Highly specific characterization and discrimination of monosodium urate crystals in gouty arthritis based on aggregation-induced emission luminogens

Existing technologies used to detect monosodium urate(MSU)crystals for gout diagnosis are not ideal due to their low sensitivity and complexity of operation.The purpose of this study was to explore whether aggregation-induced emission luminogens(AIEgens)can be used for highly specific imaging of MSU crystals to assist in the diagnosis of gout.First,we developed a series of luminogens(i.e.,tetraphenyl ethylene(TPE)-NH_(2),TPE-2NH_(2),TPE-4NH_(2),TPE-COOH,TPE-2COOH,TPE-4COOH,and TPE-Ketoalkyne),each of which was then evenly mixed with MSU crystals.Next,optimal fluorescence imaging of each of the luminogens was characterized by a confocal laser scanning microscope(CLSM).This approach was used for imaging standard samples of MSU,hydroxyapatite(HAP)crystals,and mixed samples with 1:1 mass ratio of MSU/HAP.We also imaged samples from mouse models of acute gouty arthritis,HAP deposition disease,and comorbidities of interest.Subsequently,CLSM imaging results were compared with those of compensated polarized light microscopy,and we assessed the biosafety of TPE-Ketoalkyne in the RAW264.7 cell line.Finally,CLSM time series and three-dimensional imaging were performed on MSU crystal samples from human gouty synovial fluid and tophi.As a promising candidate for MSU crystal labeling,TPE-Ketoalkyne was found to detect MSU crystals accurately and rapidly in standard samples,animal samples,and human samples,and could precisely distinguish gout from HAP deposition disease.This work demonstrates that TPE-Ketoalkyne is suitable for highly specific and timely imaging of MSU crystals in gouty arthritis and may facilitate future research on MSU crystal-related diseases.

Therapeutic Mechanism of Santeng Dingtong Recipe (三藤定痛方) on Monosodium Urate Crystal-Induced Rabbit Arthritis

Objective: To study the therapeutic mechanism of Santeng Dingtong recipe (STDT) on monosodium urate crystal (MSU) induced rabbit arthritis Methods: Forty-two rabbits were randomly divided into six groups, 7 in each group. Group 1 received 0.9% saline 2. 5 ml/kg per day by gastrogavage (ig) for 10 days; Group 2, 3 and 4 received STDT 0.125 g/kg, 1.0 g/kg and 8.0 g/kg per day respectively by ig for 10 days; Group 5 received colchicine 4. 5 mg/kg per day by ig for 4 days; and Group 6 was untreated. MSU crystals 10 mg /500ul containing polymyxin B 10 u/ml was injected into the knee joints of Group 1-5 to make rabbit arthritis models. Leukocytes in synovial lavage fluids was then counted and differentiated; pathological injury of synovial membranes was observed under HE staining; interleukin-1 beta (IL-1B), tumor necrosis factor alpha (TNFa) and leukotriene B4 (LTB4) content in synovial lavage fluids were determined by ELISA. Results: MSU caused a rapid leukocyte infiltration and increased production of IL-1B, TNFa and LTB4 2 hrs after intra-articular injection. STDT inhibited neutrophil infiltration in synovial fluids dose-dependently, protected the synovial membrane against pathological injury and reduced the production of IL-1B, TNFa and LTB4; while colchicine did not decrease the level of TNFa, but significantly inhibited neutrophil infiltration in synovial fluid and reduced the production of IL-11B and LTB4. Conclusion: STDT exerts an anti-inflammatory effect in rabbit model of acute MSU arthritis, its mechanism being probably due to the decrease of XL-1B, TNFa and LTB4 synthesis.

