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肾小球系膜细胞(glomerularmesangialcel,GMC)是肾小球中功能最为活跃的细胞。GMC收缩可影响肾小球滤过率。血小板活化因子(platelet-activatingfactor,PAF)是一种重要的免疫炎性介质,通过多种途径参与了... 相似文献
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为探讨血小板活化因子(PAF)对肾小球系膜细胞(GMC)产生活性氧的调节作用,本研究观察了PAF对体外培养的大鼠GMC产生超氧化物阴离子(O)和过氧化氢(H-2O-2)的影响。结果表明10 ̄(-9)M/L的PAF已能诱导GMC产生O和H2O2(1.14±0.42nmol/10 ̄5GMC、0.97±0.16nmol/10-5GMC),这一效应呈剂量依赖性;溶PAF无诱导GMC产生活性氧作用;特异性PAF受体拮抗剂BN52021抑制PAF的刺激作用,抑制作用也呈剂量依赖性,5×10 ̄(-5)M/LBN52021完全抑制GMC合成活性氧。本研究说明PAF可诱导GMC合成活性氧。 相似文献
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细胞因子对肾小球系膜细胞增殖的影响 总被引:2,自引:0,他引:2
应用^3H-TdR掺入法和MTT比色法,观察了人重组IL-1β,IL-6和TNFα对体外培养的成人肾小球系膜细胞增殖的影响,结果表明,在无血清培养条件下,HrIL-1β,HrIl-6和HrTNFα在一个较大的浓度范围内,均不能刺激系膜细胞的增殖;而有少量胎牛血清存在时,HrIL-6和HrTNFα可以促进系膜细胞增殖,但HrIL-1β无此作用另外,在实验中还发现,雷公藤多甙(TⅡ)对系膜细胞生长具有 相似文献
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目的: 探讨高糖高脂对系膜细胞产生细胞外基质(ECM)和血小板活化因子(PAF)的影响。方法:人系膜细胞单独培养,分为对照组、甘露醇组、高糖高脂组、PAF受体拮抗剂BN52021+高糖高脂组, ELISA法检测各组细胞上清液中纤维连接蛋白(Fn)、IV型胶原(Col IV)、血小板活化因子(PAF)含量,实时荧光定量检测系膜细胞血小板活化因子受体(PAF-R) mRNA表达。结果:高糖高脂可引起系膜细胞培养上清液Fn和Col IV含量升高(均P<0.05),BN52021可抑制高糖高脂引起的Fn和Col IV含量升高(P<0.05);高糖高脂可引起系膜细胞培养上清液PAF含量升高(P<0.05);高糖高脂可上调系膜细胞PAF-R mRNA表达(P<0.05)。结论: 高糖高脂刺激系膜细胞Fn、Col IV、PAF产生,高糖高脂刺激的ECM分泌增加部分与PAF有关; 高糖高脂可促进系膜细胞PAF-R基因表达,使PAF的生物效应进一步放大。 相似文献
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探讨细胞因子和地塞米松对肾小球系膜细胞表面表达细胞间粘附分子-1(ICAM-1)的影响。通过体外系膜细胞培养的方法,分别加入不同的细胞因子,观察其浓度及作用时间产生的影响;并用ELISA法检测不同浓度的地塞米松对系膜细胞表达ICAM-1的影响。结果显示系膜细胞表达ICAM-1的强弱与细胞因子的作用呈剂量、时间依赖关系;随着地塞米松浓度的增加,ICAM-1表达逐渐减弱,与未加地塞米松组相比相关均非常 相似文献
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目的观察血小板活化因子 (PAF)对大鼠肾小球系膜细胞 (GMC)作用 ,探讨其作用机制 ,并寻找由PAF诱导GMC损伤的保护方法。方法Wistar大鼠 ,160~200g,用改良的Hapar方法培养肾小球系膜细胞。分别置于硅化试管中 ,随机分为5组。①PAF组 ,调整RPML1640培养液中含PAF含量和45CaCl2,浓度为10 -7mmol/L和1.25mmol/L,分别测定GMC存活率、细胞内蛋白漏出量、细胞丙二醛 (MDA)、测定45Ca 的放射活性。②MnSO4 组 :调整细胞悬液中MnSO4 浓度为5×10 -4mmol/L ,测定内容同PAF组。③Verapamil(异博定 )组调整细胞悬液中Verapamil浓度为2×10 -4mmol/L,测定内容同PF组。④银杏皂甙组 :调整细胞血液中银杏皂甙提取物为浓度为100μg/ml外 ,测定内容同PF组。⑤对照组 :细胞悬液中不含PAF,测定内容同PF组。结果经PAF刺激后GMC死亡率显著升高 ,细胞外钙内流、细胞pr漏出、细胞脂质过氧化显著加强 (P<0.01)。应用Verapamil可显著减少细胞外钙内流 ,减少GMC死亡率 (P<0.01)。结论应用银杏叶醇提取物可显著减少GMC死亡率细胞脂质过氧化 相似文献
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肾小球系膜细胞增生与系膜硬化机制的初步探讨 总被引:17,自引:0,他引:17
我们应用细胞培养及免疫组化的方法,探讨了肾小球系膜细胞增生及系膜硬化的机制,并进而观察了它们在肾小球硬化听作用。结果显示:系膜细胞有自分泌功能,肿瘤坏死因子和血管内皮素可促进系膜细胞增生。大量增多的系膜基质不但含有Ⅳ型胶原,也有Ⅲ型胶原。系膜细胞增生,系膜基质增多最终导致肾小球硬化。 相似文献
