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1.
目的:探讨副干酪乳酸杆菌(Lactobacillus paracasei,L.para)与嗜酸性乳酸杆菌(Lactobacillus acidophilus,L.acid)对脂多糖(LPS)诱导THP-1细胞分泌肿瘤坏死因子(TNF)-α、白细胞介素(IL)-12 p40、转化生长因子(TGF)-β的调节作用。方法:先将THP-1细胞在佛波酯(PMA)的刺激作用下活化、分化为巨噬细胞样细胞,再进行LPS诱生炎性细胞因子实验,实验分为空白对照组、LPS处理组、单独L.para处理组、L.para与LPS共处理组、单独L.acid处理组及L.acid与LPS共处理组。以RT-PCR法检测THP-1细胞TNF-αmRNA水平的表达;以ELISA法检测细胞培养上清中TNF-α、IL-12 p40和TGF-β水平的变化;以Western blot方法检测胞内未磷酸化及磷酸化IκB-α蛋白水平的表达;细胞免疫荧光技术观察NF-κB p65的亚细胞定位,TransAM核蛋白定量分析法检测THP-1细胞核内NF-κB(p65/p50)的水平。结果:①LPS能显著刺激PMA诱导分化的THP-1细胞分泌炎性细胞因子TNF-α、I...  相似文献   

2.
目的 探讨miR-155在THP-1巨噬细胞炎性反应中的作用及调控机制。方法 使用脂多糖(LPS)处理THP-1来源的巨噬细胞构建体外巨噬细胞炎性模型,分为control组、LPS组、miR-155 mimics+LPS组和mimics NC+LPS组;采用实时荧光定量PCR(RT-qPCR)检测上述各组中miR-155、炎性细胞因子(TNF-α、IL-1β)mRNA、STAT3 mRNA表达水平,酶联免疫吸附法(ELISA)检测细胞上清中炎性细胞因子(TNF-α、IL-1β)含量,免疫印迹(Western blot)法检测细胞中STAT3蛋白及STAT3磷酸化(p-STAT3)蛋白水平。结果 LPS诱导THP-1巨噬细胞中miR-155表达增加,同时炎性细胞因子(TNF-α、IL-1β)的表达增加;转染miR-155 mimics组进一步促进LPS诱导的巨噬细胞中上述炎性细胞因子的表达;Western blot法分析显示miR-155 mimics+LPS组中p-STAT3蛋白表达水平较LPS组进一步增加。结论 miR-155可上调THP-1巨噬细胞炎症中TNF-α、IL-1β的表达,其调控机制是增加转录因子STAT3的磷酸化活化,进而靶向JAK/STAT3信号通路促进LPS诱导的THP-1巨噬细胞炎性反应。  相似文献   

3.
目的探讨脂多糖(LPS)对THP-1细胞CD14诱导表达的影响。方法应用不同质量浓度(0.1~50 mg/L)的LPS刺激培养48 h的THP-1细胞,并测定其CD14mRNA、肿瘤坏死因子α(TNFα)和白细胞介素6(IL-6)表达的变化。结果正常培养条件下,THP-1细胞CD14表达量极低,而LPS对单核巨噬细胞THP-1具有显著的诱导激活作用,在较低质量浓度剂量刺激下即可被活化,表现为CD14mRNA表达量迅速增加,同时,LPS可诱导THP-1细胞产生TNF-α和IL-6,并在一定范围内呈剂量依赖关系。结论LPS可导致THP-1细胞活化,显著上调CD14mRNA的表达,并诱导产生炎症介质TNF-α和IL-6。  相似文献   

4.
目的:探讨视黄酸诱导基因Ⅰ(RIG-Ⅰ)和Toll样受体3(TLR3)两受体在呼吸道合胞病毒(RSV)感染A549细胞后诱导的炎症和抗病毒反应中的作用。方法:应用Real-timePCR方法检测RSV感染A549细胞后0,2,4,8,24hRIG-Ⅰ,TLR3及细胞因子信号转导抑制因子1/3(SOCS1/3)的mRNA的表达水平;用特异性的si-RNA分别抑制RIG-Ⅰ和TLR3两个受体基因的表达,应用Real-timePCR方法检测RSV感染抑制后的A549细胞不同时间点SOCS1/3的mRNA的表达水平。结果:RSV上调RIG-Ⅰ/TLR3两受体及SOCS1/3mRNA的表达(P<0.05);抑制RIG-Ⅰ及TLR3后,SOCS1/3mRNA的表达与对照组相比无差异性(P>0.05)。结论:RSV感染A549细胞后上调负性调节因子SOCS1/3的表达,其表达不依赖RIG-Ⅰ及TLR3受体途径,可能由病毒复制直接诱导。  相似文献   

