Expression of IL-1β and TNF-α in MCMV Myocarditis and Its Role

Autoi mmune myocarditis in Coxsackievirus B3(CB3)-infected mice is associated with infiltrationof the heart by inflammatory cells that secreteTNF-αand IL-1[1].In this study we examine theexpression of cytokines , TNF-αand IL-1βpro-duced locallyinthe heart inresponse to MCMVin-fectionin order to determine their potential role inthe development of acute myocarditis .1MATERIALS AND METHODS1 .1Reagents and AntibodiesMonoclonal goat anti-mouse IL-1β, TNF-αanti-body and SABC …     

Effect of Interleukin-1β on the Elevation of Cytoplasmic Free Calcium of the Cultured Hippocampal Neurons Induced by L-Glutamate

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Effect of IL-1βand TNF-αon the Expression of MonocyteChem otactic Protein-1 in Endom etriotic Cells

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Effect of TGF-β1 on the Expression of IL-12, IL-15, IL-18, IL-4 and IL-10 in Heart Transplantation Rejection in Rats

To investigate the effect of TGF-β1 on the expressions of IL-12, IL-15, IL-18, IL-4 and IL-10 in heart transplantation rejection in rats, a model of rat cervical heterotopic heart transplantation was set up and the model rats were randomly divided into three groups： control group, transplant group and TGF-β1 group. The mRNA expression levels of IL-12, IL-15, IL-18, IL-4 and IL-10 were determined by RT-PCR at the 5th day after the transplantation. The mRNA expression levels of IL- 12, IL-15, IL-18 were increased obviously and those of IL-4, IL-10 were significantly decreased in the transplant group as compared with the control group （P〈0.01）. In the TGF-β1 group, the mRNA ex- pression levels of IL- 12, IL- 15, IL- 18 were significantly decreased and those of IL-4, IL- 10 were significantly increased as compared with the transplant group （P〈0.01）. The immunosuppressive effect of TGF-β1 on heart transplantation rejection was related to its inhibition of the expressions of Th1-type cytokines （IL-12, IL-15, IL-18 etc） and its promotion of the expressions of Th2-tpye cyto- kines （IL-4, IL-10）.     

Shp-2／NF-κB Pathway Mediates the Inhibition of Lipoxin A4 onIL-1β-induced Synthesis of IL-6 in Glomerular Mesangial Cells

Objective: To examine whether lipoxin A4 (LXA4) has an antagonistic effect on IL-1β-induced synthesis of IL-6 in glomerular mesangial cells, and to explore the molecular mechanisms of signal pathway in LXA4 actions. Methods: The glomerular mesangial cells of rat were cultured and treated with IL-1β with or without preincubation with LXA4 at different concentrations. The amount of IL-6 in the supernatant of cells was analyzed by enzymelinked immunosorbent assay(ELISA). The expressions of mRNA of IL-6 were determined by RT-PCR. The expressions of Src homology 2( SH2 ) containing protein-tyrosine phosphatase 2(Shp-2) were assessed by immunoprecipitation and immunoblotting. Activities of DNA-binding of nuclear factor-kappa B(NF-κB) were measured by electrophoretic mobility shift assay(EMSA). Results:IL-1β- snulated secretion of protein and expression of mRNA of IL-6 in mesangial cells were inhibited by LXA4 in a dose-dependent manner. LXA4 antagonizes the phosphorylation of Shp-2 and activities of NF-κB induced by IL-1β Conclusion: LXA4 antagonists IL-1β-induced synthesis of IL-6 in glomerular mesangial cellsthrough the mechanism of Shp-2/NF-κB pathway-dependent signal transduction.     

Effect of Interleukin-1β on IA and IK Currents in Cultured Murine Trigeminal Ganglion Neurons

To investigate the effect of interleukin-1β (IL-1β) on IA and IK currents in cultured murine trigeminal ganglion (TG) neurons, whole-cell patch clamp technique was used to record the IA and IK currents before and after 20 ng/mL IL-1β perfusion. Our results showed that 20 ng/mL IL-1β inhibited IA currents (18.3±10.7)% (n=6, P<0.05). IL-1β at 20 ng/mL had no effect on G-V curve of IA but moved the H-infinity curve V0.5 from -36.6±6.1 mV to -42.4±5.2 mV (n=5, P<0.01). However, 20 ng/mL IL-1β had effect on neither the amplitude nor the G-V curve of IK. IL-1β was found to selec- tively inhibit IA current in TG neurons and the effect may contribute to hyperalgesia under various in- flammatory conditions.     

