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1.
罗容  吴霞  李静宜  崔湖荣  张楠  张贵君 《生物磁学》2012,(32):6228-6233
目的:研究枳实提取物及其药效组分橙皮苷和新橙皮苷对氧化低密度脂蛋白(oxidized lowd ensity lipoprotein,Ox—LDL)损伤的人脐静脉内皮细胞(human umbilical vein endothelial cells line,HUVEC)细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)表达和一氧化氮(nitric oxide,NO)释放的影响。方法:体外培养HUVEC,50μg/mLOX—LDL制造HUVEC损伤模型。以MTS染色法检测细胞毒性确定用药浓度。细胞ELISA法测定细胞表面ICAM-1的含量,试剂盒测定细胞培养上清液中NO含量。结果:①枳实提取物小于等于2mg/mL时,橙皮苷浓度小于等于0.03125mg/mL时,新橙皮苷浓度小于等于0.25mg/mL时,HUVEC存活率分别大于80%。②2.0mg/mL和1.0mg/mL两个浓度的枳实提取物、15.625μg/mL的橙皮苷和0.2500mg/mL新橙皮苷对OX—LDL诱导的HUVEC的ICAM-1表达有显著抑制作用。③2.0mg/mL枳实提取物显著提高OX—LDL诱导的HUVEC和正常HUVEC培养液中的NO含量;7.813ixg/mL、15.625μg/mL和31.250μg/mL 3个浓度的橙皮苷能显著提高OX—LDL诱导的HUVEC培养液中的NO含量,31.250μg/mL的橙皮苷能促进正常HUVEC的NO释放;0.2500mg/mL和0.1250mg/mL 2个浓度的新橙皮苷能显著提高OX—LDL诱导的HUVEC培养液中的NO含量。结论:枳实提取物及其药效组分橙皮苷、新橙皮苷能抑制Ox-LDL诱导的HUVEC的ICAM-1表达,促进Ox-LDL诱导的HUVEC的NO释放。  相似文献   

2.
目的:研究并建立紫外分光光度法(UV)和高效液相色谱法(HPLC)分别测定温胆汤物质基准中总黄酮、柚皮苷和新橙皮苷含量。方法:以柚皮苷为对照品,采用紫外-可见分光光度法测定总黄酮的含量并采用HPLC法测定柚皮苷和新橙皮苷的含量。结果:采用直接测定法测定总黄酮类成分在284 nm处有较强吸收,柚皮苷在20.22~80.88μg/mL范围内与吸光度呈良好的线性关系,平均加样回收率为98.33%。柚皮苷和新橙皮苷分别在0.01~0.51 mg/mL和0.01~0.50 mg/mL范围内有良好的线性关系,精密度、稳定性、重复性的RSD均小于5.0%,加样回收率分别为97.11%和102.72%;温胆汤物质基准中总黄酮的平均含量为15.03%,柚皮苷、新橙皮苷的平均含量分别为22.78 mg/g和9.75 mg/g。结论:建立的温胆汤物质基准总黄酮、柚皮苷和新橙皮苷含量测定方法简单、稳定、重复性好,为温胆汤及其相关制剂进一步研究和开发提供实验基础。  相似文献   

3.
利用ox-LDL对培养的人脐静脉内皮细胞(HUVECs)造成脂质过氧化损伤模型,探讨车前子多糖(PSP)对HUVECs的保护作用及其可能机制。用MTT和化学方法,从细胞水平观察ox-LDL对细胞增殖活性、细胞上清液中一氧化氮(NO)含量、胞内一氧化氮合酶(NOS)活性、丙二醛(MDA)含量、SOD活力;用ELISA和RT-PCR技术,从分子水平观察ICAM-1和凋亡相关基因mRNA的表达,探讨不同浓度的PSP(25 mg/L、50 mg/L、100 mg/L)对上述指标的影响。结果表明ox-LDL导致HUVECs损伤,而伴随PSP浓度的增加,HUVECs增殖活性呈明显升高趋势(P0.05);而PSP使HUVECsMDA含量显著下降(P0.05),SOD、NO和NOS水平明显升高(P0.05),ICAM-1、c-myc mRNA和p53 mRNA表达降低,表明PSP对受损的内皮细胞具有保护作用,其机制可能与抑制ox-LDL诱导的ICAM-1、c-myc mRNA和p53 mRNA的表达有关。  相似文献   

