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1.
目的观察没食子酸对脂多糖(LPS)诱导RAW264.7巨噬细胞Toll样受体4/核因子κB(TLR4/NF-κB)通路的影响。方法将巨噬细胞分为空白对照组、LPS组、LPS联合没食子酸组、LPS联合NF-κB抑制剂吡咯二硫代甲酸(PDTC)组和LPS联合地塞米松(DM)组。处理后的细胞培养24 h,ELISA检测肿瘤坏死因子α(TNF-α)、白细胞介素1(IL-1)、IL-6水平,实时定量PCR检测TLR4、NF-κB mNRA水平,Western blot法检测p65、p-p65、TLR4、磷酸化的NF-κB抑制蛋白α(IκBα)的蛋白表达水平。结果 LPS诱导RAW264.7巨噬细胞后TNF-α、IL-1、IL-6水平升高,没食子酸可降低LPS诱导引起的TNF-α、IL-1和IL-6表达水平升高。LPS刺激后TLR4 mRNA及蛋白表达增加,NF-κB活化,没食子酸可拮抗以上作用,阻止NF-κB活化。结论没食子酸可通过TLR4/NF-κB通路抑制LPS诱导的RAW264.7巨噬细胞炎性反应。  相似文献   

2.
目的基于PPARγ/p38MAPK通路探讨二甲双胍对结核分枝杆菌感染巨噬细胞炎症反应的影响。方法以牛型结核分枝杆菌减毒株(M.bovis)诱导THP-1源性巨噬细胞建立感染模型。实验分4组,即对照组、M.bovis组、M.bovis+二甲双胍(metformin,Met)组、M.bovis+Met+GW9662(PPARγ抑制剂)组。分别采用RT-PCR和Western blot检测PPARγ、磷酸化p38MAPK(p-p38MAPK)基因及蛋白表达;酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)检测各组巨噬细胞TNF-α、IL-6及IL-10水平;检测各组巨噬细胞荷菌量。结果M.bovis感染巨噬细胞中PPARγ表达较对照组细胞显著升高(P<0.05),同时M.bovis感染显著增加巨噬细胞p-p38MAPK表达及TNF-α、IL-6、IL-10水平(P<0.05);二甲双胍具有激活PPARγ的功能,与单纯M.bovis组相比,二甲双胍处理后,进一步升高了PPARγ表达,但显著抑制了M.bovis感染所致的p-p38MAPK表达,降低了TNF-α、IL-6水平,升高IL-10水平(P<0.05);特异性PPARγ拮抗剂GW9662阻断PPARγ通路后,二甲双胍的上述作用被逆转;相关性分析结果显示,二甲双胍处理后,PPARγ表达与TNF-α、IL-6呈负相关,与IL-10呈正相关(P<0.05),而p-p38MAPK表达则与TNF-α、IL-6呈正相关,与IL-10呈负相关(P<0.05)。与M.bovis组相比,M.bovis+Met及M.bovis+Met+GW9662组细菌负荷量均明显降低(P<0.05),而M.bovis+Met组和M.bovis+Met+GW9662组细菌负荷量差异无统计学意义(P>0.05)。结论二甲双胍能抑制结核分枝杆菌感染巨噬细胞炎症反应,其部分机制可能与其激活PPARγ通路,进而抑制p38MAPK活化有关。  相似文献   

3.
目的探讨降钙素基因相关肽(CGRP)抑制巨噬细胞炎性反应的机制。方法使用不同浓度CGRP处理LPS活化/未活化的小鼠巨噬细胞(RAW264.7),实时荧光定量PCR(qRT-PCR)检测巨噬细胞内促炎细胞因子白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和还原型辅酶Ⅱ氧化酶-活性氧簇-NOD样受体蛋白3(NLRP3)mRNA表达水平;酶联免疫吸附法(ELISA)定量检测细胞上清分泌型IL-1β和TNF-α蛋白表达量;蛋白免疫印迹法(Western blot)检测细胞内NLRP3蛋白表达水平。结果 CGRP显著降低LPS活化/未活化RAW264.7细胞内炎性因子IL-1β、TNF-α、NLRP3 m RNA水平与蛋白水平,同时,NLRP3下游调节分子Caspase-1的蛋白水平被显著下调。在mRNA水平,NLRP3随CGRP剂量变化而变化的趋势与IL-1β、TNF-α非常相似。结论CGRP抑制巨噬细胞炎性激活,其机制可能与CGRP抑制NLRP3表达与活性有关。  相似文献   

