赤芍总苷抗氧化活性及对大鼠肝星状细胞增殖的影响

目的：研究赤芍总苷体外抗氧化活性及对大鼠肝星状细胞HSC-T6细胞增殖的影响。方法：采用DPPH法测定赤芍总苷体外清除DPPH自由基及抗氧化活性,根据清除率曲线,确定EC50值;采用MTT法考察赤芍总苷对大鼠肝星状细胞HSC-T6细胞增殖的影响,确定起效剂量,并考察其量效关系。结果：赤芍总苷对DPPH自由基具有较强清除作用,其对DPPH的清除率的回归方程为y=7.4432x＋0.6111（r=0.9967）,EC50值为6.64 mg/L;赤芍总苷对HSC-T6的增殖具有明显的抑制作用,给药浓度为1.152mg/mL时,其对HSC-T6细胞增殖的抑制率可达39.240%。结论：赤芍总苷具有较强的抗氧化活性,对肝星状细胞的增殖有一定抑制作用。     

3'-大豆苷元磺酸钠的体外抗氧化研究

为研究3'-大豆苷元磺酸钠(DSS)的体外抗氧化活性,测定了DSS对超氧自由基(O2·)、羟基自由基(·OH)和1,1-二苯基-2.苦苯肼自由基(DPPH·)的清除能力,及其对Fe2 诱导的脂质过氧化反应和β-胡萝卜素/亚油酸自氧化体系的抑制作用.结果表明,DSS对DPPH·有一定的清除能力,对超氧自由基的清除能力较强,能很好地清除·OH自由基,对脂质过氧化有一定的抑制作用,对β-胡萝卜素/亚油酸自氧化体系有明显的抑制作用.说明DSS的药理作用可能与其较强的抗氧化能力有关.     

不同提取方法对桔梗多糖体外抗氧化性的影响

本实验考查热水浸提法、超声波提取法、微波提取法和复合酶提取法在提取桔梗多糖时对其活性所产生的影响。在模拟体外抗氧化条件下,从总还原能力、清除超氧阴离子能力、清除羟自由基能力、清除DPPH·自由基能力和体外抗肝组织自发性脂质过氧化能力等五个方面,对不同方法提取的桔梗多糖的抗氧化活性进行了分析和评估。结果表明:微波法提取的桔梗多糖体外抗氧化效果最好且差异显著(P0.01)。此方法提取的桔梗多糖对羟自由基、DPPH·自由基的清除和对脂质过氧化作用的抑制均具有较强效果,其最大多糖浓度的清除率或抑制率均接近或超过50%,但对超氧阴离子自由基的清除能力相对较弱,最大清除率仅有25%。     

绞股蓝多糖体外抗氧化活性研究

研究绞股蓝多糖的单糖组成及抗氧化活性.采用气相色谱(GC)分析绞股蓝多糖的单糖组成,通过体外抗氧化评价体系研究绞股蓝粗多糖(GPMPP)和精制多糖(GPMP)的总还原力、清除DPPH自由基、羟自由基的活性以及抗脂质过氧化作用.结果显示,绞股蓝多糖由鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖和半乳糖组成,物质的量比为1.39∶3.76∶1.00∶1.64∶4.98∶5.88.绞股蓝多糖具有较好的还原能力,对DPPH自由基和·OH具有较强的清除能力,并且对小鼠肝匀浆自发性脂质过氧化和Fe2+-H2O2诱导的小鼠肝匀浆脂质过氧化具有较好的抑制作用.以上结果表明,绞股蓝多糖具有明显的抗氧化活性.     

兖州卷柏生物碱的抗氧化活性研究

从兖州卷柏中提取生物碱类化合物,用沉淀法和分光光度法对生物碱进行鉴定,并检测兖州卷柏生物碱对DPPH自由基清除活性以及对大鼠肝脏匀浆丙二醛(MDA)含量、总抗氧化(T-AOC)和一氧化氮合酶(NOS)活性的影响。结果表明,从兖州卷柏中分离出的生物碱类化合物具有较强体外抗氧化活性。兖州卷柏生物碱具有清除DPPH自由基活性、提高大鼠肝脏匀浆的T-AOC活性、降低NOS活性以及清除MDA含量的功能。     

