AM3 inhibits HBV replication through activation of peripheral blood mononuclear cells

In this report, we have analyzed the effect of AM3, a glycoconjugate of natural origin with immunomodulatory properties, which is available under the commercial name of Inmunoferon, on hepatitis B virus (HBV) replication in HBV-transfected cells. We found that AM3 inhibited HBV RNA expression as well as DNA synthesis and viral antigen expression by an indirect mechanism. We found that AM3 lacked intrinsic antiviral properties, and that the antiviral effect of the glycoconjugate was due to stimulation of secretion of molecules with antiviral properties by peripheral blood mononuclear cells. Our data indicate that the employment of AM3 as an adjuvant administered simultaneously with conventional antiviral drugs may potentiate the endogenous response against viral infection.     

Polydatin ameliorates DSS-induced colitis in mice through inhibition of nuclear factor-kappaB activation

Nuclear factor- κB (NF- κB) plays a pivotal role in the regulation of immune and inflammatory responses. The real-time expression level of NF- κB reflects the development of ulcerative colitis (UC). Polydatin has vast pharmacological activities, including inhibiting the production of inflammatory mediators, inducing the production of antioxidants, regulating immune function, etc. The purpose of this study was to investigate the potential inhibitory effects of polydatin on NF- κB pathway activation in a mouse UC model. The results showed that polydatin treatment downregulated NF- κB p65 activity and expression, blocked the expression of TNF- α, IL-6 and IL-1 β at both mRNA and protein levels, decreased myeloperoxidase (MPO) activity, and alleviated inflammatory damage of colitis in mice with UC (p < 0.05), suggesting that the anti-inflammation effects of polydatin can be attributed, at least partially, to the blocking of the NF- κB pathway.     

Mibefradil-induced inhibition of proliferation of human peripheral blood mononuclear cells

To evaluate the role of intracellular calcium and particularly Ca2+-uptake in the initiation of lymphocyte mitogenesis, the effect of mibefradil, which blocks both L- and T-type calcium channels with a more selective blockade of T-type channels, on the proliferation of human peripheral blood mononuclear cells (PBMCs) is compared with the effect of nifedipine, which blocks only the L-type calcium channel. The rate of [3H]thymidine incorporation into control and concanavalin A-stimulated PBMCs in the presence or absence of the calcium channel blockers mibefradil or nifedipine (1, 10, or 50 microM), and of the intracellular calcium antagonist 3,4,5-trimethoxybenzoic acid 8-(diethylamino) octyl ester (TMB-8; 1, 10, 25, or 50 microM) was assayed in the cells cultured for 3 days. The cellular cytotoxicity and the cell number in growing cultures also was determined in mibefradil- or nifedipine-treated control or stimulated cells. Restoration of the proliferative response in mibefradil- or nifedipine-treated cells was investigated by addition of exogenous interleukin-2. Interleukin-2-receptor expression in the cells was monitored by using anti-activated T-cell antigen (Tac) antibody, and the interleukin-2 production in the cell supernatants of the cultures was determined by an enzyme-amplified sensitive immunoassay. Mibefradil and nifedipine concentration-dependently reduced the cell number and the [3H]thymidine incorporation or the de novo DNA synthesis in control and concanavalin A-stimulated human PBMCs. Mibefradil exhibited a more pronounced inhibition of the proliferation of human PBMCs than did nifedipine. The inhibitory effect of mibefradil or nifedipine on DNA synthesis was dependent on the timing of treatment with the drugs. The inhibitory effect of mibefradil or nifedipine on the lymphoproliferative response was nearly abolished if the drugs were added 20 h after cell stimulation. A markedly reduced inhibitory effect was found when mibefradil or nifedipine was added 1-7 h after cell stimulation. However, regardless of time of addition, TMB-8 caused a persistent inhibition of the proliferation of human PBMCs. The inhibitory effect of mibefradil or nifedipine on the proliferation of human PBMCs is nearly abolished by addition of the calcium channel activator Bay K 8644. The proliferative response of mibefradil- or nifedipine-treated cells is restored by addition of exogenous interleukin-2. The normal expression of interleukin-2 receptors was preserved, whereas the interleukin-2 production was blocked in the presence of mibefradil or nifedipine. Our data show that mibefradil has a more pronounced inhibitory effect on the proliferation of human PBMCs than nifedipine and that this inhibitory effect on DNA synthesis is dependent on the timing of treatment with both drugs.     

