膝骨关节炎患者全血白细胞线粒体DNA复制数量和血浆炎性细胞因子的相关性研究（英文）

目的:本实验研究全血白细胞线粒体DNA复制数量(mtDNACN)和血浆炎性细胞因子在膝骨关节炎患者中的变化和其相关性。创新点:探讨了老年(50~80岁)膝骨关节炎患者白细胞mtDNACN和血浆炎性细胞因子水平及二者的关系。方法:分别收集膝骨关节炎组和对照组血液样本并对膝关节评分(Kellgren-Lawren grading)。使用实时定量聚合酶链反应(qRT-PCR)检测相对mtDNACN;使用多重免疫分析(multiplex immunoassay)测定血浆中10种炎性细胞因子水平;应用线性相关、Logistic回归和主成分分析(PCA)揭示骨关节炎白细胞mtDNACN和血浆炎性细胞因子的相关性。结论:血浆中白介素4(IL-4)、IL-6和全血白细胞mtDNACN可能是膝骨关节炎有效的生物标志物;IL-5则对mtDNACN减少具有潜在的影响。     

IL-6对再生障碍性贫血骨髓造血的影响

骨髓正常造血是在骨髓造血微环境的支持下,通过造血干细胞的自我维持以及造血祖细胞的大量增殖及分化为基础的活动过程.近年来研究证实骨髓造血微环境的损伤及造血细胞因子分泌紊乱是再生障碍性贫血(aplastic anemia,AA)发生的重要原因.白细胞介素-6(IL-6)是骨髓基质细胞所分泌的多种细胞因子中的一种,在免疫应答、急性期反应、造血调节等过程中都发挥重要作用.综述IL-6在再生障碍性贫血骨髓造血调控方面的研究进展.     

转染瘦素人胎盘源间充质干细胞对经X射线辐照后人脐静脉内皮细胞的成血管潜能和周围炎症的影响

目的:放创复合伤是一种以血管损伤和促炎细胞因子缺乏为特征的难愈性创伤。瘦素(leptin)的直接应用在血管生成和炎症中起着重要作用。本研究构建了一种可持续稳定的leptin表达系统——leptin修饰的人胎盘来源间充质干细胞(HPMSCs/leptin),并探究其对经X射线辐照后的人脐静脉内皮细胞(HUVECs)的成血管潜能及周围炎症的影响和潜在机制。创新点:可持续稳定的leptin表达系统(HPMSCs/leptin)促进受X射线辐照后HUVECs的成血管潜能及外周炎症反应,有助于解决放创复合伤伤口愈合过程中血管损伤和促炎因子缺乏的问题。方法:利用慢病毒载体将leptin基因转染HPMSCs获得HPMSCs/leptin。采用X射线单次照射HUVECs,剂量为20 Gy。细胞迁移侵袭实验技术(Transwell)检测照射后HUVECs的迁移情况。在Transwell体系的基础上,建立HPMSCs与受辐照HUVECs共培养体系。CCK-8比色法测定细胞增殖。酶联免疫吸附法(ELISA)检测促炎细胞因子(粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-1α(IL-1α)、IL-6和IL-8)的分泌。实时荧光定量聚合酶链式反应(RT-qPCR)检测促血管生成因子(VEGF和bFGF)mRNA的表达。蛋白免疫印迹法(westernblot)检测核因子κB(NF-κB)和JAK/STAT信号通路的相关分子表达。结论:可持续稳定的leptin表达系统(HPMSCs/leptin)具有更好的细胞增殖、迁移和成血管潜能。HPMSCs/leptin单独培养和HPMSCs/leptin与受辐照HUVECs共培养体系中,促炎细胞因子的分泌增加与NF-κB和JAK/STAT信号通路的相互作用有关。HPMSCs/leptin可能促进X射线照射后HUVECs的成血管潜能和外周炎症反应。     

