山莨菪碱及β-七叶皂苷钠对内毒素致伤HUV-ECs能量代谢的保护作用

目的：探讨内毒素（ＬＰＳ）致伤后培养人脐静脉内皮细胞（ＨＵＶＥＣｓ）能量代谢的变化及山莨菪碱、β－七叶皂苷钠干预后对ＨＵＶＥＣｓ能量代谢的影响。方法：建立体外培养的ＨＵＶＥＣｓ模型,致伤因素为ＬＰＳ;测定指标为ＡＴＰ,ＡＤＰ及能荷（ＥＣ）。结果：低浓度ＬＰＳ能激活内皮细胞,随刺激时间的延长,培养ＨＵＶＥＣ的ＥＣ降低,用山莨菪碱、β－七叶４皂苷钠治疗后各时相点能量和明显增加,其中β－七叶皂苷钠治疗效果优于山莨菪碱。结论：ＬＰＳ对体外培养的ＨＵＶＥＣ能量代谢有直接损伤作用,β－七叶皂苷钠有一定的抗损伤作用,在能量储备方面优于山莨菪碱。     

Effects of dexamethasone, cyproheptadine, anisodamine, and dinoprostone on TNF_α production in endotoxic shock

目的：研究地塞米松（Ｄｅｘ）、噻庚啶（Ｃｙｐ）、山莨菪碱（Ａｎｉ）和地诺前列酮（Ｄｉｎ）对脂多糖（ＬＰＳ）诱导的肿瘤坏死因子（ＴＮＦα）基因表达的影响和抑制ＴＮＦα产生的抗休克作用．方法：Ｗｉｓｔａｒ大鼠静脉注射ＬＰＳ（ＥｃｏｌｉＯ１１１Ｂ４,５ｍｇ·ｋｇ－１）复制内毒素休克模型．Ｎｏｒｔｈｅｒｎ印迹杂交分析肝脏ＴＮＦαｍＲＮＡ表达,放射免疫法测定血浆ＴＮＦα的含量．结果：ＬＰＳ攻击后２ｈ肝脏ＴＮＦαｍＲＮＡ表达水平显著增高（放射性自显影扫描分析３８±１０ｖｓ盐水对照组１１±８,Ｐ＜００１）;血浆ＴＮＦα水平明显升高［（２２±３）μｇ·Ｌ－１ｖｓ盐水对照组（２２±１０）μｇ·Ｌ－１,Ｐ＜００１］．静脉注射ＬＰＳ后立即静脉注射Ｄｅｘ５,Ｃｙｐ５,Ａｎｉ１０及Ｄｉｎ２ｍｇ·ｋｇ－１均能显著降低大鼠肝脏ＴＮＦαｍＲＮＡ水平和血浆ＴＮＦα含量,提高ＬＰＳ２０ｍｇ·ｋｇ－１攻击的小鼠２４ｈ的存活率．结论：Ｄｅｘ,Ｃｙｐ,Ａｎｉ和Ｄｉｎ均能显著抑制ＬＰＳ诱导的ＴＮＦα基因表达,具有较强的抗休克作用．     

地塞米松,噻庚啶,山莨菪碱和地诺前列酮对内毒素休克TNFα产？…

目的：研究地塞米松（Ｄｅｘ）噻庚啶（Ｃｙｐ）,山莨菪碱（Ａｎｉ）和地诺前列腺酮（Ｄｉｎ）对脂多糖（ＬＰＳ）诱导的肿瘤坏死因子（ＴＮＦα）,基因表达的影响和抑制ＴＮＦ,产生的抗体休克作用。方法：Ｗｉｓｔａｒ大鼠静脉注射ＬＰＳ（ＥｃｏｌｉＯ１１１Ｂ４,５ｍｇ．ｋｇ＾－１）复制内毒素休克模型,Ｎｏｒｔｈｅｒｎ印迹杂交分析肝脏ＴＮＦαｍＲＮＡ表达,放射免疫法测定血浆ＴＮＦα的含量。结果：ＬＰＳ攻击后２ｈ肝     