单个核细胞源性血管内皮生长因子在急性心肌梗死患者中的动态变化及意义

目的 动态观察急性心肌梗死后外周血单个核细胞分泌的血管内皮生长因子水平的变化,探讨VEGF水平与急性心肌梗死患者左室收缩功能的关系。方法 分别抽取28例急性心肌梗死患者发病后第1、5、10、15天及正常对照12例的外周静脉血,并对外周血单个核细胞进行分离、培养,用酶联免疫吸附法测定单个核细胞培养的上清液中VEGF的浓度。同时检测CK峰值及入院和出院时左室射血分数。结果 (1)AMI患者外周血单个核细胞分泌的VEGF在心梗后第5天即达高峰(343.2±82.5)pg/mL,显著高于对照组的(143.3±24.2)pg/mL(P<0.05);(2)外周血单个核细胞分泌的VEGF峰值与CK峰值无显著相关;(3)LVEF改善的患者,其单个核细胞分泌的VEGF水平(867.6±113.1)pg/mL显著高于LVEF未改善者(234.8±82.4)pg/mL(P<0.05)。结论 AMI患者外周血单个核细胞产生的VEGF与AMI患者左室收缩功能改善有关。     

Anti-inflammatory effect of spironolactone on human peripheral blood mononuclear cells

We evaluated the effect of alacepril, CV-11974, and spironolactone on the production of monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-alpha (TNF-alpha) in cultured human peripheral blood mononuclear cells stimulated with angiotensin (Ang) II. Alacepril, CV-11974, and spironolactone significantly reduced the enhanced production of MCP-1 and TNF-alpha induced by exogenous Ang II. Specifically, 10 muM of spironolactone significantly reduced cytokine production, compared to the same dose of alacepril or CV-11974. These findings indicate that spironolactone may contribute to ameliorate the prognosis of patients with cardiovascular diseases by reducing Ang II-induced inflammation, although further exploration including determining the mechanisms would be required.     

寻常型银屑病患者外周血单个核细胞Survivin的表达

目的检测寻常型银屑病患者外周血单个核细胞(PBMC)Survivin的表达,探讨其与寻常型银屑病临床分期间的关系。方法将寻常型银屑病患者分为进行期组、静止期组、退行期组,将健康志愿者设为健康对照组,每组10例,抽取外周静脉血3ml分离单个核细胞。应用反转录聚合酶链反应(RT-PCR)技术检测各组标本中SurvivinmRNA的表达水平。结果Survivin的相对表达:进行期组(0.936±0.010)>静止期组(0.559±0.007)>退行期组(0.079±0.004)>健康对照组(0.017±0.003),差异均有统计学意义(P<0.05)。结论Survivin参与了寻常型银屑病的发病过程,不同临床分期中Survivin的表达水平存在差异。     

芬太尼对人外周血NF-κB活性的影响

目的　验证芬太尼是否影响免疫炎症反应中的某些因素 ,如同吗啡。方法　外周血取自 7个正常志愿者 ,实验分为正常对照组、芬太尼 ( 2 0 μg·L-1和 2mg·L-1)组、模型组(脂多糖 ,LPS组 )和治疗组 (芬太尼 2 0 μg·L-1+LPS、芬太尼 2mg·L-1+LPS)。用流式细胞术检测人外周血中性粒细胞和单核细胞中核因子(NF κB)活性 ,用ELISA检测血清中肿瘤坏死因子 α(TNF α)和白介素 6(IL 6)含量。结果　芬太尼组NF κB活性及TNF α和IL 6含量与正常对照组比较 ,均无明显差异 (P >0 .0 5 )。治疗组 (芬太尼 2 0 μg·L-1+LPS、芬太尼 2mg·L-1+LPS)中NF κB的活性分别为 81 .9% ,76.1 % (中性粒细胞 )和 78.6% ,72 .6%(单核细胞) ,明显低于模型组 88.9%和 85 .1 % (P <0 .0 1 )。TNF α含量在治疗组 (芬太尼 2 0 μg·L-1+LPS、芬太尼 2mg·L-1+LPS)为 45 9和 3 5 7ng·L-1,IL 6为 796和 72 0ng·L-1,两者均低于模型组 (其中TNF α为 1 2 2 6ng·L-1,IL 6为 1 5 63ng·L-1) (P <0 .0 1 )。结论　芬太尼对NF κB的活性及TNF α和IL 6的含量无影响 ,但可抑制LPS诱导的NF κB的活性及TNF α和IL 6的含量 ,且高剂量芬太尼的抑制作用大于低剂量芬太尼的作用     

