In Vitro Effects of Calcium Fructoborate upon Production of Inflammatory Mediators by LPS-stimulated RAW 264.7 Macrophages

The present study is supported by our previous findings suggesting that calcium fructoborate (CF) has anti-inflammatory and antioxidant properties. Thus, we investigated the effects of CF on a model for studying inflammatory disorders in vitro represented by lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells. This investigation was performed by analyzing the levels of some mediators released during the inflammatory process: cytokines such as tumor necrosis factor-α (TNF-α), interleukins IL-1β and IL-6 as well as cyclooxygenase-2 (COX-2), the main enzyme responsible for endotoxin/LPS-induced prostaglandin synthesis by macrophages. We also measured production of nitric oxide (NO) that plays an important role in the cytotoxicity activity of macrophages towards microbial pathogens. After CF treatment of LPS-stimulated macrophages we found an up-regulation of TNF-α protein level in culture medium, no significant changes in the level of COX-2 protein expression and a decrease in NO production as well as in IL-1β and IL-6 release. Collectively, this series of experiments indicate that CF affect macrophage production of inflammatory mediators. However, further research is required in order to establish whether CF treatment can be beneficial in suppression of pro-inflammatory cytokine production and against progression of endotoxin-related diseases.     

Differential Regulation of Eicosanoid and Endocannabinoid Production by Inflammatory Mediators in Human Choriodecidua

An increase in intrauterine prostaglandin production is critical for the onset and progression of labor in women and indeed all mammalian species studied. Endocannabinoids can act as substrates for enzymes of the prostaglandin biosynthetic pathways and can be utilized to generate other related compounds such as prostamides. The end products are indistinguishable by radioimmunoassay. We have separated such compounds by mass spectrometry. We now show that inflammatory stimuli such as LPS and proinflammatory cytokines act differentially on these pathways in human choriodecidua and preferentially create drive through to prostaglandin end products. These findings create doubt about the interpretation of data on prostaglandin biosynthesis in intrauterine tissues from pregnant women especially in the presence of an infection. The possibility is raised that separation of these products might reduce variability in results and lead to potential uses for their measurement in the diagnosis of preterm labor.     

Synthesis of Lipid Mediators during UVB-Induced Inflammatory Hyperalgesia in Rats and Mice

Peripheral sensitization during inflammatory pain is mediated by a variety of endogenous proalgesic mediators including a number of oxidized lipids, some of which serve endogenous modulators of sensory TRP-channels. These lipids are eicosanoids of the arachidonic acid and linoleic acid pathway, as well as lysophophatidic acids (LPAs). However, their regulation pattern during inflammatory pain and their contribution to peripheral sensitization is still unclear. Here, we used the UVB-model for inflammatory pain to investigate alterations of lipid concentrations at the site of inflammation, the dorsal root ganglia (DRGs) as well as the spinal dorsal horn and quantified 21 lipid species from five different lipid families at the peak of inflammation 48 hours post irradiation. We found that known proinflammatory lipids as well as lipids with unknown roles in inflammatory pain to be strongly increased in the skin, whereas surprisingly little changes of lipid levels were seen in DRGs or the dorsal horn. Importantly, although there are profound differences between the number of cytochrome (CYP) genes between mice and rats, CYP-derived lipids were regulated similarly in both species. Since TRPV1 agonists such as LPA 18∶1, 9- and 13-HODE, 5- and 12-HETE were elevated in the skin, they may contribute to thermal hyperalgesia and mechanical allodynia during UVB-induced inflammatory pain. These results may explain why some studies show relatively weak analgesic effects of cyclooxygenase inhibitors in UVB-induced skin inflammation, as they do not inhibit synthesis of other proalgesic lipids such as LPA 18∶1, 9-and 13-HODE and HETEs.     

