Heat shock protein 70-2 and tumor necrosis factor-α gene polymorphisms in Chinese children with Henoch- Schönlein purpura

Background:Henoch-Schönlein purpura(HSP)or IgAassociated vasculitis is related to immune disturbances.Polymorphisms of the heat shock protein 70-2 gene(HSP70-2)and the tumor necrosis factor-αgene(TNF-α)are known to be associated with immune diseases.The purpose of this study was to investigate the likely association of HSP70-2(+1267A/G)and TNF-α(+308A/G)gene polymorphisms with HSP in children.Methods:The polymerase chain reaction restriction fragment length polymorphism method was used to detect the HSP70-2 and TNF-αpolymorphisms in 205 cases of children with HSP and 53 controls;and the association of these polymorphisms with HSP and HSP nephritis(HSPN)was analyzed.Results:The G/G genotypic frequencies at the+1267A/G position of HSP70-2 in the HSP group(22.9%)were signifi cantly higher than those in the healthy control group(9.4%)(χ^(2)=4.764,P<0.05).The frequencies of the A/A,A/G and G/G genotypes of HSP70-2 in patients in the nephritis-free group and the HSPN group showed no statistically significant difference.The A/A genotype frequency at the+308G/A position of TNF-αin the HSP group was 8.3%,which was higher than that in the control group(χ^(2)=6.447,P<0.05).The A allele frequency of TNF-αin the HSP group was higher than that in the control group,with a statistically significant difference(χ^(2)=7.241,P<0.05).Conclusions:The HSP70-2(+1267A/G)and TNF-α(+308G/A)gene polymorphisms were associated with HSP in children.The G/G homozygosity of HSP70-2 and the A/A homozygosity of TNF-αmay be genetic predisposing factors for HSP.

Effects of Puerarin on Plasma Endothelin and Serum Tumor Necrosis Factor-α in Type 2 Diabetes Mellitus Patients with Vascular Complications

Objective: To study the changes of endothelin (ET), tumor necrosis factor-α (TNF-α) before and after puerarin treatment in patients with diabetes mellitus vascular complications (DMVC). Methods: Ninety-eight DMVC patients were divided into 2 groups, they were given puerarin (n=68) and normal saline (n=30) respectively, 20 diabetic patients without vascular complications (NDMVC) were taken as control, who were also given puerarin. All the patients were treated on the basis of controlling blood glucose. Plasma ET and serum TNF-α were determined by radioimmunoassay (RIA) before and after treatment. Results: Plasma ET and serum TNF-α in DMVC got higher than that of NDMVC patients (P<0.05), and ET level was correlated with TNF-α (r=0.69, r=0.73, P<0.01). After treatment, the levels of ET and TNF-α were significantly lower than those before treatment of DMVC patients with puerarin (P<0.05). Conclusion: Puerarin could regulate the levels of plasma ET and serum TNF-α of DMVC patients, suggesting that it has the function of regulating endothelial cells.

Altered pattern of tumor necrosis factor-alpha production in peripheral blood monocytes from Crohn's disease

AIM To evaluate the inflammatory state in Crohn's disease(CD) patients and correlate it with genetic background and microbial spreading.METHODS By means of flow cytometry, production of tumor necrosis factor-alpha(TNF-α) was measured in peripheral blood monocytes from patients suffering from CD, ulcerative colitis(UC) and in healthy subjects after stimulation of the NOD2 and TLR pathways. CD patients were genotyped for the three most common NOD2 variants(R702W, G908 R and L1007Pfs*2) and basal production of TNF-α was correlated to NOD2 genotype. Also, production of TNF-α was correlated to plasmatic levels of LPS Binding Protein(LBP), soluble(s) CD14 and to the activity state of the disease.RESULTS The patients with CD were characterized by a significantly higher monocyte basal expression of TNF-αcompared with healthy subjects and UC patients, and after stimulation with Pam3CSK4(ligand of TLR2/1) and MDP-L18(ligand of NOD2) this difference was maintained, while other microbial stimuli(LPS, ligand of TLR4 and Poly I:C, ligand of TLR3) induced massive activation in CD monocytes as well as in UC and in healthy control cells. There was no significant difference in the production of TNF- α between patients who carried CD-associated heterozygous or homozygous variants in NOD2 and patients with wild type NOD2 genotype. Although serum LBP levels have been shown to correlate positively with the state of activity of the disease, TNF-α production did not show a clear correlation with either LBP or s CD14 levels in plasma. Moreover, no clear correlation was seen between TNF-α production and activity indices in either CD or UC.CONCLUSION Peripheral monocytes from CD express higher basal and stimulated TNF-α than controls, regardless of NOD2 genotype and without a clear correlation with disease activity.

