马齿苋提取物制备工艺、安全性及抗敏功效的研究

以透明质酸酶抑制率为考察指标,通过正交实验,确定了最佳醇提工艺:乙醇浓度为75%(体积分数),料液比1∶10,提取温度60℃,提取时间1 h,提取液过滤脱色浓缩冷冻干燥后,得到马齿苋醇提物。同浓度下的马齿苋醇提物的透明质酸酶抑制率远高于水提物,红细胞溶血实验说明马齿苋水提物和醇提物的刺激性低,安全性高。人体斑贴实验表明,马齿苋醇提物能减轻或抑制常见致敏源对皮肤的刺激作用,减弱皮肤炎症反应,增强皮肤的屏障功能。     

土牛膝醇提物提取工艺优化及体外抗人肺腺癌A549细胞活性研究CSCD

土牛膝是应用广泛的药用植物,其功效众多,为研究其醇提物抗癌功效,采用Box-Behnken响应面法优化土牛膝的醇提工艺,并对醇提物进行体外人肺腺癌A549细胞毒性及细胞划痕试验。获得了最佳提取工艺为:提取温度90℃,液料比40∶1,提取时间2 h,在此条件下,土牛膝醇提物提取率为9.23%。细胞毒性试验结果显示,随着醇提物浓度的上升,A549细胞的存活率迅速下降,当醇提物浓度为256μg/mL时,对A549细胞的抑制率达94.65%,且细胞划痕结果显示醇提物对A549细胞迁移有一定抑制作用,可为研究土牛膝药用特性提供参考。     

紫皮石斛总酚提取工艺优化及其抗氧化活性研究

目的：研究紫皮石斛总酚提取工艺及体外抗氧化活性。方法：在单因素试验的基础上，采用响应面法优化紫皮石斛总酚的超声波辅助提取工艺条件，通过测定紫皮石斛总酚对超氧阴离子、DPPH自由基及羟自由基的清除率，评价其抗氧化活性。结果：紫皮石斛总酚最优提取工艺条件为乙醇浓度69%，料液比1∶40 (g/mL)，提取时间31 min，在此条件下总酚提取量为1.56 mg/g，与理论值1.58 mg/g接近。紫皮石斛总酚对超氧阴离子、羟自由基及DPPH自由基的半数抑制浓度(IC₅₀)分别为19.40μg/mL、118.35μg/mL及30.75μg/mL。结论：优选的紫皮石斛总酚提取工艺合理可行，且其具有较强的体外抗氧化活性。     

香水莲花提取物抑制酪氨酸酶活性的研究

本实验以L-酪氨酸为底物,从马铃薯中提取酪氨酸酶,以熊果苷为阳性对照,采用比色法测定了香水莲花醇提物、水提物及总黄酮提取物对酪氨酸酶的抑制率,以探讨香水莲花醇提物、水提物及总黄酮提取物对酪氨酸酶的活性的抑制作用。结果表明,三种提取物对酪氨酸酶活性的抑制效果从高到低依次为总黄酮提取物、醇提物、水提物,说明其有效成分为醇溶性物质;总黄酮提取物对酪氨酸酶活性的抑制效果明显高于醇提物和水提物对酪氨酸酶活性的抑制效果,可以考虑将其作为一种新型的植物源美白剂,应用于化妆品行业。     

密花香薷体外对代谢综合征相关酶抑制活性研究

目的：探讨密花香薷粗提物体外对酶抑制的作用，初步阐明其对代谢综合征的药效物质基础。方法：体外测定密花香薷粗提物及其不同萃取部位对多种酶活性的抑制率，与阳性对照比较IC₅₀值，高效液相色谱法测定密花香薷中主成分含量。分子对接计算主成分与酶抑制作用相关目标基因的结合能。结果：密花香薷粗提物对α-淀粉酶、黄嘌呤氧化酶、胰脂肪酶、乙酰胆碱酯酶均有一定的抑制作用，IC₅₀值分别为0.199 mg/mL、0.254 mg/mL、0.258mg/mL、0.057 mg/mL。密花香薷乙酸乙酯部分对酶抑制作用最强，其中含量最高为绿原酸57.40 mg/g和槲皮素37.51 mg/g。分子对接发现绿原酸、槲皮素与以上代谢综合征酶抑制作用相关目标基因结合作用良好。结论：密花香薷粗提物对代谢综合征相关多种酶有一定的抑制活性，初步认为密花香薷治疗代谢综合征的药效物质基础可能为绿原酸与槲皮素。     