4.
枳实中辛弗林和橙皮苷的联合提取工艺研究   总被引:1,自引:0,他引:1  
以辛弗林和橙皮苷的收率及纯度为指标,研究了含辛弗林的枳实提取物及橙皮苷的综合制备工艺.先采用酸液渗漉法提取枳实中的辛弗林,再采用碱提酸沉法提取原料中的橙皮苷,考察盐酸浓度及用量、流速等因素的影响.辛弗林的最佳提取工艺为:药材加5倍水浸泡过夜,渗漉盐酸液的浓度为0.02 mol/mL,渗漉料液比为1:4,以5 mL/min的流速渗漉;碱提酸沉法提取橙皮苷的优化工艺为:于提取辛弗林之后的药渣中加入65%乙醇,于60℃搅拌条件下加碱调节pH 13,过滤后的滤液以稀盐酸调节pH 5,过滤后再用碱回调pH7,放置过夜.橙皮苷的收率为11.3%,HPLC测定橙皮苷的纯度为86.7%.  相似文献   

5.
采用MTT法测定不同浓度马胎盘提取物(EPE)对正常、ConA和LPS诱导的小鼠脾淋巴细胞增殖活性以及NK细胞杀伤活性的影响;并用氨基酸自动分析仪检测马胎盘提取物中氨基酸的含量。MTT实验结果表明:马胎盘提取物(187.5~750μg/mL)明显增强正常淋巴细胞的活性;23.44~1500μg/mL明显抑制ConA诱导的T淋巴细胞增殖活性,750~1500μg/mL明显抑制LPS诱导的B淋巴细胞增殖活性,其对ConA诱导的T淋巴细胞增殖的抑制作用强于对LPS诱导的B淋巴细胞增殖的抑制作用;93.75~500μg/mL明显增强NK细胞对PC-3癌细胞的杀伤活性;氨基酸含量分析表明:其含有丰富的氨基酸,其中谷氨酸(7.53%)、天门冬氨酸(5.64%)和亮氨酸(5.54%)含量较高。因此马胎盘提取物对淋巴细胞可能具有一定的免疫调节作用。  相似文献   

6.
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、Beclin1、p62的含量。不同质量浓度(0、10、20、40、60μg/mL)的ox-LDL处理HSC-T6细胞12 h后,Western blot检测Wnt5a、p-PKCδ、p-STAT3的含量。将HSC-T6细胞分为对照组(Control)、ox-LDL组、ox-LDL+si-NC组和ox-LDL+si-Wnt5a组,经相应处理后用Western blot、qRT-PCR分别检测LC3 II、Beclin1、p62、p-PKCδ、p-STAT3的蛋白和mRNA含量变化;免疫荧光检测LC3 II的变化;油红O染色观察HSC-T6脂滴含量变化;比色法检测各组细胞培养上清中羟脯氨酸(Hyp)含量;ELISA检测细胞培养上清中透明质酸(HA)和层黏连蛋白(LN)含量。PKCδ抑制剂Rottlerin预处理细胞:将HSC-T6细胞分为对照组(Control)、ox-LDL组、ox-LDL+DMSO组和ox-LDL+Rottlerin组,检测方法与敲低Wnt5a一致。经ox-LDL处理后,HSC-T6细胞中LC3 II、Beclin1含量增加(P<0.05),p62含量减少(P<0.01),且在ox-LDL质量浓度为20μg/mL、作用12 h时达到峰值。ox-LDL质量浓度为20μg/mL、作用12 h时,HSC-T6细胞中Wnt5a、p-PKCδ、p-STAT3蛋白表达显著升高(P<0.01)。敲低Wnt5a后,HSC-T6细胞中Wnt5a、LC3 II、Beclin1 mRNA和蛋白表达水平显著降低(P<0.001),p62蛋白表达增多(P<0.01),p-PKCδ、p-STAT3蛋白表达减少(P<0.05),细胞内LC3 II点状聚集减少,脂滴含量减少,细胞培养上清中Hyp、HA、LN含量也减少(P<0.05)。抑制PKCδ后,结果与敲低Wnt5a一致。ox-LDL可通过增强Wnt5a/PKCδ通路诱导HSC-T6细胞自噬。  相似文献   