4.
二甲双胍激活MIN6细胞AMPK活性并抑制其胰岛素分泌   总被引:1,自引:0,他引:1  
目的:观察二甲双胍对MIN6细胞胰岛素分泌和AMPK磷酸化水平的影响,以探讨二甲双胍影响胰岛素分泌的可能机制。方法:将MIN6细胞接种于24孔或6孔细胞培养板,用不同浓度葡萄糖和二甲双胍分别培养1h或20h后收集上清,待测胰岛素;抽提蛋白,检测蛋白磷酸化水平。结果:western blot结果显示20μM二甲双胍培养1h对MIN6细胞内AMPK磷酸化没有影响,培养20h则能增加AMPK磷酸化。MIN6在20μM二甲双胍存在条件下培养20h后,换入含25mmol/L葡萄糖和相同浓度药物的KRB缓冲液孵育1h,测定上清液胰岛素浓度,20μM二甲双胍组胰岛素分泌降低19.5%(P<0.05)。结论:抑制AMPK活性可以促进胰岛素分泌,二甲双胍可能通过激活AMPK活性抑制胰岛素分泌。  相似文献   

5.
目的探讨白细胞介素18(IL-18)、肿瘤坏死因子α(TNF-α)对3T3-L1脂肪细胞、RAW264.7巨噬细胞的白细胞介素18受体β(IL-18Rβ)表达的影响。方法采用反转录PCR和Western blot法检测不同浓度IL-18、TNF-α处理后3T3-L1脂肪细胞、RAW264.7巨噬细胞的IL-18RβmRNA及蛋白的表达。结果 10 ng/m L、100 ng/m L TNF-α显著增加3T3-L1脂肪细胞IL-18Rβ的mRNA及蛋白表达,10 ng/m L、100 ng/m L IL-18显著增加RAW264.7巨噬细胞IL-18Rβ的mRNA及蛋白表达。结论IL-18、TNF-α可能通过促进巨噬细胞、脂肪细胞IL-18R的表达参与糖尿病和肥胖的脂肪组织慢性炎症过程。  相似文献   