5.
目的探讨可溶型树突状细胞特异性的结合细胞间粘附分子的非整合素(sDC-SIGN)对脂多糖(LPS)刺激的单核细胞炎症反应的调节作用。方法以人急性单核细胞白血病细胞株(THP-1)为研究对象,分为空白组、200 ng/ml sDC-SIGN单独处理组、LPS单独刺激组、LPS刺激同时加100 ng/ml sDC-SIGN处理组和LPS刺激同时加200 ng/ml sDC-SIGN处理组,刺激后不同时间点收集上清和细胞。ELISA检测细胞上清中细胞因子含量、Q-PCR检测细胞内细胞因子的mRNA表达并利用Western blot方法检测细胞内细胞外调节蛋白激酶(ERK)和核转录因子-κB(NF-κB)信号通路的活化状况。结果不同浓度的sDC-SIGN均可抑制LPS刺激炎性细胞因子TNF-α、IL-6和IFN-α的mRNA表达和蛋白分泌,同时促进抑炎细胞因子IL-10的mRNA表达和蛋白分泌,且sDC-SIGN的抑制效果与浓度呈依赖关系;研究结果亦显示,sDC-SIGN减弱LPS刺激THP-1细胞诱导ERK和NF-κBP65的磷酸化水平。结论sDC-SIGN蛋白对LPS诱导的单核细胞炎症应答具有负向调控作用。  相似文献   

6.
目的探究miR-519d-3p在脂多糖(LPS)诱导的巨噬细胞炎症中的作用和分子机制。方法用PMA诱导THP-1细胞分化为巨噬细胞,LPS刺激细胞产生炎症反应作为后续实验的细胞模型,应用荧光定量PCR检测miR-519d-3p表达;利用antago-miR-519d-3p及antago-NC或者miR-519d-3p mimic及mimic-NC分别转染细胞,构建细胞内miR-519d-3p低表达或者过表达THP-1巨噬细胞系,si RNA构建细胞内TLR4表达敲低模型;利用ELISA检测细胞上清液炎症因子TNF-α、IL-6、IL-1β分泌水平,Western blot检测各细胞系TLR4、p-p65、T-p65、p-IκBα、T-IκBα蛋白水平。结果 LPS诱导的THP-1源巨噬细胞炎症模型miR-519d-3p表达(1.85±0.10)较对照组(1.00±0.25)明显升高(P<0.05);抑制miR-519d-3p表达显著抑制细胞炎症因子TNF-α、IL-6、IL-1β分泌[TNF-α:(1.23±0.34)比(6.33±0.61);IL-6:(1.08±0.18)比(5.75±0.54);IL-1β:(1.17±0.23)比(4.76±0.45),P均<0.05],同时降低p-p65、p-IκBα蛋白水平。抑制miR-519d-3p能明显抑制LPS刺激的巨噬细胞TLR4表达;TLR4敲低的细胞模型研究表明,miR-519d-3p通过TLR4调节炎症反应。结论 miR-519d-3p可能通过调控TLR4表达参与LPS诱导的巨噬细胞炎症反应,进而在急性肺损伤中发挥重要作用。  相似文献   

7.
目的探讨细胞因子信号传导抑制因子-1(SOCS1)基因腺病毒转染对未成熟的树突状细胞(DC)的成熟及细胞因子分泌功能的影响。方法构建SOCS1腺病毒并PCR鉴定,感染体外分离、培养的小鼠骨髓DC(阳性转染组),Western blot法检测SOCS1的表达,以阴性转染组(转染Ad-GFP)及空白对照组(未经病毒转染)为对照,采用脂多糖(LPS)刺激诱导各组DC成熟,通过流式细胞术测定LPS刺激前后各组DC表面CD40及CD80的表达水平,ELISA法检测细胞因子白细胞介素(IL)-10及IL-12的分泌水平。结果成功构建并鉴定转染腺病毒,感染DC后,阳性转染组SOCS1蛋白表达水平较阴性转染组及空白对照组显著增高。LPS刺激后,阳性转染组CD40及CD80的表达较阴性转染组及空白对照组显著降低(P<0.05),IL-12分泌水平明显较低(P<0.05),而IL-10分泌水平显著增高(P<0.05)。结论通过腺病毒转染使SOCS1在DC中过度表达,不仅能有效抑制DC的成熟,还能抑制LPS介导的DC分泌细胞因子IL-12,并诱导抑制性细胞因子IL-10的分泌。  相似文献   

