Determination of inorganic anions in ethyl acetate by in-line hollow fiber membrane extraction with ion chromatography

In this work, a novel hollow fiber membrane extractor was set up to extract inorganic anions from ethyl acetate using deionized water. Inorganic anions in slightly soluble organic solvents can be determined by the in-line hollow fiber membrane extractor coupled with ion chromatography at first time. Different aspects of the extraction procedure such as magnetic stirring speed, extraction flow rate and extraction time were optimized to achieve high extraction efficiency and good separation results. Satisfactory linear range, limits of detection and good repeatability were obtained. The procedure was applied to analyze inorganic anions in two commercial ethyl acetate samples.

Efficient Extraction and Isolation of Mangiferin from Mango Leaves by Ethyl Acetate Impurity Removal Method

[Objectives] To optimize the ethyl acetate impurity removal method for extracting and isolating mangiferin from mango leaves,and provide raw materials and technical support for development and use of mangiferin related products. [Methods]Five steps( material crushing→ ethyl acetate impurity removing → concentrated extract washing → extracting with methanol → crystallization and precipitation) were used.The single factor experiment and L9( 34) orthogonal experiment was carried out to optimize the process parameters including extraction time,ultrasonic power,extraction times,and extraction temperature.[Results] The optimum process of ethyl acetate impurity removal method for extracting and isolating mangiferin from mango leaves was as follows: the mango leaves were crushed and sieved; 3 m L/g of ethyl acetate was added,sealed and soaked for 4 h,ultrasonically shaken for 20 min( 50℃,350 W),filtered at room temperature,filtered with 100 mesh sieve,and extracted three times; added 100% methanol to the residue at 3 m L/g,extract by ultrasonic vibration for 20 min( 350 W,55℃)for four times,filtered with 100 mesh sieve when it was still hot; mixed the extract of each time,condensed by vacuum decompression to get the extract; added 100% methanol at 4 m L/g,mixed and washed for 5 min at room temperature,placed for 10 min,filtered with 100 mesh sieve,washed 3 times repeatedly,and dried the filter residue at 60℃ to obtain the crude mangiferin; added 100% methanol at 4 m L/g,mixed and washed at 50℃ for 5 min,placed at 6℃ for 8 h,dried the filter residue at 60℃,and repeatedly crystallized two times. According to the above process,crude and pure mangiferin products could be obtained,the purity of mangiferin of the crude product was higher than 64. 00%,the total recovery rate was 83. 90%,and the purity of mangiferin of the pure product was higher than 98. 00%,and the total recovery rate was about 66. 40%. [Conclusions] The optimized ethyl acetate impurity removal method is easy in operation,low in cost,and high in efficiency for extracting and isolating mangiferin,and can be applied for actual production of mangiferin.

Screening of ethyl acetate extract of Bridelia micrantha for hepatoprotective and anti-oxidant activities on Wistar rats

Objective:To explore the hepatoprotective and anti-oxidant activities of the methanolic leaf extract of Bridelia micrantha(B.micrantha) on paracetamol induced liver damage in Wistar rats. Methods:Parameters were measured including alanine aminotransaminase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),bilirubin and total protein.The anti-oxidant effects were studied using the 1,l-Diphenynl-2-Picrylhydrazyl(DPPH) and Ferric Reducing Antioxidant Power(FRAP) assay methods.Results:B.micrantha extract decreased the level of AST in the rats given PCM from(129.47±0.921) IU/L to(57.78±1.71) IU/L(P【0.05).This was lower than the value for Silymarin which was(59.92±1.41) IU/L.ALT concentration was reduced from (150.18±2.23) IU/L to(79.10±2.01) IU/L(P【0.05).ALP was reduced from(49.86±0.85) IU/L to(29.64±1.53) IU/L(P【0.05).Total bilirubin was reduced from(2.14±0.10 mg/dL) to(0.18±0.07) mg/dL (P【0.05) while total protein was increased from(4.26±0.30) mg/dL to(6.20±0.19) mg/dL(P【0.05). Concentrations ranging from 10 - 400μg/mL of B.micrantha were assayed for antioxidant activities.The DPPH assay showed 98%antioxidant activity at concentration of 400μg/mL. The FRAP values were 0.016,0.39,0.455,0.601 and 1.382μM at 10.50,100,200 and 400μg/ mL respectively.Conclusions:Results suggest that B.micrantha has hepatoprotective and anti oxidant potentials.However,further work involving fractionation needs to done to isolate the active compound responsible for the hepatoprotective activity.