Chitosan Enriched Three-Dimensional Matrix Reduces Inflammatory and Catabolic Mediators Production by Human Chondrocytes

This in vitro study investigated the metabolism of human osteoarthritic (OA) chondrocytes encapsulated in a spherical matrix enriched of chitosan. Human OA chondrocytes were encapsulated and cultured for 28 days either in chitosan-alginate beads or in alginate beads. The beads were formed by slowly passing dropwise either the chitosan 0.6%–alginate 1.2% or the alginate 1.2% solution through a syringe into a 102 mM CaCl2 solution. Beads were analyzed histologically after 28 days. Interleukin (IL)-6 and -8, prostaglandin (PG) E₂, matrix metalloproteinases (MMPs), hyaluronan and aggrecan were quantified directly in the culture supernatant by specific ELISA and nitric oxide (NO) by using a colorimetric method based on the Griess reaction. Hematoxylin and eosin staining showed that chitosan was homogeneously distributed through the matrix and was in direct contact with chondrocytes. The production of IL-6, IL-8 and MMP-3 by chondrocytes significantly decreased in chitosan-alginate beads compared to alginate beads. PGE₂ and NO decreased also significantly but only during the first three days of culture. Hyaluronan and aggrecan production tended to increase in chitosan-alginate beads after 28 days of culture. Chitosan-alginate beads reduced the production of inflammatory and catabolic mediators by OA chondrocytes and tended to stimulate the synthesis of cartilage matrix components. These particular effects indicate that chitosan-alginate beads are an interesting scaffold for chondrocytes encapsulation before transplantation to repair cartilage defects.     

二氢青蒿素对超高分子量聚乙烯颗粒诱导的巨噬细胞源性炎性因子释放的影响

目的:研究二氢青蒿素(Dihydroartemisinin,DHA)对超高分子量聚乙烯(Ultra highmolecularweightpolyethylene,UHMWPE)颗粒诱导的小鼠巨噬细胞系RAW264.7细胞源性炎性因子释放的影响。方法:建立UHMWPE颗粒诱导的小鼠巨噬细胞系RAW264.7细胞源性炎性因子释放模型;施加不同浓度的二氢青蒿素观察药物对细胞的影响,酶联免疫分析法(Enzyme-linked immuno sorbent assay,ELISA)检测细胞培养液上清中TNF-α,IL-1β,IL-6和IL-10含量,MTT法检测细胞毒性反应。结果:酶联免疫分析方法结果表明,二氢青蒿素可以显著抑制由UHMWPE颗粒诱导的小鼠RAW264.7细胞促炎细胞因子TNF-α,IL-1和IL-6的表达,并显著促进抗炎因子IL-10的释放,其效应具有剂量依赖性。结论:二氢青蒿素具有显著的抗炎作用,可以抑制UHMWPE颗粒诱导的巨噬细胞炎症反应,其在预防人工关节置换术后假体无菌性松动的药物治疗方面具有潜在的作用。     

The Anti-Parkinson Potential of Gingko biloba-Supplement Mitigates Cortico-Cerebellar Degeneration and Neuropathobiological Alterations via Inflammatory and Apoptotic Mediators in Mice

Neurochemical Research - Activation of nuclear factor erythroid 2 related factor 2 (Nrf2) associated with the suppression of various oxido-inflammatory pathways and the controller of several gene...     

葡寡糖对LPS诱导小鼠氧化应激与炎症反应调节作用的研究

本文研究了低、中、高三种不同聚合度葡寡糖(LGOS、MGOS、HGOS)对小鼠氧化应激与炎症反应的调节,并探讨其可能的作用机制。在采食正常日粮基础上,各组小鼠每日分别灌胃生理盐水、低聚果糖(FOS)、LGOS、MGOS、HGOS。21 d后,腹腔注射脂多糖(LPS)诱导小鼠建立氧化应激与炎症反应模型,分别测定LPS刺激后4 h和18 h时血清和肝脏中活性氧(reactive oxygen species,ROS)水平、总抗氧化能力(total antioxidant ca-pacity,T-AOC)、过氧化氢酶(catalase,CAT)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性、丙二醛(Malondialdehyde,MDA)含量、肝脏中一氧化氮(NO)含量、一氧化氮合酶(nitric oxide synthase,NOS)活性、血清中白细胞介素1(Interleukin-1,IL-1)和肿瘤坏死因子(tumor necrosis factor,TNF-a)含量。结果表明:三种聚合度葡寡糖均能显著降低LPS刺激小鼠血清、肝脏中ROS水平(P<0.05),降低肝脏中NO含量、T-NOS、iNOS活性、血清中炎性细胞因子(IL-1、TNF-a)含量(P<0.05),显著提高总抗氧化能力(T-AOC)和抗氧化酶(CAT、GSH-Px)活性(P<0.05)。葡寡糖具有保护机体免受氧化损伤,减缓炎症反应的作用,并随着平均聚合度(degree ofpolymerization,DP)的增加,其调节功能逐渐增强。     