平喘宁对哮喘豚鼠IL-4、TXB2、6-Keto-PGF1α的影响

目的:探讨平喘宁治疗哮喘的作用机制。方法:用放射性免疫法(RIA)测定豚鼠支气管肺泡灌洗液(BALF)中IL-4、血浆中TXB2、6-Keto-PGF1α在平喘宁治疗哮喘前后的变化。结果:平喘宁能显著降低豚鼠BALF中IL-4、血浆TXB2水平,提高血浆6-Keto-PGF1α的水平。结论:平喘宁通过降低豚鼠BALF中IL-4、血浆中TXB2水平,提高血浆6-Keto-PGF1α的水平,从而减轻气道炎症反应,缓解气道平滑肌痉挛而发挥治疗哮喘的作用。     

Effect of low frequency pulsed electromagnetic field on the expression and secretion of IL-1β and TNF-α in rabbit's knee chondrocytes

目的:测定在低频脉冲磁场(Low frequency pulsed electromagnetic field,LFPEMF)刺激的情况下,兔膝关节软骨细胞白介素-1β(Interleukin-1β,IL-1β)与肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)表达与分泌的变化.方法:分离培养由兔骨髓干细胞(Marrow stem cells,MSCs)诱导分化形成的软骨细胞,分别使用不同强度的LFPEMF进行电磁辐射刺激,用免疫印迹法测细胞中IL-1β与TNF-α的表达状况,并用EIISA检测细胞培养液中IL-13与TNF-α的水平.结果:施加LFPEMF的兔膝关节软骨细胞中IL-Iβ与TNF-α的表达无明显差异,但其培养液中IL-1β与TNF-α的水平有明显降低,并且随着施加脉冲磁场刺激强度的增加,IL-1β与TNF-α的降低程度增强.结论:LFPEMF可以抑制体外兔膝关节软骨细胞IL-13与TNF-α的分泌.     

Effect of HIV-1 Tat on Secretion of TNF-α and IL-1β by U87 Cells in AIDS Patients with or without AIDS Dementia Complex

Objective To explore the role of HIV-1 tat gene variations in AIDS dementia complex(ADC) pathogenesis.Methods HIV-1 tat genes derived from peripheral spleen and central basal ganglia of an AIDS patient with ADC and an AIDS patient without ADC were cloned for sequence analysis. HIV-1 tat gene sequence alignment was performed by using CLUSTAL W and the phylogentic analysis was conducted by using Neighbor-joining with MEGA4 software. All tat genes were used to construct recombinant retroviral expressing vector MSCV-IRES-GFP/tat. The MSCV-IRES-GFP/tat was cotransfected into 293T cells with pCMV-VSV-G and pUMVC vectors to assemble the recombinant retrovirus. After infection of gliomas U87 cells with equal amount of the recombinant retrovirus, TNF-α, and IL-1β concentrations in the supernatant of U87 cells were determined with ELISA. Results HIV-1 tat genes derived from peripheral spleen and central basal ganglia of the AIDS patient with ADC and the other one without ADC exhibited genetic variations. Tat variations and amino acid mutation sites existed mainly at Tat protein core functional area(38-47aa). All Tat proteins could induce U87 cells to produce TNF-α and IL-1β, but the level of IL-1β production was different among Tat proteins derived from the ADC patient's spleen, basal ganglia, and the non-ADC patient's spleen. The level of Tat proteins derived from the ADC patient's spleen, basal ganglia, and the non-ADC patient's spleen were obviously higher than that from the non-ADC patient's basal ganglia. Conclusion Tat protein core functional area(38-47aa) may serve as the key area of enhancing the secretion of IL-1β. This may be related with the neurotoxicity of HIV-1 Tat.     