Effects of Wenglian Jiedu decoction on expression of serum IL-18, TNF-α and MUC2 in colon tissue of ulcerative colitis rats

Objective:To study the effect of Wenglian jiedu decoction on the expression of serum interleukin-18(IL-18),tumor necrosis factor-α(TNF--α)and colonic mucin-2(MUC2)in rats with ulcerative colitis of turbid toxin type,and to explore its mechanism.Methods:60 Wistar male rats were randomly divided into model group,Wenglian jiedu decoction low,middle and high dose groups,mesalazine group and blank group.Except for the blank group,the rat model of ulcerative colitis was established by trinitrobenzenesulfonic acid(TNBS)/ethanol compound method.The general condition,body weight,fecal properties and occult blood of rats were observed 2 weeks after administration.The disease activity index(Diseaseactivityindex,DAI)was scored,the content of MUC2 protein was detected by immunohistochemical method,and the contents of IL-18 and TNF-αin serum were detected by enzyme-linked immunosorbent assay(Elisa).Results:compared with the blank group,the DAI score increased,the contents of IL-18 and TNF-αincreased significantly,and the content of MUC2 protein decreased in the model group.Compared with the model group,the DAI score,the content of IL-18 and TNF-αdecreased and the content of MUC2 protein increased in each treatment group.Compared with the mesalazine group,the DAI score and the contents of IL-18 and TNF-αin the high and middle dose groups had no significant difference.Compared with the low dose group,the DAI score in the mesalazine group and the high and middle dose group decreased,and the contents of IL-18 and TNF-αdecreased.Conclusion:Wenglian jiedu decoction can regulate the immune system of intestinal epithelium by regulating the levels of serum IL-18 and TNF-αand increasing the expression of MUC2 protein,so as to achieve the purpose of repairing intestinal mucosa.

Fibrinogen-like protein 2 expression correlates with microthrombosis in rats with type 2 diabetic nephropathy

Fibrinogen-like protein 2 （fgl2）, a novel prothrombinase, is involved in microthrombosis. We examined fgl2 expression in the glomerular and tubulointerstitial capillaries and its correlation with microthromsis in rats with streptozocin-induced type 2 diabetic nephropathy. Our RT-PCR and immunoblotting analysis showed that fgl2 mRNA and protein levels were increased in microvascular endothelial cells of the glomeruli and renal interstitia at week 19 and became significantly elevated with the development of diabetic nephropathy （P 〈 0.01）. Fgl2 was not or only weakly expressed in the renal tissues of normal rats. Furthermore, a direct significant correlation （r = 0.543, P 〈 0.01） was found between fgl2 expression and microthrombotic capillaries in the renal tissues. Enzyme linked immunosorbent assays （ELISA） additionally showed that circulating TNF-α levels in rats with type 2 diabetes were significantly elevated and closely correlated with fgl2 expression （r = 0.871, P 〈 0.01）. Our results suggest that fgl2 may activate renal microthrombosis, thus contributing to glomerular hypertension and renal ischemia.

Expression and Significance of fgl2 Prothrombinase in Cardiac Microvascular Endothelial Cells of Rats with Type 2 Diabetes

Microthrombosis may be involved in the pathogenesis of cardiac microangiopathy due to diabetes.Recent studies have shown that fibrinogen-like protein 2 (fgl2) plays a pivotal role in microthrombosis in viral hepatitis, acute vascular xenograft rejection and cytokine-induced fetal loss syndrome.The current study was designed to examine the expression of fgl2 in microvascular endothelial cells and investigate the effects of microthrombi due to fgl2 on cardiac function and structure in rats with type 2 diabetes.Following induction of type 2 diabetes, 24 rats were observed dynamically.Fgl2 expression and related cardiac microthrombosis were examined.Local or circulating TNF-α was measured.Coronary flow (CF) per min was calculated as an index of cardiac microcirculation.Cardiac function and morphology were evaluated.It was found that Fgl2 was highly expressed in cardiac microvascular endothelial cells of rats with type 2 diabetes, which was promoted by local or circulating TNF-α.The Fgl2 expression was associated with cardiac hyaline microthrombosis.In parallel with the fgl2 expression, CF per min, cardiac diastolic or systolic function and cardiac morphology were aggravated to some extent.It was concluded that in rats with type 2 diabetes, microthrombosis due to fgl2 contributes to the impairment of cardiac diastolic or systolic function and morphological changes.