Comprehensive Analysis of Inflammatory Immune Mediators in Vitreoretinal Diseases

Inflammation affects the formation and the progression of various vitreoretinal diseases. We performed a comprehensive analysis of inflammatory immune mediators in the vitreous fluids from total of 345 patients with diabetic macular edema (DME, n = 92), proliferative diabetic retinopathy (PDR, n = 147), branch retinal vein occlusion (BRVO, n = 30), central retinal vein occlusion (CRVO, n = 13) and rhegmatogenous retinal detachment (RRD, n = 63). As a control, we selected a total of 83 patients with either idiopathic macular hole (MH) or idiopathic epiretinal membrane (ERM) that were free of major pathogenic intraocular changes, such as ischemic retina and proliferative membranes. The concentrations of 20 soluble factors (nine cytokines, six chemokines, and five growth factors) were measured simultaneously by multiplex bead analysis system. Out of 20 soluble factors, three factors: interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemoattractant protein-1 (MCP-1) were significantly elevated in all groups of vitreoretinal diseases (DME, PDR, BRVO, CRVO, and RRD) compared with control group. According to the correlation analysis in the individual patient''s level, these three factors that were simultaneously increased, did not show any independent upregulation in all the examined diseases. Vascular endothelial growth factor (VEGF) was significantly elevated in patients with PDR and CRVO. In PDR patients, the elevation of VEGF was significantly correlated with the three factors: IL-6, IL-8, and MCP-1, while no significant correlation was observed in CRVO patients. In conclusion, multiplex bead system enabled a comprehensive soluble factor analysis in vitreous fluid derived from variety of patients. Major three factors: IL-6, IL-8, and MCP-1 were strongly correlated with each other indicating a common pathway involved in inflammation process in vitreoretinal diseases.     

A CB2 Receptor Agonist Reduces the Production of Inflammatory Mediators and Improves Locomotor Activity in Experimental Autoimmune Encephalomyelitis

Background:Cannabinoids (CBs) have been found to regulate the immune system, affect innate and adaptive immune responses, and reduce inflammatory reactions. This study assessed the therapeutic effects of GW-405833 synthetic CB2 agonist on inflammatory factors as well as locomotor activity in experimental autoimmune encephalomyelitis (EAE).Methods:In this experimental study, 48 adult male C57BL/6 mice were randomly and equally assigned to eight groups. By injecting 250 mg of MOG35-55 peptide, EAE was induced. Every other day for 17 days after EAE onset, EAE-afflicted mice in groups 1–3 received an intraperitoneal injection of GW-405833 at a dose of 3, 10, and 30 mg/kg, respectively. Clinical status and locomotor activity, measured using the beam walking assay, were assessed every other day during the first 17 days after EAE onset. Mice were euthanized in day 17th of treatment and the serum levels of the IL-1β, IL-12, CRP, and TNF-α proinflammatory cytokines as well as IL-4 and TGF-β anti-inflammatory cytokines were measured by ELISA method.Results:Clinical manifestations of EAE in groups 2 and 3 were significantly milder than group 4 and locomotor activity in groups 1–3 was significantly better than group 4 in days 5–17 (p< 0.05). GW-405833 also significantly decreased the levels of IL-12, TNF-α, and CRP and significantly increased the levels of IL-4 and TGF-β but had no significant effects on the level of IL-1β. GW-405833 was not associated with significant side effects.Conclusion:The CB2 receptor agonist GW-405833, improves clinical conditions and reduces inflammation in mice with EAE.Key Words: Clinical evaluation, Experimental autoimmune encephalomyelitis, GW-405833, Locomotor activity, Multiple sclerosis, Proinflammatory cytokines     

氯化镧对雄性小鼠精子质量及睾丸酶活力的影响

探讨氯化镧对小鼠精子质量及睾丸细胞酶的影响。40只成年昆明种雄性小鼠随机分成对照组、低(25mg·kg-1)、中(50mg·kg-1)、高(100mg·kg-1)剂量组,腹腔注射1次/4d,饲养35d。测定睾丸和附睾脏器指数,检测并计算精子数量、活精率、活动率和畸形率以及睾丸碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、乳酸脱氢酶(LDH)、一氧化氮合酶(NOS)活力。结果显示,高剂量氯化镧降低了小鼠睾丸AKP活力,抑制了精子数量和质量;中剂量氯化镧能促进NOS活力,使精子数量减少,对精子质量造成损伤。     

