Cytokine-induced cell surface expression of adhesion molecules in vascular endothelial cellsIn vitro

Summary  Regulation of the adhesion molecules expression by cytokine in vascular endothelial cells was investigated. Human umbilical vein endothelial cells (HUVEC) were stimulated with cytokines, TNF-α (1–250 U/ml) or IL-1? (0. 1–50 U/ml) for 24 h. HUVEC were also cultured with cytokines, TNF-α (100 U/ml) or IL-1? (10 U/ml), for 4–72 h, cell surface expression of adhesion molecules (ICAM-1 and VCAM-1) were detected and quantitated by immunocytochemical methods and computerized imaging analysis technique. Adhesion molecules expression were up-regulated by TNF-α, IL-1? in a concentration- and time-dependent manner. Some significant differences were observed between the effects of cytokines on the ICAM-1 and on VCAM-1 expression. Cytokines might directly induce the expression of ICAM-1 and VCAM-1 in vascular endothelial cells. Our observations indicate differential functions of the two adhesion molecules during the evolution of inflammatory responses in stroke.     

Molecular basis of the adhesive process of polymorphonuclear neutrophils to vascular endothelial cells under endotoxin condit

ＭｏｌｅｃｕｌａｒｂａｓｉｓｏｆｔｈｅａｄｈｅｓｉｖｅｐｒｏｃｅｓｓｏｆｐｏｌｙｍｏｒｐｈｏｎｕｃｌｅａｒｎｅｕｔｒｏｐｈｉｌｓｔｏｖａｓｃｕｌａｒｅｎｄｏｔｈｅｌｉａｌｃｅｌｌｓｕｎｄｅｒｅｎｄｏｔｏｘｉｎｃｏｎｄｉｔｉｏｎＺｈｏｕＸｉａｎｇｄｏｎ...     

ICAM—1在缺氧—再氧化引起的白细胞与内皮细胞粘附中的作用

目的探讨细胞间粘附分子（ＩＣＡＭ－１）是否介导缺氧－再氧化引起的中性粒细胞（ＰＭＮ）和血管内皮细胞（ＶＥＣ）的粘附反应。方法血管内皮细胞（ＶＥＣ）经缺氧再氧化（Ｈ／Ｒ）处理后,加入ＰＭＮ,用计数法检测粘附率,以细胞免疫化学及原位杂交法检测ＩＣＡＭ－１及ＩＣＡＭ－１ｍＲＮＡ表达。结果ＶＥＣ经Ｈ／Ｒ处理后,ＰＭＮ与其粘附率增高１倍（Ｐ＜０．０１）,ＩＣＡＭ－１单克隆抗体（ｍＡｂ）与ＣＤ１１ａ／ＣＤ１８ｍＡｂ可明显降低粘附率的增高,Ｈ／Ｒ能增加ＶＥＣ的ＩＣＡＭ－１及ＩＣＡＭ－１ｍＲＮＡ表达。结论ＩＣＡＭ－１介导Ｈ／Ｒ后的ＰＭＮ－ＶＥＣ间粘附反应。     

人促血液血管细胞生成素对内皮细胞粘附功能的影响

目的研究人促血液血管细胞生成素(HAPO)对人脐静脉内皮细胞(HUVEC)粘附功能的影响。方法采用结晶紫粘附实验、流式细胞术和半定量RT-PCR方法检测HUVEC的粘附性和粘附分子CD54、CD102、CD106、CD31、CD62E和CD62P的表达。结果HAPO以剂量依赖性方式增加HUVEC的总粘附性。HAPO能显著促进HUVEC表面粘附分子CD106和CD62E表达,并呈时间依赖性。HAPO作用HUVEC6h后,CD106和CD62E的阳性率分别为(80·83±2·22)%和(22·77±2·59)%,是各自对照组的2·10倍和5·84倍,且转录水平和蛋白质水平一致。CD106和CD62E的最高转录水平分别为对照组的1·86倍和6·16倍。结论HAPO可能对造血干/祖细胞归巢有一定促进作用。     