6.
黄浩  黄文成  刘金凤  黄昭东  梁艺华 《免疫学杂志》2020,36(10):829-837,845
目的探讨髓系细胞触发受体-l(triggering receptor expressed oil myeloid cells-1,TREM-1)对脂多糖(lipopolysaccharide,LPS)诱导的小鼠心肌功能障碍的作用以及对巨噬细胞RAW264.7迁移的影响。方法将健康C57BL/6J雄性小鼠随机分为对照组、LPS组、LPS+LP17组、LPS+TAK-242组并进行相应处理,6 h后超声心电图检测小鼠心功能指标,12 h取心肌组织HE染色观察病理改变,免疫组化染色观察巨噬细胞表面分子F4/80阳性表达,ELISA检测血清中炎症因子TNF-α、IL-6、IL-1β及IFN-γ的表达情况,实时荧光定量PCR检测TREM-1、BNP以及炎症因子TNF-α、IL-6、IL-1β、IFN-γ的mRNA水平;培养巨噬细胞RAW264.7,将细胞随机分为正常组、LPS组、LP17+LPS组、TAK-242+LPS组并进行对应处理,实时荧光定量PCR检测TREM-1与TLR4的mRNA水平,ELISA检测细胞培养液中炎症因子TNF-α、IL-6、IL-1β及IFN-γ的表达情况,Transwell小室观测RAW264.7细胞迁移情况,蛋白免疫印迹检测RAW264.7中TLR4/NF-κB信号通路相关蛋白水平。结果与对照组相比,超声心电图检测结果显示LPS组小鼠的EF与FS下降,LVD增大,IVS减小,LVPW变薄,变化均显著(P0.05),实时荧光定量PCR结果显示TREM-1 mRNA与BNP mRNA表达水平上升(P0.05),免疫组化染色法显示F4/80有大量阳性表达(P0.01),ELISA检测显示炎症因子TNF-α、IL-6、IL-1β、IFN-γ的含量增加(P0.05),实时荧光定量PCR检测也表明这些炎性因子mRNA水平升高(P0.05);与LPS组小鼠相比,注射TREM-1抑制剂LP17使EF、FS上升,有效抑制了LVD增大、IVS减小以及LVPW变薄的现象(P0.05),TREM-1 mRNA与BNP mRNA表达水平均降低(P0.05),心肌损伤减轻,F4/80阳性表达减少(P0.01),炎症因子水平也明显下降(P0.05),与注射TLR4抑制剂TAK-242的效果一致。与对照组相比,LPS诱导RAW264.7细胞后,细胞中TREM-1 mRNA与TLR4 mRNA水平升高(P0.05),炎症因子TNF-α、IL-6、IL-1β、IFN-γ的含量增加(P0.05),细胞大量迁移(P0.01),细胞中TLR4、MyD88、NF-κBp65、p-IκBα蛋白水平升高(P0.05);与LPS组相比,TREM-1抑制剂LP17预处理后再给予LPS诱导,细胞中TREM-1 mRNA与TLR4 mRNA水平降低(P0.05),炎症因子的含量下降(P0.05),RAW264.7细胞迁移被抑制(P0.01),TLR4、MyD88、NF-κBp65、p-IκBα蛋白水平也明显降低(P0.05)。结论给小鼠注射TREM-1抑制剂可有效减轻LPS诱导的心肌功能障碍,抑制RAW264.7细胞TREM-1表达可抑制细胞的迁移,TLR4/NF-κB途径可能参与了这一过程。  相似文献   

7.
目的:探讨三百棒醇提物(TAAE)对脂多糖(LPS)诱导的小鼠单核-巨噬细胞RAW264.7极化趋向以及其对炎症反应的影响。方法:用LPS诱导RAW264.7细胞建立炎症模型。CCK-8法检测细胞活力;ELISA检测细胞因子(TNF-α、IL-1β、IL-6和IL-10)的水平;Griess法检测细胞培养上清液中一氧化氮(NO)的分泌情况;荧光染色双标法观察巨噬细胞的极化状态;免疫荧光染色检测NF-κB定位及表达;Transwell检测TAAE对RAW264.7细胞迁移和趋化性的影响;RT-qPCR检测TNF-α、IL-1β、IL-6、IL-10、Arg-1、NF-κB和TLR4的mRNA水平;Western blot检测iNOS、COX-2、TLR4、NF-κB、p-NF-κB、IκBα和p-IκBα蛋白水平。使用TLR4通路抑制剂TAK-242进一步验证TAAE对TLR4/NF-κB信号通路的影响。结果:与模型组相比较,TAAE降低LPS诱导的炎症模型RAW264.7细胞中M1型促炎因子(TNF-α、IL-1β和IL-6)及NO水平,促使M2型抑炎因子(IL-10和Arg-1)分泌...  相似文献   

