from the leaves of Micromelum integerrimum, with proliferative activities on NIH3T3 cells. This study was to investigate whether DM could mitigate retinal degeneration of rd10 mice, a well-characterized mouse model of RP. Materials and method: Rd10 mice were treated with DM daily by intraperitoneal injection from postnatal day 12 (P12) to P26. Electroretinography (ERG) reflects the mass response of photoreceptor cells and was used to test the outer retinal function after DM treatment. Haematoxylin and Eosin staining was used to show the retinal morphology and evaluate the rod photoreceptor cell loss. TUNEL assay was used to detect the apoptosis-positive cells. Inflammatory factors were measured by ELISA to show the inflammatory response. Real-time PCR and western blot were applied to measure the gene and protein change to explore the underlying mechanisms. Results: Results showed that DM significantly improved the retinal function by increasing the ERG amplitude, preserving the retinal morphology, reducing photoreceptor cell apoptosis, decreasing inflammatory response, and inhibiting endoplasmic reticulum stress in rd10 mice. Conclusion: This is the first time when the protective effects of DM against photoreceptor cell degeneration of rd10 mice have been demonstrated, providing scientific rationale to develop DM as a potential agent to treat RP.     

Cryptorchidism and endocrine disrupting chemicals

Prospective clinical studies have suggested that the rate of congenital cryptorchidism has increased since the 1950s. It has been hypothesized that this may be related to environmental factors. Testicular descent occurs in two phases controlled by Leydig cell-derived hormones insulin-like peptide 3 (INSL3) and testosterone. Disorders in fetal androgen production/action or suppression of Insl3 are mechanisms causing cryptorchidism in rodents. In humans, prenatal exposure to potent estrogen diethylstilbestrol (DES) has been associated with increased risk of cryptorchidism. In addition, epidemiological studies have suggested that exposure to pesticides may also be associated with cryptorchidism. Some case-control studies analyzing environmental chemical levels in maternal breast milk samples have reported associations between cryptorchidism and chemical levels. Furthermore, it has been suggested that exposure levels of some chemicals may be associated with infant reproductive hormone levels.     

Natural killer cells ameliorate liver fibrosis by killing activated stellate cells in NKG2D-dependent and tumor necrosis factor-related apoptosis-inducing ligand-dependent manners

BACKGROUND & AIMS: Viral hepatitis infection, which is a major cause of liver fibrosis, is associated with activation of innate immunity. However, the role of innate immunity in liver fibrosis remains obscure. METHODS: Liver fibrosis was induced either by feeding mice with the 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) diet or by injecting them with carbon tetrachloride. The Toll-like receptor 3 ligand, polyinosinic-polycytidylic acid, was used to activate innate immunity cells and mediators, including natural killer cells and interferon gamma. RESULTS: In the mouse model of DDC-induced liver fibrosis, natural killer cell activation by polyinosinic-polycytidylic acid induced cell death to activated hepatic stellate cells and attenuated the severity of liver fibrosis. Polyinosinic-polycytidylic acid treatment also ameliorated liver fibrosis induced by carbon tetrachloride. The observed protective effect of polyinosinic-polycytidylic acid on liver fibrosis was diminished through either depletion of natural killer cells or by disruption of the interferon gamma gene. Expression of retinoic acid early inducible 1, the NKG2D ligand, was undetectable on quiescent hepatic stellate cells, whereas high levels were found on activated hepatic stellate cells, which correlated with the resistance and susceptibility of quiescent hepatic stellate cells and activated hepatic stellate cells to natural killer cell lysis, respectively. Moreover, treatment with polyinosinic-polycytidylic acid or interferon gamma enhanced the cytotoxicity of natural killer cells against activated hepatic stellate cells and increased the expression of NKG2D and tumor necrosis factor-related apoptosis-inducing ligand on liver natural killer cells. Blocking NKG2D or tumor necrosis factor-related apoptosis-inducing ligand with neutralizing antibodies markedly diminished the cytotoxicity of polyinosinic-polycytidylic acid-activated natural killer cells against activated hepatic stellate cells. CONCLUSIONS: Our findings suggest that natural killer cells kill activated hepatic stellate cells via retinoic acid early inducible 1/NKG2D-dependent and tumor necrosis factor-related apoptosis-inducing ligand-dependent mechanisms, thereby ameliorating liver fibrosis.