Ethyl acetate extract of Smilax glabra Roxb roots and its major active compound astilbin promote osteoblastogenesis in vitro by upregulating bone cell differentiation-associated genes

Objective:To investigate the osteoblastogenic activity of the ethyl acetate(EtOAc)extract of Smilax glabra Roxb roots and its major active compound astilbin.Methods:Astilbin was isolated from EtOAc extract using silica gel chromatography combined with fraction crystallization.Chemical structure of astilbin was determined by analysis of the spectroscopic data in comparison with the literature.MTT method was used to detect the toxicity.Alkaline phosphatase(ALP)activity was determined by the spectrophotometric method at 405 nm using p-nitrophenyl phosphate as a substrate.Calcium deposition was stained with alizarin red-S,distained with cetylpyridium chloride,and quantified at 562 nm.In silico model for astilbin-ALP interaction was analyzed using AutoDock 4.2.6.The changes in expression of osteoblast differentiation related genes were determined using quantitative real-time PCR.Results:Both the EtOAc extract and astilbin had no toxicity toward osteoblast MC3T3-E1 cells at 5.0,10,25,and 50μg/mL.At 25μg/mL,they enhanced ALP activity and mineralization of osteoblasts up to 30%and 55%for the EtOAc extract and 22%and 41%for astilbin,respectively.Molecular docking analysis of astilbin-ALP interaction revealed Arg167,Asp320,His324,and His437 were key residues participating in hydrophobic interaction;meanwhile,His434 and Thr436 residues were involved in hydrogen bond formation in the active site of human tissue-nonspecific ALP.Moreover,the expression level of genes opn,col1,osx,and runx2 were up-regulated in astilbin treated samples with the fold changes as 2.2;3.7;4.1;2.3,respectively at 10μg/mL(P<0.05).Conclusions:The EtOAc extract and its major compound astilbin exhibit osteoblastogenic activity by up-regulating important markers for bone cell differentiation.It could be a new and promising osteogenic agent with dual actions for therapeutic applications.

Quantitative variation of bioactive phyto compounds in ethyl acetate and methanol extracts of Pergularia daemia(Forsk.)Chiov.

Dear Editor： Pergularia daemia Forsk （Asclepiadaceae） is a perennial twining herb commonly known as veliparuthi in Tamil. The plant has anthelmintic, laxative, antidiabetic, hepatoprotective and anti-inflammatory activities. The pharmacological properties of this plant come from bioactive phytochemicals such as alkaloids, triterpenes, saponins and flavonoids. Phytochemically, the plant has been investigated for the presence of cardenolides, alkaloids, saponins and steroidal compoundst. In the present study, we developed a rapid method for identification and quantitative determination of putative phyto compounds in the crude extracts of ethyl acetate and methanol from whole plant of Pergularia daemia.

Effects of Ethyl Acetate Extract of Phyllanthus reticulatus Leaves on Autophagy-related Proteins Beclin-1,ATG5 and LC3 in Nude Mice Xenograft Model of HCC