Monosodium urate crystals trigger Nrf2- and heme oxygenase-1-dependent inflammation in THP-1 cells

Gouty arthritis is an inflammatory disease that is caused by an accumulation of monosodium urate （MSU） crystals in the joints. MSU is capable of activating the nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3 （NLRP3） inflammasome, leading to interleukin-lβ （IL-lβ） secretion. Reactive oxygen species （ROS） are major mediators of the NLRP3/IL-lβ interaction. Although nuclear factor E2-related factor 2 （Nrf2） is recognized as a transcription factor that is involved in the response to oxidative stress, the effect of MSU on Nrf2 and on Nrf2-mediated antioxidant enzymes remains unclear. The treatment of THP-1 monocytes using phorbol 12-myristate 13-acetate （PMA） was shown to initiate inflammatory responses. Here, we showed that THP-1 cells, following treatment with MSU crystals, significantly increased IL-1β release, NLRP3 inflammasome activation and ROS production. MSU also promoted the nuclear translocation of Nrf2 and activated lysosomal destabilization. Moreover, the levels of heme oxygenase-1 （HO-1） in gene and protein expressions were upregulated by MSU. MSU-induced IL-lβ secretion and NLRP3 inflammasome activation were inhibited by the knockdown of Nrf2 and via the HO-1 inhibitor zinc （11） protoporphyrin IX （ZnPP）. In addition, HO-1 inhibition increased the level of superoxide anion production and the consumption of glutathione. These findings suggest that Nrf2 and HO-1 mediate redox homeostasis and interact with pro-inflammatory factors in MSU-challenged THP-1 cells, thereby providing new insight into how MSU-induced gouty inflammation is mediated by specific mechanisms that are involved in the Nrf2/Ho-1 antioxidant signaling pathway.

Anti-inflammatory effects of traditional mixed extract of medicinal herbs(MEMH)on monosodium urate crystal-induced gouty arthritis

Korean oriental medicine prescription is widely used for the treatment of gouty diseases. In the present study, we investigated anti-inflammatory effects of modified Korean herbal formulation, mixed extract of medicinal herbs(MEMH), and its modulatory effects on inflammatory mediators associated with gouty arthritis. Both in vitro and in vivo studies were carried out to assess the anti-inflammatory efficacy of MEMH on monosodium urate(MSU) crystals-induced gouty inflammation. MSU crystals stimulated human chondrosarcoma cell line, SW1353, and human primary chondrocytes were treated with MEMH in vitro. The expression levels of pro-inflammatory mediators and metalloproteases were analyzed. The effect of MEMH on NFκB signaling pathway in SW1353 cells was examined. Effect of MEMH on the mR NA expression level of pro-inflammatory mediators and chemotactic factor from human monocytic cell line, THP-1, was also analyzed. The probable role of MEMH in the differentiation process of osteoblast like cells, SaO S-2, after MSU treatment was also observed. To investigate the effects of MEMH in vivo, MSU crystals-induced ankle arthritic model was established. Histopathological changes in affected joints and plasma levels of pro-inflammatory mediators(IL-1β and TNFα) were recorded. MEMH inhibited NFκB signaling pathway and COX-2 protein expression in chondrocytes. MSU-induced mR NA expressions of pro-inflammatory mediators and chemotactic cytokines were suppressed by MEMH. In MSU crystals-induced ankle arthritic mouse model, administration of MEMH relieved inflammatory symptoms and decreased the plasma levels of IL-1β and TNFα. The results indicated that MEMH can effectively inhibit the expression of inflammatory mediators in gouty arthritis, demonstrating its potential for treating gouty arthritis.

Suppression of monosodium urate crystal-induced inflammation by inhibiting TGF-β-activated kinase 1-dependent signaling:role of the ubiquitin proteasome system