Effect of Jianpi Huoxue decoction (健脾活血方)-containing serum on tumor necrosis factor-α secretion and gene Expression of endotoxin receptors in RAW264.7 cells induced by lipopolysaccharide

Objective:To evaluate the effect of Jianpi Huoxue decoction(健脾活血方,JHD)-containing serum on tumor necrosis factor-α(TNF-α) secretion and endotoxin receptor gene expression in RAW264.7 cells induced by lipopolysaccharide(LPS).Methods:The cytotoxicity of blank-control serum and JHD-containing serum at different concentrations were evaluated through the lactate dehydrogenase(LDH) assay in RAW264.7 cells.RAW264.7 cells were divided into six groups:5%blank-control serum group(C1,n=3),5%blank-control serum plus...     

Effect of IL-Iβ and TNF-α on the expression of monocyte chemotactic protein-1 in endometriotic cells

Summary To investigate the clinical significance of monocyte chemotactic protein-1 (MCP-1) produced by endometriotic tissues, the endometriotic tissues were taken from 15 patients with endometriosis. MCP-1 mRNA and MCP-1 protein were determined by dot blot analysis and enzyme linked immunosorbent assay (ELISA) in endometriotic cells cultured with or without interleukin-1β (IL-lβ, 2 μg/L), tumor necrosis factor-α (TNF-α, 20 g/L). After exposure to IL-1β or TNF-α, the expression of MCP-1 mRNA in the endometriotic cells (8.635 ±0.826, 7.031 ±0.970, respectively) were significantly higher than that in the control group (4.482±0.435, P<0.05); The expression of MCP-1 protein in IL-lβ and TNF-α group was 4.52±0.09 μg/L,2.87±0.27 μg/L, respectively, which were significantly higher than 1.74±0.16 μg/L in control (P<0.01). The results suggested that IL-l\ and TNF-α could up-regulate the expression of MCP-1 in endometriotic cells, which might be related to the development of endometriosis.     

Antitumor effects of the fibroblasts transfected TNF-α gene and its mutants

Summary To compare the anti-tumor effects of transmembrane TNF-α (TM-TNF) and secreted TNF-α (S-TNF)in vivo, mouse fibroblasts NIH3T3 were transfected separately with three types of retrovirus containing wild type TNF-α (Wt-TNF), TM-TNF mutant (TM-TNFm), S-TNF mutant (S-TNFm). Southern blot, RT-PCR, FACS and bioassay were used to investigate TNF-α gene integration, expression and its biological activity. It was found that both fixed cells and supernatant of NIH3T3/Wt-TNF, the fixed cells of NIH3T3/TM-TNFm and the supernatant of NIH3T3/S-TNFm could express high level of TNF-α or its mutants and effectively kill H22in vitro. The trans-fected NIH3T3 were separately injected into the mice at the sites of H22 tumor cell inoculation according to a ratio of 5∶1 or 1∶1 (effector/target cells, E/T) after the third day of H22 challenge, respectively. At the E/T=5∶1, the NIH3T3/TM-TNFm induced the highest tumor regression, while NIH3T3/S-TNFm exerted the strongest tumor depressing effect at the E/T=1∶1in vivo. No obvious side effects were noted throughout the course of treatment. The results suggest that both TM-TNF and S-TNF could cause tumor regression. The anti-tumor effect of TM-TNF would be more powerful and safe than that of S-TNF at the proper E/T ratio. LI Qingfen, female, born in 1965, Associate Professor     

Up-regulation of HIF-1α and VEGF Expression by Elevated Glucose Concentration and Hypoxia in Cultured Human Retinal Pigment Epithelial Cells