[Objectives]To explore the effects of ethyl acetate extract of Phyllanthus reticulatus leaves on autophagy-related proteins Beclin-1,ATG5 and LC3 by immunohistochemistry,and to preliminarily explore their effects on autophagy.[Methods]BEL-7404 Hepatocellular Carcinoma(HCC)nude mice model was established,and blank group(same volume of pure water),positive control group(20 mg/kg fluorouracil),high dose drug group(600 mg/kg),and medium dose drug group(300 mg/kg),and low dose drug group(150 mg/kg)were set up.After 2 weeks of intragastric administration,the nude mice were sacrificed,and the tumor tissues were taken out,processed by immunohistochemistry,and then made into paraffin sections.Photos were taken under an optical microscope(10×40),and evaluation and analysis were performed with the aid of the Image-Pro Plus 6.0 image analysis software.Differences were calculated using SPSS 20.0 software.The effects of drugs on autophagy-related proteins LC3,Beclin-1 and ATG5 were observed.[Results]Compared with the blank group,the medium and high dose groups of ethyl acetate extract of P.reticulatus leaves had the effect of promoting the increase of autophagy-related proteins LC3,Beclin-1 and ATG5(P<0.05).However,there was no significant difference between the low dose group of ethyl acetate extract of P.reticulatus leaves and the blank group(P>0.05).[Conclusions]The ethyl acetate extract of P.reticulatus leaves has a promoting effect on autophagy-related proteins LC3,Beclin-1,and ATG5.

Effects of pomegranate ethyl acetate extract on rat islet cells

Objective The present study investigated the effects of pomegranate ethyl acetate extract(PEE)on cell viability of primary rat islet cells.Methods PEE was extracted by using 95% ethanol and a series of organic solvents.Primary isolated islet cells of rat were cultured in high glucose and/or high fat conditions with or without PEE.Cell viability was determined by MTT assay.Insulin secretion was assessed by using radioimmunoassay.Results Insulin levels were significantly higher in cells treated with high glucose+PEE,palmitic acid+PEE,and olein acid+PEE compared to the cells treated with high glucose,palmitic acid,or oleic acid alone,in which the insulin levels were markedly less than that of the normal control group.Conclusion PEE promotes islet cell growth and insulin secretion,which is suggested that PEE can protect islet cell function in both high glucose and high fatty acid environments.

天茄子乙酸乙酯部位化学成分研究

采用脂多糖诱导小鼠单核巨噬细胞RAW 264.7炎症模型导向研究天茄子(Ipomoea turbinata seeds)的抗炎活性成分。采用硅胶、Sephadex LH-20、MCI、ODS柱色谱及半制备型高效液相等方法对乙酸乙酯活性部位进行分离纯化,结合现代波谱技术与理化性质确定化合物结构。从中共分离鉴定了15个化合物,分别为华佗豆甲碱(1)、华佗豆丙碱-4′-O-β-D-(6-O-反式香豆酰基)葡萄糖苷(2)、1,8,15,22-四氮杂环二十八烷-2,9,16,23-四酮(3)、1,8,15,22,29-五氮杂环三十五烷-2,9,16,23,30-五酮(4)、绿原酸甲酯(5)、绿原酸(6)、3,5-二咖啡酰基奎宁酸(7)、3,4-二咖啡酰基奎宁酸(8)、咖啡酸(9)、3,5-二羟基桂皮酸(10)、4-甲氧基肉桂酸(11)、6-氧-(E)-对羟基桂皮酰基-1-β-乙基-葡萄糖甙(12)、E-3-(3,4-二羟基苯亚甲基)-5-(3,4-二羟基苯基)二羟基呋喃-2-酮(13)、丁香脂素-4-O-β-D-吡喃葡萄糖苷(14)和松脂素-4-O-β-D-葡萄糖苷(15)。化合物3~8和10~15为首次从该植物中分离得到,其中8个化合物(3~5、10~13和15)为首次从旋花科中分离得到。活性筛选结果表明,咖啡酰奎宁酸类(5~8)和苯丙酸类成分(9~12)显示有较好的抑制NO释放活性,在50μmol/L浓度下抑制率为29.07%~40.30%。