7.
脂多糖直接损伤血管内皮细胞后内皮细胞膜脂的修饰   总被引:1,自引:1,他引:0  
Wang X  Cai SX  Luo XD  Wang P  Luo Q  Liang GP  Yang ZC 《生理学报》2001,53(6):419-424
我们前期的研究显示,脂多糖(lipopolysaccharide,LPS)直接损伤人脐静脉血管内皮细胞(human umbilical vein endothelial cell,HUVEC)后,HUVEC膜粘度随LPS浓度的增加而增大,这暗示LPS可能改变HUVEC的膜结构和组成。本文旨在研究LPS直接损伤HUVEC后内皮细胞膜脂的修饰。用高效毛细管电泳(high performance capillary electrophoresis,HPCE)污测定HUVEC膜磷脂组成,HUVEC用不同浓度(O、0.3125、0.625、1.25、2.5、5、7、8.5、9、10μg/ml)的LPS无血清培养液直接损伤3h;用含LPS(0.625μg/ml)的无血清培养液直接损伤1、3、6、12、24、48h。结果显示:在3h LPS的作用下,HUVEC的总磷脂含量随LPS浓度的增加而增加,在0.625μg/ml LPS的浓度条件下,HUVEC的总磷脂含量随LPS作用时间的延长而降低;LPS的作用浓度和作用时间对磷脂酰鞘磷脂(SM)、磷脂酰乙醇胺(PE)以及磷脂酰丝氨酸(PS)的含量影响不大。LPS激活膜磷脂代谢的反应特性表现出典型的酶促动力学特性。实验结果表明,LPS可直接激活血管内皮细胞膜磷脂代谢,诱导HUVEC膜磷脂的修饰,提示LPS直接损伤血管内皮细胞HUVEC的途径可能与膜结构组成及膜脂代谢有关。  相似文献   

8.
为了评价人面果叶子、根部、果实提取物体外抗糖尿病活性,相应测定了其石油醚提取物(PFr.)、乙酸乙酯提取物(EFr.)、正丁醇提取物(BFr.)、水提取物(WFr.)的α-葡萄糖苷酶与α-淀粉酶抑制活性,以及HepG2细胞的促葡萄糖消耗能力。果实乙酸乙酯提取物(IC50=17.81±1.09μg/mL)、叶子乙酸乙酯提取物(IC50=18.60±1.56μg/mL)、根部乙酸乙酯提取物(IC50=14.05±0.24μg/mL)、根部正丁醇提取物(IC50=13.01±0.38μg/mL)显示了较好的α-葡萄糖苷酶抑制活性(acarbose IC50200μg/mL)。而根部乙酸乙酯与正丁醇提取物在600μg/mL的浓度下就显示了90%的α-葡萄糖苷酶抑制率,在1.5 mg/mL的浓度下显示了90%的α-淀粉酶抑制率。在促葡萄糖消耗试验中,果实乙酸乙酯提取物在浓度为7.5~30 mg/mL时显示了很好的促HepG2细胞葡萄糖消耗能力(P0.001),叶子乙酸乙酯提取物、根部正丁醇与乙酸乙酯提取物的促葡萄糖消耗率达到了3.08、3.12、1.93,仅次于果实乙酸乙酯提取物(3.91)。此次研究为人面果抗糖尿病活性开发提供一定理论基础。  相似文献   

