Antihyperlipidaemic activity of swertiamarin,a secoiridoid glycoside in poloxamer-407-induced hyperlipidaemic rats

We have investigated antihyperlipidaemic effect of swertiamarin (50 mg/kg, oral once) isolated from the perennial herb Enicostemma littorale Blume in poloxamer 407 (P-407)-induced hyperlipidaemic rats. Rats were made hyperlipidaemic by intraperitoneal administration of P-407 (400 mg/kg). Serum lipid levels such as total cholesterol, triglycerides and low-density lipoprotein cholesterol increased significantly (P < 0.001) compared with normal control rats. All these changes were significantly prevented in the rats treated with swertiamarin. Serum high-density lipoprotein (HDL) cholesterol was found to be reduced in the P-407 control rats. However, administration of swertiamarin significantly (P < 0.01) increased HDL levels and it showed a significant lipid-lowering effect, as well as a high antiatherogenic potential. Overall swertiamarin is an effective lipid-lowering lead compound and can be useful for preventing atherosclerosis.     

Antioxidant activities of the flaxseed lignan secoisolariciresinol diglucoside, its aglycone secoisolariciresinol and the mammalian lignans enterodiol and enterolactone in vitro.

The flaxseed lignan secoisolariciresinol diglucoside (SDG) and mammalian lignans enterodiol (ED) and enterolactone (EL) were previously shown to be effective antioxidants against DNA damage and lipid peroxidation. Others reported inhibition of activated cell chemiluminescence by supra-physiological concentrations of secoisolariciresinol (SECO), ED and EL. Thus, we evaluated the antioxidant efficacy of potential physiological concentrations of SDG, SECO, ED and EL against 1,1-diphenyl-2-picrylhydrazyl (DPPH()), and 2,2'-azo-bis(2-amidinopropane) dihydrochloride (AAPH)-initiated peroxyl radical plasmid DNA damage and phosphatidylcholine liposome lipid peroxidation. SDG and SECO were effective (p<0.01) antioxidants against DPPH() at 25-200muM; whereas, ED and EL were inactive. Efficacy of lignans and controls against AAPH peroxyl radical-induced DNA damage was: SDG>SECO=17alpha-estradiol>ED=EL>genistein>daidzein. Lignan efficacy against AAPH-induced liposome lipid peroxidation was: SDG>SECO=ED=EL. Plant lignan antioxidant activity was attributed to the 3-methoxy-4-hydroxyl substituents of SDG and SECO, versus the meta mono-phenol structures of ED and EL. Benzylic hydrogen abstraction and potential resonance stabilization of phenoxyl radicals in an aqueous environment likely contributed to the antioxidant activity of the mammalian lignans. These represent likely extra- and intracellular antioxidant activities of flax-derived lignans at concentrations potentially achievable in vivo.     

贻贝提取物抗高血脂作用的观察

本实验采用食饵性高血脂模型观察了胎贝提取物的调节血脂作用。结果表明：贻贝提取物可显著降低食饵性高血脂大鼠血浆胆固醇和甘油三酯。     

Evaluation of antihyperlipidemic effect of Polygonum minus,a South East Asian salad plant in poloxamer 407-induced acute hyperlipidemic rats