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目的:探讨高糖高脂条件下内皮细胞与系膜细胞相互作用对PAF产生、系膜细胞paf-R基因表达的影响及阿托伐他汀的干预作用。方法:内皮细胞与系膜细胞共培养和系膜细胞单独培养后随机,分为对照组、甘露醇组、高糖高脂组、阿托伐他汀组,培养24h后,ELISA法检测各组细胞上清液PAF含量,实时荧光定量检测系膜细胞paf-RmRNA表达。结果:(1)高糖高脂促进共培养组和单培养组PAF升高(P0.05),共培养组PAF均较单培养组升高(P0.05);(2)高糖高脂可上调系膜细胞paf-R mRNA表达(P0.05);(3)共培养和单培养条件下,阿托伐他汀可抑制高糖高脂引起的PAF升高(P0.05),并可抑制高糖高脂引起的系膜细胞paf-RmRNA表达上调(P0.05)。结论:(1)高糖高脂环境下,系膜细胞和内皮细胞存在异常的相互作用,这种作用促进PAF产生;(2)高糖高脂促进系膜细胞paf-R基因表达,使PAF的生物效应进一步放大;(3)阿托伐他汀可影响高糖高脂条件下内皮细胞与系膜细胞之间的相互作用。 相似文献
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细胞因子诱导不同来源的贴壁细胞分化为树突状细胞的研究 总被引:11,自引:1,他引:10
通过贴壁的方法分离胚肝、脐带血、造血动员后的外周血及正常人外周血中的树突状细胞 (DC )的前体细胞 ,体外采用细胞因子诱导 ,并对其进行形态学观察和细胞表型分析 ;同时比较了不同来源DC的增殖、分泌IL 12的能力及激发同种异体T细胞和脐带血幼稚型T细胞 (naiveTcell)的作用。实验结果表明 ,经过造血动员后的外周血细胞产生的DC数量和纯度均较高 ,而且能经体外细胞因子诱导为功能性DC ,是较理想的DC来源 相似文献
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Guillermo A. Herrera 《Ultrastructural pathology》2013,37(6):471-479
In the last two decades, the ability of mesangial cells to respond to various stimuli or injurious agents by altering their phenotype and function has become recognized. The plasticity of these mesangial cells has been linked to the morphological and functional alterations responsible for the pathologic findings. Many of the glomerular disorders target the mesangium as the primary and/or initial site of injury. Understanding how mesangial cells are altered in the various conditions provides a platform for conceptualizing pathologic mechanisms and defining key steps amenable to therapeutic intervention. The present paper reviews the normal and altered mesangium with an emphasis on mechanisms involved in alterations of mesangial homeostasis. Mesangial cells and matrix are very important in maintaining normal glomerular structure, and function and the plasticity of these cells is responsible for pathological manifestations, repair, and scarring. Our more sophisticated understanding of mesangial cell behavior and matrix biology provides very useful information to help design new therapeutic approaches to the treatment of renal diseases. The potential for bone marrow-derived cells to differentiate into mesangial cells and repopulate damaged mesangium, thus “healing” what is today considered to be irreversible damage represents an exciting new area of research. 相似文献
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IL-13抑制人肾小球系膜细胞IL-12的表达 总被引:2,自引:0,他引:2
