高效液相色谱法测定桦褐孔菌4种成分含量及不同部位含量比较

目的 评价桦褐孔菌药材质量,建立高效液相色谱(HPLC)法测定桦褐孔菌药材中4种成分含量,并比较不同部位4种成分含量的差异。方法 采用HPLC法测定桦褐孔菌中栓菌酸、桦褐孔菌醇、麦角甾醇、羊毛甾醇含量,并对桦褐孔菌药材黑褐色外壳和内部黄色菌肉中4种成分含量进行比较。色谱柱为Agilent 5 TC-C₁₈(250 mm×4.6 mm, 5μm),柱温30℃;以乙腈-水为流动相,梯度洗脱,流速为1.0 mL·min^-1;检测波长为203,282 nm;进样量：20μL。采用SPSS23.0版统计软件对不同产地桦褐孔菌药材进行聚类分析,对内外两部位含量进行统计学分析。结果 4种成分在各自浓度范围内线性关系良好(r≥0.999 7);平均加样回收率分别为101.79%,102.96%,101.12%和101.37%,RSD分别为2.35%,2.25%,2.27%和2.13%(n=6);精密度、重复性、稳定性(24 h)实验RSD均<3.0%(n=6);23批桦褐孔菌药材中栓菌酸、桦褐孔菌醇、麦角甾醇、羊毛甾醇含量范围分别为0.383 0～3....     

桦褐孔菌中的活性化合物桦褐孔菌醇

本文对桦褐孔菌醇自问世以来的大量国内外文献资料进行分析、归纳和综合评价,介绍了桦褐孔菌醇的发现过程、结构研究、药理活性、结构修饰和半合成,以及菌丝体的体外培养等,为桦褐孔菌醇的进一步研究开发提供参考。     

桦褐孔菌不同提取物小鼠急性毒性实验研究

目的研究桦褐孔菌不同提取物对小鼠急性毒性作用,为临床安全用药提供依据。方法采用经典的急性毒性实验方法,测定桦褐孔菌水提物和70%乙醇提取物的半数致死量(LD50),并观察其毒性反应。结果随着给药量的增大,桦褐孔菌水提物和醇提取物对小鼠毒性作用逐渐增强,死亡率逐渐升高,空白组没有异常变化。按生药量计算水提物的LD50是17.49g·kg-1,95%可信区间为16.36～18.70g·kg-1;醇提取物LD50是61.97g·kg-1,95%可信区间为53.73～71.46g·kg-1。结论桦褐孔菌水提物和70%乙醇提取物均有一定的毒性,且水提物毒性起效量比70%乙醇提取物的起效量小。     

桦褐孔菌甲醇提取物抗氧化及抗炎作用研究

目的:观察桦褐孔菌甲醇提取物的抗氧化及抗炎作用,为更合理有效地开发该多糖真菌提供科学依据。方法:通过大鼠足肿胀及大鼠棉球肉芽组织增生模型考察其抗炎活性,并通过清除DPPH自由基活性的清除作用评价其抗氧化作用。结果:100mg/kg以上的桦褐孔菌甲醇提物对角叉菜胶所致大鼠足肿、大鼠棉球肉芽组织增生有抑制作用;100mg/kg的桦褐孔菌甲醇提物可以清除DPPH自由基。结论:桦褐孔菌甲醇提取物具有良好的抗氧化及抗炎作用。     

珍稀药用菌-桦褐孔菌的研究进展

目的介绍药用菌桦褐孔菌的研究进展。方法以国内外发表的文献为依据,从其学名、分类学地位、形态、液体培养特性、化学成分、药理学研究方面综述了桦褐孔菌的研究进展。结果桦褐孔菌化学成分主要是三萜类化合物．其菌核水提物具有良好的抑制肿瘤,抗氧化,抗病毒作用。结论桦褐孔菌是一种很有研究价值的药用菌。     

