丹参多酚酸盐对ApoE-/-小鼠血清IL-6的影响

摘要目的：观察中药丹参多酚酸盐（salvianolate）对C57BIM6J ApoE-/-小鼠血清白细胞介素_6（Interleu—kin-6,IL-6）的影响,以探讨其抗动脉粥样硬化的可能机制。方法：给4周龄雄性C57BIM6J ApoE-/-小鼠喂食高脂饮食,腹腔注射丹参多酚酸盐,随机将其分为模型组（仅腹腔注射生理盐水）、丹参多酚酸盐低剂量（60mg／kg）组、中剂量（120mg／kg）组、高剂量（240mg／kg）组,每组6只,共24只;正常对照组（即C57BIM6J野生型小鼠）5只。24周末时处死各组小鼠,留取血清,采用特异性放射免疫分析各组小鼠血清IL-6水平。结果：24周末,随丹参多酚酸盐剂量的增加,ApoE-/-小鼠血清浓度逐渐降低,丹参多酚酸盐各浓度组血清IL-6水平均明显低于模型组（P均〈0．05）;丹参多酚酸盐120mg／kg组血清IL-6水平明显低于60mg／kg组（P〈0．05）;丹参多酚酸盐240mg／kg组血清IL-6水平明显低于120mg／kg组和60mg／kg组（P〈0．05）,丹参多酚酸盐240mg／kg组IL-6浓度仍明显高于正常对照组水平。结论：丹参多酚酸盐各剂量组均能够降低ApoE-/-小鼠血清IL-6水平,随剂量增加,减低越明显,说明丹参多酚酸盐能够抑制动脉粥样硬化炎症反应,可能是其抗动脉粥样硬化的作用机制之一。     

不同年龄段人血清肿瘤坏死因子－α（TNF－α）值的变化

我们用放免分析测定了不同年龄人血清ＴＮＦ－α值。结果显示，正常新生儿脐血、新生儿、幼儿及成人之间血清ＴＮＦ－α值有显著性差异（Ｐ＜０．０１），其值分别为８．０９±３．３９、１７．０９±３．２９、２４．５３±２．７９、１０．０１±４．１０ｐｍｏｌ／Ｌ；新生儿败血症患者血清ＴＮＦ－α值显著高于成人肺炎患者（Ｐ＜０．０１）．结果提示不同年龄人血清ＴＮＦ－α值的差异，可能反映了机体免疫状态的差异，从而引起不同年龄的人发病情况存在差异，值得进一步研究。     

慢性充血性心力衰竭患者血清TNF-α、IL-6 RIA的临床意义

目的：研究慢性充血性心力衰竭（chronic congestive heart failure,CHF）患者血清中肿瘤坏死因子-α（TNF—α）、白细胞介素-6（IL-6）放射免疫分析（radioimmunoassay,RIA）的价值。方法：用放射免疫分析测定176例CHF患者和30例健康人血清TNF-α、IL-6水平。比较分析CHF患者与正常人以及不同心功能程度的CHF患者之间TNF—α、IL-6水平的变化。结果：CHF患者血清TNF—α、IL-6水平明显高于正常对照组（P〈0．05）,且均随着心衰程度的加重而增高,呈高度正相关。不同程度心衰患者之间TNF—α、IL-6水平均有明显差异（P〈0．01）。结论：CHF患者存在免疫激活和心肌炎症反应,心衰程度越重,炎症反应也越明显。放射免疫分析测定TNF—α、IL-6时CHF病程及治疗疗效、预后判断具有重要的价值。     

中国汉族人群中TNF-α基因多态性与血脂关系的研究

目的研究肿瘤坏死因子-α(TNF-α)基因多态性与健康人群血脂谱改变的关系.方法随机选择182例湖北地区无血缘关系健康汉族人群,用聚合酶链反应限制性片段长度多态性(PCR-RFLP)技术检测其TNF-α基因多态性,分析各基因型对血脂、脂蛋白、栽脂蛋白的影响.结果TNF-α-238位点基因型在中国正常人群中的分布仅与日本人群中的分布较为接近,TNF-α-863位点基因型的分布与日本人、高加索人中的分布差异无显著性.结论在汉族人群中存在TNF-α基因多态性,且与血脂水平无相关性(P＞0.05).     