8.
目的 研究白念珠菌磷脂甘露聚糖( PLM)诱导人急性单核细胞白血病细胞系细胞(THP-1)产生的炎症反应是否依赖Toll样受体(TLR)2.方法 实时荧光定量逆转录PCR分析PLM体外刺激THP-1细胞TLR2、TLR4、前炎症因子[白介素(IL)-6]和趋化因子(IL-8)的mRNA表达水平.酶联免疫吸附法检测IL-6、IL-8分泌含量.免疫印迹法分析TLR2的蛋白表达.结果 PLM可升高THP-1细胞的IL-6和IL-8 mRNA表达和分泌水平(均P=0.0000).PLM上调THP-1细胞的TLR2 mRNA和蛋白表达水平(P=0.0000),但对TLR4 mRNA表达无影响.PLM经β-D-甘露糖苷水解酶处理后,不能诱导上述受体及因子的表达.TLR2中和抗体能抑制PLM诱导的IL-6和IL-8产生(P =0.0003,P=0.0010).结论 白念珠菌胞壁PLM依赖TLP2介导激活人THP-1细胞产生炎症反应.  相似文献   

9.
目的:探讨呼吸道合胞病毒(RSV)感染上皮细胞中抗病毒信号激活以及细胞因子负调节系统对细胞因子分泌的影响。方法:应用ELISA法检测RSV感染人喉癌上皮细胞(Hep2)培养上清中0,1,2,4,8,12,24 h干扰素α(IFNα-)的浓度;RT-PCR检测病毒感染后各时段上皮细胞内细胞因子信号传导抑制因子(SOCS)1,3和Toll样受体(TLR)3,4的mRNA表达水平。结果:RSV感染上调Hep2细胞SOCS3 mRNA的表达(P<0.05),SOCS1的表达先增高后减低,但无统计学意义;TLR3,TLR4 mRNA的表达增高表现为时间依赖性(P<0.05);IFNβ-的表达在RSV感染后短暂升高而后迅速降低并低于基础表达量(P<0.05)。结论:RSV感染Hep2细胞后上调TLR3,TLR4和SOCS3 mRNA的表达,抑制IFN-α的分泌,提示病毒复制可能通过上调SOCS3而拮抗TLR途径激活的宿主抗病毒免疫应答。  相似文献   

10.
目的 探讨HT-29结肠癌细胞系经细胞因子TNF-а、IL-1β、IFN-γ及脂多糖(LPS)刺激后,Toll样受体4(TLR4)、髓样分化蛋白2(MD2)及人β防御素2(HBD2)在转录和蛋白水平的表达变化及其与NF-kB激活的关系.方法 HT-29细胞分为8组,分别加入RPMI 1640、TNF-а(20 ng/mL)、IL-1β(20 ng/mL)、IFN-7(20 ng/mL)、LPS(50rig/mL)、TNF-а(20 ng/mL)+LPS(50 ng/mL)、IL-1β(20 ng/mL)+LPS(50 ng/mL)、IFN-γ(20 ng/mL)+LPS(50 ng/mL)干预.ELISA测定各组HT-29细胞上清液IL-8水平,RT-PCR检测各组细胞TLR4、MD2、HBD2 mRNA水平,免疫印迹检测各组细胞TLR4和NF-kB蛋白表达.结果 细胞因子和细胞因子加LPS组上清液IL-8的表达高于对照组(P<0.01);HT-29细胞与细胞因子预孵育后再以LPS刺激,IL-8的表达进一步升高(P<0.01).细胞因子和细胞因子加LPS均能上调HT-29细胞TLR4和MD2 mRNA的水平(P<0.01);HBD2 mRNA水平在IL-1β,LPS协同TNF-а、IL-1β、IFN-γ组升高(P<0.05).细胞因子和细胞因子加LPS均能上调HT-29细胞TLR4和NF-kB蛋白表达(P<0.01);HT-29细胞与细胞因子预孵育后再以LPS刺激,NF-kB蛋白表达进一步增高(P<0.05).结论 细胞因子(TNF-а、IL-1β、IFN-γ)能够增加肠上皮细胞(IEC)的TLR4和MD2表达、增强其对LPS的反应性,造成IEC对常驻菌的过度反应,从而启动或加重肠道炎症;炎症信号(IL-1β、LPS+TNF-а、LPS+IFN-γ)刺激可诱导IEC的HBD2基因转录表达,增强肠道获得性免疫反应,影响肠道炎症进程.  相似文献   