Jeju seaweeds suppress lipopolysaccharide-stimulated proinflammatory response in RAW 264.7 murine macrophages

Objective:To investigate the anti-inflammatory effects of Jeju seaweeds on macrophage RAW264.7 cells under lipopolysaccharide(LPS)stimulation.Methods:Ethyl acetate fractions were prepared from five different types of Jeju seaweeds,Dictyopteris divaricata(D.divaricata),Dictyopteris prolifera(D.prolifefa),Prioutis cornea(P.comea,Grateloupia laceolata(G,lanceolate,and Cralcloupia filicina(G.filicina)They were screened for inhibitory effects on proinflammatory mediators and cytokines such as nitric oxide(NO),prostaglandin E,,tumor necrosis factor-a(TNF-a),and interleukin-6(11.-6).Results:Our results revealed that D.divaricata,D.prolifera,P.cornea,G.lanceolata,and G.filicina potently inhibited I.PS-stimulaled NO production(IC_(50),values were 18.0,38.36,38.43,32.81 and 37.14μg/mL,respectively).Consistent with these findings,D.divtricata,D.prolifera,P.cornea,and G.fdicina also reduced the IPS-induced and prostaglandin E,production in a concentration-dependent manner.Expectedly,they suppressed the expression of inducible NO synthase and cyclooxygenase-2 at the protein level in a dose-dependent manner in the RAW264.7 cells,as detennined by western blotting.In addition,the levels of TNF-a and IL-6,released into the medium,were also reduced by D.divaricata,D.prolifera,P.cornea,G,lanceolata,and G.fdicina in a dose-dependent manner(IC_(50)values for TNF-a were 16.11,28.21,84.27,45.52 and74.75μg/mL,respectively;IC_(50),values for IL-6 were 37.35,80.08,103.28,62.53 and 84.28μg/mL,respectively).The total phlorotannin content was measured by the Folin-Ciocalteu method and expressed as phloroglucinol equivalents.The content was 92.0μg/mg for D.divaricata,151.8μg/mg for D.prolifera,57.2μg/mg for P.cornea,53.0 pg/mg for G.lanceolata,and 40.2μg/mg for G.fdicina.Conclusions:Thus,these findings suggest that Jeju seaweed extracts have potential therapeutic applications for inflammatory responses.

Dual therapy of rosiglitazone/pioglitazone with glimepiride on diabetic nephropathy in experimentally induced type 2 diabetes rats

Diabetic nephropathy is a major cause of end-stage renal disease （ESRD） in the general population. It is estimated that diabetic nephropathy will eventually develop in about 40% of all patients with diabetes; therefore, prevention is critical for delaying the development and progression of diabetic kidney disease. Despite extensive efforts, medical advances are still not successful enough to prevent the progression of the disease. In the present study, we focused on the comparison of combination therapies and whether they offered additional renoprotection. Type 2 diabetes mellitus was induced by intraperitoneally administering streptozotocin （90 mg/kg） in neonatal rats and then these rats were treated with rosiglitazone （1.0 mg/kg） in combination with glimepiride （0.5 mg/kg） or with pioglitazone （2.5 mg/kg） in combination with glimepiride （0.5 mg/kg）. Diabetic nephropathy markers were evaluated by biochemical and ELISA kits and renal structural changes were examined by light microscopy and transmission electron microscopy. Results show that the combination of pioglitazone with glimepiride is more effective in amelioration of diabetic nephropathy than rosiglitazone with glimepiride drug therapy due to glycemic control, suppressing albumin excretion rate, total protein excretion rate and augmented TNF-α signaling during the development of streptozotocin induced type 2 diabetic nephropathy.