雷公藤多甙片对狼疮性肾炎外周血单个核细胞多药耐药蛋白表达的影响

目的研究雷公藤多甙片(TG)对狼疮性肾炎外周血单个核细胞(PBMCs)多药耐药蛋白表达的影响。方法选择2015年11月~2017年5月我院收治的58例LN患者为研究对象,分为接受TG治疗的TG组(29例)和接受常规激素治疗的对照组(29例),分别提取两组治疗前后PBMCs,采用反转录-聚合酶链反应(RT-PCR)法检测TG对PBMCs多药耐药相关蛋白蛋白(MRP)和多药耐药相关基因(MDR1)和P-糖蛋白170(P-gp 170)表达的影响。结果治疗3个月后,两组PBMCs MRP、MDR1 m RNA相对表达量均明显提高(P0.05),且TG组PBMCs MRP、MDR1 m RNA相对表达量显著高于对照组(P0.05);两组PBMCs P-gp 170阳性表达率均明显提高(P0.05),且TG组PBMCs P-gp 170阳性表达率显著高于对照组(P0.05)。结论 TG可有效提高LN患者PBMCs MRP、MDR1及P-gp 170表达,提高患者免疫力,改善预后,LN患者治疗中应检测多药耐药蛋白表达水平以确定其耐药性。     

The immunosuppressive effect of Gamisanghyulyunbueum through inhibition of mitogen-activated protein kinase and nuclear factor activation in MOLT-4 cells

Gamisanghyulyunbueum (GSHYBE) has been used clinically to treat skin related disease in South Korea. We investigated GSHYBE-mediated changes in downstream T cell signal transduction. To determine the mechanism of inhibition, we have studied many of the major pathways in phytohemagglutinin (PHA)-activated T cell. We show that among the mitogen-activated protein kinase family activation of phosphorylation of extra cellular signal-regulated kinase 1/2 (ERK1/2, p44/42) and p38, but not c-jun NH2-terminal kinase is inhibited. In activated MOLT-4 cells, the nuclear localization of nuclear factor of activated T cells (NFATc) was blocked by GSHYBE (1 mg/ml). Also, degradation of inhibitor kappaB-alpha and transactivation by nuclear factor-kappaB (NF-kappaB)/Rel A were impaired by GSHYBE (1 mg/ml). Furthermore, interlukin (IL)-2, IL-4 and Interferen (IFN)-gamma secretion by PHA activated MOLT-4 cells and peripheral blood mononuclear cells (PBMC) were significantly diminishes following GSHYBE treatment (1 mg/ml). Also, oral administration of GSHYBE inhibited IL-2 secretion in skin allergic reaction. In conclusion, our data indicate that GSHYBE treatment of T cells inhibits ERK1/2 and p38 activation and nuclear translocation of NFATc, NF-kappaB, resulting in diminished secretion of IL-2.     