Monosodium urate(MSU)crystals activate inflammatory pathways that overlap with interleukin-1β(IL-1β)signaling.However,the post-translational mechanisms involved and the role of signaling proteins in this activation are unknown.In the present study,we investigated the intracellular signaling mechanisms involved in MSU-induced activation of THP-1 macrophages and human nondiseased synovial fibroblasts(NLSFs)and the in vivo efficacy of an inhibitor of tumor grovArth factor-β(TGF-β)-activated kinase 1(TAK1),5Z-7-oxozeaenoI,in MSU-induced paw inflammation in C57BL76 mice.THP-1 macrophage activation with MSU crystals(25-200 ng/ml)resulted in the rapid and sustained phosphorylation of interleukin-1 receptor-activated kinase 1(IRAK1 Thr^(209))and TAK1(Thr^(184/187))and their association with the E3 ubiquitin ligase TRAF6.At the cellular level,MSU inhibited the deubiquitinases A20 and UCHL2 and increased 20s proteasomal activity,leading to a global decrease in K^(63)-linked ubiquitination and increase in K^(48)-linked ubiquitination in THP-1 macrophages.While MSU did not stimulate cytokine produaion in NLSFs,it significantly amplified IL-1β-induced IL-6,IL-8,and ENA-78/CXCL5 production.Docking studies and MD simulations followed by TAK1 in vitro kinase assays revealed that uric acid molecules are capable of arresting TAK1 in an active-state conformation,resulting in sustained TAK1 kinase activation.Importantly,MSU-induced proinflammatory cytokine production was completely inhibited by 5Z-7-oxozeaenol but not IRAK1/4 or TRAF6 inhibitors.Administration of 5Z-7-oxozeaenol(5 or 15 mg/kg;orally)significantly inhibited MSU-induced paw inflammation in C57BL/6 mice.Our study identifies a novel post-translational mechanism of TAK1 activation by MSU and suggests the therapeutic potential of TAK1 in regulating MSU-induced Inflammation.

痛风基因表达谱的差异基因表达及免疫细胞浸润分析

目的:研究痛风患者的差异基因表达及免疫细胞浸润情况,寻找痛风发病的关键基因及免疫细胞,探究免疫细胞与痛风的关系。方法:从GEO数据库下载痛风的芯片GSE160170,借助R语言筛选差异基因,随后采用STRING数据库对差异基因进行分析,并借助Cytoscape软件筛选关键基因,再对其进行富集分析,同时对免疫细胞浸润情况进行分析。结果:研究发现IL-6、IL-1β、TNF、CCL3、CXCL8和CXCL1为痛风发病的关键基因,主要通过IL-17、Toll样受体、NOD样受体、NF-κB等信号通路发挥细胞对脂多糖、细菌来源分子及生物刺激的反应等过程导致疾病发生;免疫浸润结果表明记忆性B细胞、活化的NK细胞、活化的树突状细胞、活化的肥大细胞和嗜酸性粒细胞在痛风患者中表达显著;免疫细胞间的相关性分析结果表明滤泡辅助性T细胞与活化的肥大细胞表达呈正相关,未活化的NK细胞与单核细胞表达呈负相关。结论:关键基因和差异表达的免疫细胞与痛风发病机制密切相关,为痛风的免疫机制研究提供了新思路。

基于xCell算法研究痛风的免疫生物标志物及相关靶向中药的筛选

目的研究痛风致病的关键基因以及免疫生物标志物,挖掘潜在的靶向中药,为痛风的临床治疗提供新方向。方法从GEO数据库下载痛风芯片数据集,使用R软件进行差异基因的筛选,并对这些差异基因进行GO及基因通路KEGG富集分析。应用STRING数据库对差异基因进行蛋白质互作网络分析,利用Cytoscape筛选关键基因。使用ELISA方法对关键基因的蛋白表达水平进行实验验证。进一步使用xCell估计免疫细胞在痛风中的相对表达情况及相关性,最后通过Coremine Medical数据库对显著富集的免疫相关生物学过程及关键基因中药进行预测。结果筛选出痛风差异基因共计852个。在GO富集分析中,与免疫反应密切相关的主要生物学过程包括白细胞的趋化性、IL(白细胞介素)-2的产生以及血管生成的调控;KEGG通路主要富集于IL-17信号通路、趋化因子信号通路和肿瘤坏死因子(TNF)信号通路等。筛选出TNF、IL-6、IL-1β、趋化因子配体8(CXCL8)、FOS原癌基因(FOS)和血管内皮生长因子A(VEGFA)等10个关键蛋白,实验验证关键基因蛋白表达水平与芯片数据的表达趋势一致。免疫分析显示,单核细胞、记忆B细胞、初始CD8+T细胞和树突状细胞与痛风关系密切。中药预测发现,人参、三七、黄芩、三白草和丹参等为治疗痛风的潜在药物。结论本研究揭示了痛风致病的可能关键基因以及免疫机制,挖掘出潜在的靶向中药。这些关键基因和中药有望为痛风的治疗提供新的思路和方法。