TNF-α基因多态性在中国人群中的分布及其与冠心病相关性的研究进展

除单核巨噬细胞可以分泌肿瘤坏死因子α（TNF-α）外,心肌细胞也可以合成并分泌TNF—α,当心肌缺血时可诱导心肌细胞表达TNF—α增强。TNF-α可以直接或间接损伤血管内皮细胞,产生促炎症、促凝血、抗纤溶反应,提示其参与冠心病的发生发展过程。TNF-α基因启动子区的多态性位点影响TNF—α基因的表达,TNF—α基因在不同种族和不同地区人群间的多态性分布使冠心病在不同种族、不同地区间具有不同的发病率和临床特点。     

大鼠重症急性胰腺炎及白藜芦醇治疗时血清IL-6及TNF-α测定的意义

目的:探讨重症急性胰腺炎(SAP)时大鼠IL-6、TNF-α的变化及白藜芦醇对其影响。方法:将实验大鼠分为假手术组、SAP组、白藜芦醇治疗1组(5mg/kg)、治疗2组(10mg/kg)、治疗3组(20mg/kg),制作SAP的动物模型后,3h、6h、12h剖杀大鼠,记录腹水量和性质,应用放射免疫分析测定血清IL-6、TNF-α浓度,取胰腺组织,观察病理变化。结果:①术后12h假手术组、治疗2组、3组生存率为100%,与SAP组(41.7%)生存率比较有显著差异(P<0.05),治疗1组生存率为50%;②治疗组2、3各时段腹水量与SAP组和治疗1组比较明显减少(P<0.05);③治疗组2、3各时段IL-6水平与SAP组和治疗1组比较明显降低(P<0.01),治疗组1与SAP组比较IL-6水平下降不明显;④治疗组2、3与SAP组各时段比较,TNF-α水平明显降低(P<0.05);⑤治疗组2、3术后6h、12h胰腺细胞坏死,评分较SAP组降低(P<0.05)。结论:①SAP时血清IL-6、TNF-α水平均显著增高,其在SAP进展中有重要作用;②白藜芦醇可明显降低,SAP大鼠IL-6、TNF-α水平,抑制炎症介质的释放和作用。     

南蛇藤素对ApoE基因敲除小鼠主动脉粥样硬化斑块内CIMO配体表达、巨噬细胞和平滑肌细胞数量的影响

目的：探讨南蛇藤素对高脂饲养ApoE基因敲除小鼠（ApoE^-/-）主动脉粥样硬化斑块内CD40配体表达、巨噬细胞和平滑肌细胞数量的影响。方法：8周龄雄性ApoE^-/-小鼠12只,随机分为南蛇藤素组或二甲基亚砜（DMSO）溶剂对照组,每组各6只。均给以高脂饲养8周,在高脂饲养的后4周,分别给予南蛇藤素2mg&#183;kg^-1&#183;d^-1或相当剂量的DMSO腹腔注射（ip）4周。麻醉处死小鼠后,取小鼠主动脉,以石蜡包埋,行主动脉根部连续切片。免疫组化法检测主动脉粥样硬化斑块内CD40配体、CD68和平滑肌α-actin表达水平,以Image ProPlus6．0软件进行图像分析。结果：与对照组相比,南蛇藤素组主动脉粥样硬化斑块内CD40配体表达显著减少（P〈0．05）;巨噬细胞的阳性率显著降低（P〈0．05）;而两组问动脉粥样硬化斑块内平滑肌细胞的阳性率没有显著差异（P〉0．05）。结论：南蛇藤素可能通过减少ApoE^-/-小鼠粥样斑块内CD40配体的表达和巨噬细胞的聚集,抑制动脉粥样硬化斑块中炎症反应,而发挥稳定动脉粥样硬化斑块的作用。     