血清TNF-α IL-1β OPG与2型糖尿病合并冠心病的关系探讨

目的探讨肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)、血清骨保护素(OPG)与2型糖尿病合并冠心病的关系。方法入选健康个体(对照组)和2型糖尿病合并冠心病患者(冠心病组)各30例,用ELISA方法测定TNF-α、IL-1β、OPG,生化法测定血糖及糖化血红蛋白,并观察组间各指标的变化及相互关系。结果 2型糖尿病合并冠心病组血清空腹血糖、糖化血红蛋白、OPG明显高于对照组(P均<0.01),TNF-α、IL-1β高于对照组(P均<0.05)。Spearman相关分析显示,OPG与TNF-α、IL-1β呈显著正相关(P均<0.01)。结论血糖、糖化血红蛋白、TNF-α、IL-1β、OPG在2型糖尿病合并冠心病患者中明显增高,OPG与TNF-α、IL-1β改变有关。

Retinal ganglion cell death in a DBA/2J mouse model of glaucoma Microglial activation and intraocular pressure

BACKGROUND： Retinal microglia has been shown to reactivate in a murine model of pigmentary glaucoma. However, the relationship between microglial activation and intraocular pressure （lOP） elevation and retinal ganglion cell （RGC） death is still unclear. OBJECTIVE： To verify that microglial activation and tumor necrosis factor alpha （TNF-α） expression is involved in RGC death with elevated lOP and prolonged time of glaucomatous optic nerve lesion in a DBA/2J mouse model of glaucoma. DESIGN, TIME AND SETTING： This randomized, controlled, animal experiment was performed at the Peking University Third Hospital, Peking University Eye Center, China between December 2006 and May 2008.MATEFIiALS： DBA/2J mice and C57BL/6J mice （Jackson Laboratory, USA）, rat anti-mouse CD11 b monoclonal antibody （Serotec, UK）, and goat anti-TNF-α polyclonal antibody （Sigma, USA） were used in this study.METHODS： A total of 100 female, DBA/2J mice at 3, 6, 9, 12, and 14 months of age （20 mice per age group） were used for the glaucoma model, and 18 C57BL/6J mice at 3, 9, 14 months of age （6 mice per age group） were used as normal controls. The anterior segment of the eye was observed using a slit-lamp biomicroscope, lOP was measured using a microneedle system. Morphology and number of retinal microglia were observed using immunohistochemistry. RGCs were quantified using Nissl staining. Co-localization of TNF-α and microglia was observed using double-labeling immunofluorescence. Excavation of the optic nerve head was observed utilizing hematoxylin-eosin staining. MAIN OUTCOME MEASURES： The following parameters were measured： lOP levels, numbers of RGCs and activated microglia, and TNF-α expression. RESULTS： In 6-month-old DBA/2J mice, dispersed pigment was observed, and some mice developed increased IOP. At 9 months of age, lOP levels reached a peak. In 3-month-old DBA/2J mice, microglia were activated. In 6-month-old DBA/2J mice, the number of activated microglia was significantly increased and migrated to the outer retinal layer. In 9-month-old mice, TNF-a expression was co-localized with microglia. Significant RGC loss occurred in mice aged 9 to 14 months, with the presence of optic nerve fiber loss and optical nerve head excavation, lOP returned to normal levels at 12 months of age, but microglia remained activated, which was consistent with RGC loss. CONCLUSION： Retinal microglial activation was partially attributed to increased lOP. Activated microglia might be mainly responsible for RGC loss. TNF-α expression was evident in the inner retinal layer. However, the relationship between TNF-α and RGC loss remains poorly understood.

浙江台州地区老年2型糖尿病患者TNF-α基因多态性检测与分析

目的研究TNF-α基因多态性在浙江台州地区2型糖尿病患者中的分布与相关性。方法利用DNA测序法对100例老年2型糖尿病患者(观察组)及50例健康体检者(对照组)的TNF-。α基因-308位点和-238位点进行基因检测与分析。结果观察组FPG、TNF-α、TG、TC、胰岛素抵抗指数(IRI)分别为(10.68±2.75)mmol/L、(2.01±0.25)ug/L、(2.43±1.15)mmol/L、(5.49±1.63)mmol/L及4.65±0 97,均高于对照组,差异均有统计学意义(均P<0.05),HDL为(1.23±0.28)mmol/L,低于对照组,差异有统计学意义(P<0.05)。而BMI、LDL[(25.34±2.41)kg/m^2、(3 26±1 3)mmol/L]与对照组比较,差异无统计学意义(P>0.05)。TNF-α基因-308位点在观察组和对照组GA+AA基因频率分别为31%和10%,A等位基因频率分别为16%和5%,差异有统计学意义(P<0.05)。而-238位点未见突变,两组间比较无明显差异。结论 TNF-α基因G-308A单核苷酸多态性与2型糖尿病存在相关性。