转染瘦素人胎盘源间充质干细胞对经X射线辐照后人脐静脉内皮细胞的成血管潜能和周围炎症的影响（英文）

目的:放创复合伤是一种以血管损伤和促炎细胞因子缺乏为特征的难愈性创伤。瘦素(leptin)的直接应用在血管生成和炎症中起着重要作用。本研究构建了一种可持续稳定的leptin表达系统——leptin修饰的人胎盘来源间充质干细胞(HPMSCs/leptin),并探究其对经X射线辐照后的人脐静脉内皮细胞(HUVECs)的成血管潜能及周围炎症的影响和潜在机制。创新点:可持续稳定的leptin表达系统(HPMSCs/leptin)促进受X射线辐照后HUVECs的成血管潜能及外周炎症反应,有助于解决放创复合伤伤口愈合过程中血管损伤和促炎因子缺乏的问题。方法:利用慢病毒载体将leptin基因转染HPMSCs获得HPMSCs/leptin。采用X射线单次照射HUVECs,剂量为20 Gy。细胞迁移侵袭实验技术(Transwell)检测照射后HUVECs的迁移情况。在Transwell体系的基础上,建立HPMSCs与受辐照HUVECs共培养体系。CCK-8比色法测定细胞增殖。酶联免疫吸附法(ELISA)检测促炎细胞因子(粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-1α(IL-1α)、IL-6和IL-8)的分泌。实时荧光定量聚合酶链式反应(RT-q PCR)检测促血管生成因子(VEGF和b FGF)m RNA的表达。蛋白免疫印迹法(westernblot)检测核因子κB(NF-κB)和JAK/STAT信号通路的相关分子表达。结论:可持续稳定的leptin表达系统(HPMSCs/leptin)具有更好的细胞增殖、迁移和成血管潜能。HPMSCs/leptin单独培养和HPMSCs/leptin与受辐照HUVECs共培养体系中,促炎细胞因子的分泌增加与NF-κB和JAK/STAT信号通路的相互作用有关。HPMSCs/leptin可能促进X射线照射后HUVECs的成血管潜能和外周炎症反应。     

细胞因子与重症肌无力的关系

重症肌无力是由乙酰胆碱受体抗体(AChRAb)引起的自身免疫性疾病。根据分泌细胞因子的不同，将辅助性T细胞分为Th1和Th2细胞两个亚类，Th1分泌IFN-γ，IL-2为主，参与细胞免疫；Th2分泌IL-4、IL-5、IL-6、IL-10等，参与体液免疫。研究表明细胞因子在重症肌无力发病机制中起重要作用。     

IL-17在沙眼衣原体呼吸道感染中的早期表达及其与机体防御关系

目的:探讨炎症性细胞因子IL-17在沙眼衣原体呼吸道感染中的早期表达及与机体防御的关系.方法:用沙眼衣原体小鼠肺炎株(MoPn)通过鼻腔感染小鼠,用IFA检测衣原体在肺组织的生长;通过ELISA检测小鼠肺组织IL-17、IL-6、IL-8、TNF-α、IL-1α和IL-1β等炎症性细胞因子表达.结果:MoPn感染后第1天,肺组织有衣原体生长,于感染后第8天达高峰;感染后第2天,IL-17在小鼠肺组织中的表达达峰值,并很快下降;TNFα、IL-1α、IL-1β和IL-6在感染后第3天达到峰值,而IL-8则在第8天出现高峰.结论:IL-17在衣原体肺炎早期出现,能提高局部IL-6,IL-8,TNF-α,IL-1α,IL-1β的表达.这些结果显示IL-17早期表达可能作为固有免疫反应成分在起动宿主抵御细胞内病原体感染中发挥重要的作用.     