Production by Cultured Spleen Cells of Inflammatory Substances and Other Lymphokines that Mediate Delayed-Type Hypersensitivity in Mice

In vitro cultivation of normal mouse spleen cells with human serum albumin generated effector cells that mediate the delayed-type hypersensitivity (DTH) reaction. The cultured cells, when incubated in a serum-free medium for a further 24 hr, released substances (FPRF) which caused a footpad inflammatory reaction at a maximum of 6 hr after injection into normal syngeneic or allogeneic strains of mice, as well as macrophage migration inhibition factor (MIF) and macrophage activating factor (MAF). The DTH-effector cells in the culture were fractionated in the low density layers by discontinuous bovine serum albumin density gradient centrifugation. The effector cells in the low density layers were further enriched in the Lyt 1 subpopulation of T cells when fractionated on a fluorescence activated cell sorter. Cells capable of producing the inflammatory substances (FPRF), MIF and MAF were also enriched in the same fraction containing DTH-effector cells. These results suggest that low density, Lyt 1-positive T cells mediating the DTH reaction produce FPRF as well as MIF and MAF.     

Effect of Specific Immune Mouse Serum on the Growth of Salmonella enteritidis in Nonvaccinated Mice Challenged by Various Routes

Salmonella enteritidis was injected intravenously, intraperitoneally, or subcutaneously into specific pathogen-free mice. The number of organisms in the blood, liver, spleen, peritoneal cavity, and draining inguinal lymph node was determined by daily enumeration. Opsonization of the organism with hyperimmune serum increased the rate of phagocytosis, resulting in rapid blood clearance together with an alteration in the relative numbers of organisms accumulating in the liver and spleen. Serum treatment also brought about a substantial increase in the number of bacteria killed during the first 60 min of the infection. However, the survivors of this initial period of inactivation then multiplied rapidly in the liver and spleen, ultimately resulting in the death of the animal from a generalized infection. Attempts to passively protect mice with hyperimmune serum were uniformly negative. The effects of treatment of the virulent S. enteritidis with hyperimmune serum were consistent with the general thesis that cellular rather than humoral factors play the major role in the expression of an effective antibacterial immunity against salmonella infections.     

硝酸镧对马铃薯试管苗生长的影响

用含0、1.0、1.5、2.0、2.5、3.0、3.5、4.0mg·L~(-1)8种浓度硝酸镧的MS培养基处理马铃薯脱毒试管苗的结果表明,2mg·L~(-1)的硝酸镧对试管苗株高有显著促进,2.5mg·L~(-1)的硝酸镧对其茎粗和生物产量有极显著的促进;过高的硝酸镧浓度对试管苗有一定的抑制。     

抑制性寡脱氧核苷酸对实验性肝损伤细胞因子的影响

探讨抑制性寡脱氧核苷酸（suppressive oligonucleotides,Sup ODN）对刀豆蛋白A（Con A））诱导的小鼠实验性肝损伤的保护作用．实验采用Balb／C小鼠40只,随机分4组．分别为生理盐水（NS）组、对照寡脱氧核苷酸（control oligonucleotides,Con ODN）组、环孢菌素A（cyclosporin A,CsA）组、Sup ODN组．Con A（20mg,kg）尾静脉注射Ba1b/C小鼠,2h时腹腔给药,给药8h时观察小鼠的血清丙氨酸氨基转移酶（ALT）活性、死亡率;检测血清TNF-α、IFN-γ、TIL-4水平;RT-PCR检测肝组织TNF-α、IFN-γ mRNA的表达;并观察肝组织的病理改变．发现Sup ODN组与NS组、Con ODN组相比,ALT活性明显降低（P〈0．01）;与NS组、Con ODN组和CsA组比较,IFN-γ明显降低（P〈0．01）,IL-4明显升高（P〈0．01）,TIFN-γ mRNA表达亦显著减少：而且Sup ODN可明显减轻肝脏炎性细胞浸润和肝细胞坏死（P〈0．05）．实验表明Sup ODN对Con A小鼠实验性肝损伤有明显保护作用．