9.
张志伟  李瑜  陈雪莲  尹婵  英智  曹建国 《生物磁学》2009,(13):2436-2439
目的:观察白花蛇舌草醇提取物对乳腺癌细胞T-47D生长的影响。方法:采用平皿克隆形成实验、软琼脂集落形成实验和BrdUrd(溴脱氧尿苷)掺入实验,观察不同浓度的白花蛇舌草醇提取物(1.0μg/mL,3.0μg/mL,10μg/mL,30μg/mL,100μg/mL)对T-47D细胞锚定依赖性生长和软琼脂集落形成能力及核酸合成的抑制作用。结果:①随着白花蛇舌草醇提取物浓度的升高,对T-47D细胞的生长呈现明显抑制作用;②白花蛇舌草醇提取物可呈浓度依赖性抑制对数生长期T-47D细胞核酸合成。结论:白花蛇舌草醇提取物可能通过影响肿瘤细胞核酸合成而抑制T-47D细胞生长。  相似文献   

10.
通过复制LPS诱导的BEAS-2B炎症模型,探讨连翘提取物对炎症反应中炎性因子分泌的作用。采用MTT法检测BEAS-2B细胞活力,筛选连翘乙醇提取物(FSEE)、60%连翘酯苷A(60%FTA)、90%连翘酯苷A(90%FTA)最佳给药浓度;采用倒置显微镜观察连翘提取物对细胞作用后的细胞形态;采用Griess Reagent法检测细胞培养上清NO含量;流式细胞术检测细胞内活性氧(ROS)的水平。毒性试验结果表明,连翘提取物浓度小于128μg/m L时,对BEAS-2B细胞没有产生毒性;浓度在128~256μg/m L时,60%FTA与90%FTA均表现出对细胞的毒性。抗炎结果表明,在给药浓度设置为25、50、100μg/m L时,三种提取物均表现出减少NO分泌的效果(P0.01),90%FTA组存在剂量依赖关系;三种提取物可显著降低LPS诱导的BEAS-2B细胞胞内活性氧的水平(P0.05),且呈量效关系。另外,FSEE、60%FTA、90%FTA组间的治疗效果也逐渐增强。由此得出,90%FTA为连翘提取物抗炎作用的主要活性部位,其抗炎活性成分可能为连翘酯苷A,这将为连翘抗炎活性的进一步研究提供基础。  相似文献   

11.
目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。  相似文献   

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国内外蝗害治理技术现状与展望   总被引:3,自引:0,他引:3  
张龙 《昆虫知识》2011,48(4):804-810
本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...  相似文献   

15.
The molecular characterisation of species and genotypes of Cryptosporidium and Giardia is essential for accurately identifying organisms and assessing zoonotic transmission. Results of recent molecular epidemiological studies strongly suggest that zoonotic transmission plays an important role in cryptosporidiosis epidemiology. In such cases the most prevalent zoonotic species is Cryptosporidium parvum. Genotyping and subtyping data suggest that zoonotic transmission is not as prevalent in the epidemiology of giardiasis. Molecular characterisation of Cryptosporidium and Giardia is a relatively recent application that is evolving as new genes are found that increase the accuracy of identification while discovering a greater diversity of species and yet unnamed taxa within these two important genera. As molecular data accumulate, our understanding of the role of zoonotic transmission in epidemiology and clinical manifestations is becoming clearer.  相似文献   