重组血管内皮生长因子的研究进展

血管内皮生长因子（Ｖａｓｃｕｌａｒ ｅｎｄｏｔｈｅｌｉａｌ ｇｒｏｗｔｈ ｆａｃｔｏｒ,ＶＥＧＦ）是１９８９年Ｆｅｒｒａｒａ等在牛垂体滤泡星状细胞体外培养液中首先纯化出来的。到目前为止已经发现了六个ＶＥＧＦ异构体,包括ＶＥＧＦ１２１,ＶＥＧＦ１４５,ＶＥＧＦ１６５,ＶＥＧＦ１８３,ＶＥＧＦ１８９,ＶＥＧＦ２０６。已经发现高亲和力的ＶＥＧＦ受体有两类,分别为Ｆｌｔ－１（Ｆｍｓ－ｌｉｋｅ ｔｙｒｏｓｉ     

氧化甾醇诱导血管平滑肌细胞凋亡及其作用特点的研究

目的研究３β,５α,６β－胆甾烷二醇（Ｔｒｉｏｌ）诱导血管平滑肌细胞（ＶＳＭＣｓ）凋亡及其与２５－羟胆固醇（２５－ＯＨ）比较的特点。方法体外培养ＶＳＭＣｓ、光镜、透射电镜及ＴＵＮＥＬ技术。结果ＶＳＭＣｓ经Ｔｒｉｏｌ处理后,贴壁层细胞呈现核染色质浓集,核碎裂和凋亡小体形成等凋亡的超微结构变化;ＴＵＮＥＬ显示ＶＳＭＣｓ凋亡细胞数随Ｔｒｉｏｌ浓度的增加而增多;剂量为 ３０ μｍｏｌ·Ｌ－１的 Ｔｒｉｏｌ和 ２５－ＯＨ诱导 ＶＳＭＣｓ的凋亡作用,前者不能而后者能被 ５０ μｍｏｌ·Ｌ－１的胆固醇所抑制。结论Ｔｒｉｏｌ能诱导ＶＳＭＣｓ凋亡,不同氧化甾醇可能有不同的作用通道和机制;氧化甾醇诱导ＶＳＭＣｓ凋亡可能是引起动脉粥样硬化斑块破溃,导致急性心血管事件发生的机制之一。     

炊事工人血浆VitC,VitE,β—胡萝卜素,LPO,红细胞SOD值的检？…

对３６名烹调油烟接触者和３０名非接触者血浆ＶｉｔＣ、ＶｉｔＥ、β胡萝卜素,ＬＰＯ、红细胞ＳＯＤ水平进行研究,结果表明,烹调油烟接触组血ＶｉｔＣ、ＶｉｔＥ、β－胡萝卜素的含是细胞ＳＯＤ的活怀较非接触组的低,ＬＰＯ较非接触组的高,并与接触ＣＯＦ浓度呈一定程度的直线相关;接触ＣＯＦ浓度越高,血浆中ＶｉｔＣ、ＶｉｔＥ、β－胡萝卜素的含量,红细胞ＳＯＤ的活性平均值越低,血浆ＬＰＯ含量平均值越高,提示烹调油烟     

炊事工人血浆VitC、VitE、β-胡萝卜素、LPO、红细胞SOD值的检测及相关分析

对３６名烹调油烟接触者和３０名非接触者血浆ＶｉｔＣ、ＶｉｔＥ、β－胡萝卜素、ＬＰＯ、红细胞ＳＯＤ水平进行研究,结果表明,烹调油烟接触组血浆中ＶｉｔＣ、ＶｉｔＥ、β－胡萝卜素的含量、红细胞ＳＯＤ的活性较非接触组的低,ＬＰＯ较非接触组的高（Ｐ＜０．０１）,并与接触ＣＯＦ浓度呈一定程度的直线相关;接触ＣＯＦ浓度越高,血浆中ＶｉｔＣ、ＶｉｔＥ、β－胡萝卜素的含量、红细胞ＳＯＤ的活性平均值越低,血浆ＬＰＯ含量平均值越高。提示烹调油烟可能损害机体内活性氧防卫生系统,导致体内氧自由基反应及脂质过氧化反应加剧。     

Tumor necrosis factor mediated release of platelet-derived growth factor from bovine cerebral microvascular endothelial cells