探讨IL-34对慢性根尖炎中RANKL/OPG平衡的影响

该文研究之目的:慢性根尖炎主要是以根尖炎性骨破坏为表现的口腔常见疾病,RANKL/OPG调节慢性根尖周病中的骨吸收。该文主要研究炎性因子IL-34对RANKL/OPG的平衡的影响,从而进一步探索IL-34在该疾病中的致病机理。之方法:实验组选用18组根尖周炎病例的肉芽组织,对照组选用正畸拔牙中的健康牙周组织8例。应用免疫组化检测IL-34、RANKL及OPG的表达情况,并进行数据测量及定量分析三者之间的相关性。之结果:在慢性根尖周炎中IL-34在根尖周炎和对照组的表达量分别为0.206±0.038和0.131±0.023,差异有统计学意义(P<0.05)。在根尖周炎组IL-34与RANKL呈负相关(r=-0.578,P=0.039);IL-34与OPG呈正相关(r=0.778,P=0.000),差异均有统计学意义。之讨论:本次实验证明了IL-34与根尖周炎的发生有很大关系,但IL-34可能只是很多影响RANKL/OPG平衡的炎性因子的一种。     

Rock抑制剂-Fasudil对EAE外周免疫的调节作用

目的探讨Rock抑制剂Fasudil对实验性自身免疫性脑脊髓炎（EAE）发病起始环节的外周免疫调节机制。方法将雌性C57BL/6小鼠分为CFA组、EAE组和Fasudil组。Fasudil组动物免疫后第7 d腹腔给予Fasudi（l50mg/kg.d）,连续给药14 d。免疫后隔天观察各组小鼠临床评分和体质量变化。在免疫后28 d处死动物,取脊髓进行HE染色和髓鞘染色,制备脾脏单个核淋巴细胞（MNC）,测定细胞增殖和细胞因子IL-4,IL-10,IL-1β,IFN-γ的含量,流式细胞术观察CD4,CD25调节性T细胞的比例,部分脾脏组织和MNC涂片行免疫组化荧光染色观察Rock的表达。结果Fasudil在EAE外周免疫组织和细胞通过下调Rock的表达,抑制Rock的激活,明显改善EAE的临床症状评分（P〈0.05）;减轻EAE外周炎性细胞向中枢的浸润,缓解神经髓鞘的脱失;经Fasudil干预可以下调EAE外周特异性MOG35-55反应性T淋巴细胞增殖能力,下调炎性细胞因子IFN-γ和IL-1β水平,升高保护性细胞因子IL-10水平,促进CD4,CD25调节性T细胞比例增加。结论 Rho/Rock信号通路的激活在EAE的发病中起着重要作用,应用Rock抑制剂--Fasudil抑制EAE免疫病理过程中外周免疫细胞激活这一起始重要环节,减少炎性细胞的中枢浸润,缓解EAE的临床症状。     

莲心碱可减轻LPS诱导的小鼠脓毒症模型中脾脏组织炎症和氧化应激反应（英文）

本研究基于脂多糖（LPS）诱导的小鼠脓毒症模型,探讨莲心碱（LIE）对脓毒症中脾损伤的潜在保护作用。脓毒症常伴有炎症、氧化应激和细胞凋亡,并将导致身体器官功能障碍,对脾脏损伤尤甚。将30只小鼠随机分为5组（n=6）：对照组、LPS(10 mg/kg)、LIE(10 mg/kg)+LPS、LIE(20 mg/kg)+LPS和LIE(40 mg/kg)+LPS。脾脏损伤通过苏木精–伊红染色（H&E）确定;采用定量聚合酶链反应（q PCR）检测脾脏组织中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-10、IL-1β和一氧化氮合酶（i NOS）的转录水平。同时对包括丙二醛（MDA）、过氧化氢酶（CAT）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）在内的氧化应激指标进行测定;采用TUNEL法检测细胞凋亡水平。结果显示,LIE可减轻脾脏组织病理学损伤并抑制细胞凋亡,显著降低促炎因子TNF-α、IL-6、IL-1β和i NOS的转录水平,并增加抗炎因子IL-10的表达。此外,LIE预处理后可降低MDA脂质过氧化指标,增强CAT、SOD和GSHPx的抗氧化活性...     