Summary： In order to explore the effect of high glucose concentration and high glucose concentration with hypoxia on the production of hypoxia-inducible factor-1α （HIF-1α） and vascular endothelial growth factor （VEGF）, human RPE cells were cultured in 5,56 mmol/L glucose （control group）, 5.56 mmol/L glucose with 150 !a mol/L COCl2 （hypoxic group）, 25 mmol/L glucose （high glucose group） and 25 mmol/L glucose with 150 μmol/L COCl2 （combination group）. RT-PCR was used to detect the expression of HIF-1α and VEGF mRNAs. Western blot analysis was used to measure the levels of HIF-1α and VEGF proteins. Although the small amount of HIF-1α protein was able to be detected in high glucose group but not in control group, there was no significant difference between the expression of HIF-1α mRNA of RPE cells in high glucose group and that of RPE cells in control group. As compared with RPE cells in control group, the mRNA expression and the protein synthesis of VEGF in high glucose group were up-regulated. As compared with RPE cells in hypoxic group, the expression of HIF-1α mRNA of RPE cells in combination group was not different, but the protein synthesis of HIF-1α, the mRNA expression and the protein synthesis of VEGF were more obviously up-regulated. In conclusion, high concentration glucose mainly influence the protein synthesis of HIF-1α of RPE cell, and HIF-1α protein is able to be accumulated in high concentration glucose. Under hypoxia, the HIF-1α protein induced by high concentration glucose is more stable, and the expression of VEGF is obviously increased. It is suggested that high concentration glucose may play a role in retinal neovascularization, especially at ischemia stage of diabetic retinopathy.     

Effects of Baicalin on Expression of Inducible Nitric Oxide Synthase in Cultured Fibroblasts Stimulated by Cytokines

Objective: To investigate the effects of baicalin on expression of inducible nitric oxide synthase (iNOS) in fibroblasts and its mechanisms in treating psoriasis. Methods: Fibroblasts cultured in vitro were stimulated with tumor necrosis factor-α(TNF-α), interferon-γ (IFN-γ), interleukin-8 (IL-S) in different groups. iNOS was detected by western blot and immunocytochemistry assay, and in addition, the effects of baicalin on its expression were investigated. Results: Fibroblasts did not express iNOS without cytokine stimulation. When treated for 24 h with 1. 0× 106 U/L TNF-α, 0.2× 106U/L IFN-γ, 0.2× 106 pg/L IL-8 alone or in combinations indicated, fibroblasts produced iNOS when stimulated by TNF-α alone while neither IFN-γ nor IL-8 could induce the production of iNOS. The combination of TNF-α and IL-8 induced a strong expression of iNOS, the combined exposure of three kinds of cytokines showed an even stronger effects. The strongly stained area was in the cytoplasm near the nuclei. Expression of iNOS induced by TNF-α and IL-8 was inhibited by 50 μg/ mi of baicalin. Conclusion: Fibroblasts might express iNOS when stimulated by certain cytokines. Baicalin decreased production of nitric oxide through inhibiting the expression of iNOS, furthermore it reduced inflammation, which might be part of its mechanisms in treating psoriasis.     

ICAM-1在缺氧-再氧化引起的白细胞内皮细胞粘附中的作用

目的探讨细胞间粘附分子(ICAM-1是否介导缺氧-再氧化引起的中性粒细胞(PMN)和血管内皮细胞(VEC)的粘附反应.方法血管内皮细胞(VEC)经缺氧再氧化(H/R)处理后,加入PMN,用计数法检测粘附率,以细胞免疫化学及原位杂交法检测ICAM-1及ICAM-1mRNA表达.结果VEC经H/R处理后,PMN与其粘附率增高1倍(P＜0.01),ICAM-1单克隆抗体(mAb)与CD11a/CD18mAb可明显降低粘附率的增高,H/R能增加VEC的ICAM-1及ICAM-1mRNA表达.结论ICAM-1介导H/R后的PMN-VEC间粘附反应.     