OBJECTIVE To investigate the antihyperlipidemic effect of methanol and aqueous extracts of the leaves of Polygonum minus in acute hyperlipidemic rat model.METHODS Acute hyperlipidemia was chemically induced in Sprague Dawley rats by using poloxamer 407(500mg·kg-1 of body weight;intraperitoneal).Increase in total cholesterol was confirmed after six hours of induction.The normal and hyperlipidemic control groups were administered with 1mL carboxymethylcellulose(CMC),the two test groups received aqueous and methanol extract of leaves of P.minus respectively(1000mg·kg-1;orally;suspended in 1% CMC)whereas reference standard treated group received atorvastatin(60mg·kg-1;orally;suspended in 1% CMC)once daily for 3consecutive days.Blood samples were collected at 10 th and 24 th hour of the study for total cholesterol and triglycerides determination,while terminal blood samples were collected at58 th hour for full lipids profile analysis.RESULTS In the present study,both methanol and aqueous extracts lowered the serum total cholesterol and triglycerides significantly(P<0.001 and P<0.05,respectively)when compared with the hyperlipidemic control,similar to the standard drug atorvastatin.Varying effects were observed for both extracts on the other lipid parameter studied.Methanol extract showed significant reduction in LDL(P<0.05),VLDL(P<0.01)and atherogenic index(AI;P<0.001)and it showed a significant elevation in HDL levels(P<0.05).On the other hand,the aqueous extract showed significant reduction only in VLDL and AI(P<0.05)but no increase in HDL levels.CONCLUSION Present study confirmed the antihyperlipidemic effect of leaves of P.minus in acute hyperlipidemic rat model.The study also suggested that the methanol extract possess higher antihyperlipidemic effect than aqueous extract.Currently a study is ongoing to evaluate the antihyperlipidemic effect of the methanol extract in high-fat diet-induced chronic hyperlipidemic rat model.     

短尾蝮蛇毒纤溶酶原激活剂的分离纯化及活性测定

目的从短尾蝮蛇毒中分离纯化纤溶酶原激活剂(plasminogen activator of Gloydius brevicaudus venom,GBV-PA),对其理化性质及部分生物活性进行研究。方法应用苯甲脒-琼脂糖凝胶(Benzamidine Sepharose6B)亲和层析及Lichrospher C18反相层析柱进行分离纯化,应用SDS-PAGE测定分子量、聚丙烯酰胺凝胶盘状电泳测定等电点,以发色底物法测定生物活性。结果通过亲和层析、反相层析等方法可从短尾蝮蛇毒中分离纯化出纤溶酶原激活剂至电泳纯以上,其相对分子质量约为32.6×103,等电点约为5.2,能特异激活人纤溶酶原为纤溶酶,其比活性为2.87t-PA IU.mg-1;该酶为丝氨酸蛋白酶,对纤维蛋白无亲和性。结论应用亲和层析和反相层析可从短尾蝮蛇毒获得一种纤溶酶原激活剂;该酶为丝氨酸蛋白酶,对纤维蛋白无亲和性。     

茶薪菇子实体多糖的分离纯化和抗氧化活性的测定

烘干研磨成粉的茶薪菇(Agrocybe cyclindracea)子实体用热水提取，乙醇沉淀，Sevage法去蛋白，分级沉淀，得多糖PⅠ、PⅡ、PⅢ。Pm用DEAE纤维素、SephadexG-200柱层析进一步纯化，得PⅢ’，聚丙烯酰胺凝胶电泳和葡聚糖凝胶柱层析证明PⅢ’为均一组分，分子质量为94400u。抗氧化活性的测定实验表明，PⅠ、PⅡ、PⅢ均有抗氧化活性。     

Sex does not seem to influence the formation of aortic lesions in the P-407-induced mouse model of hyperlipidemia and atherosclerosis

Coronary heart disease secondary to atherosclerosis is the leading cause of death for men in the United States. Using a new, nontransgenic, non-fat-fed mouse model of hyperlipidemia and atherosclerosis developed in our laboratory, we investigated the effect of sex on lipid profiles and subsequent aortic atherosclerotic lesion formation. Female and male C57BL/6 mice, which consumed a low-fat diet, were treated with either normal saline or poloxamer 407 (P-407), a triblock copolymer comprised of poly(oxyethylene) and poly(oxypropylene) units, for 4 months. Blood samples were obtained at 0, 1, 2, 3, and 4 months, whereas hearts and livers were harvested only at 4 months, because this model requires approximately 4 months for significant atheroma formation. P-407-treated mice of either sex demonstrated a profound increase in plasma cholesterol and triglyceride; at 3 and 4 months the plasma lipids were significantly (p < 0.05) higher for male mice compared with female mice. Aortas retrieved from P-407-treated mice of either sex after 4 months demonstrated a significant (p < 0.001) increase in the mean atherosclerotic lesion size compared with their respective saline-treated controls, but there was no significant (p > 0.05) difference between lesion sizes for P-407-treated male mice (1.02 +/- 0.074 x 10(5) microm(2)) compared with P-407-treated female mice (1.14 +/- 0.28 x 10(5) microm(2)). Livers harvested at 4 months from either sex of P-407-treated mice displayed no damage to hepatocytes but increased proliferation of macrophages (Kupffer cells), which contained sequestered lipids. Thus, male C57BL/6 mice form atherosclerotic lesions as extensive as female mice in the P-407 mouse model of atherosclerosis.     