胰岛β细胞凋亡与一氧化氮

胰岛β细胞对各种促凋亡刺激非常敏感,β细胞凋亡在1、2型糖尿病的发病过程中起了重要作用.近年研究发现,β细胞凋亡过程涉及诸多方面,目前人们的研究主要集中在自身免疫、Fas/FasL系统、Caspase、细胞因子、过氧化等方面.一氧化氮作为一个新发现的免疫因子,参与多种因素诱导的β细胞凋亡的过程,其具体作用机制还不十分清楚.     

白细胞介素-6在神经系统内的作用

在机体内部错综复杂的细胞因子网络中，白细胞介素-6（interleukin-6，IL-6）具有重要的生物学活性．除参与机体免疫反应外，最近研究发现，IL-6参与神经系统痰患的炎症病理过程，神经系统的营养、修复和保护功能，抑制学习记忆等认知功能，因此，IL-6在神经系统内具有病理和正常生理的双重作用．IL-6与神经系统内的神经肽神经递质相互作用共同调节神经-内分泌-免疫网络．     

大鼠佐剂关节炎血清细胞因子水平与炎症及病理变化的相关性研究

目的为进一步探讨细胞因子在类风湿关节炎中的作用．方法５０只ＳＤ大鼠以完全弗氏佐剂（ＦＣＡ）形成ＡＡ动物模型．随机分为Ⅰ组（２周）、Ⅱ组（３周）、Ⅲ组（４周）、Ⅳ组（８周）以及Ⅴ组（正常对照）．分别对各组大鼠血清前炎症细胞因子ＴＮＦ－α、ＩＬ－６以及抑制性细胞因子 ＩＬ－４、 ＩＬ－ １０水平进行测定（ＥＬＩＳＡ法）．同时检查炎症指标血沉（ＥＳＲ）、关节肿胀度、病理切片，所有结果进行对比并做动态分析．结果ＡＡ大鼠Ⅰ－Ⅲ组ＥＳＲ水平及关节肿胀度与对照组均有显著差异（Ｐ＜０．０１）；ＴＮＦ－α组水平各组均高于正常（Ｐ＜０．０５，Ｐ＜０．０１，Ｐ＜０．００１）；ＩＬ－６水平Ⅱ组（Ｐ＜０．０５）、Ⅲ组高于正常；ＩＬ－４水平Ⅰ组、Ⅱ组低于正常组，Ⅲ组、Ⅳ组高于正常组，但均无显著性意义；与正常组比较，ＩＬ－１０水平Ⅰ组（Ｐ＜０．００１）、Ⅱ组（Ｐ＜０．０５）、Ⅲ组均低于正常，Ⅳ组高于正常组并有非常显著性意义（Ｐ＜０．０１）；细胞因子动态观察结果，ＴＮＦ－α、ＩＬ－ ６水平随急性期（第２周）开始而升高，高峰水平随急性期过去（第８周）而下降．而调节性因子ＩＬ－４、ＩＬ－１０高峰时间滞后于炎症因子，并随慢性期延长而升高．结论ＡＡ大?     

Up-regulation interleukin-6 and interleukin-8 by activated protein C in lipopolysaccharide-treated human umbilical vein endothelial cells