TNF-α-238G/A基因多态性与原发性肝癌的关系

目的 探讨肿瘤坏死因子-α（TNF-α）基因启动子-238G/A多态性与原发性肝癌（PHC）的相关关系.方法 应用聚合酶链反应-限制性片段长度多态性法（PCR-RFLP）检测100例PHC患者和150例健康对照者TNF-α基因启动子-238G/A多态性.结果 TNF-α-238基因型GG、GA频率在PHC组和对照组分别为95.3%和4.7%,88%和12%;等位基因G、A频率在PHC组和对照组分别为97.7%和4.7%,94%和6%.两组差异均有显著性（P＜0.05）,携带GA基因型个体患PHC的风险约是GG基因型的2.786倍（OR=2.786,95%CI=1.057～7.343）.结论 TNF-α-238G/A基因多态性与PHC的发病有相关性,A等位基因可能是PHC的遗传易感基因.     

南蛇藤素对ApoE基因敲除小鼠主动脉粥样硬化斑块内CD40配体表达、巨噬细胞和平滑肌细胞数量的影响

目的:探讨南蛇藤素对高脂饲养ApoE基因敲除小鼠(ApoE-/-)主动脉粥样硬化斑块内CD40配体表达、巨噬细胞和平滑肌细胞数量的影响。方法:8周龄雄性ApoE-/-小鼠12只,随机分为南蛇藤素组或二甲基亚砜(DMSO)溶剂对照组,每组各6只。均给以高脂饲养8周,在高脂饲养的后4周,分别给予南蛇藤素2 mg·kg-1·d-1或相当剂量的DMSO腹腔注射(ip)4周。麻醉处死小鼠后,取小鼠主动脉,以石蜡包埋,行主动脉根部连续切片.。免疫组化法检测主动脉粥样硬化斑块内CD40配体、CD68和平滑肌α-actin表达水平,以Image Pro Plus 6.0软件进行图像分析。结果:与对照组相比,南蛇藤素组主动脉粥样硬化斑块内CD40配体表达显著减少(P＜0.05);巨噬细胞的阳性率显著降低(P＜0.05);而两组间动脉粥样硬化斑块内平滑肌细胞的阳性率没有显著差异(P＞0.05)。结论:南蛇藤素可能通过减少ApoE-/-小鼠粥样斑块内CD40配体的表达和巨噬细胞的聚集,抑制动脉粥样硬化斑块中炎症反应,而发挥稳定动脉粥样硬化斑块的作用。     

Wortmannin对急性肺损伤小鼠肺组织IL-1β和TNF-α表达的影响

目的研究Wortmannin对急性肺损伤模型小鼠肺组织白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)表达的影响。方法 30只昆明小鼠随机分为正常对照组、急性肺损伤组和Wortmannin处理组。采用腹腔注射LPS(10 mg/kg)建立小鼠急性肺损伤模型,对照组腹腔注射同体积的生理盐水,Wortmannin处理组则于造模前2 h腹腔注射Wortmannin(1.4 mg/kg)。LPS注射后6 h处死大鼠,计算肺组织湿/干重(W/D)比值,Western blot方法检测三组小鼠肺组织内IL-1β和TNF-α蛋白的表达变化,RT-PCR方法检测三组小鼠肺组织内IL-1βmRNA和TNF-αmRNA的表达变化。结果急性肺损伤组小鼠肺组织IL-1β和TNF-α蛋白及mRNA表达水平显著上升,显著高于正常对照组(0.05);相比于急性肺损伤组小鼠,Wortmannin处理组小鼠肺组织IL-1β和TNF-α蛋白及mRNA表达水平显著降低(0.05)。结论 Wortmannin能抑制急性肺损伤小鼠肺组织IL-1β和TNF-α表达。     

Effects of Tumor Necrosis Factor-Alpha on Human Trophoblast Cell Adhesion and Motility