桦褐孔菌的药理作用研究进展

桦褐孔菌属于真菌门、担子菌亚门、层菌纲、非褐菌目、多孔菌科、褐卧孔菌属，学名：Fuscoporia obliqua（Pers．Fr．）Aoshi或Inonotus obliquus（Fr．）Pilat，俄语名：Chaga。     

桦褐孔菌总三萜提取物的纯化及其体外抗肿瘤活性研究

目的 优化桦褐孔菌总三萜提取物的纯化工艺,考察其纯化物的抗肿瘤活性。方法 以桦褐孔菌醇为对照品,建立桦褐孔菌总三萜的含量测定方法。采用单因素实验筛选大孔吸附树脂种类、上样体积、上样液质量浓度、上样流速、洗脱剂体积分数、洗脱剂用量、洗脱流速,确定并验证该粗提物的纯化工艺。通过细胞增殖实验、迁移实验、流式细胞术和AO/EB试剂盒检测桦褐孔菌总三萜纯化物对人宫颈癌细胞HeLa增殖、迁移及凋亡的影响。结果 桦褐孔菌总三萜粗提物的最佳纯化工艺为：AB-8型大孔吸附树脂,上样液质量浓度为2.0 mg/mL,上样体积为140 mL,上样流速为1.0 mL/min;洗脱时先用50%的乙醇40 m L除杂,再用95%的乙醇160 m L洗脱,洗脱流速为3.0 mL/min。纯化后桦褐孔菌总三萜的质量分数从34.36%升至73.39%。桦褐孔菌总三萜纯化物可抑制HeLa细胞增殖,半数抑制浓度为184.20μg/mL;与对照组相比,该纯化物可显著抑制HeLa细胞的迁移,促进其凋亡（P<0.05或P<0.01）。结论 成功优化了桦褐孔菌总三萜提取物的纯化工艺,该纯化物可抑制HeLa细胞增殖和迁移,...     

桦褐孔菌活性组分提取分析方法研究进展

桦褐孔菌Inonotus obliquus属于多孔菌科褐卧孔菌属,是珍贵的药用真菌,分布于中国、芬兰、波兰、俄罗斯及日本等国家。桦褐孔菌具有抗衰老和抑制病毒的药理作用,主要用于治疗各种癌症、糖尿病及心脏病,主要成分为多糖、三萜类化合物、木质素及黑色素,其提取分离方法主要涉及水提法、超声法、微波辅助提取法、酶诱导结合负压空气化提取法,分析方法主要为薄层色谱法、高效液相法和紫外-分光光度法。天然产物在新药研发及新药先导化合物发现中起重要作用,综述了桦褐孔菌活性成分的提取及分析方法研究进展,以期为其深入研究及新药研发提供参考。     

桦褐孔菌多糖的提取工艺优化

目的研究桦褐孔菌多糖提取工艺优化。方法以多糖含量为指标,在单因素研究基础上,采用4因素3水平的响应面分析法对桦褐孔菌多糖的提取工艺进行优化研究,并检测重金属砷的含量。结果桦褐孔菌多糖提取的优化工艺条件为：乙醇浓度30％、提取温度95℃、提取时间2．5h、液料比30：1。在该条件下,粗多糖得率为33．6％,样品中多糖含量为6．177％。桦褐孔菌多糖实际提取含量为5．993％,重金属含量降低。结论优化工艺提取率高,快速,重金属含量低,可用于桦褐孔菌的多糖提取。     

药用真菌白桦茸--桦褐孔菌

白桦茸是一种药用菌,研究表明其活性物质成分中的氧化三萜化舍物和生物碱类化舍物及其桦褐孔菌醇等。对肿瘤细胞有明显的抑制作用,并对治疗糖尿病、爱滋病、高血压、过敏性皮肤等都有明显效果。本文通过对白桦茸的生物学特性、化学成分及其药理作用进行阐述。     