11.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

12.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

13.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

14.
Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.  相似文献   

15.
Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.  相似文献   

16.
A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care(~'4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.  相似文献   

17.
Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.  相似文献   

18.
Journal of Nanjing Medical University (English Edition) JNMU, sponsored by Nanjing Medical University, was established in 1987. It is a bimonthly comprehensive English medical journal published locally and abroad.Since 2007, Journal of Nanjing Medical University (English Edition )was granted Elsevier the full publishing and distribution rights worldwide for the Electronic Edition, excluding the People's Republic of China.  相似文献   

19.
Objective To measure Derpl and Blot5 allergen levels in asthmatics' homes in Hongkong.Methods Seventy houses were enrolled for a mite indoor environment study. Dust samples were obtained from two sites of each patients' house: bed and floor. Derpl and Blot5 levels were quantified by a two-site monoclonal antibody-based ELISA technique.Results The levels of Derpl allergens found in bed (geometric mean (GM) 3.43 μg/g of dust; 95%CI, 1.89-4.96 μg/g)and on the floor (GM 1.12 μg/g of dust; 95%CI, 0.71-1.53 μg/g) indicated significant differences (P=0.005). However, the levels of Blot5 allergens found in bed (GM 19.00 μg/g of dust; 95%CI, 0.89-38.90 μg/g) and on the floor (GM 6.14 μg/g of dust; 95%CI, 0.40-11.90 μg/g) showed no statistically significant difference. In addition, in regards to the exposure index for Derpl and Blot5 allergens found in bed and on the floor, 17.6% in bed and 8.6% on the floor had levels of Blot5 ≥ 10 μg/g of dust, higher than those obtained for Derp1 (7.2% and 0% in bed and on the floor respectively, P< 0.05); higher percentages in bed and on the floor (25.0% and 35.7%) were observed for levels of Blot5 =0 μg/g of dust as compared with Derpl in bed and on the floor (4.3% and 14.5% respectively, P< 0.05).Conclusions Derpl and Blot5 are the major allergens found in this regional study, Blot5 is a more potent allergen in Hongkong, probably reflecting the high level of exposure to Blomia tropicalis (Bt). Bt and Dermatophagoides pteronyssinus (Dp) allergens should be included for precise diagnosis and effective immuno-therapeutic treatment of mite allergy in Hongkong.  相似文献   

20.
ANTI-PROLIFERATION EFFECT OF ORIDONIN ON HL-60 CELLS AND ITS MECHANISM   总被引:4,自引:0,他引:4  
Objcetive To investigate the anti-proliferation effect of oridonin on leukemic HL-60 cells and its mechanism. Methods HL-60 cells in vitro in culture medium were given different concentrations of oridonin. The inhibitory rate of cells were measured by microculture tetrazolium (MTT) assay, cell apoptotic rate was detected by flow cytometry (FCM), morphology of cell apoptosis was observed by hoechst 33258 fluorescence staining, and the activity of telomerase was detected using telomere repeat amplification protocol (TRAP) PCR-ELISA before and after apoptosis occurred. Results Oridonin could decrease telomerase activity, inhibit growth of HL-60 cells, and cause apoptosis significantly. The suppression was both in time- and dose-dependent manner. Marked morphological changes of cell apoptosis including condensation of chromatin and nuclear fragmentation were observed clearly by hoechst 33258 fluorescence staining especially after cells were treated 48-60 hours by oridonin. Conclusions Oridonin has apparent anti-proliferation and apoptotic effects on HL-60 cells in vitro, decreasing telomerase activity of HL-60 cells may be one of its most important mechanisms. These results will provide strong laboratory evidence of oridonin for clinical treatment of acute leukemia.  相似文献   

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