研究ＴＮＦ介导的ＢＣＭＥＣ释放ＰＤＧＦ及药物对ＢＣＭＳＭＣ增殖的保护作用．方法：体外培养ＢＣＭＥＣ和ＢＣＭＳＭＣ，用结晶紫染色法观察ＴＮＦ引起的ＢＣＭＥＣ释放ＰＤＧＦ继而促进ＢＣＭＳＭＣ增殖作用．结果：ＴＮＦ不能促进无血清培养的ＢＣＭＳＭＣ增殖，但ＴＮＦ（５－２０μｇ·Ｌ－１）剂量依赖地促进ＢＣＭＥＣ释放ＰＤＧＦ．ＴＮＦ（２０μｇ·Ｌ－１）促进ＢＣＭＳＭＣ增殖的百分率为３４±４％．药物Ｉｍｐｅｒａｔｏｒｉｎ，ｉｓｏｉｍｐｅｒａｔｏｒｉｎ和ＰＭＤＰ不影响ＴＮＦ引起ＢＣＭＥＣ释放ＰＤＧＦ，但能剂量依赖地（１－１００μｍｏｌ·Ｌ－１）抑制ＰＤＧＦ促ＢＣＭＳＭＣ增殖的作用．结论：ＴＮＦ促进ＢＣＭＥＣ释放ＰＤＧＦ继而引起ＢＣＭＳＭＣ的增殖．药物能抑制ＰＤＧＦ引起的ＢＣＭＳＭＣ增殖．     

白芍总甙对大鼠腹腔巨噬细胞产生肿瘤坏死因子的调节机制

目的：研究不同浓度白芍总甙（ＴＧＰ）调节大鼠腹腔巨噬细胞（ＭΦ）产生肿瘤坏死因子（ＴＮＦ）作用。方法：在ＭΦ培养系统中有或无环氧酶抑制剂和钙调蛋白抑制剂等工具药，测定４５Ｃａ内流、ＰＧＥ２和ＴＮＦ含量。结果：ＴＧＰ（０．５～１０ｍｇ·Ｌ－１）明显促进ＬＰＳ诱导ＭΦ的４５Ｃａ内流和ＴＮＦ产生。线性回归分析表明，４５Ｃａ内流和ＴＮＦ产生呈明显正相关。三氟拉嗪（４０μｍｏｌ·Ｌ－１）可阻断ＴＧＰ促进ＬＰＳ诱导ＭΦ产生ＴＮＦ。ＴＧＰ－ＬＰＳ的ＴＮＦ释放曲线呈钟罩形，而ＴＧＰ－ＬＰＳ的ＰＧＥ２产生曲线呈浓度依赖性升高。当ＴＧＰ在低浓度（０．５～１２．５ｍｇ·Ｌ－１）时，ＴＮＦ与ＰＧＥ２产生明显正相关，而高浓度（１２．５～２５０ｍｇ·Ｌ－１）两者呈明显负相关。吲哚美辛（１０μｍｏｌ·Ｌ－１）可使ＴＮＦ量效曲线下降支消失，而Ｎω亚硝基－Ｌ－精氨酸（１５μｍｏｌ·Ｌ－１）对此无明显影响。结论：低浓度ＴＧＰ对ＴＮＦ产生上调作用可能与促进４５Ｃａ内流，提高钙调蛋白活性从而促进ＰＧＥ２分泌等有关，而高浓度下调作用是可能与ＭΦ自身产生大量ＰＧＥ２介导有关。     

银杏叶制剂对心绞痛患者的抗氧化和抗脂质过氧化作用

目的：探讨银杏叶制剂对心绞痛患者的抗氧化和抗脂质过氧化作用。方法：检测了７８例心绞痛患者经银杏叶制剂“天宝宁”治疗前后的血浆维生素Ｃ（Ｐ－ＶＣ）、维生素Ｅ（Ｐ－ＶＥ）、β－胡萝卜素（Ｐ－β－ＣＡＲ）、过氧化脂质（Ｐ－ＬＰＯ）以及红细胞超氧化物歧化酶（Ｅ－ＳＯＤ）、过氧化氢酶（Ｅ－ＣＡＴ）、谷胱甘肽过氧化物酶（Ｅ－ＧＳＨ－ＰＸ）、过氧化脂质（Ｅ－ＬＰＯ）值。结果：与治疗前比较,治疗后的Ｐ－ＶＣ、Ｐ－     