Pharmacokinetics of clomipramine, an antidepressant, in poloxamer 407-induced hyperlipidaemic model rats

Objective This study was undertaken to investigate the effects of hyperlipidaemia on the pharmacokinetics of clomipramine, an antidepressant, particularly addressing the change of clomipramine distribution to plasma components in poloxamer 407‐induced hyperlipidaemia model rats. Methods Clomipramine pharmacokinetic studies in hyperlipidaemic rats were performed with clomipramine continuous infusion. Furthermore, clomipramine protein binding and distribution to the brain and plasma components such as lipoproteins were investigated. Key findings Mean plasma concentration of clomipramine at steady state during continuous infusion (17.5 µg/min/kg) in hyperlipidaemic rats (0.45 ± 0.01 µg/ml) was significantly higher than that in the control rats (0.30 ± 0.02 µg/ml). However, the amount of clomipramine in the brain in hyperlipidaemic rats (0.31 ± 0.06 µg/g) was dramatically lower than in the control rats (1.89 ± 0.13 µg/g). However, the plasma unbound fraction in hyperlipidaemic rats (0.98 ± 0.05%) was significantly lower than that of the control rats (6.51 ± 0.62%). Conclusions Lower distribution to the brain and lower plasma clearance of clomipramine in hyperlipidaemic rats resulted from lower plasma unbound fraction because of higher lipid‐rich protein contents in blood. Results of this study provide useful information for dosage adjustment of clomipramine in hyperlipidaemia.     

猴头菌多糖的分离纯化及活性探讨

目的 :猴头菌多糖的分离纯化及活性探讨。方法 :人工栽培的猴头菌子实体经热水提取、去蛋白、超滤、醇化、真空干燥得多糖粗品 ,经 DEAE- 5 2、Sephacryl S- 4 0 0柱层析得到精品多糖 HEP,并对 HEP进行免疫活性研究。结果 :精品多糖HEP均一、分子量为 5 .5× 10 4 ,可明显增强细胞因子 IL- 2的生物活性。结论 :猴头菌多糖分离纯化工艺的建立及其免疫活性研究结果为以后进一步研究和开发猴头菌多糖提供了便利条件。     

Isolation of the Lignan 6-Methoxypodophyllotoxin from Linum boissieri. and Biosynthesis of Lignans in this Species

Abstract

We were able to establish a suspension culture of Linum boissieri. that produces 6-methoxypodophyllotoxin (6MPT). As a first step to gain insight into the lignan biosynthesis in L. boissieri. cell cultures, we were able to measure phenylalanine ammonia-lyase (PAL) activity in raw protein extracts. PAL is a key enzyme in the early part of the general phenylpropanoid pathway, leading (beside others) to the precursors for lignan biosynthesis.     

珠母贝氨基多糖的分离纯化及其抗肿瘤活性的初步研究

探讨珠母贝氨基多糖的分离纯化及其抗肿瘤活性。马氏珠母贝全脏器经胰酶和枯草杆菌蛋白酶双酶水解、醇沉后得到氨基多糖粗品(CPG)．再经DEAE-52-纤维素柱纯化，并采用MTT法检测其体外抗肿瘤活性。结果表明：CPG和经DEAE-52-纤维素柱纯化级分GAG-1，GAG-2中总氨基己糖的质量分数分别为45．9％，56．5％和59.2％，级分GAG-1硫酸基的质量分数高达13.8％；体外抗肿瘤试验表明CPG，GAG-1和GAG-2具有抑制肿瘤细胞生长的作用，对HL-60细胞的抑制率分别为54．6％。50.0％和35．4％，且能显著增敏化疗药物5-Fu抑制肿瘤细胞的作用，其中GAG-1为主要的活性成分。     