灵芝复合剂乙酸乙酯提取物的化学组成及其抗肿瘤活性

该文研究了灵芝复合剂乙酸乙酯提取物(Ganoderma lucidum Complex Ethyl Acetate Extract,GCE)的化学组成及抗肿瘤活性。GCE由灵芝、菟丝子、五味子、党参(质量比50.3:25.2:15.7:8.8)组成复合剂后再经乙酸乙酯萃取所得。试验采用超高压液相色谱飞行时间质谱联用仪(UPLC-Q-TOF-MS)分析其物质组成;设空白组、模型组、阳性对照组、GCE低、中、高剂量组。检测其抑瘤率,脏器指数,脾淋巴细胞增殖能力,血清细胞因子。结果表明:GCE主要由11种化合物组成:L-焦谷氨酸、DL-亮氨酸腺苷、D-赤酮酸内酯、L-正亮氨酸、邻苯三酚、2-羟基-3-甲基吡喃-4-酮、绿原酸、3-(3,4,5-三甲氧基苯基)丙酸、槲皮素、五味子素;GCE低、中、高剂量组(50、100、200 mg/kg)H22荷瘤小鼠的抑瘤率分别为35.14%、38.25%、50.87%;与模型组相比,GCE低、中、高剂量均能显著促进小鼠脾淋巴细胞的增殖(P<0.01);GCE高剂量能显著促进小鼠的IL-6、TNF-α和IL-12分泌(P<0.05)。结论,GCE对H22荷瘤小鼠肿瘤有一定抑制作用,该研究结果可为抗肿瘤功能的产品研发提供新思路。

南方红豆杉果乙酸乙酯部位对抑郁症模型大鼠表型和炎症介质的影响

目的:探讨南方红豆杉果乙酸乙酯部位对抑郁症模型大鼠的影响及可能作用机制。方法:采用随机数字法将20只雌性斯泼累格·多雷(SD)大鼠随机分为对照组、模型组、红豆杉果组和氟西汀组,每组5只。除对照组外,其余各组采用慢性不可预知温和应激(CUMS)法4周制备抑郁症模型。造模后红豆杉果组给予南方红豆杉果乙酸乙酯提取物200 mg/(kg·d)灌胃,对照组和模型组给予等体积的生理盐水灌胃,共给药4周。在造模结束后对对照组和模型组进行糖水实验和旷场实验,给药结束后对各组大鼠进行糖水实验和旷场实验,检测各组大鼠血清中白细胞介素-1β(IL-1β)、IL-6、IL-18的含量。结果:在造模结束后与对照组比较,模型组大鼠的糖水消耗率明显减少(P<0.01),活动距离明显降低(P<0.01)。实验结束后,与模型组比较,红豆杉果组和氟西汀组大鼠的糖水消耗率明显增加(P<0.01),活动距离明显增加(P<0.01);与对照组比较,模型组血清中IL-1β、IL-6、IL-18的含量均明显升高(P<0.05,P<0.01);与模型组比较,红豆杉果组和氟西汀组血清中IL-1β、IL-6、IL-18的含量均明显下降(P<0.05,P<0.01)。结论:南方红豆杉果乙酸乙酯部位有抗抑郁的作用,同时有抗炎的作用,可能通过降低抑郁症大鼠炎症反应起到缓解大鼠抑郁样行为。

香椿枝叶的化学成分及其抗氧化活性研究

采用大孔树脂、葡聚糖凝胶Sephadex LH-20、硅胶、ODS及制备型HPLC等色谱技术对香椿枝叶乙酸乙酯部位进行分离纯化,通过NMR技术并结合文献对分离得到的化合物进行结构鉴定。从香椿枝叶分离鉴定了15个化合物,分别鉴定为:去氢吐叶醇(1)、(6R,9R)9-hydroxy-4-megastigmen-3-one(2)、blumenol A(3)、corchoionol C(4)、megastigm-5-en-3,9-diol(5)、badounoid B (6)(3S,5R,6S,7E,9R)-3,6-dihydroxy-5,6-dihydro-β-ionol(7)、3S,5R-dihydroxy-6R,7-mestigmadien-9-one (8)、3S,5R-dihydroxy-6S,7-mestigmadien-9-one (9)、foliasalaciol A(10)、loliolide(11)、(6R,9S)-9,10-dihydroxy-4-megastigmen-3-one(12)、嗪皮啶(13)、东莨菪亭(14)、杜鹃醇(15)。化合物2、12具有较弱的DPPH、ABTS自由基清除能力,化合物13~14具有明显DPPH、ABTS自由基清除能力。其中化合物1-12、15为首次从香椿属分离得到。表明化合物2、12-14具有一定的抗氧化活性。