为了探讨白细胞介素 13(IL 13)对体外培养的人肾小球系膜细胞产生白细胞介素 12 (IL 12 )的影响。我们用脂多糖(LPS 10 μg/ml) ,不同浓度的IL 13对系膜细胞培养 ,分别采用ELISA法和半定量RT PCR法检测细胞上清液的IL 12和系膜细胞IL 12p4 0mRNA表达。结果提示 5 %NCSRPMI 16 4 0基础培养条件下的系膜细胞未检测到IL 12蛋白分泌及其mRNA表达。在LPS刺激下系膜细胞的IL 12p4 0mRNA表达加强 ,并分泌出大量的IL 12。IL 13在 1~ 10 0ng/ml浓度范围内对LPS诱导的系膜细胞IL 12分泌及IL 12p4 0mRNA表达的抑制作用呈剂量依赖趋势。本研究认为IL 13可能通过抑制IL 12的产生 ,而调整了体内Th1/Th2细胞因子平衡 ,作为抗炎性细胞因子在肾小球肾炎发病机制中发挥一定作用 相似文献
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Lupus nephritis (LN) is a prototypic autoimmune disease, however, the precise immuno-pathogenesis of LN remains to be elucidated. In our previous studies, autoantibodies against mesangial cells had been identified in sera from patients with lupus nephritis and could bind the membrane proteins of human mesangial cells (HMC) directly through antigen-antibody interaction without DNA bridge. The current study is to investigate whether the autoantibodies were associated with anti-DNA antibodies and their target antigens distribution in different cell types. Sera from nine patients with renal biopsy proven lupus nephritis with positive anti-dsDNA antibodies and four healthy subjects were collected. IgG was isolated by Protein G affinity chromatography and then non-DNA-binding IgG fractions were obtained after deletion of anti-DNA antibodies using a DNA-cellulose affinity column. Membrane proteins, obtained from HMC, human umbilical vein endothelial cells (HUVEC), peripheral mononuclear cells by sonication and sequential centrifugation, were solubilized and applied in Western-blot analysis to characterize the target antigens. In results, the non-DNA-binding IgG fractions from sera of patients with lupus nephritis could blot the protein(s) of HMC membrane at 74, 63, and 42 kD. However, only a similar 74-kD protein could be blotted on membrane of HUVEC, and the target antigens on membranes of mononuclear cells were heterogeneous. In conclusion, our preliminary study had demonstrated that non-DNA binding autoantibodies against mesangial cells could be found in sera from patients with lupus nephritis. Although the target antigens might not be cell specific, the roles of these autoantibodies in the pathogenesis of lupus nephritis need further investigation. 相似文献
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自由基对培养的鼠肾小球系膜细胞膜系统损伤的研究 总被引:1,自引:0,他引:1
目的 观察自由基对肾小球系膜细胞 (GMC)膜系统的影响。方法 用黄嘌呤氧化 酶黄嘌呤生成自由基 ,作用于培养的鼠GMC ,通过TBA比色法检测脂质过氧化产物丙二醛 (MDA)含量及透射电镜观察超氧化物歧化酶 (SOD)对细胞膜系统的保护作用。结果 未加SOD组GMC的MDA含量明显增加 ,电镜下出现细胞膜系统的损伤性改变。加SOD组MDA含量未见明显增加。电镜下细胞膜系统未见明显改变。结论 自由基可引起GMC膜系统脂质过氧化损伤 ,SOD具有保护作用 相似文献