土藤草颗粒对微晶型尿酸钠致家兔急性痛风性关节炎的药效学研究

目的观察土藤草颗粒对微晶型尿酸钠(monosodium urate,MSU)致家兔急性痛风性关节炎模型的药效学作用。方法选用取雄性日本大耳白兔,随机分为正常对照组,模型对照组,阳性药秋水仙碱组(0.18 mg·kg^(-1)),土藤草颗粒高、中、低剂量组分别为600、300、150 mg·kg^(-1)。通过家兔左后肢膝关节腔内注入MSU的方式建立急性痛风性关节炎模型,给药5天后收集关节液,检测关节腔积液中的白细胞数,测定关节液中炎症细胞因子白介素-6(interleukin-6,IL-6)、白介素-8(interleukin-8,IL-8)、肿瘤坏死因子-α(tumour necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)、前列腺素E2(prostaglandin E2,PGE2)含量;采血检测血尿酸值,给药前后血清中尿素(urea,UREA)、胆固醇(cholesterol,CHO)、血肌酐(serum creatinine,Cre)、血糖(glucose,GLU)及甘油三酯(triglyceride,TG)含量。结果土藤草颗粒高、中、低剂量组给药5 d后关节腔内白细胞数均有所下降,中剂量组与模型对照组比较有显著性差异(P<0.01);高、中剂量组有降低血尿酸的趋势;土藤草颗粒对家兔血清中UREA、CHO、Cre、GLU及TG含量均无影响;与模型对照组比较,土藤草颗粒高、中、低剂量组IL-6、IL-8、TNF-α及PGE2含量均明显降低,其中高、中、低剂量组的IL-6、TNF-α含量,高、中剂量组IL-8含量及中剂量组PGE2含量与之比较有显著性差异(P<0.01);土藤草颗粒高、中、低剂量组可以一定程度上降低关节液中IL-1β含量。结论土藤草颗粒对MSU致急性关节炎家兔模型有明显的治疗作用,可以减少动物关节腔内白细胞数量、抑制炎症细胞因子的表达,降低血清中尿酸含量。

能谱CT检测下肢关节周围尿酸盐结晶沉积的应用研究

目的:探讨应用能谱CT成像技术检测下肢关节周围尿酸盐(MSU)结晶沉积的临床价值。方法:选取90例拟诊断为痛风性关节炎(GA)的患者作为研究对象,均接受下肢关节能谱CT成像技术及常规实验室检查,以后者检查结果为准,分析能谱CT成像技术对GA的诊断价值。结果:应用能谱CT成像技术检查,有77例见MSU结晶沉积,其与常规实验室检查的一致性较强(Kappa=0.732)。77例GA患者中检查出膝关节44例(218处MSU结晶沉积区),足踝关节48例(162处MSU结晶沉积区)。77例GA患者重度MSU沉积组(n=11)、中度MSU沉积组(n=36)、轻度MSU沉积组(n=30)间的MSU结晶数积分和临床症状积分均有统计学差异(P<0.05),均表现为重度MSU沉积组>中度MSU沉积组>轻度MSU沉积组(P<0.05)。Pearson分析显示,GA患者MSU结晶计数与临床症状正相关(r=0.739,P<0.05)。结论:能谱CT成像技术对下肢关节周围MSU结晶沉积有较高显示率,在GA早期诊断中具有重要价值。