榆干玉蕈子实体及其提取物的免疫调节作用

为研究榆干玉蕈(Hypsizygusulmarius)子实体粉末及提取物对小鼠的免疫调节作用,分别采用榆干玉蕈子实体粉末(高、低剂量分别为1.2、0.3g·kg^(-1))、石油醚提取物(高、低剂量分别为1.392、0.348mg·kg^(-1))、乙酸乙酯提取物(高、低剂量分别为3.306、0.826 mg·kg^(-1))、水提取物(高、低剂量分别为28.78、7.19 mg·kg^(-1))灌胃大黄水煎液建立的模型小鼠,测定其体质量、脾脏指数及血清中白细胞介素2(interleukin-2,IL-2)、白细胞介素6(interleukin-6,IL-6)、免疫球蛋白M(immunoglobulin M,IgM)、干扰素γ(interferon-γ,IFN-γ)、D-木糖(D-xylose)、白蛋白(albumin,ALB)、胰脂肪酶(pancrelipase,PL)含量,采用苏木素-伊红染色观察小鼠脾脏组织显微结构,采用RT-qPCR法检测维生素D结合蛋白(D binding protein,DBP)和紧密连接蛋白(occludin,OCLN)的基因表达情况,并采用Westernblotting方法检测DBP和OCLN蛋白表达情况。结果表明:与模型组相比,第14天,各给药组小鼠体质量均显著增加。粉末高剂量组脾脏指数显著增加。所有给药组D-xylose、IFN-γ、ALB、IL-2、IgM、PL含量均显著增加,IL-6含量显著降低。各给药高剂量组DBP基因相对表达量显著升高,所有低剂量组显著降低;粉末低剂量组、石油醚提取物高和低剂量组、水提物高和低剂量组的OCLN基因相对表达量均显著升高。各给药组DBP和OCLN的蛋白表达量均显著升高。榆干玉蕈子实体及各提取物均通过免疫调节发挥作用,研究结果可为榆干玉蕈进一步开发利用提供参考。

侧柏叶乙酸乙酯提取物缓解小鼠糖尿病心肌病

目的:研究侧柏叶乙酸乙酯提取物(EAEPOL)缓解糖尿病心肌病(DCM)的作用及其机制。方法:将健康成年C57BL/6小鼠随机分为正常对照组、DCM组和EAEPOL组。以小鼠心脏超声心动图为指标观察心脏结构和功能。以心肌羟脯氨酸含量、心肌组织Masson染色和天狼星红染色及心肌I型胶原(Col I)和III型胶原(Col III)免疫组化染色评价心肌纤维化程度。试剂盒检测心肌丙二醛(MDA)、超氧化物歧化酶(SOD)和总抗氧化能力(T-AOC),以及qRT-PCR和Western blot检测心肌血红素加氧酶1(HO-1)和核因子E2相关因子2(Nrf-2)表达水平,评价心肌氧化应激状态。Western blot检测CD31和α-平滑肌肌动蛋白(α-SMA)表达,qRT-PCR检测钙黏素5(CDH5)和成纤维细胞特异性蛋白1(FSP1)的mRNA水平,以及心肌石蜡切片α-SMA免疫荧光染色,评价心肌内皮-间充质转化(EndMT)程度。结果:(1)小鼠心脏超声心动图显示,与正常对照组相比,DCM组小鼠心率(HR)和射血分数(EF)显著下降(P<0.05或P<0.01),收缩期和舒张期左心室前壁厚度(LVAWs和LVAWd)及收缩期和舒张期左心室后壁厚度(LVPWs和LVPWd)均显著增大(P<0.05);与DCM组相比,EAEPOL组小鼠HR和EF显著增大(P<0.01);LVAWs、LVAWd、LVPWs和LVPWd显著减小(P<0.05)。(2)与正常对照组相比,DCM组小鼠心肌羟脯氨酸含量、天狼星红和Masson染色所示胶原面积比及Col Ⅰ和Col Ⅲ免疫组化阳性面积比均显著增加(P<0.01);与DCM组相比,EAEPOL组小鼠以上心肌纤维化指标均显著下降(P<0.05或P<0.01)。(3)与正常对照组相比,DCM组小鼠心肌MDA含量和Nrf-2表达显著增加,SOD活性、T-AOC水平和HO-1表达显著下降(P<0.01);与DCM组相比,EAEPOL组小鼠心肌MDA含量显著减少,SOD、T-AOC、Nrf-2和HO-1水平显著增加(P<0.05或P<0.01)。(4)与正常对照组相比,DCM组小鼠心肌CD31和CDH5的表达显著减弱,α-SMA和FSP1的表达显著增强(P<0.05或P<0.01),α-SMA免疫荧光阳性面积显著增大(P<0.01);与DCM组相比,EAEPOL显著上调CD31和CDH5的表达,下调α-SMA和FSP1的表达,α-SMA免疫荧光阳性面积显著下降(P<0.05或P<0.01)。结论:EAEPOL可能通过抑制氧化应激,减轻EndMT,从而抑制DCM小鼠心肌纤维化,改善心功能。