PROBLEM: Adhesive interaction between trophoblast cells and uterine endometrial basement membrane is one of the critical processes in embryo implantation. This interaction is directly or indirectly regulated by hormones, growth factors, and cytokines. Since tumor necrosis factor-alpha (TNF-α) is synthesized by both decidual and trophoblast cells, we hypothesized that TNF-α may play a regulatory role in trophoblast cell invasion. To test this hypothesis, we have used in vitro models to determine the effect of TNF-α on human trophoblast cell adhesion and motility, two major steps in trophoblast invasion. METHODS: The effect of TNF-α on the motility of extended-lifespan first trimester trophoblasts (HTR) and JEG-3 choriocarcinoma cells was tested using the phagokinetic track motility assay. An in vitro adhesion assay was used to determine the effect of TNF-α on the adhesion of HTR and JEG-3 cells to laminin, a major basement membrane component. In addition, the effect of TNF-α on the surface expression of the laminin receptor β1 integrin subunit was examined using flow cytometry. RESULTS: HTR or JEG-3 cells were strongly adherent to laminin which was not significantly altered by TNF-α treatment. We also measured the effect of TNF-α on the surface expression of β1 integrin on HTR and JEG-3 cells; no difference was observed between control and treatment groups. Interestingly, the motility of both HTR and choriocarcinoma JEG-3 cells was significantly inhibited by TNF-α. CONCLUSIONS: The role of TNF-α in human embryo implantation is currently unknown. Our data demonstrate that TNF-α does not alter trophoblast cell adhesion to laminin, but significantly inhibits trophoblast cell motility in vitro, suggesting that TNF-α may play a regulatory role in trophoblast cell invasion.     

重组腺病毒载体转染Apo~(E-/-)小鼠的安全性评价和在主动脉血管组织的表达情况

腺病毒介导的脂联素过表达对Apo E-/-小鼠动物的安全性检测。将48只普通饮食饲养的12周龄雄性Apo E-/-小鼠分为对照组和脂联素干预组,每组24只。对照组给予一次性尾静脉注射生理盐水100μl;干预组则同法注射100μl重组腺病毒(Ad-APN-e GFP,3.0×108p.f.u)。分别在注射后0天、1周、2周、及4周处死小鼠,取全血和主动脉血管组织。用ELISA法测定血清中脂联素浓度;用WB法检测血管组织中标识绿色荧光蛋白(GFP)以确定重组腺病毒的转染率。全自动生化仪检测血常规指标、肝功能(ALT、AST)、肾功能指标(CR、BUN)、心肌酶(CK、CKMB)、以确定重组腺病毒转染的安全性。腺病毒对血常规指标和心肌酶的表达没有显著影响,对肝功能指标谷丙转氨酶(ALT)(1周时组间差异P0.01和2周时组间差异P0.05)、肾功能指标尿素氮BUN(1周时组间差异P0.01)有一过性影响。重组腺病毒可在体内高效介导外源基因脂联素的表达,血清中的脂联素浓度在病毒转染后1周达到了最高值,约是正常对照组的4倍(78.28±15.02 vs 23.42±5.35)μg/ml,2周内稳定表达。1周时GFP蛋白在血管组织中表达的量最高,1周时GFP/GAPDH为83.63±1.50%,随后逐渐降低,2周时GFP/GAPDH为54.00±4.00%,4周时GFP/GAPDH为22.00±3.99%。重组腺病毒注射存在一过性的肝毒性和肾毒性,但是对动物的生理状态没有显著影响。重组腺病毒对Apo E-/-小鼠的血管组织有高效转染、2周内安全、稳定表达的特性。     

High Serum Interleukin-10 and Tumor Necrosis Factor Alpha Levels in Chronic Paracoccidioidomycosis

In patients with chronic paracoccidioidomycosis (n = 10), levels of tumor necrosis factor alpha, interleukin-10, and interleukin-2 in serum, measured by enzyme-linked immunosorbent assay (in picograms per milliliter, as mean ± standard error of the mean), were higher than in normal controls (n = 8): 186 ± 40 versus 40 ± 7 (P < 0.05), 203 ± 95 versus 20 ± 8 (P = 0.001), and 96.3 ± 78.57 versus 1.19 ± 1.19 (P = 0.045), respectively. Gamma interferon and interleukin-4 levels were similar in patients and controls.     