TNF-α对原代培养大鼠心肌细胞β_3-AR表达的影响及意义

目的探讨肿瘤坏死因子-α(TNF-α)对心脏β3-肾上腺素能受体(β3-AR)表达的影响及其在心衰治疗中的意义。方法培养生后2 d SD大鼠心肌细胞,取第4天的细胞加入含不同浓度TNF-α的培养液,检测不同时间作用下β3-AR mRNA及蛋白表达。结果心肌细胞受TNF-α刺激后,β3-AR mRNA及蛋白随着TNF-α作用时间的延长、作用浓度的增加,表达量增加,至作用浓度为1.6 ng/mL、作用时间为12 h达峰值。结论TNF-α作用下心肌β3-AR上调,且有时间及浓度依赖性,这可能是引起心衰的原因之一。     

灯盏花素对血小板活化因子致肺微血管内皮细胞损伤的影响

目的　探讨血小板活化因子 (PAF)诱导肺微血管内皮细胞损伤的机制及灯盏花素的干预作用。方法　观察PAF对体外培养大鼠肺微血管内皮细胞 (RPMVEC)的形态、单层通透性、F 肌动蛋白 (F actin)的影响和灯盏花素干预后的变化 ,F actin用流式细胞仪测定。结果　PAF浓度大于 0 1mg·L-1作用 4 8h内观察到细胞脱落和破裂。 10mg·L-1PAF 12 0min内可使RPMVEC单层通透性增高、F actin解聚 ,灯盏花素可抑制上述变化。结论　①PAF引起RPMVEC脱落和破裂呈时间和剂量依赖性。②PAF引起内皮单层通透性的增高的机制与F actin解聚密切相关。③灯盏花素可抑制PAF引起RPMVEC单层通透性的增高和F actin下降 ,对PAF引起的RPMVEC损伤有一定保护作用。     

Australasian Society of Clinical and Experimental Pharmacologists and Toxicologists, 1994: SIGNALLING PATHWAYS IN CARDIAC FAILURE

1. Cardiac failure in humans and in animal models is associated with a marked desensitization of the catecholamine signalling pathway. 2. β₁- and β₂- and possibly β₃-adrenoceptors (β-AR) are found in the hearts of humans and common laboratory animals such as rats and guinea-pigs. In rats and guinea-pigs chronic stimulation of cardiac β-AR leads to a rapid loss of β₂-AR whereas heart failure in humans is associated with a loss of β₁-AR or β₁-AR and β₂AR. 3. Desensitization is also associated with phosphorylation of β-AR by β-AR kinase β-ARK) and uncoupling of receptors from the signalling pathway. β-ARK but not β-arrestin activity and mRNA are markedly increased in heart failure. 4. Chronic β-AR stimulation and heart failure are associated with increases in Giα but little if any change in Gsα. 5. The roles of βm? subunits of G-proteins, adenylate cyclase subtypes and cAMP dependent protein kinase A in heart failure are unclear at present.     

Effects of calycosin on the impairment of barrier function induced by hypoxia in human umbilical vein endothelial cells

The purpose of the present study was to examine the effects of calycosin, an isoflavonoid isolated from Astragali Radix, on the impairment of barrier function induced by hypoxia in cultured human umbilical vein endothelial cells. Hypoxia induced an increase in endothelial cell monolayer permeability, indicating endothelial cell barrier impairment. Endothelial barrier dysfunction induced by hypoxia was accompanied by decreases in cytosolic ATP concentration and cAMP level, the development of actin stress fibers and intercellular gap formation, suggesting that the decreases in cytosolic ATP and cAMP levels and rearrangements of F-actin could be associated with an increase in permeability of endothelial monolayers. Application of calycosin inhibited the hypoxia-induced increase in endothelial permeability in a dose-dependent fashion, which is compatible with inhibition of lactate dehydrogenase release, decrease of the fall in ATP and cAMP contents, and improvement of F-actin rearrangements. These findings indicate that calycosin protected endothelial cells from hypoxia-induced barrier impairment by increasing intracellular energetic sources and promoting regeneration of the cAMP level, as well as improving cytoskeleton remodeling.     