The anti-inflammatory and pro-resolution effects of aspirin-triggered RvD1 (AT-RvD1) on peripheral blood mononuclear cells from patients with severe asthma

Asthma is an inflammatory disease that is characterized by a predominance of eosinophils and/or neutrophils in the airways. In the resolution of inflammation, lipid mediators such as resolvin D1 (RvD1) and its epimer aspirin-triggered RvD1 (AT-RvD1) are produced and demonstrate anti-inflammatory and pro-resolution effects. In experimental models such as airway allergic inflammation induced by ovalbumin in mice, RvD1 and AT-RvD1 alleviate some of the most important phenotypes of asthma. Here, we demonstrated the effects of AT-RvD1 on peripheral blood mononuclear cells (PBMCs) from healthy individuals and patients with severe asthma stimulated with lipopolysaccharide (LPS) or Dermatophagoides pteronyssinus (DM). AT-RvD1 (100 nM) reduced the concentration of TNF-α in PBMCs from healthy individuals and patients with severe asthma stimulated with LPS or DM. In addition, AT-RvD1 lowered the production of IL-10 only in PBMCs from patients with severe asthma stimulated with LPS. These effects were associated in part with decreasing NF-κB activation. Moreover, AT-RvD1 significantly increased phagocytosis of apoptotic neutrophils by monocytes from patients with severe asthma. In conclusion, AT-RvD1 demonstrated both anti-inflammatory and pro-resolution effects in PBMCs from patients with severe asthma and could become in the future an alternative treatment for asthma.     

肝细胞癌患者外周血单个核细胞端粒酶的表达

目的 探讨外周血单个核细胞(PBMC)端粒酶活性在肝细胞癌(HCC)患者中的意义.方法 采用PCR-TRAP-ELISA法检测HCC、慢性肝炎和健康人PBMC端粒酶活性.结果 HCC患者PBMC端粒酶活性与肿瘤组织学分化程度、肿瘤直径、血清AFP水平无显著相关性.但与门静脉侵犯和术后1年内复发显著相关.结论 PBMC端粒酶活性的高表达是HCC术后复发的指标.     

高压氧治疗对于急性脑梗死患者外周血单核细胞 Toll 样受体4的表达及预后的影响

目的 观察高压氧治疗对于急性脑梗死患者外周血单核细胞(PBMCs) Toll样受体4 (TLR-4)mRNA的表达及神经功能康复的影响。方法 入选58例脑梗死患者,随机分为对照组和高压氧治疗组,两组患者均予常规药物治疗,高压氧治疗组在常规治疗基础上予以高压氧治疗。另入选25例健康体检者为健康对照组(正常组),高压氧治疗前、治疗第5、10天分离两组患者PBMCs,荧光定量RT-PCR法检测TLR-4mRNA表达。高压氧治疗前、治疗第5、10天及起病21 d患者随访进行美国国立卫生院神经功能缺损评分(NIHSS)及日常生活能力(ADL)评分。结果 脑梗死患者(对照组,高压氧治疗组)起病时外周血单核细胞TLR-4mRNA的表达均显著高于正常组,治疗后两组患者外周血TLR-4mRNA表达均下降,高压氧治疗组显著低于对照组。高压氧治疗组治疗第10天及起病21 d后NIHSS评分均显著低于对照组,ADL评分则高于对照组。结论 高压氧治疗显著降低急性脑梗死患者外周血TLR-4mRNA的表达,减轻脑损伤后继发性炎症反应,促进神经功能的恢复,改善预后。     

白癜风患者外周血单个核细胞IL-2 IL-10和IFN-γ的表达特点

目的探讨白癜风患者中Th1/Th2型细胞因子表达的情况。方法采用细胞培养技术对白癜风患者和正常对照者的外周血单个核细胞(PBMC)在刀豆素A(ConA)作用下进行体外培养;用酶联免疫吸附试验(ELISA)检测培养上清中白细胞介素(IL)2、IL10,干扰素(IFN)γ含量。结果①白癜风患者PBMC产生IL2水平高于正常对照组(P<0.05),IL10水平低于正常对照组(P<0.01)、IFNγ水平高于正常对照组(P<0.05)。②患者和对照组PBMC加ConA刺激孔中IL2高于相应自然增殖孔(P<0.05,P<0.01)。结论白癜风患者Th1型细胞因子的表达增高,Th2型细胞因子的表达降低,可能在发病机制中起一定作用。ConA可刺激PBMC分泌细胞因子。     

肾病综合征患儿外周血单个核细胞白细胞介素13的变化

研究小儿原发性肾病综合征时白细胞介素１３的变化。方法对５０例肾病综合征患儿和３０例正常健康儿童外周血单个核细胞进行分离，培养，并应用双抗体夹心ＥＬＩＳＡ法检测培养上清中由Ｔｈ２细胞来源的细胞因子ＩＬ－１３的水平。     