Expression and Significance of NF-κB, IL-1β and COX-2 in the Murine Model of Estrogen-dependent Experimental Vulvovaginal Candidiasis

Objective To investigate the possible role of estrogen in the pathogenesis of vulvovaginal candidiasis(VVC). Methods Estrogen-dependent experimental murine model of C. albicans vaginal infection was established by injecting subcutaneously with estradiol benzoate and then 5×106 stationary-phase C. albicans blastoconidia was inoculated intravaginally to mice (group EI), and other 3 groups were set up: estrogen-treated but not infected (group E); estrogen-untreated but infected (group I); normal control (group C). The dynamic change of colony-forming unit (CFU) of cervivovaginal lavage fluid was observed. Vaginal tissues at different time points (d 2, d 4, d 7 and d 14) after inoculation of C. albicans were obtained. In situ hybridization staining was used to detect expression of cyclooxygenase-2 (COX-2) mRNA and expression of nuclear factor-κB (NF-κB) was examined by immuohistochemistry. ELISA was applied to determine the interlenkin-1β (IL-1β) level. Results The constitutional high level expression of COX-2 mRNA in the vaginal tissue of group E was significantly higher than that in group C on d 4 and d 7 (P<0.01), and the optical density (OD) in group EI was higher than that in the other 3 groups (P<0.05). There were higher IL-1a levels in vaginal tissues from d 4 to d 7 postin- oculation in group EI and group I than group C (P<0.01). Furthermore, IL-1β in group EI was markedly elevated on d 4 and d 7 compared with group I (P<0.01). Compared with group C, the expression of NF-κB in group E was increased obviously on d 4 (P<0.01), and there was significant difference between group EI and group Ion d 4 and d 7 (P<0.01). Conclusions In the murine model of estrogen-dependent experimental VVC, estrogen promotes the infection establishment by up-regulating expression of COX-2 via activating NF-κB signal pathway, and the high expression of COX-2 promoted by the interaction of IL-1β and NF-êB after infection formation was involved in persistence of infection.     

Effect of Tumor Necrosis Factor-αon Resistin Expression in 3T3-L1 Adipocytes and Its Mechanism

Insulin resistance plays an important role inthe pathogenesis, development and complication indiabetes mellitus. Obesity is one important cause,but its mechanism is complicated. Steppan foundthat resistin had direct effect on insulin in mouseadipose cells in 2001 and believed that it might bethe key molecule of diabetes mellitus[1]. TNFαis apolypeptide which has extensive biological actionand effects in the development of anti tumourmechanism, immunoloregulati…     

Construction of Antisense Transforming Growth Factorβ_1 Gene and Its Effect on the Proliferation by Expression in Osteosarcoma Cells

Summary:To construct the antisense transforming growth factorβ1(TGFβ1)gene and investigatethe effect of TGFβ1 autocrine loop blockage on the proliferation of osteosarcoma cells.TGFβ1 cDNAwas cloned by RT-PCR from human osteosarcoma cells(MG-63)and inserted into pcDNA_3 to con-struct an antisense expression vector,which was dubbed pcDNA_3-TGFβ1(-).MTT was used to de-tect the proliferation of osteosarcoma cells transfected by antisense TGFβ1 gene.Our results showedthat the proliferation of the transfected osteosarcoma cells was suppressed markedly.It is concludedthat TGFβ1 autocrine loop blockage in osteosarcoma cells could inhibit cell proliferation,which mightbe helpful for gene therapy of osteosarcoma.     

Effect of remifentanil on toll-like receptor 4, NF-κB and IL-6 in rabbit myocardial ischemia/reperfusion model

Objective:To investigate whether remifentanil induced cardioprotecting effect is associated with expression of toll-like receptor 4 (TLR4).nuclear factor κB (NF-κB) and serum interleukin -6 (IL-6).Meth...     

Effect of Quercetin on Atherosclerosis Based on Expressions of ABCA1, LXR-α and PCSK9 in ApoE-/- Mice

Objective: To investigate the effect of quercetin on ATP binding cassette transporter A1(ABCA1), liver X receptor(LXR), and proprotein convertase subtilisin/kexin type 9(PCSK9) expressions in apo E-knockout(Apo E~(-/-) ) mice. Methods: The high-fat diet-induced atherosclerosis(AS) in Apo E~(-/-) mice was established. Thirtysix mice were divided into 3 groups using random number table method: model group(n=12), quercetin group(n=12), and atorvastatin group(n=12), with C57 BL/6 J mice of the same strain and age as the control group(n=12). Quercetin group and atorvastatin group were administrated with quercetin and atorvastatin by oral gavage, with doses of 12.5 and 4 mg/(kg·d), respectively. Animals in the control and model groups were given an equal volume of distilled water by oral gavage once per day for a total of 12 weeks. Western blot and immunohistochemical methods were employed to determine the aortic ABCA1, LXR-α and PCSK9 protein expression. Enzyme linked immunosorbent assay method was used to detect the expression of serum total cholesterol(TC), triglyceride(TG), high density lipoprotein-cholesterol(HDL-C), low density lipoproteincholesterol(LDL-C), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and IL-10, combined with tissue pathological examination. Results: Apo E~(-/-) mice fed with a high-fat diet had notable atherosclerosis lesions, with reduced ABCA1, LXR-α and IL-10 levels(all P0.01), elevated PCSK9, TNF-α and IL-6 expression, and increased TC and LDL-C contents(all P0.01). After quercetin intervention, the areas of AS plaques and the expressions of PCSK9, TNF-α and IL-6 were significantly reduced(all P0.01), while the expressions of ABCA1 and LXR-α were increased significantly(all P0.01). Conclusion: Quercetin effectively interfered with AS development by regulating the expressions of ABCA1, LXR-α and PCSK9 in Apo E~(-/-) mice.     