巴戟天中大黄素甲醚的提取、分离和纯化方法研究

目的研究获得高纯度大黄素甲醚的可行性方法。方法先用稀硫酸溶液将蒽醌苷水解成苷元,再用热氯仿将苷元提取出来,用2%NaOH反复萃取将大黄素甲醚从氯仿液中提取出来,用薄层色谱法及层析柱法将大黄素甲醚进行分离纯化,经用丙酮反复多次重结晶可得纯度较高的大黄素甲醚。结果本方法提纯得到的大黄素甲醚的纯度较高。结论该方法操作简单、实用性强、成本低,有推广价值。     

Isolation,characterization and anticancer activity of secondary metabolites from Verbascum speciosum

Herein, two iridoid glucosides aucubin (1) and ajugol (2), and two phenyl ethanoids, verbascoside (3) and poliumoside (4) were isolated from the methanol extract of the aerial parts of Verbascum speciosum and used to study about their anticancer activity for the first time. The structures of all compounds were elucidated using spectroscopic data (IR, 1D and 2D NMR, LC-TOF/MS). Antiproliferative activities of Aucubun ( 1 ) and Verbascoside (3) were tested against A-549 (human colon cancer), MDA-MD-453 (human breast cancer) and 3T3-L1 (mouse fibroblast)cell lines by XTT assay. In addition, the anticarcer mechanism of action of aucubin (1) was investigated on MDA-MB-453 cells for the first time. XTT result showed that both applied compounds exhibited antiproliferative effect at different dose ranges depending on the cancer type, as well as selectivity between cancer and healty cell lines. Flow cytometry analyzes revealed that aucubin (1) exerts its cytotoxic effect in MDA-MB-453 cells by directing cells to early apoptosis and inhibiting the P13K/AKT signaling pathway.     

Isolation and characterization of bulgecins, new bacterial metabolites with bulge-inducing activity

Bulgecins A, B and C, new bacterial metabolites which induce the formation of bulges by cooperation with beta-lactam antibiotics, were isolated from the culture broth of Pseudomonas mesoacidophila. The three components, separated by column chromatography on QAE-Sephadex A-25, are water-soluble acidic compounds containing a sulfate group in the molecule. Acid hydrolysis showed that D-glucosamine and a new proline derivative are common constituents of the three components. In addition, taurine and beta-alanine are constituents of bulgecins A and B, respectively.     

决明子中大黄素的提取、分离和纯化方法的研究

目的寻求获得高纯度大黄素的可行性。方法先用稀硫酸溶液将蒽醌苷水解成苷元，再用热氯仿将苷元提取出来，然后用PH8．0缓冲液将大黄酸从氯仿液中除去，再用0．2％NaOH反复萃取将大黄素从氯仿液中提取出来，用层析柱法将大黄素分离纯化，经用丙酮反复多次重结晶可得纯度较高的大黄素。结果本方法提纯得到的大黄素的纯度达到98．1％以上。结论该方法操作简单、实用性强、成本低、可以推广应用。     

Isolation of a new aloe-emodin dianthrone diglucoside from senna and its potentiating effect on the purgative activity of sennoside A in mice

Two aloe-emodin dianthrone diglucosides (I and II) were isolated from the leaves of Cassia angustifolia Vahl by successive column chromatography with Amberlite XAD-2, silica gel, Polyamide C-200 and Sephadex LH-20. The stereostructures of I and II were elucidated as trans and meso isomers at 10-10', respectively, from the patterns of the ultraviolet absorption spectra and circular dichroism curves. This is the first report of isolation of diglucoside I from senna. Despite the lack of purgative activity, diglucoside I exerts a potentiating effect of about 1.3 times on the purgative activity of sennoside A in mice when even 15% is included in the mixture. The difference between I and a third active glycoside based on aloe-emodin is also discussed.     