P-glycoprotein function in peripheral blood mononuclear cells of myasthenia gravis patients treated with tacrolimus

Tacrolimus hydrate (FK506) reduces the symptoms of myasthenia gravis (MG) due to its immunosuppressive properties. A drug efflux pump P-glycoprotein (P-gp) actively transports FK506 out of target cells, thereby reducing their efficacy. We investigated the influence of FK506 therapy on the P-gp function of peripheral-blood mononuclear cells (PBMCs) in MG patients. Six MG patients treated with FK506 (MG(FK+)), four MG patients treated without FK506 administration (MG(FK-)), and 18 healthy subjects were included in this study. P-gp function was estimated by transporter activity that was inferred from a decrease in fluorescent P-gp substrate Rhodamine 123 (Rh123) and its inhibition by cyclosporine A (CsA). The P-gp efflux function in MG (FK+) patients assessed by the Kolmogorov-Smirnov (KS) statistic D was lower than in the healthy subjects (p=0.0084). However, PBMC sensitivity to FK506 in MG (FK+) patients was significantly higher compared to that of the healthy subjects (p=0.02). There was a significant correlation between the Rh123 efflux activity and PBMC sensitivity to FK506 in vitro (p=0.011). The data raise the possibility that FK506 treatment attenuated P-gp function in the PBMCs of the MG patients.     

2-Acetylaminofluorene suppresses immune response through the inhibition of nuclear factor-kappaB activation during the early stage of B cell development

2-Acetylaminofluorene (AAF), an arylamide carcinogen, has been known to inhibit humoral and cell-mediated immune response by lipopolysaccharide (LPS). In the current study we demonstrate that AAF induced the down-regulation of protein kinase C (PKC) that is a key enzyme in the pathways leading to LPS-induced B-cell proliferation, while having no inhibitory effect on intracellular cAMP in spleen cells. Additionally, to identify the mechanism of action of AAF during B-cell development, we determined the effects of AAF on LPS-induced nuclear factor-kappaB (NF-kappaB) activation in 70Z/3 murine pre-B cells, CH12 murine mature B cells and S194 murine plasmacytoma cells. LPS-induced NF-kappaB activation, which is dependent on PKC, was inhibited by pretreatment with AAF for 2 h in the nuclei of 70Z/3 murine pre-B cells by detection of NF-kappaB specific DNA-protein binding. Conversely, AAF barely inhibited the constitutive NF-kappaB binding activity in mature B-cells, S194 and CH12. To confirm the effect of AAF on NF-kappaB activation, a chloramphenicol acetyl transferase (CAT) expression vector containing multiple copies of the NF-kappaB element (pCAT(kappaB)(3)) was transiently transfected into 70Z/3 or S194 cells, and assessed for inducible CAT activity. AAF treatment of 70Z/3 cells resulted in a significant inhibition of CAT activity induced by LPS. However, AAF exhibited no inhibitory effect on constitutive CAT activity in mature B cells, S194, indicating that AAF no longer has suppressive effects on the immune response in differentiated B cells. Taken together, these results suggest that AAF may act to suppress immune response by blocking the activation of PKC and nuclear expression of NF-kappaB at the early stage of B cell development.     

电刺激小脑顶核对脑梗死患者局部脑血流的影响

目的 :研究FNS对脑梗死患者局部脑血流变化与临床预后关系。方法 :应用SPECT半定量分析技术测定10例脑梗死患者经FNS前、后的局部脑血流量 ,以临床神经功能缺损评分判定其情况。结果 :10例脑梗死患者电刺激前皆有不同程度局部脑血流减低 ,FNS后原脑血流减低区域和对侧大脑半球镜像区域皆有不同程度改善 ,差异有显著性 (P<0.05) ,脑血流功能改善以病灶周边较明显 ;神经功能缺损评分相应改善 ,差异有显著性 (P<0.05)。结论 :FNS对脑梗死患者的脑血流功能改善有肯定作用 ,随着脑血流功能改善其临床症状亦得到改善。