血管紧张素Ⅱ对肾小球内皮细胞骨架和单层通透性的影响

目的观察血管紧张素Ⅱ(AngⅡ)对培养肾小球内皮细胞(GENC)形态、单层通透性、肌动蛋白骨架的影响,探讨AngⅡ对GENC炎性损伤的机制。方法倒置显微镜观察AngⅡ对体外培养大鼠GENC的形态影响;用二室弥散系统检测AngⅡ对GENC单层通透性的影响;用免疫细胞化学的方法观察AngⅡ对GENCF-肌动蛋白(F-actin)分布的影响。结果AngⅡ浓度大于0.1mg·L-1作用48h内观察到细胞脱落和破裂。10mg·L-1AngⅡ6、12h可使GENC单层通透性增高、F-actin解聚。结论AngⅡ引起GENC脱落和破裂呈时间和剂量依赖性;AngⅡ引起内皮单层通透性的增高的机制可能与F-actin解聚相关;     

蛋白激酶Cδ亚型抑制剂对脂多糖诱导肺微血管内皮细胞损伤的影响

目的研究蛋白激酶C(PKC)的亚型PKCδ在大鼠肺微血管内皮细胞(RPMVEC)中的表达及其抑制剂Rottlerin对脂多糖(LPS)导致RPMVEC单层通透性增高的影响。方法取体外培养的RPMVEC进行PKCδ的W estern b lot和免疫组化检测,用针头式滤器检测LPS及LPS+Rottlerin干预后RPMVEC单层滤过系数(K f)的变化。结果PKCδ在RPMVEC中有广泛的表达,其表达部位主要位于胞质。Rot-tlerin能减低LPS导致的RPMVEC单层通透性增高。结论LPS导致RPMVEC损伤的机制与PKC的激活有关,PKCδ亚型抑制剂可减轻LPS诱导的RPMVEC单层通透性增高。     

Cilostazol inhibits the redistribution of the actin cytoskeleton and junctional proteins on the blood-brain barrier under hypoxia/reoxygenation.

Cilostazol is a selective inhibitor of cyclic nucleotide phosphodiesterase 3 (PDE3), which induces a vasodilatoric antiplatelet effect. In the present study, we investigated the impact of cilostazol on the blood-brain barrier (BBB), while focusing on the actin cytoskeleton (F-actin), the permeability of endothelial cells, and the junctional proteins under hypoxia/reoxygenation (H/R). Cilostazol was thus found to inhibit the cytoskeletal reorganization under H/R, in which F-actin decrease at the cell periphery. Accordingly, cilostazol was able to attenuate the hyperpermeability of endothelial cells in H/R to the level of the permeability in normoxia. However, the adherens junction (AJ) protein VE-cadherin was not preserved in the presence of cilostazol under H/R. On the other hand, beta-catenin was slightly retained by cilostazol. In contrast to the redistribution of these proteins, immunoblotting demonstrated the total amount of AJ and tight junction (TJ) proteins (occludin, ZO-1 and ZO-2) to not show any significant change under H/R stress in either the presence or absence of cilostazol. Taken together, cilostazol potently displayed a protective effect against acute ischemia by preventing an increase in the endothelial permeability through the preservation of the actin cytoskeleton and the redistribution of junctional proteins.     

Exposure to cigarette smoke downregulates β2-adrenergic receptor expression and upregulates inflammation in alveolar macrophages

Abstract

Cigarette smoke-triggered inflammation is important in the pathophysiology of chronic obstructive pulmonary disease (COPD). β₂-Adrenergic receptor (β₂-AR) is abundantly expressed on inflammatory cells, which is associated with inflammation regulation. To observe alterations in inflammation, pathological changes in lung tissues, and detect changes in β₂-AR expression, rats were exposed for 4 months to cigarette smoke. Pathological changes were observed in lung tissue sections. The levels of inflammatory mediators tumor necrosis factor (TNF)-α, interleukin (IL)-1β in bronchoalveolar lavage fluid (BALF), and lung tissues were measured using enzyme-linked immunosorbent assay (ELISA). Nuclear factor (NF)-κB activity was detected by electrophoretic mobility shift assay (EMSA). Exposure to this regimen of cigarette smoke induced peribronchial and perivascular lymphocytic aggregates and parenchymal accumulation of macrophages in rats. EMSA demonstrated that smoke exposure enhanced NF-κB activation in rats’ alveolar macrophages (AMs). Compared with the control group, smoke exposure induced a notable increase in TNF-α and IL-1β in BALF, lung tissues, and a decrease of β₂-AR expression of AMs. The expression of β₂-AR from AMs was inversely correlated with TNF-α and IL-1β levels of BALF. These data demonstrated that chronic smoke-triggered lung inflammation was accompanied by down-regulation of β₂-AR in rat lungs’ AMs.