纤维素酶辅助提取桦褐孔菌多糖的研究

目的优化桦褐孔菌提取工艺,提高桦褐孔菌多糖得率。方法首先对桦褐孔菌多糖提取的主要影响因素纤维素酶用量、酶解温度、溶剂倍量、酶解时间、pH值进行单因素条件筛选。然后采用正交设计法对影响纤维素酶辅助提取多糖的条件进行优化。结果最佳提取条件为:料液比1:40、酶解时间60 min、酶解温度50℃、加酶量0.15 g、酶解pH=5.0。结论纤维素酶对桦褐孔菌多糖进行辅助提取,相比传统的热水浸提,能够显著提高桦褐孔菌多糖的提取率,比水浸提高29.07%,是一种较为有效的提高多糖得率的方法。     

桑黄水煎液对D-半乳糖衰老模型小鼠的抗氧化作用

目的 观察桑黄水煎液对D-半乳糖诱导小鼠氧化损伤的保护作用,并探讨其保护作用的可能机制。方法 ①ICR小鼠分为正常对照组,桑黄水煎液低、中、高剂量（2,6,12 g·kg^-1）组,连续给药15 d。②ICR小鼠分为正常对照组、模型组、维生素E组（50 mg·kg^-1）,桑黄水煎液低、中、高剂量（2,6,12 g·kg^-1）组,小鼠颈背部皮下注射D-半乳糖（120 mg·kg^-1）造成亚急性衰老模型,按分组连续给药42 d。比色法测定小鼠肝脏及血清中丙二醛（MDA）含量和抗氧化酶活力,如超氧化合物歧化酶（SOD）、总抗氧化能力（T-AOC）、过氧化酶（POD）等。Western blot检测肝脏中核因子E2相关因子2（Nrf2）和血红素氧合酶1（HO-1）的表达。结果 桑黄水煎液对正常小鼠和D-半乳糖诱导小鼠体质量及脏器指数均无明显影响。桑黄水煎液可显著提高正常小鼠肝脏中T-AOC和SOD酶活力（P<0.01,P<0.05）。各剂量桑黄水煎液可显著提高衰老模型小鼠血清中T-AOC,降低MDA含量（P<0.01,P<0.05）,同时中剂量增强肝脏中SOD （P<0.05）和POD酶活力（P<0.01）,中、高剂量均上调肝脏中Nrf2和HO-1蛋白表达水平（P<0.01）。结论 桑黄水煎液具有提高正常小鼠抗氧化能力和改善D-半乳糖致小鼠氧化损伤作用,此作用可能是通过调节Nrf2/HO-1途径增加体内抗氧化酶的活性,减少过氧化脂质的生成,进而提高机体抗氧化能力。     

Hepatoprotective effects of Juglans regia extract against CCl4-induced oxidative damage in rats

Abstract

Context: Different parts of the walnut [Juglans regia L. (Juglandaceae)] have been used in folk medicine for protection against liver injury, although its actual efficacy remains uncertain.

Objective: The present study investigated the protective effect of walnut leaf extract against carbon tetrachloride (CCl₄)-induced liver damage in rats.

Materials and methods: The rats were randomly divided into seven groups: control, CCl₄ (i.p., 0.5?mL/kg b.w., 50% CCl₄ in olive oil), walnut extract (at dose level of 0.2?g/kg b.w.) alone, walnut extract (at dose levels of 0.05, 0.1, 0.2 and 0.4?g/kg b.w.) with CCl₄, and treatment was carried out accordingly. On the 28th day, rats were sacrificed and blood was withdrawn by cardiac puncture. Liver damage was assessed by serum biochemical parameters (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and albumin), antioxidant enzymes (superoxide dismutase and catalase) and histopathological observation.

Results: Administration of walnut leaf extract (ranging from 0.2 to 0.4?g/kg b.w.) significantly lowered serum alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase levels in CCl₄-treated rats. Walnut leaf extract increased antioxidant enzymes, including superoxide dismutase and catalase. Histopathological examination of livers showed that walnut leaves extract reduced fatty degeneration, cytoplasmic vacuolization and necrosis in CCl₄-treated rats.