16.
The synthesis and turnover of cerebrosides and phospholipids was followed in microsomal and myelin fractions of developing and adult rat brains after an intracerebral injection of [U-14C]serine. The kinetics of incorporation of radioactivity into microsomal and myelin cerebrosides indicate the possibility of a precursor-product relationship between cerebrosides of these membranes. The specific radioactivity of myelin cerebrosides was corrected for the deposition of newly formed cerebrosides in myelin. Multiphasic curves were obtained for the decline in specific radioactivity of myelin and microsomal cerebrosides, suggesting different cerebroside pools in these membranes. The half-life of the fast turning-over pool of cerebrosides of myelin was 7 and 22 days for the developing and adult rat brain respectively. The half-life of the slowly turning-over pool of myelin cerebrosides was about 145 days for both groups of animals. The half-life of the rapidly turning-over microsomal cerebrosides was calculated to be 20 and 40 h for the developing and adult animals respectively. The half-life of the intermediate and slowly turning-over microsomal cerebrosides was 11 and 60 days respectively, for both groups of animals. The amount of incorporation of radioactivity into microsomal cerebrosides from L-serine was greatly decreased in the adult animals, and greater amounts of the precursor were directed towards the synthesis of phosphatidylserine. In the developing animals, considerable amounts of cerebrosides were synthesized from L-serine, besides phosphatidylserine. The time-course of incorporation indicated that a precursor-product relationship exists between microsomal and myelin phosphatidylserine. The half-life of microsomal phosphatidylserine was calculated to be about 8 h for the fast turning-over pool in both groups of animals.  相似文献   

17.
Enterococcus faecalis was the most frequently isolated enterococcal species from anal swabs and tonsils of dogs and cats, although in the anal samples from dogs Ent. hirae was found almost as often as Ent. faecalis. Most Ent.faecium strains from dog tonsils differed from those associated with humans and other animals in that they fermented sorbitol. Typical Ent. avium as well as atypical Ent. avium -like strains were seen in dogs, while the related species Ent. raffinosus was associated with cat tonsils. Enterococcus cecorum also occurred mainly in cats. Certain atypical strains, presumptively identified as Ent. cecorum , shared characteristics with Ent. columbae.
The most frequent streptococcal species in tonsils of cats and dogs were Streptococcus suis and Strep. canis. Streptococcus canis and Strep. bovis predominated in anal swabs. The canine Strep. suis differed from the common porcine strains in fermenting mannitol.
Forty-seven of the 288 isolates examined could not be identified or related to known species. The characteristics of two groups of these bacteria, provisionally called 'Ton 31 group' and 'O7 group' are described.  相似文献   

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The nanomechanical properties of the coiled-coils of myosin are fundamentally important in understanding muscle assembly and contraction. Force spectra of single molecules of double-headed myosin, single-headed myosin, and coiled-coil tail fragments were acquired with an atomic force microscope and displayed characteristic triphasic force-distance responses to stretch: a rise phase (R) and a plateau phase (P) and an exponential phase (E). The R and P phases arise mainly from the stretching of the coiled-coils, with the hinge region being the main contributor to the rise phase at low force. Only the E phase was analyzable by the worm-like chain model of polymer elasticity. Restrained molecular mechanics simulations on an existing x-ray structure of scallop S2 yielded force spectra with either two or three phases, depending on the mode of stretch. It revealed that coiled-coil chains separate completely near the end of the P phase and the stretching of the unfolded chains gives rise to the E phase. Extensive conformational searching yielded a P phase force near 40 pN that agreed well with the experimental value. We suggest that the flexible and elastic S2 region, particularly the hinge region, may undergo force-induced unfolding and extend reversibly during actomyosin powerstroke.  相似文献   

20.
The concentration of carnitine in liver increased 28-fold and urinary carnitine excretion 5-fold in alloxan-diabetic sheep. In contrast there were no similar increases in alloxan-diabetic rats. The creatine content of liver decreased 3-fold and creatine excretion decreased 2-fold in diabetic sheep. In contrast the creatine content of liver increased nearly 4-fold in diabetic rats with no change in creatine excretion. The marked increased in production of carnitine by the liver of the diabetic sheep appears possible because of decreased production and excretion of creatine.  相似文献   

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