含羞草根乙酸乙酯部位对急性髓系白血病小鼠的影响

目的 研究含羞草根乙酸乙酯部位(简称为乙酸乙酯部位)对急性髓系白血病小鼠的影响。方法 以不同质量浓度(0.062 5、0.125、0.25、0.5 mg/mL)乙酸乙酯部位作用于急性粒-单核细胞白血病细胞WEHI-3,考察其对该细胞活力的影响。将50只BALB/C小鼠随机分为空白对照组、模型组、阳性对照组(5-氟尿嘧啶,13 mg/kg)和乙酸乙酯部位低、高剂量组(50、200 mg/kg),每组10只。除空白对照组外,其余各组小鼠均腹腔注射WEHI-3细胞悬液进行造模,并在造模第2天起灌胃相应药物/水,每天1次,连续14 d。末次给药后,检测小鼠肝、脾指数,并进行肝组织病理学形态观察和血液学分析以及白细胞分化检测;检测小鼠血清中细胞因子[白细胞介素2(IL-2)、IL-3、干扰素γ(IFN-γ)、肿瘤坏死因子α(TNF-α)]水平;检测小鼠全血中白细胞表面标志物[分化簇3(CD3)、CD19、CD11b、分化簇107b(即Mac-3)]抗体水平。结果 经0.062 5~0.5 mg/mL的乙酸乙酯部位作用后,WEHI-3细胞增殖抑制率均显著升高(P<0.05)。经高剂量的乙酸乙酯部位干预后,小鼠的肝、脾指数,血清中TNF-α水平,全血中白细胞表面CD11b、Mac-3抗体水平均显著降低(P<0.05),血清中IL-2、IL-3、IFN-γ水平和全血中白细胞表面CD3、CD19抗体水平均显著升高(P<0.05);肝实质偶有淋巴细胞浸润,几乎无炎症细胞浸润;血液学指标改善且白细胞分化减弱。结论 含羞草根乙酸乙酯部位可抑制WEHI-3细胞增殖,改善急性髓系白血病小鼠症状,其作用机制与增强机体免疫功能有关。

基于质控与药效统一的扁蕾颗粒标准提升探讨

采用薄层色谱法,对扁蕾颗粒中齐墩果酸进行定性鉴别;采用萃取重量法测定扁蕾颗粒中乙酸乙酯提取物;采用高效液相色谱法测定药效成分木犀草素的含量。色谱柱为Agilent C18柱(250mm×4.6mm,5μm),流动相为甲醇-0.1%磷酸溶液(50:50),V/V;流速1.0mL/min;检测波长为350nm;柱温:25℃。薄层色谱图谱获得斑点较多,分离效果好,阴性对照无干扰;乙酸乙酯提取物为2.56mg/g~2.87mg/g;木犀草素在0.11μg~1.06μg范围内呈良好的线性关系,平均加样回收率为98.10%,RSD为2.05%(n=6)。该方法科学合理,简单易行,可较为全面的评价扁蕾颗粒质量。