Role of P2X_7 receptors in the development of diabetic retinopathy

The P2X7 receptor is one of the members of the family of purinoceptors which are ligand-gated membrane ion channels activated by extracellular adenosine 5'-triphosphate. A unique feature of the P2X7 receptor is that its activation can result in the formation of large plasma membrane pores that allow not only the flux of ions but also of hydrophilic molecules of up to 900 Da. Recent studies indicate that P2X7-mediated signaling can trigger apoptotic cell death after ischemia and during the course of certain neurodegenerative disorders. Expression of the P2X7 receptor has been demonstrated in most types of cells in the retina. This purinoceptor mediates the contraction of pericytes and regulates the spatial and temporal dynamics of the vasomotor response through cell-to-cell electrotonic transmission within the microvascular networks. Of potential clinical significance, investigators have found that diabetes markedly boosts the vulnerability of retinal microvessels to the lethal effect of P2X7 receptor activation. This purinergic vasotoxicity may result in reduced retinal blood flow and disrupted vascular function in the diabetic retina. With recent reports indicating an association between P2X7 receptor activation and inflammatory cytokine expression in the retina, this receptor may also exacerbate the development of diabetic retinopathy by a mechanism involving inflammation.

Differential mRNA Expression of <i>COX</i>-2 and Proinflammatory Mediators in Patients with Rotator Cuff Tears and Osteoarthritis of the Hip

Purpose: Bursal inflammation is thought to be a major cause of pain in degenerative rotator cuff tears (RCTs). While the expression of proinflammatory mediators, such as COX-2, TNF-α, IL-1β, and IL-6, is crucial for the pathophysiology of osteoarthritis (OA), their role in degenerative RCTs remains unknown. The aim of this study was to determine the expression of COX-2 and proinflammatory mediators in the development of RCT-induced pain by comparing their levels in patients with hip OA or RCTs. Methods: We included samples obtained from 31 shoulders of 31 patients with RCTs and samples from 30 hips of 27 patients with hip OA. The mRNA levels of COX-2, TNF-α, IL-1β, and IL-6 were determined using RT-PCR, and were compared between the subacromial bursa and hip joints. We also analyzed IL-1β-induced COX-2 expression in the subacromial bursa and synovial blast of the hip. Results: COX-2, IL-1β, and IL-6 expression levels were significantly lower in the subacromial bursa of RCTs than in hip OA samples, while no significant difference was observed for TNF-α. No significant difference in the fold increase was observed between subacromial bursa and hip OA samples, even though IL-1β-induced COX-2 expression increased in both samples. Conclusion: Our findings suggest that the main mechanism underlying pain development differs between patients with RCTs and those with hip OA.

The gene expression patterns of BMPR2,EP300,TGFβ2,and TNFAIP3 in B-Lymphoma cells

Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor, type II(BMPR2), E1 A binding protein p300(EP300), transforming growth factor-β2(TGFβ2), and tumor necrosis factor, and alpha-induced protein 3(TNFAIP3) gene expression patterns in B-cell malignancies were studied. Methods: The relative expression levels of BMPR2, EP300, TGFβ2, and TNFAIP3 mRNA in B-lymphoma cell lines, myeloid cell lines, as well as in cells from healthy volunteers, were determined by real-time quantitative reverse transcriptpolymerase chain reaction(qRT-PCR) with SYBR Green Dye. Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) was used as reference. Results: The expression level of TGFβ2 mRNA in B-lymphoma cell lines was significantly higher than those in the cells from the healthy control(P<0.05). However, the expression level of TNFAIP3 mRNA in B-malignant cells was significantly lower than that of the healthy control(P<0.05). The expression levels of BMPR2 and EP300 mRNA showed no significant difference between B-malignant cell lines and the healthy group(P>0.05). In B-lymphoma cell lines, correlation analyses revealed that the expression of BMPR2 and TNFAIP3(r=0.882, P=0.04) had significant positive relation. The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in cell lines from myeloid leukemia were significantly lower than those in the cells from the healthy control(P<0.05). The expression levels of TGFβ2 mRNA showed no significant difference between myeloid leukemia cell lines and the healthy control or B-malignant cell lines(P>0.05). The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in B-lymphoma cells were significantly higher than those of the myeloid leukemia cells(P<0.05).Conclusion: Different expression patterns of BMPR2, EP300, TGFβ2, and TNFAIP3 genes in B-lymphoma cells exist.