The mechanical study of vascular endothelial growth factor on the prevention of restenosis after angioplasty

Restenosis following percutaneous transluminalcoronary angioplasty ( PTCA) is one of the majorre-search subjects of coronary heart disease( CHD) .The mechanism is quite complicated and not yetclear,buttheimpairmentofendothelium isknown asthe primary evocative factor of the progress ofrestenosis.It is supposed to prevent the restenosisfollowing PTCA by accelerating restoration of en-dothelial integrity and function.In the experimentalstudies,it was proved that exogenic vascular en-dothelial…     

Anti-Inflammatory mechanism of total glycosides of Acanthopanax Giraldii

Objective:To study the anti-inflammatory mechanisms of total glycosides of Acanthopanax Giraldii (TGA).Methods:The changes of prostaglandin E_2(PGE_2),tumor necrosis factor(TNF-α),nitric oxide(NO), and expressions of COX-1 mRNA and COX-2 mRNA in BALB/c mouse macrophages were observed by the radioimmunoassay,ELISA and nitric acid reduction and RT-PCR in the presence or absence of TGA.Results: (1) TGA could significantly decrease the production of PGE_2 and NO in mouse peritoneal macrophages.The inhibitory...     

同型半胱氨酸对人脐静脉内皮细胞表达ICAM-2及与树突状细胞粘附的影响

目的 探讨同型半胱氨酸（Hcy）对人脐静脉内皮细胞（HUVECs）分泌和表达细胞间粘附分子-2（1-CAM-2）以及内皮细胞与树突状细胞（DC）粘附的影响。方法 在人脐静脉内皮细胞培养基中加入浓度为0．5mmol／L的Hey作用不同时间，以及加入不同浓度的Hey作用24h。用ELISA法检测内皮细胞培养基中HU-VECs分泌ICAM-2的蛋白含量，用Western blot法检测HUVECs表达ICAM-2的蛋白含量，用分子探针标记的DC与HUVECs共孵育，共聚焦显微镜下计数粘附于HUVECs的DC数量，以检测Hey对HUVECs与DC粘附的影响。结果用0．5mmol／L Hey干预后，HUVECs分泌ICAM-2蛋白在12h开始高于空白组（P〈0．05），24h和48h均增加显著（均P〈0．01）；而ICAM-2蛋白的表达在6h开始高于空白组（P〈0．05），24h达峰值（P〈0．01），48h下降显著，且与空白组相比无统计学差异。用不同浓度的Hey干预24h后，ICAM-2蛋白的分泌和表达及HUVECs与Dc粘附均呈剂量依赖性增加，当Hey浓度高于0．5mmol／L时增加更显著。结论 Hey能刺激HUVECs分泌和表达ICAM-2蛋白，从而促进HUVECs与DC的粘附，这可能是其诱导动脉粥样硬化早期形成的重要机制之一。     

Protective effect and mechanism of sodium tanshinone II A sulfonate on microcirculatory disturbance of small intestine in rats with sepsis

To explore the protective effect of sodium tanshinone ⅡA sulfonate(STS) on microcirculatory disturbance of small intestine in rats with sepsis,and the possible mechanism,a rat model of sepsis was induced by cecal ligation and puncture(CLP).Rats were randomly divided into 3 groups:sham operated group(S),sepsis group(CLP) and STS treatment group(STS).STS(1 mg/kg) was slowly injected through the right external jugular vein after CLP.The histopathologic changes in the intestinal tissue and changes of mesenteric microcirculation were observed.The levels of tumor necrosis factor-α(TNF-α) in the intestinal tissue were determined by using enzyme-linked immunoabsorbent assay(ELISA).The expression of intercellular adhesion molecule-1(ICAM-1) in the intestinal tissue was detected by using immunohistochemisty and Western blot,that of nuclear factor κB(NF-κB) and tissue factor(TF) by using Western blot,and the levels of NF-κB mRNA expression by using RT-PCR respectively.The microcirculatory disturbance of the intestine was aggravated after CLP.The injury of the intestinal tissues was obviously aggravated in CLP group as compared with S group.The expression levels of NF-κB p65,ICAM-1,TF and TNF-α were upregulaed after CLP(P<0.01).STS post-treatment could ameliorate the microcirculatory disturbance,attenuate the injury of the intestinal tissues induced by CLP,and decrease the levels of NF-κB,ICAM-1,TF and TNF-α(P<0.01).It is suggested that STS can ameliorate the microcirculatory disturbance of the small intestine in rats with sepsis,and the mechanism may be associated with the inhibition of inflammatory responses and amelioration of coagulation abnormality.     