毛菊苣乙酸乙酯萃取物对大鼠溃疡性结肠炎合并急性肝损伤防治作用研究

目的研究毛菊苣根乙酸乙酯萃取物(CGEA)对大鼠急性肝损伤合并溃疡性结肠炎的保护作用。方法将SD大鼠36只随机分为正常组(Control)、模型组(Model)、柳氮磺吡啶肠溶片阳性对照组(Sulfasalazine,100 mg·kg^(-1))和CGEA高、低剂量组(150、100 mg·kg^(-1)),每组6只,除正常组外其余各组给予30%TNBS-40%乙醇溶液灌肠(60 mg·kg^(-1)),同时皮下注射含体积分数40%CCl4的橄榄油溶液2 mL·kg^(-1),一周2次,制备急性肝损伤模型合并溃疡性结肠炎模型,造模成功后将CGEA制备成水混悬液,按照CGEA高剂量组(150 mg·kg^(-1)),CGEA低剂量组(100 mg·kg^(-1))连续灌胃大鼠14 d,每日1次。14 d后,处死大鼠并取血清及肝脾组织,计算肝脾指数,检测大鼠血清相关指标;采用HE染色法观察肝、肠组织病理变化。结果与正常组比较,模型组大鼠肝脾指数明显上升(P<0.01);血清中AST、ALT、AKP、γ-GT和MDA水平显著升高(P<0.05或P<0.01),SOD水平显著降低(P<0.05);肝组织遭到破坏,炎性细胞浸润;结肠组织结构杂乱,出现炎症细胞浸润现象。与模型组比较,给药组肝脾指数显著降低(P<0.01);血清中AST、ALT、AKP、γ-GT和MDA指标水平显著降低(P<0.05或P<0.01);给药组肝肠组织破坏程度减轻,炎性细胞浸润减少。结论CGEA可以减轻大鼠溃疡性结肠炎合并急性肝损伤程度,对其起到保护作用。

空心莲子草乙酸乙酯部位体内外抗H9N2亚型禽流感病毒活性的研究

[目的]探究空心莲子草乙酸乙酯部位(ethyl acetate extract from Alternanthera philoxeroides,AETAC)体内外抗H9N2亚型禽流感病毒(H9N2 subtype Avian influenza virus, AIV-H9N2)活性及其作用机制。[方法]利用CCK-8法检测不同浓度AETAC作用不同时间对鸡胚成纤维细胞(DF-1)的抑制作用,通过预防感染、影响复制、阻止吸附及直接杀灭4种方式作用后检测AETAC对DF-1细胞的毒性,选择体外最佳抗AIV-H9N2作用方式。尿囊腔接种不同浓度AETAC确定其对鸡胚的最大安全浓度;将AETAC以3种不同给药方式作用于AIV-H9N2感染后鸡胚,选择体内最佳给药方式。经尿囊腔同时接种AETAC和AIV-H9N2,统计各组鸡胚存活数,测定血凝效价,并观察胚胎的发育情况和鸡胚法氏囊病理组织变化。[结果]在一定范围内,随着AETAC浓度升高或作用时间延长,AETAC对DF-1细胞的抑制率升高。与空白对照组相比,4种抗AIV-H9N2作用方式下,病毒对照组和药物组细胞存活率均显著降低(P<0.05);与病毒对照组相比,AETAC浓度为5~30μg/mL时,4种抗AIV-H9N2作用方式下,药物组细胞存活率均显著升高(P<0.05)。4种作用方式下,DF-1细胞存活率分别为8.15%~64.96%(预防感染)、8.56%~83.83%(影响复制)、1.82%~5.90%(阻止吸附)和6.90%~40.12%(直接杀灭)。AETAC对鸡胚的最大安全浓度为16.41 mg/mL,对感染鸡胚的最佳给药方式为AETAC和病毒混合后接种。以最佳体内作用方式给药后,AETAC中、低浓度组鸡胚存活率为40%~60%,高浓度组鸡胚全部存活,胚胎喙部发育明显,鸡胚法氏囊淋巴滤泡发育完整,滤泡大小和数量正常,充血量少,组织病理状态改善。[结论]AETAC在体内外对AIV-H9N2均具有显著的抑制作用,且主要与影响病毒复制作用方式相关。