波叶大黄多糖对胰腺五种酶的抑制作用

 本文报道波叶大黄多糖(Rheum hotaoense polysaccharide,RHP)对胰腺五种酶的抑制作用。结果表明。(1)RHP对胰蛋白酶、胰脂肪酶、胰淀粉酶、胰弹性蛋白酶和胰激肽释放酶均有很明显的抑制作用。IC_(50)分别为250μg/mL、21μg/mL、18μg/mL,189μg/mL和300μg/mL。(2)动力学研究表明,RHP对胰蛋白酶和胰脂肪酶的抑制均是非竞争性的,Km值分别为1.2×10~(-4)μmol/mL和1.2mg/mL,Ki值分别为355.0μg/mL和63.85μg/mL。(3)牛血清白蛋白(BSA)对RHP抑制胰蛋白晦和胰脂肪酶具有拮抗作用。当BSA浓度达72.0mg/mL时,对胰蛋白酶活性的恢复率为71.54％。当BSA浓度达20.0mg/mL时,对胰脂肪酶活性的恢复率为63.64％。上述结果表明,RHP对胰腺五种酶的抑制作用,可能是大黄治疗急性胰腺炎作用的生化机制之一,预示RHP在临床上将有重要应用前景。     

植物来源的脂肪酶抑制剂的筛选

目的：随着人们生活水平的不断提高,肥胖问题日趋严重。胰脂肪酶抑制剂是药物治疗肥胖的有效途径之一,本文主要通过筛选一些植物来源的化合物来寻找具有胰脂肪酶抑制作用的天然活性物。方法：以4-甲基伞形酮油酸酯作为底物,用荧光检测的方法测定对胰脂肪酶（triacylglycerol lipase,EC 3.1.1.3）的抑制活性。结果：皂苷类化合物可有效地抑制胰脂肪酶的活性,其中薯蓣皂苷,毛地黄皂苷和皂树皂苷的活性较强,IC50值分别为3.5μM、2.7μM和3.0μg/mL。黄酮类化合物也有抑制脂肪酶活性的作用,在浓度为100μM时,对胰脂肪酶活性的抑制率在35.7%到68.2%之间。相比之下,相同浓度的苷元类化合物和咖啡酸衍生物对胰脂肪酶活性的抑制作用低于50%,而氨基糖类化合物的抑制率则低于15%。结论：皂苷这类植物的次级代谢产物具有良好的抑制脂肪酶活力的作用,黄酮次之,并且这两类化合物是从植物中提取的,毒性小,安全性高,可以为进一步研发治疗肥胖药物提供理论依据。     

米胚芽中胰脂肪酶抑制剂的提取工艺

以米胚芽为原料,提取具有抑制胰脂肪酶活力的胚芽蛋白。经工艺优化得到提取条件为:原料颗粒60目,料液比1∶6,pH 10.18,温度55℃,提取时间4h,水提后离心速度4 456r.min-1,等电点沉降pH 5.1,等电点沉降后离心速度3 692r.min-1。在此条件下提取得到的胰脂肪酶抑制剂的蛋白质纯度为60.7%。15.18g/L的胚芽蛋白对30.1U/mg的猪胰脂肪酶的平均抑制率达71.17%。     

Extraction of Pancreatic Lipase Inhibitor from Rice Germ

以米胚芽为原料,提取具有抑制胰脂肪酶活力的胚芽蛋白.经工艺优化得到提取条件为:原料颗粒60目,料液比1:6,pH 10.18,温度55℃,提取时间4h,水提后离心速度4 456 r·min-1,等电点沉降pH5.1,等电点沉降后离心速度3 692r·min-1.在此条件下提取得到的胰脂肪酶抑制剂的蛋白质纯度为60.7％.15.18 g/L的胚芽蛋白对30.1 U/mg的猪胰脂肪酶的平均抑制率达71.17％.     