Optimization of the extraction conditions and quantification by RP-LC analysis of three alkaloids in Zanthoxylum nitidum roots

Context: A classic traditional Chinese medicine, Zanthoxylum nitidum (Roxb.) DC. widely used in China, exhibits anticancer, anti-inflammatory and antianalgesic activities. Alkaloids are one of the main bioactive components. It is urgent to develop a simple and reliable method to determine the main alkaloids in Z. nitidum roots.

Objective: To determine the three alkaloids in Z. nitidum roots, a reversed-phase liquid chromatographic (RP-LC) method combined with an optimum extraction condition was established.

Materials and methods: A method involving four-factor-three-level orthogonal array design including the extracting solvent and the RP-LC condition was assayed. Twenty batches were collected from different areas of the Guangxi Province at different harvesting times. The determined alkaloids were nitidine chloride (NC, 1), ethoxychelerythrine (2) and liriodenine (3). The stable mobile phase was a C₁₈ packing, and the mobile phase was acetonitril-aqueous phosphoric acid-triethylamine-buffer solution.

Results: The optimum extraction and detection conditions have been determined in the process of quantification of Z. nitidum root alkaloids. The three alkaloids were detected simultaneously in the 20 batches of samples. The results clearly showed that alkaloid concentrations differed significantly among Z. nitidum collected from various collection areas.

Discussion and conclusion: We have established an optimum extraction and detection conditions in the process of quantification the three alkaloids in Z. nitidum roots. From this research, the most influenced factor on Z. nitidum roots was the collecting location, and the next factor was the harvesting time. The collecting location and the harvesting time should be considered as the high-quality medicinal herbs factors.     

Pharmacological studies on the bungarotoxins: separation of the fractions and their neuromuscular activity

A commercial sample of Bungarus multicinctus venom was separated into its constituent fractions by column chromatography on CM-Sephadex C-50. The fractions were tested on the responses of the chick biventer cervicis preparation to nerve stimulation, and to added acetylcholine and carbachol.13 fractions were obtained: 1 was inactive, 4 exhibited weak postjunctional neuromuscular blocking activity, 3 had potent postjunctional blocking activity, and the remaining 5 fractions exhibited predominately prejunctional blocking activity. Use of higher concentrations of the prejunctional toxins revelead that 2 of these fractions also possessed some postjunctional blocking activity.Because small differences in separation procedures may result in differences in the elution pattern of the venom, and because some fractions may possess nonspecific actions only obvious at high concentrations, it is suggested that, when using venom components as experimental tools, the separation method is detalied and the fractions thoroughly tested.     

藻类脂肪酸分离提纯及检测

报道了藻类脂肪酸用索氏提取器提取脂肪，甲酯化后用气相色谱仪测定的方法，从单细胞藻类中提取脂肪酸必须破细胞壁后再用乙醚浸泡，提取的时间要达12h以上，大型藻类不用破壁，室温下用乙醚浸泡24h，再用索氏法提取5h左右即可，脂溶性色素含量较高的样品，用乙醚提取的脂肪要在检测之前用活性炭和硅藻土进行脱色，两种物质的量要适宜，否则将影响脂肪酸的含量，硅胶G－CMC混合薄层层析（正己烷，乙酸乙酯85：15为展层剂，10％磷钼酸乙醇液为显色剂），是快速检测脂肪酸的方法，计算脂肪酸的含量时，仅能计算加入石油醚和苯的上层体积，因为下层不含有脂肪酸。     

Isolation and characterization of some potential impurities in ropinirole hydrochloride

Three impurities in ropinirole hydrochloride drug substance at levels approximately 0.06-0.15% were detected by reverse-phase high performance liquid chromatography (HPLC). These impurities were isolated from the drug substance. These impurities were analyzed using reverse-phase HPLC. Based on the spectral data (IR, NMR and MS), structures of these impurities were characterized as 4-[2-(propylamino) ethyl]-1,3-dihydro-2H-indol-2-one hydrochloride (impurity-A), 5-[2-(diropylamino) ethyl]-1,4-dihydro-3H-benzoxazin-3-one hydrochloride (impurity-B) and 4-[2-(diropylamino) ethyl]-1H-indol-2,3-dione hydrochloride (impurity-C). Synthesis of these impurities is discussed.