六种食用菌体外抗氧化及抗细胞增殖活性研究

以海鲜菇、香菇、茶树菇、杏鲍菇、平菇及金针菇6种食用菌为试验材料,比较各食用菌的抗氧化及抗细胞增殖活性。通过测定总抗氧化、总还原能力,以及对自由基的清除能力评价各食用菌体外抗氧化活性,Alamar blue法检测各提取物的抗细胞增殖活性。结果表明,各食用菌的总酚和总黄酮含量分别在155.76 mg/L-283.54 mg/L和5.19 mg/L-49.79 mg/L之间。其中,金针菇、茶树菇和香菇的总酚和总黄酮含量较高,海鲜菇和平菇含量最低。茶树菇和香菇的总抗氧化能力和总还原力(分别是22.94 U/L和20.10 U/L,0.26和0.233)优于其他几种食用菌。金针菇对DPPH·、·OH清除能力高于其他食用菌(分别是48.8%和75.12%),各食用菌对O2-·清除能力没有明显差别。6种食用菌提取物对HepG2、SGC-7901、NCI-H460、MDA-MB-231、LO2细胞均具有一定的增殖抑制作用,其中茶树菇对HepG2和NCI-H460细胞抑制作用最高(IC50=4.62±2.13,4.96±1.84 mL/L),而金针菇对SGC-7901和MDA-MB-231细胞增殖抑制作用最强(IC50=5.01±1.03,6.95±1.03 mL/L),而各提取物对LO2细胞的增殖抑制活性都最低。各食用菌均具有一定的抗氧化和抑制肿瘤增殖能力,其中茶树菇、香菇、金针菇的抗氧化和抑制肿瘤增殖活性最强。     

地中海柏挥发油抗氧化、抑肿瘤及其化学表征

为研究地中海柏(Cupressus sempervirens L.)挥发油(CEO)的化学表征,寻找抗氧化、抑肿瘤的物质基础.利用分光光度法研究了CEO总抗氧化能力,及对活性氧自由基、DPPH·的清除作用和对肝组织脂质过氧化产物(MDA)形成的抑制作用;用GC-MS法结合人工解析鉴定化学成分,并用归一化法测定组分相对含量;用MTY与SRB法检测CEO对人鼻咽癌细胞(CNE)的抑制作用,采用形态学观察以及流式细胞术(FCM)检测了CEO体外抑肿瘤的效应.结果发现CEO能很好地清除自由基,显著抑制肝组织MDA的产生;在体外能抑制CNE细胞的增殖;初步推断CEO中占总量82.91%的烯类可能与抗氧化、抗肿瘤的活性有关.     

芦笋提取液体外抗氧化、细胞毒作用及抑制α-葡萄糖苷酶活性研究

本文研究了芦笋提取液的抗氧化活性、对结肠癌细胞的细胞毒作用及对α-葡萄糖苷酶活性的抑制作用。通过研究其对DPPH自由基、羟基自由基及亚硝酸盐的清除能力来评价芦笋提取液的抗氧化活性,通过MTT法研究了芦笋提取液对结肠癌细胞株的细胞毒作用,通过α-葡萄糖苷酶体外抑制试验观察了芦笋提取液对大鼠小肠α-葡萄糖苷酶活性的影响。研究结果表明,芦笋提取液清除羟基自由基和亚硝酸盐的能力强于清除DPPH自由基的能力,一定浓度的芦笋提取液对结肠癌细胞株具有较强的细胞毒作用,并能够有效抑制大鼠小肠α-葡萄糖苷酶活性。     

眼镜蛇毒细胞毒素-4N的分离纯化及其对HSC-T6细胞的作用

为了得到高纯度眼镜蛇毒细胞毒素-4N,探索眼镜蛇毒中细胞毒素-4N(cytototxin-4N,CTX-4N)对大鼠肝星状细胞(hepatic stellate cells,HSC-T6)的增殖抑制作用。本研究采用DEAE-Sepharose CL-6B阴离子交换柱、Spehadex G-50凝胶层析柱、Macro-prep High S阳离子交换柱结合的方法对眼镜蛇毒蛋白进行分离纯化。在每一步分离纯化过程中,采用CCK-8法检测各蛋白峰组分对HSC-T6细胞的增殖抑制作用活性,收集增殖抑制作用最强的CTX-4N峰。经SDS-PAGE电泳鉴定蛋白纯度,Nano-LC-ESI-MS/MS质谱方法鉴定其组分,Cell Counting Kit-8(CCK-8)试剂检测CTX-4N对肝星状细胞(HSC-T6)的增殖抑制作用,从而确定其药理作用。经DEAE-Sepharose CL-6B阴离子交换柱、Spehadex G-50凝胶层析柱、Macro-prep High S阳离子交换柱分离纯化后得到一个电泳纯度的蛋白,蛋白质谱鉴定为CTX-4N,分子量约为9.605 k D。不同浓度的CTX-4N作用HSC-T6细胞24 h后,随着其浓度的增大对HSC-T6细胞增殖抑制作用越强,呈剂量-效应关系,IC_(50)为(12.836±0.045)μg/m L。因此,本研究建立一种眼镜蛇毒细胞毒素-4N的分离纯化方法,并确定了其对HSC-T6细胞的增殖抑制的药理作用随浓度的增加而增强,为进一步研究其药理作用提供一定的理论依据。     