7株野生虫草菌的鉴定及其菌丝体醇提取物对HepG2细胞的抑制活性

结合形态学与ITS序列分析对7株野生虫草真菌进行分类鉴定。MTT法分析它们的菌丝体醇提取物对肝癌HepG2细胞增殖的抑制活性。鉴定结果表明菌株MF7、MF9、MF14为细脚棒束孢Isaria tenuipes,菌株MF11、MF12、MF13为蝉棒束孢Isaria cicadae,菌株MF10为球孢白僵菌Beauveria bassiana;MTT结果显示分离到的3株细脚棒束孢和3株蝉棒束孢的菌丝体醇提取物对HepG2的抑制活性较差,IC₅₀均大于500μg/mL;球孢白僵菌MF10对HepG2细胞有一定抑制作用,IC₅₀值为221.6μg/mL,略强于蝙蝠蛾拟青霉发酵菌丝粉产品金水宝胶囊（IC₅₀=364μg/mL）和中华被毛孢发酵菌丝粉产品百令胶囊（IC₅₀=268.7μg/mL）。另外,发现供对比试验的3株蛹虫草菌株（MF1、MF5、MF15）对HepG2细胞均有较好的抑制作用,其中MF15的发酵菌丝体醇提取物活性最强,IC₅₀为55.56μg/mL,暗示蛹虫草发酵菌丝体具有重要的研究价值。     

Impact of stress hormone on adipogenesis in the 3T3-L1 adipocytes

Stress hormone is known to play a vital role in lipolysis and adipogenesis in fat cells. The present study was carried out to evaluate the effect of epinephrine on adipogenesis in the 3T3-L1 cells. The investigation on adipogenesis was done in both mono and co-cultured 3T3-L1 cells. 3T3-L1 preadipocytes and C2C12 cells were grown independently on transwell plates and transferred to differentiation medium. Following differentiation, C2C12 cells transferred to 3T3-L1 plate and treated with medium containing 10 μg/ml of epinephrine. Adipogenic markers such as fatty acid binding protein 4, peroxisome proliferator activating receptor, CCAAT/enhancer-binding protein, adiponectin, lipoprotein lipase and fatty acid synthase mRNA expressions were evaluated in the 3T3-L1 cells. Epinephrine treatment reduced adipogenesis, evidenced by reducing adipogenic marker mRNA expression in the 3T3-L1 cells. In addition, glycerol accumulation and oil red-O staining supported the reduced rate of adipogenesis. Taking all together, it is concluded that the stress hormone, epinephrine reduces the rate of adipogenesis in the mono and co-cultured 3T3-L1 cells. In addition, the rate of adipogenesis is much reduced in the co-cultured 3T3-L1 cells compared monocultured 3T3-L1 cells.     

Endothelin-1 inhibits adipogenic differentiation of 3T3-L1 preadipocytes

The effect of endothelin (ET)-1 on the adipogenic differentiation of 3T3-L1 preadipocytes was examined. Cellular morphology and lipoprotein lipase activity were used as differentiation markers. ET-1 inhibited the hormone-induced adipogenic differentiation of 3T3-L1 preadipocytes morphologically and biochemically in a dose-dependent manner. These findings promote ET-1 as a potent inhibitor of adipogenic differentiation, playing an important role in cellular differentiation of preadipocytes and making it a significant regulator of lipid metabolism.     

Optimization of pancreatic lipase inhibition by Cudrania tricuspidata fruits using response surface methodology

The fruits of Cudrania tricuspidata (Carr.) Bur. (Moraceae) significantly inhibited pancreatic lipase, which plays a key role in fat absorption. Optimization of extraction conditions with minimum pancreatic lipase activity and maximum yield was determined using response surface methodology with three-level-three-factor Box–Behnken design (BBD). Regression analysis showed a good fit of the experimental data and the optimal condition was obtained as ethanol concentration, 74.5%; temperature 61.9 °C and extraction time, 13.5 h. The pancreatic lipase activity and extraction yield under optimal conditions were found to be 65.5% and 54.0%, respectively, which were well matched with the predicted value of 65.8% and 47.1%. Further fractionation of C. tricuspidata extract resulted in the isolation of compound 1, which was identified as 5,7,4′-trihydroxy-6,8-diprenylisoflavone. It inhibited pancreatic lipase activity with IC₅₀ value of 65.0 μM. HPLC analysis suggested positive correlation between pancreatic lipase inhibition and 5,7,4′-trihydroxy-6,8-diprenylisoflavone of C. tricuspidata fruits.     