莱菔硫烷降低尿酸钠诱导成骨细胞的炎症和氧化应激反应

目的观察尿酸钠盐(monosodium urate,MSU)对成骨细胞的损伤,并探讨莱菔硫烷(sulforaphane,SFN)对MSU诱导的成骨细胞损伤的机制及治疗作用。方法用CCK-8试剂盒检测成骨细胞活力,用流式细胞仪检测成骨细胞凋亡,用ROS敏感荧光探针测定成骨细胞ROS水平。在成骨细胞培养基中分别加入MSU(10、50、100、300、500μg/mL)和SFN(1、5、10、20、50μmol/L)后0、12、24、48、72 h分别检测,确定药物的作用浓度和作用时间,用蛋白质印迹法检测成骨细胞Nrf2、HO-1、HIF-1、IL-6和caspase-3。结果MSU对成骨细胞的毒性作用随着加入作用浓度和作用时间的增加而增加。当MSU作用浓度为300μg/mL,作用时间超过48 h,细胞活力低于50%。当SFN在作用浓度为1μmol/L和10μmol/L时对成骨细胞的活力没有影响,但作用浓度为20μmol/L和50μmol/L时细胞活力明显降低。确定MSU作用浓度为300μg/mL,SFN作用浓度为10μmol/L,作用时间为48 h。MSU增加了成骨细胞内的ROS水平,SFN降低了MSU增加的成骨细胞内ROS水平和凋亡率。SFN降低了MSU诱导的成骨细胞的HIF-1、IL-6和caspase-3的表达水平,但增加了Nrf2、HO-1的表达水平。结论SFN可减少MSU对成骨细胞的损伤。

痛风频发的有效预测因素——尿酸盐沉积负荷

目的探讨影响痛风频繁发作的危险因素。方法收集2019年12月—2020年12月南京医科大学第一附属医院风湿免疫科就诊的痛风患者的基线临床资料,同时行发作关节的双能CT检查。临床随访患者1年,患者通过网络在线填写每次痛风发作的时间、部位及临床表现。依据患者痛风是否频发,将患者分为痛风非频发组(发作次数<2次)和痛风频发组(发作次数≥2次)。探讨关节内单钠尿酸盐晶体沉积体积与痛风再发的关系。结果129例痛风患者纳入本研究,其中118例患者完成1年随访,11例患者失访。与痛风非频发组相比,痛风频发组患者的病程更长,合并高血压患者占比和血肌酐水平更高,发作关节内尿酸盐沉积体积更大,差异有统计学意义(P<0.05)。关节内尿酸盐沉积体积大是痛风频发的独立危险因素,尿酸盐沉积体积每增加1 cm3,痛风频繁发作的风险增加2倍(OR=2.0,95%CI:1.00~3.97,P<0.05)。受试者工作特征(ROC)曲线分析显示,以基线期发作关节内尿酸盐沉积体积为0.21 cm3作为截断值,可以预测痛风频发情况,其敏感度为59%,特异度为90%。结论关节内尿酸盐沉积负荷是痛风频发的独立危险因素,以双能CT检测关节内尿酸盐沉积负荷为0.21 cm3作为标准,更易早期识别痛风频发患者。