金樱子总黄酮体外抗氧化活性的研究

本文研究了金樱子总黄酮(RLTF)体外清除自由基的能力,并观察其对过氧化氢诱导损伤的人脐静脉内皮细胞(HUVEC)抗氧化作用的影响。采用邻苯三酚自氧化法、水杨酸法、DPPH法分别测定RLTF对超氧阴离子自由基(O-·2)、羟自由基(·OH)、二苯代苦味酰基自由基(DPPH·)的清除率;Hoechst染色法分析各组细胞凋亡情况;MTT法测定细胞活力;分光光度法检测各组细胞中SOD、CAT、GSH-Px活性。结果显示,在20~200μg范围内,随着样品加入量增大,RLTF对·OH和DPPH·的清除率逐渐增大,而对O-·2的清除率则是先增大后减少;RLTF对O-·2的清除作用偏低,对·OH的清除率与芦丁相当,对DPPH·的清除能力则强于芦丁而弱于Vc。Hoechst染色观察结果表明RLTF预处理细胞后,能明显提高细胞的抗凋亡作用。与H2O2损伤组比较,不同浓度RLTF预处理组的细胞活力均显著升高(P0.01);不同浓度RLTF预处理组细胞中的SOD、CAT和GSH-Px活性均明显增加(P0.01),且呈剂量依赖性。上述结果表明,RLTF具有较强的自由基体外清除能力,能有效提高氧化损伤HUVEC细胞中SOD、CAT、GSH-Px的酶活性,其可能通过清除细胞内自由基和提高细胞抗氧化酶活性来提高细胞的抗氧化活性。     

阿达木单抗注射液联合白芍总苷治疗甲氨蝶呤不耐受型类风湿关节炎的临床疗效

目的:探讨阿达木单抗注射液联合白芍总苷治疗甲氨蝶呤不耐受风湿关节炎患者的临床疗效。方法:收集我院治疗的86例甲氨蝶呤不耐受的类风湿关节炎患者,随机分为实验组和对照组,每组43例。对照组患者给予阿达木单抗注射液治疗,实验组患者在对照组基础上给予白芍总苷胶囊治疗。观察并比较两组患者的晨僵时间、血沉(ESR)、类风湿因子(FR)以及临床疗效进行检测并比较。结果:与治疗前相比,治疗后两组患者的晨僵时间、血沉(ESR)、类风湿因子(FR)水平均下降(P0.05);与对照组相比,实验组患者的晨僵时间、血沉(ESR)、类风湿因子(FR)水平较低(P0.05),临床治疗有效率较高(P0.05)。结论:阿达木单抗注射液联合白芍总苷能够降低甲氨蝶呤不耐受的类风湿关节炎患者的ESR、FR水平,改善患者的临床症状,临床疗效较好。     

眼镜蛇毒神经生长因子抑制肝星状细胞增殖并诱导其凋亡

目的:从眼镜蛇毒中分离纯化神经生长因子(Nerve Growth Factor,NGF),观察眼镜NGF对肝星状细胞HSC-T6增殖、凋亡活性的影响,进一步为蛇毒NGF在抗肝纤维化治疗提供依据。方法:采用shephadex G-75和CM Sepharose CL-6B二步柱色谱对眼镜蛇毒NGF进行纯化分离;PC12细胞测定各洗脱峰的活性,再用SDS-PAGE鉴定具有NGF活性洗脱峰的纯度和相对分子质量。实验设立空白对照和NGF处理组,分别作用于HSC-T6,培育相应时间,MTT检测眼镜蛇毒NGF对HSC-T6细胞活力影响;HE染色、紫外激光显微镜与透射电镜观察HSC-T6细胞的形态学变化;TUNEL、流式细胞技术检测眼镜蛇毒NGF对HSC-T6细胞凋亡的影响。结果:眼镜蛇毒经PC-12细胞鉴定第Ⅵ峰具有NGF活性;SDS-PAGE检测为电泳纯,相对分子质量为22.3KD;NGF对HSC-T6细胞增殖具有明显抑制作用(2μg/ml NGF的抑制率为49.66%±6.50%,P<0.05;6.25μg/ml NGF的抑制率为71.33%±1.53%,P<0.05);TUNEL法检测发现NGF干预组的凋亡率28.71%±1.59%(2ug/ml NGF)和34.4%±2.49%(5μg/mlNGF)明显高于对照组的15.85%±1.58%(P<0.05);流式细胞仪也有同样的发现,NGF干预组的凋亡率16.12%±3.02%(2 ug/mlNGF)和21.15%±3.31%(5μg/ml NGF)明显高于对照组的2.7%±1.55%(P<0.05)。结论:眼镜蛇毒NGF能抑制肝星状细胞HSC-T6增殖并诱导其凋亡。     