Objective： To investigate the effect of activated protein C （APC） on inflammatory responses in human umbilical vein endothelial cells （HUVEC） stimulated with lipopolysaccharide （LPS）. Methods： The second passage of collagenase digested HUVEC was divided into the following groups： serum free medium control group （SFM control）, phosphate buffer solution control group （PBS control）, LPS group with final concentration of 1 μg/ml （LPS group）, APC group with final concentration of 7 μg/ml, Pre-APC group （APC pretreatment for 30 min prior to LPS challenge）, and Post-APC group （APC administration 30 min after LPS challenge）. Supernatant was harvested at 0, 4, 8, 12 and 24 h after LPS challenge. Interleukin-6 （IL-6） and Interleukin-8 （IL-8） levels were analyzed with ELISA. Cells were harvested at 24 h after LPS challenge, and total RNA was extracted. Messenger RNA levels for IL-6 and IL-8 were semi-quantitatively determined by RT-PCR. Results： Compared with control group, IL-6 and IL-8 levels steadily increased 4 to 24 h after LPS stimulation. APC treatment could increase LPS-induced IL-6 and IL-8 production. The mRNA levels of IL-6 and IL-8 exhibited a similar change. Conclusion： APC can further increase the level of IL-6 and IL-8 induced by LPS. The effect of these elevated cytokines is still under investigation.     

Study of androgen and atherosclerosis in old-age male

Objective: To observe the difference of androgen and inflammatory cytokines level in atherosclerosis and analyse their relations. Method: Both carotid arteries and arteries of lower extremity were subjected to ultrasonic examination by Doppler's method. Those with much atheromatous plaque formation were ranged into case group, and those with normal result formed control group. Total, free testosterone and estradiol were assayed by radioimmunoassay. C reactive protein (CRP) was assayed by nepheloturbidity. Tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin- 18 (IL- 18), soluble intercellular adhesion molecule- 1 (sICAM- 1) and soluble vascular cell adhesion molecule- 1 (sVCAM- 1)were assayed by ELISA. The mean difference between two groups and the correlation between free testosterone and cytokines were analysed. Results: Free testosterone was (6.337±3.371) pg/L in case group and (11.375±4.733) pg/L in control group, P＜0.01.No differences were found in total testosterone and estradiol. CRP was (27.294±10.238) mg/L in case group and (12.843±6.318)mg/L in control group, P＜0.01. IL-6 was (41.700±31.385) pg/L in case group and (25.396±20.772) pg/L in control group, P＜0.05.IL-8 was (89.249±58.357) pg/L in case group and (67.873±31.227) pg/L in control group, P＜0.05. sICAM-1 was (470.491±134.078) pg/L in case group and (368.487±97.183) pg/L in control group, P＜0.01. sVCAM-1 was (537.808±213.172)pg/L in case group and (457.275±157.273) pg/L in control group, P＜0.05. There were no differences in TNF-α, IL-10 and IL-18.Correlation analysis showed that FT (free testosterone) had negative correlation with CRP, IL-6 and sICAM-1. Among them FT had well correlation with CRP, correlation index was -0.678. Conclusion: Free testosterone was in negative correlation with atherosclerosis in old-age male. Free testosterone may have the role of anti-atherosclerosis, and this effect was not achieved by its transformation to estradiol. Low free testosterone level was followed by increased level of inflammatory cytokines. Low free testosterones coexist with inflammation and they both affect the process of atherosclerosis in old-age male.     

A corn straw-based diet increases release of inflammatory cytokines in peripheral blood mononuclear cells of dairy cows

Recent studies have shown that diet can affect the body’s immunity. Roughage of dairy cows consists of a variety of plant materials which make different contributions to health. This study investigated the effect of different roughages on the immunity of dairy cows. Serum, peripheral blood mononuclear cells (PBMCs), and milk samples were collected from 20 multiparous mid-lactation cows fed mixed forage (MF)- or corn straw (CS)-based diets. Expression profile analysis was used to detect the differentially expressed genes (DEGs) from PBMCs. The results showed that milk protein in the MF group increased to 3.22 g/100 ml, while that of the CS group milk was 2.96 g/100 ml; by RNA sequencing, it was found that 1615 genes were differentially expressed between the CS group and the MF group among the 24 027 analyzed probes. Gene ontology (GO) and pathway analysis of DEGs suggested that these genes (especially genes coding cytokines, chemokine and its receptors) are involved in the immune response. Results were confirmed at the protein level via detecting the levels of interleukin-2 (IL-2), IL-6, IL-10, IL-12, leptin (LEP), interferon-γ (IFN-γ), transforming growth factor-β1 (TGF-β1), and tumor necrosis factor-α (TNF-α) in peripheral blood by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay analysis. Our data supported the conclusions that the protein content in milk of the MF group was higher than that of the CS group, the CS-based diets induced more release of cytokines than the MF-based diets in dairy cows’ PBMCs, and milk protein content may be affected by cytokines.     