N-9,10-蒽醌-2-甲酰基苯丙氨酸治疗大鼠痛风性关节炎的作用及机制

目的探究N-9,10-蒽醌-2-甲酰基苯丙氨酸(NAY)对尿酸钠(MSU)所致痛风性关节炎(GA)大鼠的作用及机制。方法将雄性SD大鼠随机分为7组,每组8只,即正常组、模型组、NAY低(20 mg·kg^(-1))、中(40 mg·kg^(-1))、高剂量组(80 mg·kg^(-1))、秋水仙碱组(0.8 mg·kg^(-1))和别嘌呤醇组(10 mg·kg^(-1)),通过关节腔内注射MSU晶体建立大鼠急性GA模型,各组灌胃给药5 d,给药第3天造模。将原代培养的大鼠腹部巨噬细胞分成空白组、模型组、NAY组、MCC950组和联合给药组。软尺测量大鼠踝关节周长并计算大鼠关节肿胀度;检测大鼠血清尿酸(UA)、肌酐(CRE)和尿素氮(BUN)含量;HE染色观察大鼠踝关节滑膜组织病理切片;检测各组血清、关节滑膜和细胞上清液中肿瘤坏死因子-α(TNF-α)和白介素-1β(IL-1β)的含量;并检测各组关节滑膜和细胞中NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)和半胱氨酸天冬氨酸蛋白酶-1(caspase-1)的蛋白表达。结果动物实验发现,与模型组相比,NAY给药组在造模12、24和48 h均可不同程度改善GA大鼠踝关节肿胀度,且NAY各个剂量组均显著降低GA大鼠血清UA、CRE和BUN水平,同时显著降低GA大鼠血清中TNF-α和IL-1β水平及GA大鼠关节滑膜中炎症因子TNF-α和IL-1β水平,而NAY能显著降低GA大鼠关节滑膜中NLRP3、ASC和caspase-1的蛋白表达,HE染色结果显示NAY可减轻GA模型大鼠踝关节滑膜组织炎性细胞浸润,改善细胞形态。细胞实验发现,NAY、MCC950和联合给药组均可显著提高MSU诱导大鼠巨噬细胞活力,降低上清液TNF-α和IL-1β的含量,并显著降低模型组NLRP3、ASC和caspase-1蛋白表达,但是联合给药组与MCC950组相比,上述指标没有显著性差异。结论NAY可在体内和体外有效预防MSU晶体诱导的GA,机制可能与其降尿酸和抑制NLRP3炎症小体表达有关。

加味四妙方通过调控蛋白多糖降解治疗大鼠痛风性关节炎的机制研究

目的:基于蛋白多糖降解探讨加味四妙方对单钠尿酸盐(MSU)诱导的大鼠痛风性关节炎(GA)的作用及其作用机制。方法:将6只SD大鼠随机分为空白组和模型组,每组3只,模型组向关节腔内注射0.1 mL MSU溶液(100 mg/mL)建立GA模型,对照组注射等体积生理盐水。24 h后对两组大鼠的滑膜组织进行转录组测序,筛选出差异表达基因。体外培养家兔膝关节软骨细胞,甲苯胺蓝染色、RT-PCR法检测不同代数软骨细胞中蛋白多糖(PGs)的表达。采用Western blot与qRT-PCR法检测不同浓度MSU(200、500、1000µmol/L)处理后MMP-3的蛋白及mRNA表达。将33只SD大鼠随机分为空白组、模型组、给药组,每组11只。膝关节注射0.1 mL MSU溶液(100 mg/mL)制备大鼠痛风性关节炎模型,免疫组化检测滑膜组织中MMP-3的表达,番红固绿染色法检测大鼠关节软骨细胞蛋白多糖的表达。结果:MMP-3是显著上调的差异表达基因,与Jak/STAT信号通路、TGF-β信号通路、趋化因子信号通路、IL-17信号通路等密切相关。体外验证发现,与空白组比较,经250、500、1000µmol/L MSU刺激24 h后,MMP-3蛋白表达水平明显升高(P<0.05);与空白组比较,MMP-3 mRNA表达水平经250µmol/L MSU刺激后明显升高(P<0.01),经500、1000µmol/L MSU刺激后其表达显著升高(P<0.001)。与空白组比较,经250、500µmol/L MSU刺激后,各组软骨细胞蛋白多糖表达均显著降低(P<0.05);与250µmol/L MSU组比较,10%和20%加味四妙方含药血清(MSM)组均能有效逆转上述结果(P<0.001)。与500µmol/L MSU组比较,20%MSM能够有效上调蛋白多糖的表达(P<0.05)。与空白组比较,模型组软骨细胞蛋白多糖mRNA表达显著降低(P<0.001);与模型组相比,10%MSM与20%MSM均能有效增加其mRNA表达(P<0.001,P<0.05)。与空白组比较,模型组大鼠膝关节肿胀明显(P<0.05,P<0.01);与模型组比较,各给药组能明显抑制其关节肿胀(P<0.05,P<0.01)。与空白组比较,模型组MMP-3表达增加且关节软骨表层蛋白多糖丢失;与模型组相比,各给药组对上述现象均有明显改善作用。结论:加味四妙方通过抑制MMP-3生成调控蛋白多糖降解从而治疗痛风性关节炎。