基于Aspen的乙酸乙酯-乙醇-水体系恒沸精馏和萃取精馏模拟

利用AspenPlus软件模拟乙酸乙酯-乙醇-水体系先恒沸精馏,再用水作为萃取剂,通过三次萃取精馏,最终得到质量分数大于99.5%乙酸乙酯成品,总回收率约89%~90%,萃取剂通过提馏塔提纯,循环使用。研究思路和方法适用于二元或三元共沸物体系的精馏提纯。

Chemical Composition of Rubus pirifolius Smith

[Objectives]This paper aimed to study the chemical composition of Rubus pirifolius Smith[Methods]A preliminary experiment was carried out using the qualitative reactions of various chemical constituents,in order to roughly detect the types of chemical constituents that may be contained in R.pirifolius Smith The ethanol extract of R.pirifolius Smith were separated and purified using silica gel column chromatography,silica gel thin-layer chromatography and recrystallization,and the chemicals obtained were identified using mass spectrometry,nuclear magnetic resonance and literature reference.[Results]The main constituents of R.pirifolius Smith are saponins,tannins,flavonoids,phytosterols,triterpenes and anthraquinones.A total of five compounds were isolated from the ethanol extract of R.pirifolius Smith,and they were identified asβ-sitosterol(1),arjunolic acid(2),stigmasterol-3-O-β-D-glucopyranoside(3),euscaphic acid(4)and 3,3′,4-trimethyle llegic acid(5).[Conclusions]Compounds 1,2,3,4 and 5 were isolated from R.pirifolius Smith for the first time.

In vitro cytotoxicity of Indonesian stingless bee products against human cancer cell lines

Objective:To screen crude extracts of propolis,bee pollen and honey from four stingless bee species[Trigona incisa(T.incisa)],Timia apicalis,Trigona fuso-baltata and Trigona filscibasis)native to East Kalimantan.Indonesia for cytotoxic activity against five human cancer cell lines(HepG2,SW620,ChaGo-1,KATO-Ⅲand BT474).Methods:All samples were extracted with methanol,and then subpartitioned with n-hexane and ethyl acetate.Each crude extract was screened at 20μg/mL for in vitro cytotoxicity against the cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.Tn addition,four previously shown bioactive components from propolis(apigenin,cafieic acid phenyl ester,kaempferol and naringenin)and two chemotherapeutic drugs(doxorubicin and 5-fluorouracil)were used to evaluate the sensitivity of the cell lines.Results:Overall,crude extracts from propolis and honey had higher cytotoxic activities than bee pollen,but the activity was dependent upon the extraction solvent,bee species and cell line.Propolis extracts from T.incisa and Tarda apicalis showed the highest and lowest cytotoxic activity,respectively.Only the HepG2 cell line was broadly sensitive to the honey extracts.For pure compounds,doxorubicin was the most cytotoxic,the four propolis compounds the least,but the ChaGo-I cell line was sensitive to kaempferol at 10μg/mL and KATO-Ⅲwas sensitive to kaempferol and apigenin at 10μg/mL,.All pure compounds were effective against the BT474 cell line.Conclusions:Propolis from f,incisa and Trigona fusco-balteata contain an in vitro cytotoxic activity against human cancer cell lines.Further study is required,including the isolation and characterization of the active antiproliferative agent(s).