腰椎间盘突出症患者血清MMP-3、PGE2、IL-1β与疼痛的相关性分析

目的：分析腰椎间盘突出症患者血清基质金属蛋白酶3(MMP-3)、前列腺素E₂(PGE₂)、白介素-1β(IL-1β)与疼痛的相关性。方法：选择2019年5月-2021年5月于乐平市中医医院骨科诊治的腰椎间盘突出症患者80例作为观察组,选择同期于本院进行体检的57例健康成年人作为对照组。比较两组血清MMP-3、PGE₂、IL-1β水平;根据视觉模拟评分(VAS)将80例腰椎间盘突出症患者分为轻度疼痛组(≤3分)、中度疼痛组(4～6分)、重度疼痛组(7～10分),比较不同疼痛程度腰椎间盘突出症患者血清MMP-3、PGE₂、IL-1β水平,并进一步分析三项指标与VAS评分的相关性。结果：与对照组相比,观察组MMP-3、PGE₂、IL-1β水平均显著升高(P<0.01)。与轻度疼痛组相比,中、重度疼痛组MMP-3、PGE₂、IL-1β均显著升高(P<0.05);与中度疼痛组相比,重度疼痛组MMP-3、PGE₂、IL-1β均显著升高...     

The Effect of Vitamin A on Secretion of IFN-γ and IL-4 in A549 Cells Induced by Mycoplasma Pneumoniae

In order to investigate the effect of vitamin A (VA) on the secretion of IFN-γ and IL-4 in Mycoplasma Pneumoniae (MP)-induced A549 cells, A549 cells were co-cultured with MP for different time lengths and then the levels of IFN-γ and IL-4 in the cell culture supernatants were detected before and after treatment with different concentrations of VA by using the enzyme-linked immu-nosorbent assay ( ELISA). The results showed that the level of IFN-γ and IL-4 in the supernatants of MP-induced A549 cells was much higher than that in non-induced cells (P<0.01). After application of VA, IL-4 level was not increased until the concentration of VA was up to 0.5×10-5 mol/L (P<0.01). However, with concentration of VA increased up to 1×10-4 mol/L, IL-4 was significantly suppressed (P<0.01). It was concluded that MP could induce the secretion of IFN-γ and IL-4 in A549 cells. VA could inhibit the secretion of IFN-γ and increase the IL-4 level in MP-induced A549 cells. However, high concentration of VA had an inhibitory effect on the secretion of IL-4 as well as on the IFN-γ. These data provided a theoretical basis for the application of VA in MP pneumonia in the clinical practice.     

MSCs通过促分泌TGF-β1、IL-6和IL-10抑制干燥综合征患者CD4+T细胞的活化增殖

目的 体外探讨人脐带间充质干细胞(mesenchymal stem cells, MSCs)对原发性干燥综合征(primary Sjgren syndrome, pSS)患者外周血活化CD4⁺ T细胞的免疫抑制效应,分析MSCs促分泌的转化生长因子-β1(transforming growth factor-β1, TGF-β1)、IL-6及IL-10在免疫抑制效应中的作用。方法 原代分离脐带MSCs,并经流式细胞术鉴定。分选成年健康者和pSS患者外周血CD4⁺T细胞,分为对照组活化CD4⁺T细胞(CD3、CD28抗体共刺激72h)、MSCs共培养组(活化CD4⁺T细胞与MSCs共培养72h)和干扰素-γ(IFN-γ)预刺激后MSCs共培养组(活化CD4⁺T细胞与IFN-γ预刺激后的MSCs共培养72h),活化CD4⁺T细胞组为对照。ELISA法检测培养上清液中TGF-β1、IL-6、IL-10因子水平。结果 相对于活化CD4⁺T细胞组,MSCs共培养组培养上清中TGF-β1、IL-6、IL-10浓度均显著升高(P≤0.01),并且活化的CD4⁺T细胞增殖受到抑制。结论 MSCs通过促分泌TGF-β1、IL-6、IL-10抑制活化的CD4⁺T细胞增殖,起到免疫抑制作用。     