瑞巴派特对MSU诱导的腹膜炎腹腔巨噬细胞极化表型的影响

目的 观察瑞巴派特对单钠尿酸盐(MSU)诱导的小鼠痛风性急性腹膜炎腹腔巨噬细胞极化表型及细胞因子表达的影响。方法 18只雄性BALB/c小鼠随机分为对照组(MSU组,生理盐水0.2 mL/10 g灌胃)、秋水仙碱组(0.75 mg/kg的秋水仙碱0.2 mL/10 g灌胃)、瑞巴派特组(150 mg/kg的瑞巴派特0.2 ml/10 g灌胃),每组6只,灌胃给药2次后3组小鼠均以MSU腹腔注射1次制备急性腹膜炎模型,16 h后,收集腹腔巨噬细胞,分析巨噬细胞表型;检测腹腔灌洗液上清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和IL-10细胞因子;腹膜组织苏木素-伊红(HE)染色分析炎性细胞浸润情况。结果 秋水仙碱组和瑞巴派特组M2型标志CD206阳性比例分别为(19.36±2.80)%和(21.22±1.22)%,均显著高于MSU组的(15.80±1.32)%,差异均有统计学意义(P均<0.05)。秋水仙碱组、瑞巴派特组腹腔灌洗液上清促炎因子TNF-α、IL-6浓度分别为(138.73±14.06)、(122.89±6.66)pg/mL,(119.91±4.99)、(115.32±6.73)pg/mL,较MSU组的(238.89±16.16)、(272.99±11.35)pg/mL显著降低,差异均有统计学意义(P均<0.05);秋水仙碱组、瑞巴派特组抗炎因子IL-10的浓度分别为(73.11±4.77)、(70.69±3.72)pg/mL,较MSU组的(32.23±3.51)pg/mL显著升高,差异均有统计学意义(P均<0.05)。小鼠腹膜组织HE染色结果显示,MSU组腹膜组织松散,细胞排列紊乱,可见大量的核大、深染、无胞浆的炎症细胞浸润;秋水仙碱组和瑞巴派特组细胞水肿,炎症细胞浸润较MSU组少。结论 瑞巴派特通过增加腹腔巨噬细胞增加M2极化,减轻MSU诱导的小鼠痛风性腹膜炎的炎性反应。

TLR2/TLR4-NF-κB信号通路在痛风中的研究进展

痛风是由于血尿酸浓度升高,尿酸钠晶体在关节、肌腱和周围组织中沉积,从而导致炎症性关节炎间歇性发作。Toll样受体是先天免疫系统中模式识别受体的一种。痛风性关节炎是由于过饱和的尿酸钠晶体在血液中析出,并与巨噬细胞产生作用,导致中性粒细胞的趋化、聚集,从而激活TLR2/TLR4-NF-κB信号通路,释放IL-1β、IL-6、TNF-α等炎性因子和蛋白酶,导致痛风性关节炎的发生、发展。该文以痛风及TLR2/TLR4-NF-κB信号通路为主线,综述了TLR2/TLR4-NF-κB信号通路与痛风的相关性以及以该信号通路为靶点的治疗方法。

双能CT在诊断痛风心血管损害中的应用价值

痛风是由尿酸钠结晶沉积在关节或其他组织中引起的一种常见的疾病,其发病率和患病率呈逐年上升趋势,高尿酸血症是发展为痛风最重要的危险因素。近年来许多流行病学和实验研究均证实痛风与高血压、冠心病和心肌梗死等心血管疾病的发生和死亡密切相关。而双能CT作为一种新型的无创性影像学检查手段,可以特异性并定量显示尿酸盐结晶,近年来已成为诊断痛风的一个有效工具,有研究发现其在痛风心血管损害的诊断中也有着很好的临床价值。该综述分析痛风与心血管疾病之间复杂的相关性,以及双能CT在诊断痛风心血管损害中的优势和应用价值。