In vitro study of immunosuppressive effect of apoptotic cells

INTRODUCTION Apoptosis plays an important role in differen- tiation, development and pathophysiological proc- esses such as inflammation, neoplasia and autoim- mune diseases. For a long time, apoptotic cells per se and the clearance of apoptotic cells had been viewed as neutral in immune response. Recently many in- vestigations suggested that apoptotic cells actively regulate the immune response (Voll et al., 1997; Fadok et al., 1998; 2000; Byrne and Reen, 2002). Apoptotic cells release …     

Study of androgen and atherosclerosis in old-age male

Objective: To observe the difference of androgen and inflammatory cytokines level in atherosclerosis and analyse their relations. Method: Both carotid arteries and arteries of lower extremity were subjected to ultrasonic examination by Doppler's method. Those with much atheromatous plaque formation were ranged into case group, and those with normal result formed control group. Total, free testosterone and estradiol were assayed by radioimmunoassay. C reactive protein (CRP) was assayed by nepheloturbidity. Tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-18 (IL-18), soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) were assayed by ELISA. The mean difference between two groups and the correlation between free testosterone and cytokines were analysed. Results: Free testosterone was (6.337±3.371) pg/L in case group and (11.375±4.733) pg/L in control group,P<0.01. No differences were found in total testosterone and estradiol. CRP was (27.294±10.238) mg/L in case group and (12.843±6.318) mg/L in control group,P<0.01. IL-6 was (41.700±31.385) pg/L in case group and (25.396±20.772) pg/L in control group,P<0.05. IL-8 was (89.249±58.357) pg/L in case group and (67.873±31.227) pg/L in control group,P<0.05. sICAM-1 was (470.491±134.078) pg/L in case group and (368.487±97.183) pg/L in control group,P<0.01.sVCAM-1 was (537.808±213.172) pg/L in case group and (457.275±157.273) pg/L in control group,P<0.05. There were no differences in TNF-α, IL-10 and IL-18. Correlation analysis showed that FT (free testosterone) had negative correlation with CRP, IL-6 and sICAM-1. Among them FT had well correlation with CRP, correlation index was −0.678. Conclusion: Free testosterone was in negative correlation with atherosclerosis in old-age male. Free testosterone may have the role of anti-atherosclerosis, and this effect was not achieved by its transformation to estradiol. Low free testosterone level was followed by increased level of inflammatory cytokines. Low free testosterones coexist with inflammation and they both affect the process of atherosclerosis in old-age male. Project (No. 2003B045) supported by the Health Bureau of Zhejiang Province, China     

In vitro study of immunosuppressive effect of apoptotic cells

Recent studies revealed that apoptotic cells are actively involved in immunosuppression and anti-inflammation. After being phagocytosed by macrophages, apoptotic cells can actively regulate cytokines secretion from lipopolysaccharide (LPS)-stimulated macrophages in which the secretion of immunosuppressive cytokines such as interleukin-10 (IL-10) is increased while the pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNFα), interleukin-1beta (IL-1β) and leukin-8 (IL-8) are suppressed. In this paper, we first present evidence that phagocytosed apoptotic cells regulate cytokine secretion of LPS-stimulated macrophages, but also inhibit the activation of T lymphocytes stimulated by ConA. These data suggest that apoptotic cells can alter the biological behavior of macrophages which gain immunosuppressive property. Project supported by the National Basic Research Program (973) of China (No. 2003CB515500) and the National Natural Science Foundation of China (No. 30371358)