Standardized extract of Centella asiatica ECa 233 inhibits lipopolysaccharide-induced cytokine release in skin keratinocytes by suppressing ERK1/2 pathways

Objective:To evaluate the effect of standardized extract of Centella asiatica ECa 233 on inflammatory mediator production through cyclooxygenase-2(COX-2),extracellular signal-regulated kinase 1/2(ERK1/2)and nuclear factor-κB(NF-κB)pathway in keratinocyte Ha Ca T cells.Methods:Ha Ca T cells were treated with 0.1,1,10 and 100μg/m L ECa 233 in the presence of lipopolysaccharide(LPS).Proinflammatory cytokines and prostaglandin E2 were assessed with ELISA.Western blotting was used to determine the inhibition of COX-2,ERK1/2 and NF-κB protein expression.Results:ECa 233 suppressed LPS-induced release of interleukin-1β,tumor necrosis factor-α,and prostaglandin E2.ECa 233 also inhibited COX-2,phosphorylation of ERK1/2 and the activation of NF-κB.Moreover,the formation of reactive oxygen species(ROS)was decreased in response to LPS-inflamed keratinocytes.Conclusions:ECa 233 inhibits LPS-stimulated production of inflammatory mediators in keratinocytes via suppressing ERK1/2 and NF-κB pathways.The suppressive effect of ECa 233 may be related to an inhibition of ROS production.

Cinnamon extract suppresses experimental colitis through modulation of antigen-presenting cells

AIM:To investigate the anti-inflammatory effects of cinnamon extract and elucidate its mechanisms for targeting the function of antigen presenting cells.METHODS:Cinnamon extract was used to treat murine macrophage cell line(Raw 264.7),mouse primary antigen-presenting cells(APCs,MHCII+) and CD11c+dendritic cells to analyze the effects of cinnamon extract on APC function.The mechanisms of action of cinnamon extract on APCs were investigated by analyzing cytokine production,and expression of MHC antigens and co-stimulatory molecules by quantitative real-time PCR and flow cytometry.In addition,the effect of cinnamon extract on antigen presentation capacity and APC-dependent T-cell differentiation were analyzed by [H3]-thymidine incorporation and cytokine analysis,respectively.To confirm the anti-inflammatory effects of cinnamon extract in vivo,cinnamon or PBS was orally administered to mice for 20 d followed by induction of experimental colitis with 2,4,6 trinitrobenzenesulfonic acid.The protective effects of cinnamon extract against experimental colitis were measured by checking clinical symptoms,histological analysis and cytokine expression prof iles in inflamed tissue.RESULTS:Treatment with cinnamon extract inhibited maturation of MHCII+ APCs or CD11c+ dendritic cells(DCs) by suppressing expression of co-stimulatory molecules(B7.1,B7.2,ICOS-L),MHCII and cyclooxygenase(COX)-2.Cinnamon extract induced regulatory DCs(rDCs) that produce low levels of pro-inflammatory cytokines [interleukin(IL)-1β,IL-6,IL-12,interferon(IFN)-γ and tumor necrosis factor(TNF)-α] while expressing high levels of immunoregulatory cytokines(IL-10 and transforming growth factor-β).In addition,rDCs generated by cinnamon extract inhibited APC-dependent T-cell proliferation,and converted CD4+ T cells into IL-10high CD4+ T cells.Furthermore,oral administration of cinnamon extract inhibited development and progression of intestinal colitis by inhibiting expression of COX-2 and pro-inflammatory cytokines(IL-1β,IFN-γ and TNF-α),while enhancing IL-10 levels.CONCLUSION:Our study suggests the potential of cinnamon extract as an anti-inflammatory agent by targeting the generation of regulatory APCs and IL-10+ regulatory T cells.