粉萆薢降尿酸作用研究

目的研究粉萆薢的尿酸清除作用。方法①将70只小鼠随机分成7组,即空白对照组,模型组,苯溴马龙组,粉萆薢醇提物高、低剂量组和粉萆薢水提物高、低剂量组,每组10只。空白对照组和模型组均灌服0.02mL.g-10.9%氯化钠溶液,其他5组分别灌服苯溴马龙混悬液0.03 g.kg-1、粉萆薢水提物生药10和20 g.kg-1、粉萆薢醇提物生药10和20 g.kg-1。每日8:00和16:00各给药1次,连续5 d。除空白对照组外,其他各组均于最后一次给药后0.5 h腹腔注射尿酸钠混悬液制作高尿酸血症模型。并于造模后30,60和120 m in取各组小鼠摘眼球取血,离心分离血清,用尿酸测定试剂盒测定血清中尿酸的量;②将70只大鼠随机分成7组,每组10只,分组方法同上,空白对照组和模型组均灌服0.9%氯化钠溶液4 mL,其他各组给药量及测定方法同上。测定大鼠血清尿酸浓度。结果高剂量粉萆薢水提物能显著降低小鼠和大鼠血清尿酸含量,其作用效果和阳性药苯溴马龙相当。结论粉萆薢具有一定的抗痛风作用。

痛风发病机制研究进展

痛风是一种由于单钠尿酸盐(MSU)晶体与组织微环境相互作用导致的反复发作的急性炎性反应。MSU晶体启动机体的炎性级联反应涉及多种信号通路,其中包括MSU晶体直接与细胞表面Toll样受体的结合,MSU晶体与细胞膜表面脂类分子的结合以及MSU晶体以内吞的方式进入细胞。NLRP3炎性小体的活化处于炎性反应的中心环节。

急性痛风性关节炎的发病机制

急性痛风性关节炎(acute gouty arthritis,AGA)的发病机制复杂,从尿酸升高,引发尿酸盐沉积在关节或肌体其他部位,继而引起炎症来阐述急性痛风性关节炎的发病机制。

巨噬细胞诱导极化对痛风炎症反应的影响

目的:了解体外诱导极化的M1和M2型巨噬细胞对尿酸钠结晶(MSU)诱导的急性痛风模型炎症反应的影响。方法:选用雄性BALB/c小鼠作为实验对象,随机分为空白对照组、模型组、模型M1型巨噬细胞组和模型M2型巨噬细胞组。小鼠皮下气囊注射MSU结晶诱导小鼠急性痛风模型。体外将巨噬细胞诱导极化为M1和M2型巨噬细胞,并分别注射于小鼠痛风模型皮下气囊。于炎症诱导后4 h、12 h和24 h检测不同组别皮下气囊浸润的炎性细胞数量变化,并应用ELISA法测定囊内灌洗液中IL-1β、TGF-β的水平。结果:1在4 h、12 h和24 h时间点模型组、模型M1型巨噬细胞组和模型M2型巨噬细胞组皮下气囊炎性细胞均高于空白对照组(P<0.01)。在4 h时间点,模型组及模型M1型巨噬细胞组和模型M2型巨噬细胞组间皮下气囊炎性细胞数量无差异(P>0.05);而在12 h和24 h,模型M1型巨噬细胞组皮下气囊炎性细胞高于模型组,而模型M2型巨噬细胞组皮下气囊炎性细胞低于模型组(P<0.05)。2在3个时间点,模型组、模型M1型巨噬细胞组及模型M2型巨噬细胞组IL-1β均高于空白对照组(P<0.01);在3个时间点比较,IL-1β水平在模型组低于模型M1型巨噬细胞组(P<0.01),而高于模型M2型巨噬细胞组(P<0.01)。TGF-β水平在模型组高于模型M1型巨噬细胞组(P<0.01),而低于模型M2型巨噬细胞组(P<0.01)。结论:巨噬细胞的极化在痛风炎症反应不同阶段起到不同的作用。