Hydroxysafflor yellow A (HYSA) inhibited the proliferation and differentiation of 3T3-L1 preadipocytes

Hydroxysafflor yellow A (HSYA), a main component of safflor yellow, has been demonstrated to prevent steroid-induced avascular necrosis of femoral head by inhibiting primary bone marrow-derived mesenchymal stromal cells adipogenic differentiation induced by steroid. In this study, we investigate the effect of HSYA on the proliferation and adipogenesis of mouse 3T3-L1 preadipocytes. The effects of HSYA on proliferation and differentiation of 3T3-L1 cells and its possible mechanism were studied by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide spectrophotometry, Oil Red O staining, intracellular triglyceride assays, real-time quantitative RT-PCR, transient transfection and dual luciferase reporter gene methods. HSYA inhibited the proliferation of 3T3-L1 preadipocytes and cell viability greatly decreased in a dose and time dependent manner. HSYA (1 mg/l) notably reduced the amount of intracellular lipid and triglyceride content in adipocytes by 21.3 % (2.13 ± 0.36 vs 2.71 ± 0.40, P < 0.01) and 22.6 % (1.33 ± 0.07 vs 1.72 ± 0.07, P < 0.01) on days 8 following the differentiation, respectively. HSYA (1 mg/l) significantly increased hormone-sensitive lipase (HSL) mRNA expression and promoter activities by 2.4- and 1.55-fold, respectively (P < 0.01), in differentiated 3T3-L1 adipocytes. HSYA inhibits the proliferation and adipogenesis of 3T3-L1 preadipocytes. The inhibitory action of HYSA on adipogenesis may be due to the promotion of lipolytic-specific enzyme HSL expression by increasing HSL promoter activity.     

香菇多糖微波辅助提取及体外免疫学活性研究

利用微波辅助水浸提法优化影响香菇多糖提取的单因子试验(料液比、浸提温度、浸提时间、微波处理时间等)和多因子的正交试验[L9(34)]。结果表明,香菇多糖的最佳提取条件为:料液比1∶30,浸提温度90℃,浸提时间2h,微波处理3m in。在此条件下,香菇多糖提取率为4.75%。香菇多糖粗品的体外免疫学研究的结果表明,在25~1600μg/mL的浓度范围内,多糖都具有促进小鼠脾淋巴细胞体外增殖的作用;在25~400μg/mL的浓度范围内,多糖对脾淋巴细胞增殖活性呈现一定的剂量依赖性。     

Aortic carboxypeptidase-like protein is regulated by transforming growth factor beta in 3T3-L1 preadipocytes

Adipogenesis is characterized by early remodeling of the extracellular matrix, allowing preadipocytes to adopt a more spherical shape and optimize lipid accumulation as they mature. Aortic carboxypeptidase-like protein (ACLP), found in collagen-rich tissues including adipose tissue, is expressed in 3T3-L1 and 3T3-F442A preadipocytes, and is downregulated during adipogenesis. We now report that ACLP is found in medium conditioned by 3T3-L1 preadipocytes. Transforming growth factor (TGF) beta, a known modulator of fibrillar matrix protein production, increased ACLP expression by 2.4+/-0.4-fold (mean+/-SE; n=3) in 3T3-L1 preadipocytes, through a mechanism that requires p42/44 MAPK activity. Addition of TGFbeta to differentiation medium, which inhibits adipogenesis, raised ACLP levels in 3T3-L1 cells. However, sustained expression of ACLP in stable clones of 3T3-L1 or 3T3-F442A preadipocytes did not interfere with adipogenesis.