Cholinesterase inhibitory and antioxidant properties of Verbascum mucronatum Lam. and its secondary metabolites

The aqueous extract of Verbascum mucronatum Lam. along with its fractions and secondary metabolites were assessed for their antioxidant, acetylcholinesterase (AChE), and butyrylcholinesterase (BChE) inhibitory activities. The antioxidant activity was evaluated by three methods: as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferrous ion-chelating effect, and ferric-reducing antioxidant power (FRAP) tests. The AChE activity was determined by the Ellman method using an ELISA microplate reader. Phytochemical investigations revealed the presence of four iridoid glucosides, ajugol (1), aucubin (2), lasianthoside I (3) and catalpol (4), two saponins, ilwensisaponin A (5) and C (6), and a phenylethanoid glycoside, verbascoside (7), in Verbascum mucronatum. Their structures were elucidated by spectral techniques. The aqueous extract and fractions including the phenylethanoid glycoside 7 showed DPPH scavenger effect and had the best FRAP. Besides these results, one of the phenylethanoid fractions displayed the highest ferrous ion-chelating effect. While only 7 was found to possess moderate AChE inhibition, the extract, fractions, and all other tested compounds did not inhibit AChE and BChE.     

Asiatic acid inhibits liver fibrosis by blocking TGF-beta/Smad signaling in vivo and in vitro

Liver fibrosis is a major cause of liver failure, but treatment remains ineffective. In the present study, we investigated the mechanisms and anti-hepatofibrotic activities of asiatic acid (AA) in a rat model of liver fibrosis induced by carbon tetrachloride (CCl(4)) and in vitro in TGF-beta1-stimulated rat hepatic stellate cell line (HSC-T6). Treatment with AA significantly attenuated CCl(4)-induced liver fibrosis and functional impairment in a dosage-dependent manner, including blockade of the activation of HSC as determined by inhibiting de novo alpha smooth muscle actin (a-SMA) and collagen matrix expression, and an increase in ALT and AST (all p<0.01). The hepatoprotective effects of AA on fibrosis were associated with upregulation of hepatic Smad7, an inhibitor of TGF-beta signaling, thereby blocking upregulation of TGF-beta1 and CTGF and the activation of TGF-beta/Smad signaling. The anti-fibrosis activity and mechanisms of AA were further detected in vitro in HSC-T6. Addition of AA significantly induced Smad7 expression by HSC-T6 cells, thereby inhibiting TGF-beta1-induced Smad2/3 activation, myofibroblast transformation, and collagen matrix expression in a dosage-dependent manner. In contrast, knockdown of Smad7 in HSC-T6 cells prevented AA-induced inhibition of HSC-T6 cell activation and fibrosis in response to TGF-beta1, revealing an essential role for Smad7 in AA-induced anti-fibrotic activities during liver fibrosis in vivo and in vitro. In conclusion, AA may be a novel therapeutic agent for liver fibrosis. Induction of Smad7-dependent inhibition of TGF-beta/Smad-mediated fibrogenesis may be a central mechanism by which AA protects liver from injury.     

红景天苷对LPS诱导的HSC-T6细胞增殖作用及机制研究

目的：观察红景天苷（Salidroside,Sal）对大鼠肝星状细胞（hepaticstellatecell,HSC）一T6增殖的影响,并探讨一氧化氮（NO）在此过程中的作用。方法：选择脂多糖（LPS）活化HSC—T6,采用不同浓度的Sal,作用于经LPS活化的大鼠HSC-T6,24小时后进行如下实验：噻唑兰（MTT）比色法检测HSC—T6增殖;NO荧光探针（DAF-FMDA）检测细胞内NO浓度;Westem-blot检测INOS蛋白水平。结果：在LPS作用于HSC．T6细胞24小时后,与对照组相比,细胞增殖增加（P〈0．01）,在Sal作用下,HSC-T6细胞增殖与LPS组相比受到抑制（P〈0．01）,并呈浓度依赖性;在Sal处理HSC．T6细胞24小时后,细胞内NO水平有所上升,并呈浓度依赖性增加（P〈O．01）;Sal预处理后的HSC—T6细胞,iNOS蛋白表达有所升高（P〈0．01）。结论：Sal对HSC—T6细胞的增殖具有浓度依赖性的抑制作用,NO可能是抑制增殖的因素之一。     