Effect of propyl gallate on activity of cyclooxygenase 1 and 2 in mice??s peritoneal macrophages

Objective: To investigate the effect of Red Peony 801 (propyl gallate,PrG) on cyclooxygenase (COX) activity in murine peritoneal macrophages.Methods: A screening model for COX inhibitors in vitro based on murine peritoneal macrophages was used. COX-1 activity was reflected by the level of 6-ketoprostaglandin F_1α(6-keto-PGF_1α) in supernatants of cultured macrophages which were stimulated with calcium ionophore A23187 for a short-term, while COX-2 activity was reflected by the level of prostaglandin E₂ (PGE₂) in supernatants of cultured macrophages which were stimulated with lipopolysaccharide (LPS) for a long-term.Results: PrG did not affect A23187-induced, COX-1-derived 6-keto-PGF_1α synthesis at the concentrations of 1 × 10^-6, 5 × 10^-6 mol/L (P>0.05), but enhanced 6-keto-PGF_1α synthesis at the concentrations of 1 × 10^-6, 5 × 10^-7, 1 × 10^-7 mol/L (P<0.01) in vitro, and showed a good dose-dependent manner. It inhibited LPS-induced, COX-2-derived PGE₂ synthesis at the concentrations of 1 × 10^-6, 1 × 10^-6 mol/L (P< 0.05).Conclusion: Within the range of 1 × 10^-5 to 1 × 10^-7 mol/L, PrG activated COX-1 at lower concentrations and inhibited COX-2 at higher concentrations in murine peritoneal macrophages. This work is supported by the National Natural Science Foundation (item number: 90209054)     

Effects of Cyclosporin A on Proliferation of Cultured Rat Mesangial Cells

Mesangialproliferativeglomeru-lonephritis(MsPGN)isacommonpatho-logicaltypeofglomerulardiseasesinchil-dren[l].Itischaracterizedbyincreasednumbersofmesangialcells(MC)andin-creasedmesangialmatrix,finallyresultinginglomerulosclerosis.ProliferationofmesangialcellsareofgreatimportanceinpathologicalchangesandprognosisofgIomerulardiseases.Thus,attentionispaidtosearchofaninhibitorofproliferationofMCandinvestigationofitsmechanism.Cy-closporinA(CsA)isanimmunosuppressiveagentusedinthetreatmentofglome…     

The Change of Interleukin-6 and Tumor Necrosis Factor in Patients with Obstructive Sleep Apnea Syndrome

Obstructive sleep apnea syndrome( OSAS) is avery common disease,which is characterized by re-current apneas caused by occlusion of upper airwayduring sleep,leading to arterial oxygen desaturationand a decrease in tissue oxygen concentration.Pa-tients with OSAS exhibit dramatic fragmentation ofsleep,a decrease in the duration of stage3and 4sleep,decreased percentages of rapid eye movementsleep ( REMS) ,and excessive daytime somnolence.The cause and pathogenesis of OSAS are unclear.Recentl…     

巴曲酶对急性缺血性脑卒中患者外周血中黏附分子表达的影响

目的：观察巴曲酶对急性缺血性脑卒中患者外周血白细胞黏附分子和血清中可溶性黏附分子表达水平的影响。方法：治疗组18例，予巴曲酶3d（20BU）并予其他常规治疗；对照组18例，除未予巴曲酶外，余用药同巴曲酶治疗组。采用流式细胞术和酶联免疫吸附法测定患者卒中后不同时间外周血中性粒细胞和单核细胞CD11b、CD18、CD62L、CD54的表达以及血清中可溶性ICAM－1、VCAM－1的水平。结果：与对照组比，巴曲酶治疗组卒中患者在发病24h后中性粒细胞CD11b的表达下降（P＜0.05），单核细胞CD11b、中性粒细胞和单核细胞CD18、CD62L、CD54的表达及血清中可溶性ICAM－1、VCAM－1的水平均无显著性变化。结论：巴曲酶对急性缺血性脑卒中患者外周血黏附分子的表达的影响有限，提示降低外周血黏附分子的表达可是巴曲酶治疗缺血性脑卒中的主要作用机制。