Expression of urotensin Ⅱ and its relationship with pro-inflammatory cytokines of TNF-α and IL-1β in early stage of acute liver failure

目的 探讨小鼠急性肝衰竭(ALF)早期尾加压素Ⅱ(UⅡ)表达和分泌的动态变化及其对前炎症细胞因子的影响.方法 选择6周龄雄性BALB/c小鼠60只,随机分为模型组和预处理组,每组30只.两组均建立ALF动物模型,即采用脂多糖(LPS)50 μg/kg联合右旋半乳糖胺(D-GalN )800 mg/kg(LPS/D-GalN)腹腔注射;预处理组在LPS/D-GalN注射前0.5h,以UⅡ受体拮抗剂urantide 0.6 mg/kg尾静脉注射.两组均在LPS/D-GalN注射后0、0.5、1、2和6h处死动物(每一时间点各6只小鼠),其中0h作为对照(仅腹腔内注射0.2 mL无菌生理盐水);采集动物血清和肝组织标本.采用RT-PCR及ELISA方法分别检测U Ⅱ、肿瘤坏死因子α(TNF-α)及白介素1β(IL-1β)mRNA和蛋白质的表达.结果 LPS/D-GalN注射后0.5h,肝组织和血清中UⅡ的表达和分泌即快速上升并达峰值,且峰值水平持续至LPS/D-GalN攻击2h后,至6h时UⅡ的表达与分泌虽有所降低,但仍显著高于正常水平(p＜0.05);面TNF-α水平在LPS/D-GalN攻击后0.5h内无明显升高(P＞0.05),直到1h后才快速上升达峰值(P＜0.05),至6h时开始下降(P＜0.05);IL-1β的表达与分泌则直到6h后才有明显上升(P＜0.05).Urantide的应用不仅显著抑制了LPS/D-GalN攻击诱导的UⅡ高表达,也使上调的TNF-α和IL-1β表达和分泌明显受抑(P＜0.05).结论 UⅡ可刺激TNF-α的表达,有可能作为炎症级联效应的触发者在ALF的发生或启动中发挥关键性影响.     

Effect of miR-125b on expression of Cbfβ in C3H10T1/2 cells

目的 研究miR-125b对鼠间充质干细胞株C3H10T1/2中核心结合因子-β(core-binding factor beta,Cbfβ)表达的调节作用.方法 将microRNA阴性对照、miR-125b mimic和anti-miR-125b用脂质体转染小鼠间充质干细胞系C3H10T1/2,Western blot及RT-PCR测定细胞中Cbfβ蛋白及mRNA表达;以PicTar数据库预测的Cbfβ3’-非翻译区上miR-125b靶位点附近约30个碱基为基础合成3组插入序列(阳性对照、野生型序列、突变序列),连接到经HindⅢ、SpeⅠ双酶切后的PmiR-report载体质粒上,然后与miRNA阴性对照和miR-125b mimic共同转染293T细胞,双荧光素酶报告基因检测荧光素酶活性.结果 过表达miR-125b抑制Cbfβ蛋白及mRNA表达(P＜0.05),转染anti-miR-125b可促进Cbfβ蛋白及mRNA表达(P＜0.05);miR-125b抑制Cbfβ3’-非翻译区荧光素活性.结论 外源性miR-125b可作用于Cbfβ3’非翻译区从而负性调节C3H10T1/2细胞中Cbfβ的表达.     

Hemoglobin structure at higher levels of hemoglobin A1C in type 2 diabetes and associated complications

Background: Fourier transform infrared (FTIR) spectroscopy technique has not been used as a diagnostic tool for diabetes in clinical practice. This study was linked to structural changes in hemoglobin (Hb) in type 2 diabetes patients at higher levels of HbA1C using FTIR spectroscopy.Methods: Fifty-three diabetic patients from the Bahawal Victoria Hospital, Bahawalpur, Pakistan were categorized as group A (6% < HbA1C < 7%;n = 25) and group B (HbA1C ≥9%;n = 28). Another group (group N) of tw...