复方柴芩退热颗粒对发热大鼠模型退热作用的研究

目的:研究复方柴芩退热颗粒对2,4二硝基苯酚及细菌内毒素所致大鼠发热模型的退热作用,进一步明确该颗粒的退热效果,初步探讨其退热机制。方法:将SPF级SD大鼠随机分为正常对照组、模型组、酚麻美敏片组[给药剂量0.148 g/(kg·d)]、小柴胡冲剂组[给药剂量2.160 g/(kg·d)]、复方柴芩退热颗粒高、中、低剂量组[给药剂量分别为6.480、3.240、1.620 g/(kg·d)]。各给药组按相应剂量灌胃给药,正常对照组、模型组同法给予等体积的蒸馏水,每天1次,连续给药5天,第4天各组动物均开始禁食不禁水24 h,末次给药前开始造模。造模实验一:除正常对照组外,余各组大鼠均皮下注射2,4-二硝基苯酚17 mg/kg,正常对照组皮下注射同体积生理盐水;造模实验二:除正常对照组外,余各组大鼠均腹腔注射细菌内毒素80μg/kg。测大鼠各时间点的体温和血清中白细胞介素-1β(IL-1β)、前列腺素E2(PGE2)、肿瘤坏死因子-α(TNF-α)的含量。结果:实验一:与正常对照组比较,模型组大鼠造模后1 h、2 h、3 h体温上升值均显著升高,大鼠血清IL-1β、PGE2含量均显著升高(P<0.01)。与模型组比较,酚麻美敏片组、小柴胡冲剂组、复方柴芩退热颗粒高、中、低剂量组大鼠血清IL-1β、PGE2含量均明显降低(P<0.05,P<0.01);酚麻美敏片组大鼠在造模后1 h、2 h、3 h体温上升值均显著降低(P<0.05,P<0.01);小柴胡冲剂组大鼠在造模后2 h、3 h体温上升值均显著降低(P<0.05,P<0.01);复方柴芩退热颗粒高、低剂量组在造模后1 h、2 h、3 h体温上升值均明显降低(P<0.05,P<0.01);复方柴芩退热颗粒中剂量组在造模后2 h、3 h体温上升值均明显降低(P<0.05)。实验二:与正常对照组比较,模型组大鼠造模后2 h、4 h、6 h、8 h体温上升值均显著升高,大鼠血清PGE2、TNF-α含量均显著升高(P<0.01)。与模型组比较,酚麻美敏片组、小柴胡冲剂组、复方柴芩退热颗粒高、中、低剂量组大鼠血清PGE2、TNF-α含量均明显降低(P<0.05,P<0.01);酚麻美敏片组大鼠在造模后2 h、4 h、6 h体温上升值均显著降低(P<0.05,P<0.01);小柴胡冲剂组、复方柴芩退热颗粒高、中、低剂量组在造模后2 h、4 h体温上升值均明显降低(P<0.05,P<0.01)。结论:复方柴芩退热颗粒对2,4二硝基苯酚及细菌内毒素所致大鼠发热均有降温作用,能有效降低大鼠血清PGE2、IL-1β和血清TNF-α含量,对大鼠实验性高热模型有较好的解热作用。

Nitric oxide-releasing aspirin but not conventional aspirin improves healing of experimental colitis

AIM:To determine the effect of non-selective cyclooxygenase (COX) inhibitors,selective COX-2 inhibitors and nitric oxide (NO)-releasing aspirin in the healing of ulcerative colitis.METHODS:Rats with 2,4,6 trinitrobenzenesulfonic acid (TNBS)-induced colitis received intragastric (ig) treatment with vehicle,aspirin (ASA) (a nonselective COX inhibitor),celecoxib (a selective COX-2 inhibitor) or NO-releasing ASA for a period of ten days.The area of colonic lesions,colonic blood flow (CBF),myeloperoxidase (MPO) activity and expression of proinflammatory markers COX-2,inducible form of nitric oxide synthase (iNOS),IL-1β and tumor necrosis factor (TNF)-α were assessed.The effects of glyceryl trinitrate (GTN),a NO donor,and 2-(4-carboxyphenyl)-4,5-dihydro-4,4,5,5-tetramethyl-1H-imidazolyl-1-oxy-3-oxide,onopotassium salt (carboxy-PTIO),a NO scavenger,administered without and with ASA or NO-ASA,and the involvement of capsaicin-sensitive afferent nerves in the mechanism of healing the experimental colitis was also determined.RESULTS:Rats with colitis developed macroscopic and microscopic colonic lesions accompanied by a significant decrease in the CBF,a significant rise in colonic weight,MPO activity and plasma IL-1β and TNF-α levels.These effects were aggravated by ASA and 5-(4-chlorophenyl)-1-(4-methoxyphenyl)3-(trifluoromethyl)-1H-pyrazole (SC-560),but not celecoxib and counteracted by concurrent treatment with a synthetic prostaglandin E 2 (PGE 2) analog.Treatment with NO-ASA dose-dependently accelerated colonic healing followed by a rise in plasma NO x content and CBF,suppression of MPO and downregulation of COX-2,iNOS,IL-1β and TNF-α mRNAs.Treatment with GTN,the NO donor,significantly inhibited the ASA-induced colonic lesions and increased CBF,while carboxy-PTIO or capsaicin-denervation counteracted the NO-ASAinduced improvement of colonic healing and the accompanying increase in the CBF.These effects were restored by co-treatment with calcitonin gene related peptide (CGRP) and NO-ASA in capsaicin-denervated animals.CONCLUSION:NO-releasing ASA,in contrast to ASA,COX-1 inhibitors,and SC-560,accelerated the healing of colitis via a mechanism involving NO mediated improvement of microcirculation and activation of sensory nerves releasing CGRP.