Cytoglobin Exhibits Anti-Fibrosis Activity on Liver In Vivo and In Vitro

Cytoglobin, generated using genetic engineering method, is a kind of recombinant human stellate cell activation-associated protein. We speculate that it could influence the development of hepatic fibrosis like Sellate cell activation-associated protein which was discovered by Kawada et al. Therefore, we investigated its anti-fibrosis effect on liver both in vivo and in vitro. During our research, we found that cytoglobin showed obvious effect compared with the control group on Thioacetamide-induced liver fibrosis in SD rats, including significantly decrease in aspartate aminotransferase, Hyaluronic acid, laminin and collagen I(Col I) levels in serum and hydroxyproline in livers, which are the important indices reflecting the degree of hepatic fibrosis. Meanwhile, the viability of rat hepatic stellate cell line T6 (HSC-T6) cells was inhibited by cytoglobin and the apoptosis induced by cytoglobin in HSC-T6 cells was detected by Annexin V/PI double staining. Activation of the caspase cascade including caspase-3 for the intrinsic pathways was demonstrated. The results also showed that the expression of Bcl-2 protein decreased whereas that of Bax protein increased, leading to an increase of the Bax/Bcl-2 ratio. Our results demonstrated that cytoglobin exhibited anti-fibrosis activity on livers in vivo and in vitro, involving apoptosis induction.     

Modification of the B-ring during flavonoid synthesis in Petunia hybrida: Effect of the hydroxylation gene Hf1 on dihydroflavonol intermediates

The white flowering mutant W48 of Petunia hybrida is dominant for the hydroxylation gene Hf1 and homozygous recessive for the hydroxylation gene Ht1 and the anthocyanin gene An1. Flower buds of this mutant accumulate dihydrokaempferol-glucosides. Thus the effect of Hf1 being dominant is not the hydroxylation of the C15 skeleton, as is the case in mutants that are able to synthesize anthocyanins. This can be explained either by a feed-back inhibition of the hydroxylation by small amounts of dihydromyricetin (glucosides), or by a controlling effect of the gene An1 on the expression of Hf1. However, the white flowering mutant W58, which is homozygous recessive for the gene An6 and dominant for Hf1, accumulates dihydromyricetin (glucosides). This excludes a possible feed-back inhibition by dihydromyricetin and we conclude that An1 controls the expression of Hf1. Feeding of radioactive malonic acid to isolated flower limbs of mutants able to synthesize anthocyanins, leads to the incorporation of radioactivity into dihydrokaempferol (glucosides) and dihydroquercetin (glucosides). These results show that glucosylation of dihydroflavonols is a normal event in anthocyanin biosynthesis and is not induced by an inhibition of anthocyanin synthesis.     

氧化低密度脂蛋白通过Wnt5a/PKCδ信号通路诱导大鼠肝星状细胞自噬

该文旨在研究氧化低密度脂蛋白(ox-LDL)对大鼠肝星状细胞(HSC-T6)自噬的影响及机制,探讨非酒精性脂肪肝炎的发病机理.体外培养的HSC-T6细胞以不同质量浓度(0、10、20、40、60μg/mL)的ox-LDL分别处理不同时间(0、3、6、12、24 h)后,用 Western blot检测LC3 Ⅱ、Be-...     

枸杞多糖与黄芪多糖抑菌活性的研究*

目的:研究黄芪多糖和枸杞多糖的抑菌活性并探讨不同pH值对其抑菌活性的影响。方法:采用滤纸片扩散法,分析不同浓度黄芪多糖和枸杞多糖在不同pH值下对几种常见细茵和霉菌(大肠杆菌、沙门氏菌、金黄色葡萄球菌、黑曲霉、产黄青霉)的抑制效果。结果:对于细菌,枸杞多糖8mg/mL时出现抑菌圈,而黄芪多糖0.02 mg/mL时效果最佳;对于霉菌,随着枸杞多糖浓度的增大,抑菌圈的直径增大,而黄芪多糖0.02 mg/mL时效果最佳;当枸杞多糖和黄芪多糖在pH6的条件下,二者抑菌活性均最强。结论:枸杞多糖和黄芪多糖对细菌、霉菌都有一定的抑制效果,pH值可影响枸杞多糖和黄芪多糖的抑菌效果。