Discussion and conclusion: These results suggest that walnut extract has a protective effect over CCl₄-induced oxidative damage in rat liver. These results demonstrate that walnut extract acts as a good hepatoprotective and antioxidant agent in attenuating hepatocellular damage.     

Deferoxamine alleviates liver fibrosis induced by CCl4 in rats

Several chronic liver diseases can lead to the occurrence of hepatic fibrosis through the accumulation of iron, which causes induction of oxidative stress and consequently activation of fibrogenesis. The present study was designed to investigate the potential antifibrotic and anti‐oxidant effects of deferoxamine (DFO), a well‐known iron chelator in an experimental rat model of liver injury using carbon tetrachloride (CCl₄). First, the potential effective dose of DFO was screened against CCl₄‐induced acute hepatotoxicity. Then, rats were co‐treated with DFO (300 mg/kg, i.p.) for 6 weeks starting from the third week of CCl₄ induction of chronic hepatotoxicity. Liver function was assessed in addition to histopathological examination. Furthermore, oxidative stress and fibrosis markers were assessed. It was found that treatment of animals with DFO significantly counteracted the changes in liver function; histopathological lesions and hepatic iron deposition that were induced by CCl₄. DFO also significantly counteracted the CCl₄‐induced lipid peroxidation increase and reduction in antioxidant activities of superoxide dismutase and glutathione peroxidase enzymes. In addition, DFO ameliorated significantly liver fibrosis markers including hydroxyproline, collagen accumulation, and the expression of the hepatic stellate cells (HSCs) activation marker; alpha smooth muscle actin and transforming growth factor‐beta (TGF‐β). Together, these findings indicate that DFO possesses a potent antifibrotic effect due to its antioxidant properties that counteracted oxidative stress and lipid peroxidation and restored antioxidant enzymes activities as well as reducing HSCs activation and fibrogenesis.     

Resveratrol and trimethylated resveratrol protect from acute liver damage induced by CCl4 in the rat

The importance of hydroxyl groups in the antioxidant and hepatoprotective properties of resveratrol was investigated. To achieve this, resveratrol or its trimethylated analog were administered (10 mg kg(-1), p.o.) to male Wistar rats and liver damage was induced by acute administration of CCl4 (4 g kg(-1), p.o.); appropriate controls were performed. The animals were killed 24 h after CCl4 intoxication. The amount of reduced glutathione (GSH) in the liver was not modified by any treatment; interestingly, the GSH/GSSG (oxidized glutathione) ratio decreased in the groups receiving CCl4 and resveratrol associated with an increase in GSSG. In blood GSH and the GSH/GSSG ratio were decreased by CCl4; both effects were completely prevented by any of the compounds tested. Lipid peroxidation and the activity of gamma-glutamyl transpeptidase were increased significantly after CCl4. Resveratrol partially prevented these increases and surprisingly, trimethylated resveratrol completely prevented the increase of these markers. Both compounds partially but significantly prevented the increase in the activity of alanine aminotransferase; this result agrees with observations in the histological analysis. Both tested compounds administered alone produced no effect. The results of the present study suggest that OH groups are important for the antioxidant and hepatoprotective properties of the molecule of resveratrol; nevertheless, these effects can be improved by replacing hydrogen by a methyl in these groups. The differences in the antioxidant and hepatoprotective effects of these compounds could be due to the possibility that the trimethylated resveratrol acts like a prodrug, prolonging, probably, the half-life of the original compound.     

氧化应激致心室重构的研究进展

心室重构意味着血管壁中氧化应激水平的提高,当机体受到有害刺激时,氧化和抗氧化之间失衡,导致大量活性氧簇（ ROS）的产生,加速心室重构的进展。近年来,新型抗氧化剂成为治疗或预防心室重构的靶点,观察抗氧化剂治疗在心室重构患者的长期应用是当下趋势。该文就氧化应激与心室重构之间的关系及相关药物治疗进行综述。     

The MRI marker gene MagA attenuates the oxidative damage induced by iron overload in transgenic mice