Expression of matrix metalloproteinase-11 is increased under conditions of insulin resistance

AIM To investigate matrix metalloproteinase-11(MMP-11) expression in adipose tissue dysfunction, using in vitro and in vivo models of insulin resistance.METHODS Culture of mouse 3T3-L1 preadipocytes were induced to differentiation into mature 3T3-L1 adipocytes. Cellular insulin resistance was induced by treating differentiated cultured adipocytes with hypoxia and/or tumor necrosis factor(TNF)-α, and transcriptional changes were analyzed in each condition thereafter. For the in vivo studies, MMP-11 expression levels were measured in white adipose tissue(WAT) from C57BL/6J mice that underwent low fat diet or high-fat feeding in order to induce obesity and obesity-related insulin resistance. Statistical analysis was carried out with GraphP ad Prism Software.RESULTS MMP-11 m RNA expression levels were significantly higher in insulin resistant 3T3-L1 adipocytes compared to control cells(1.46±0.49vs0.83±0.21, respectively;P<0.00036). The increase in MMP-11 expression was observed even in the presence of TNF-α alone(3.79±1.11vs1±0.17, P<0.01) or hypoxia alone(1.79±0.7vs0.88±0.1, P<0.00023). The results obtained in in vitro experiments were confirmed in the in vivo model of insulin resistance. In particular, MMP-11 m RNA was upregulated in WAT from obese mice compared to lean mice(5.5±2.8vs1.1±0.7, respectively; P<3.72E-08). The increase in MMP-11 levels in obese mice was accompanied by the increase in typical markers of fibrosis, such as collagen type Ⅵ alpha 3(Col6_α3), and fibroblast-specific protein 1.CONCLUSION Our results indicate that dysregulation of MMP-11 expression is an early process in the adipose tissue dysfunction, which leads to obesity and obesity-related insulin resistance.

Anti-inflammatory effect of Heliotropium indicum Linn on lipopolysaccharide-induced uveitis in New Zealand white rabbits

AIM： To investigate the anti-inflammatory effect of an aqueous whole plant extract of Heliotropium indicum（HIE） on endotoxin-induced uveitis in New Zealand white rabbits.·METHODS： Clinical signs of uveitis including flares,iris hyperemia and miosis, were sought for and scored in1.0 mg/kg lipopolysaccharide（LPS）-induced uveitic rabbits treated orally with HIE（30-300 mg/kg）,prednisolone（30 mg/kg）, or normal saline（10 m L/kg）. The number of polymorphonuclear neutrophils infiltrating, the protein concentration, as well as levels of tumor necrosis factor-α（TNF-α）, prostaglandin E2（PGE2）, and monocyte chemmoattrant protein-1（MCP-1） in the aqueous humor after the various treatments were also determined. A histopathological study of the anterior uveal was performed.· RESULTS： The extract and prednisolone-treatment significantly reduced（P ≤0.001） both the clinical scores of inflammation（1.0-1.8 compared to 4.40 ±0.40 in the normal saline-treated rabbits） and inflammatory cells infiltration. The level of protein, and the concentrationsof TNF-α, PGE2 and MCP-1 in the aqueous humor were also significantly reduced（P ≤0.001）. Histopathological studies showed normal uveal morphology in the HIE and prednisolone-treated rabbits while normal saline-treated rabbits showed marked infiltration of inflammatory cells.· CONCLUSION： The HIE exhibits anti-inflammatory effect on LPS-induced uveitis possibly by reducing the production of pro-inflammatory mediators.