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目的研究高氨基酸血症对肾小球系膜细胞结缔组织生长因子(connective tissue growth factor,CTGF)表达的影响,探讨氧化应激在其中的作用。方法大鼠肾小球系膜细胞分为不同培养环境下的对照组(C组)、高氨基酸组(HA组)、维生素E组(HAE组)。培养24h后用荧光染料2',7'-Dichloroluores-cin diacetate(20μmol/L)测定各组细胞内活性氧产物(reactive oxygen specie,ROS)的生成情况从而反应细胞内氧化应激水平,培养24h后采用RT-PCR检测各组CTGF mRNA表达,培养48h后采用Western印迹检测各组CTGF蛋白的表达。结果HA组氧化应激水平、CTGF mRNA及蛋白的表达均明显高于C组(P<0.05);HAE组其氧化应激水平、CTGF mRNA及蛋白表达较HA组明显降低(P<0.05),且与C组比较差异无统计学意义(P>0.05)。结论高氨基酸血症可使肾小球系膜细胞氧化应激水平升高并产生过多的CTGF,抗氧化剂维生素E可减轻系膜细胞氧化应激及CTGF的表达,提示高氨基酸诱导的CTGF表达增多可能与氧化应激有关。 相似文献
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This study describes an in vitro method to evaluate a PAF acether release test (PART) from white blood cells after antigenic challenge. PAF acether activity of the supernatant was tested by platelet aggregation. The aggregating power was abolished by using SRI 63-441 (Sandoz), a PAF acether inhibitor. This method was applied to 57 patients with allergic or pseudo-allergic reactions to drugs by using different drug protein conjugates. The results of PART were evaluated in relation to the clinical history (score of imputability) and to other tests (skin tests, lymphocyte transformation tests (LTT), IgE-RAST). A good correlation was found between the release of PAF acether and a high predictability score: sensitivity 75%, specificity 83.8%. PART also correlated with skin tests (75% agreement, n = 60), with LTT (67.7% agreement, n = 74) and in 65.6% of cases with positive penicillin IgE-RAST (n = 32). This method brings a new possibility for the investigation of drug-allergic and pseudo-allergic reactions. 相似文献
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观察白细胞介素 10 (IL 10 )对体外培养的人肾小球系膜细胞 (HMC )增殖影响 ,并探讨IL 10对HMC细胞周期负调控蛋白p2 7的影响 ,旨在了解其调节HMC增殖的内在机制。HMC用含 5 %FCS的RPMI16 4 0培养 ,加入IL 10刺激后 ,应用流式细胞术分别测定HMC细胞增殖周期的变化以及p2 7的水平。结果表明IL 10可抑制血清诱导的HMC增殖 ,IL 10显著减少了HMCG0 /G1期细胞进入S期和G2 /M期 ;2 5ng/mlIL 10明显地上调细胞周期负调控蛋白p2 7水平 ,是血清刺激组的 1 7倍 (P <0 0 5 )。IL 10抑制血清诱导的HMC增殖的作用机制可能部分是通过上调细胞周期负调控蛋白p2 7来实现的 ,提示IL 10在增殖性肾小球肾炎中可能发挥重要的调节作用 相似文献
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目的:观察依维莫司对高糖诱导肾小球系膜细胞(HBZY-1)凋亡的保护作用,并探讨其意义。方法:将培养HBZY-1细胞按不同葡萄糖和依维莫司浓度分组处理72h后,用磺酰罗丹明B法测量细胞OD值,计算各组细胞存活率,用Annexin V和PI双标法检测各组细胞凋亡率。结果:30mM葡萄糖处理的细胞存活率明显降低,凋亡率明显增加(均P<0.05)。不同浓度依维莫司均能提高细胞存活率(均P<0.05),其中以200ng/ml依维莫司效果最好(P<0.05);200ng/ml依维莫司具有明显抗HBZY-1细胞凋亡作用。结论:依维莫司对高糖诱导HBZY-1细胞凋亡具有保护作用,从而为其治疗糖尿病肾病提供新的研究方向。 相似文献