Increased Levels of Soluble Tumor Necrosis Factor Receptors in Atherosclerosis: No Clear Relationship with Levels of Tumor Necrosis Factor

Inflammatory mediators, such as tumor necrosis factor alpha (TNF), may be important in the pathogenesis of atherosclerosis. Interactions between TNF and its target cell(s) requires the presence of specific receptors on the latter. Plasma levels of the two soluble forms of these receptors (tumor necrosis factor receptors, (sTNFr)) and TNF itself were measured by ELISA in 20 patients with peripheral vascular disease (PVD), 20 survivors of a myocardial infarction, and 20 age and sex matched controls. Levels of the p55 variant of the sTNFr were unchanged but levels of the p75 variant were increased in both groups of patients (ANOVA both P < 0.01). TNF was also raised in both groups of patients (both P < 0.05) but levels did not correlate with either sTNFr. In atherosclerosis, increased levels of p75 sTNFr may be further evidence of inappropriate leukocyte activation but unlikely to modulate the effect of free plasma TNF.     

Lactation Stage-Dependent Changes in Levels of Tumor Necrosis Factor/Cachectin in Milk

PROBLEM: Determination of lactation stage-dependent changes in levels of tumor neurosis factor (TNF) in milk. METHOD: Bioassay and immunoblocking assay were used to identify and assay tumor necrosis factor (TNF; mostly TNFα) in bovine milk at different stages of lactation. RESULTS: TNFα levels in milk started to increase steadily after the onset of drying-off (weaning/involution), peaked at 4 to 6 wk prior to parturition and precipitously decreased to undetectable levels at parturition (colostrum). Thereafter, TNFα reappeared and maintained midlevel concentration in the mature (normal) milk throughout the rest of the lactation cycle. Analysis of cells in mammary secretions by flow cytometry revealed that elevated TNFα levels coincided with an increase in macrophages in the secretion from the dry period. CONCLUSIONS: These lactation stage-dependent changes in TNFα levels reflect differential effects that TNFα have on involution and prepartum remodeling of the mammary gland of the dam and on gastrointestinal development and immunoregulatory function of the suckling.     

原发性高血压病患者TNF-α与IR的关联分析

目的：探讨原发性高血压病患者肿瘤坏死因子-α（TNF-α）与胰岛素抵抗（IR）的关联。方法：应用放免法测定41例原发性高血压病患者和38例正常对照组的血清TNF-α及胰岛素（fINS）水平,用胰岛素抵抗指数（HOMA-IR）评价IR程度。结果：高血压组病血清TNF-α水平显著高于正常对照组（P〈0.001）;HOMA-IR显著高于正常对照组（P〈0.001）,相关分析显示,在高血压病组及正常对照组中,TNF-α与BMI、HOMA-IR、收缩压（SBP）呈正相关（P〈0.05）。结论：高血压病患者血清TNF-α升高,且与肥胖及IR呈正相关。     

Dexmedetomidine Inhibits Tumor Necrosis Factor-Alpha and Interleukin 6 in Lipopolysaccharide-Stimulated Astrocytes by Suppression of c-Jun N-Terminal Kinases

Astrocytes play an important role in immune regulation in the central nervous system (CNS). Dexmedetomidine (DEX) has been reported to exert anti-inflammatory effects on astrocytes stimulated by lipopolysaccharide (LPS) both in vitro and in vivo studies. However, the underlying molecular mechanisms remain poorly understood. This study was designed to evaluate the effects of DEX on tumor necrosis factor-alpha (TNF-α) and interleukin 6 (IL-6) gene expressions in LPS-challenged astrocytes. Moreover, c-Jun N-terminal kinases (JNKs) and p38 mitogen-activated protein kinase (MAPK) pathways in LPS-challenged astrocytes were also investigated. In the present study, astrocytes were stimulated with LPS in the absence and presence of various concentrations of DEX. With real-time PCR assay, we found that LPS significantly increased expressions of TNF-α and IL-6 in mRNA level; however, these effects could be attenuated by DEX. Furthermore, JNK pathway might be involved in LPS-induced astrocyte activation because JNK phosphorylation was significantly increased, and the inhibition of this pathway mediated by DEX as well as SP600125 (JNK inhibitor) decreased TNF-α and IL-6 expressions. Moreover, p38 MAPK was also activated by LPS; however, this pathway seemed to have not participated in DEX-mediated LPS-induced inflammation. These results, taken together, suggest that JNK rather than p38 MAPK signal pathway, provides the potential target for the therapeutic effects of DEX for neuronal inflammatory reactions.