We aimed to create transgenic (Tg) mice engineered for magnetic resonance imaging (MRI). To ascertain if MagA expression contributes to oxidative stress and iron metabolism, we report the generation of Tg mice in which ubiquitous expression of MagA can be detected by MRI in vivo. Expression of MagA in diverse tissues of Tg mice was assessed, and iron accumulation and deposition of nanoparticles in tissues were analyzed. Levels of antioxidant enzymes, lipid peroxidation and cytokine production were determined, and iron metabolism-related proteins were also detected. MagA Tg showed no apparent pathologic symptoms and no histologic changes compared with wild-type (WT) mice. Overexpression of MagA resulted in specific alterations of the transverse relaxation rate (R₂) of water. Transgene-dependent changes in R₂ were detectable by MRI in iron-overloaded mice. We also evaluated antioxidant abilities between WT and Tg groups or two iron-overloaded groups. Together with the data of cytokines and iron metabolism-related proteins, we inferred that MagA could regulate nanoparticle production and thus attenuate the oxidative damage induced by iron overload. The novel MagA Tg mouse, which expresses an MRI reporter in many tissues, would be a valuable model of MagA molecular imaging in which to study diseases related to iron metabolism.     

Phenolics from monofloral honeys protect human erythrocyte membranes against oxidative damage

The aim of the present work was to analyze the phenolic extracts from two monofloral Cuban honeys for their in vitro total antioxidant capacity, phenolic compounds content and free radical scavenging activity. The phenolic extracts, rich in lipophilic compounds, were tested further for their ability to inhibit AAPH-induced oxidative damage (hemolysis, lipid peroxidation and cytosolic depletion of reduced glutathione and decrease of superoxide dismutase activity) in erythrocytes. Results indicate an important total antioxidant capacity measured by TEAC and ORAC assays, as well as a relevant radical scavenging activity performed by EPR. Moreover, 13 phenolic compounds were identified using HPLC–LC/MS with quercetin as the most abundant flavonoid. The results also show that both extracts were able to inhibit erythrocytes oxidative damage, and that this may likely be due to their incorporation into cell membranes and their ability to cross it and reach the cytosol. In fact, flavonoid uptake by erythrocytes was further confirmed by testing quercetin, which efficiently incorporated into erythrocytes. Overall, this study indicates that honey contains relevant antioxidant compounds responsible, at least in part, for its biological activity and that uptake of its flavonoids may provide defense and promote cell functions in erythrocytes.     

京尼平衍生物对硝普钠诱导的PC12细胞损伤的保护作用

目的探讨京尼平及其衍生物对硝普钠诱导的PC12细胞损伤的保护作用及其作用机制。方法通过62.5~1 000μmol/L硝普钠处理PC12细胞24 h建立细胞损伤模型,造模前2 h给予京尼平衍生物预处理,采用噻唑蓝比色法(MTT)考察京尼平衍生物对PC12细胞存活率的影响。Hoechst染色后观察细胞形态,以DCFH-DA探针和丙二醛(MDA)检测试剂盒分别检测PC12细胞内ROS、MDA水平,RT-PCR法检测PC12细胞抗氧化应激基因水平。结果与对照组比较,硝普钠能剂量相关性地降低PC12细胞的存活,在750μmol/L时具有统计学差异,而10μmol/L 1R-异丙基-6,7-二氢京尼平(化合物4)、1S-异丙基-6,7-二氢京尼平(化合物5)能够明显增加硝普钠诱导PC12细胞损伤后的细胞存活率(P0.05、0.01)。Hoechst染色形态学证实化合物4、5能够明显减少PC12细胞的凋亡。硝普钠诱导PC12细胞内氧化应激水平,化合物4、5能有效地降低ROS、MDA水平,并促进抗氧化酶基因GCLC、GPX、CAT m RNA表达,但对HO-1、Mn SOD基因表达水平无明显影响。结论京尼平衍生物可以抑制硝普钠诱导的PC12细胞损伤和细胞内氧化应激水平,其作用机制可能是通过影响抗氧化酶GCLC、CAT、GPX m RNA表达水平而实现。