8.
目的探讨二甲双胍(Metformin, Met)对脂多糖(Lipopolysaccharide, LPS)诱发的人主动脉内皮细胞HAEC炎性损伤的影响及作用。方法体外培养人主动脉内皮细胞株HAEC,随机分为4组:正常对照组,LPS组(1μg/mL LPS),2.5mmol/L Met预处理+LPS组,5 mmol/L Met预处理+LPS组。MTT法检测细胞生存活性;Western Blot检测细胞内cleaved-caspase-3的表达;ELISA法检测细胞上清液中炎症因子IL-1β、IL-6、TNF-α的含量。实时PCR检测IL-1βmRNA、IL-6mRNA、TNF-αmRNA表达。结果与正常对照组相比,LPS组细胞生存率明显降低,细胞内的cleaved-caspase-3蛋白表达显著升高,细胞上清液中IL-1β、IL-6、TNF-α的含量明显升高。2.5mmol/L Met、5mmol/L Met预处理12h后,能显著减轻LPS诱发HAEC细胞炎性损伤,并存在剂量依赖性。而且细胞内的IL-1βmRNA、IL-6 mRNA、TNF-αmRNA表达明显低于LPS组。结论二甲双胍拮抗LPS诱发的HAEC细胞炎性损伤,与降低细胞内IL-1βmkHA、IL-6mkHA、TNF-αmRNA的表达,从而抑制炎症因子的释放相关。  相似文献   

9.
目的探究二甲双胍(Met)对脂多糖(LPS)致炎性小鼠的抗炎效应。方法 ICR小鼠分为对照组、LPS组、二甲双胍高、中和低剂量组(400, 200和100 mg/kg)。对照组和LPS组腹腔注射0.9%氯化钠溶液,其余3组注射不同剂量的二甲双胍;0.5 h后,除对照组外,其余4组腹腔注射等体积的LPS致小鼠炎性反应,6 h后测量小鼠的脾指数;ELISA检测小鼠血清中炎性因子IL-6、TNF-α和IL-10的表达;RT-PCR检测小鼠肝脏组织IL-6、TNF-α和IL-10 mRNA表达的变化;HE染色检测小鼠肝脏组织炎性反应变化。结果相对于LPS组,二甲双胍用药组小鼠的脾指数降低(P0.05);外周血IL-6和TNF-α及肝脏组织中IL-6和TNF-αmRNA的表达明显下降(P0.05);而外周血IL-10及肝脏组织中IL-10 mRNA显示上升(P0.05);二甲双胍高剂量组可明显减轻由LPS所致小鼠肝脏的炎性反应。结论二甲双胍对于LPS致炎性小鼠具有一定的抗炎活性。  相似文献   

10.
目的 研究草木犀石油醚提取物在体外的抗炎作用.方法 采用小鼠巨噬细胞系RAW 264.7建立炎症细胞模型,加入10 μg/L的LPS培养液和不同浓度的草木犀石油醚提取物进行干预.ELISA法检测上清液中TNF-α, IL-1β, IL-6和NO的分泌量;实时荧光定量RT-PCR检测TNF-α, iNOS 和 COX-2的 mRNA表达;Western 印迹法检测COX-2 蛋白的表达.结果 草木犀提取物干预后细胞所分泌的炎性介质(TNF-α, IL-1β, IL-6和NO)与模型组相比均显著降低(P<0.01),并存在剂量依赖关系;RT-PCR结果显示干预后细胞TNF-α,iNOS和COX-2的mRNA表达水平显著降低(P<0.01),也存在剂量依赖关系;Western印迹结果显示草木犀石油醚提取物及地塞米松干预后COX-2蛋白水平明显降低(P<0.01).结论 草木犀的石油醚提取物通过下调LPS诱导的巨噬细胞表达炎性介质而发挥其体外抗炎作用, 且其下调作用呈剂量依赖性.  相似文献   

11.
Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc2 (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.  相似文献   

12.
Renal dysplasia and asplenia in two sibs   总被引:2,自引:0,他引:2  
A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.  相似文献   

13.
About 1900, modern food selection and processing caused widespread epidemics of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a panhypovitaminosis B, i.e., a mixture of substrate beriberi and substrate pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse endemic disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for primates. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.  相似文献   

14.
15.
Newton H 《Medical history》2011,55(2):153-182
Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.  相似文献   

16.
Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.  相似文献   

17.
HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.  相似文献   

18.
The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.  相似文献   

19.
Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.  相似文献   

20.
Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.  相似文献   

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