白藜芦醇抑制NLRP3炎性小体介导的细胞焦亡信号通路改善肥胖小鼠认知功能

目的 观察白藜芦醇(RES)对肥胖小鼠认知功能的保护作用,并探讨其海马组织核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎性小体—细胞焦亡信号通路机制。方法 50只小鼠随机均分为：正常对照组、模型组和白藜芦醇高(100 mg/kg)、中(50 mg/kg)、低(25 mg/kg)剂量组,喂养6个月后,检测小鼠认知功能,海马组织细胞焦亡水平,海马组织沉默信息调节因子1(SIRT1)、NLRP3、半胱氨酸天冬氨酸蛋白酶1(Caspase-1)、白细胞介素-1β(IL-1β)、IL-18和肿瘤坏死因子-α(TNF-α)等蛋白表达情况。采用单因素方差分析或重复测量方差分析进行结果分析。结果 与模型组比较,白藜芦醇高剂量组小鼠终体质量降低(t=-14.453,P=0.021),海马组织细胞焦亡水平降低(t=-17.328,P=0.011),学习记忆能力改善,海马组织SIRT1蛋白表达升高(t=32.812,P<0.001),NLRP3、Caspase-1、IL-1β、IL-18和TNF-α蛋白表达均下降(t=-28.462,P=0.003;t=-38.148,P<0.001;t=-...     

二氧化硅粉尘对J774A.1细胞NLRP3-Caspase1细胞焦亡通路的影响

目的 探讨二氧化硅粉尘对小鼠J774A.1巨噬细胞焦亡的作用及影响。方法 J774A.1细胞以DMEM（高糖）+10.0%胎牛血清置于37℃、5.0%二氧化碳浓度进行培养;二氧化硅组以终质量浓度为50μg/ml的二氧化硅粉尘悬液干预,对照组加入等体积的DMEM（高糖）培养基,各组于刺激12h后终止染尘收集细胞;Western blotting检测Nod样受体蛋白3(nod-like receptor protein 3,NLRP3)、消皮素D(gasdermin D,GSDMD)、半胱氨酸天冬氨酸特异性蛋白酶（cysteinylaspartatespecificprotease,Caspase）1、白细胞介素（interleukin,IL）-1β、IL-18蛋白表达水平;免疫荧光法检测Caspase1、TUNEL表达水平。结果 二氧化硅组NLRP3、Caspase1-p20、GSDMD-N、IL-1β、IL-18细胞白相对表达水平高于对照组,差异有统计学意义（t=3.37,P<0.01;t=6.21,P<0.001;t=2.93,P<0.05;t=2.57,P<...     

白藜芦醇减少肥胖小鼠肝细胞焦亡的初步探讨

目的 研究白藜芦醇（resveratrol,RES）减少肥胖小鼠细胞焦亡致肝损伤的机制。方法 将雄性C57BL/6小鼠随机分为标准(SCD)、高脂（HFD）及高脂+白藜芦醇（HFD+RES）饮食3组,标准或高脂饮食喂养,HFD+RES小鼠在高脂饮食的同时胃饲白藜芦醇（400 mg/（kg·d））,持续22 w。测定体重,分析血脂和interleukin-1 beta (IL-1β)、interleukin-18(IL-18)水平,HE染色检测肝脏组织的形态学变化,免疫组化检测去乙酰化酶sirtuin-1(SIRT1)、P66SHC、NLR family pyrin domain containing 3 (NLRP3)的定位和表达,实时荧光定量RT-PCR和Western blot分析SIRT1、P66SHC(66kuSrchomology2domain-containingprotein)、NLRP3、cysteinylaspartatespecificproteinase-1(caspase-1)、gasdermin-D (GSDMD)、interleukin-1 beta (IL...     

矽肺患者支气管肺泡灌洗液肺泡巨噬细胞NLRP3炎症小体表达的临床意义

收集20例不同期别矽肺患者的支气管肺泡灌洗液(BALF)和培养后的肺泡巨噬细胞(AM),用实时定量PCR法检测AM的NLRP3、ASC、Caspase-1、IL-1β及IL-18 m RNA表达水平。贰期和叁期矽肺患者NLRP3、ASC、Caspase-1炎症小体及其下游分子IL-1β及IL-18较壹期表达增高,差异有统计学意义。提示NLRP3炎症小体的活化在矽肺进行性纤维化发展中可能发挥重要作用。     

NLRP3炎症小体通路在矽肺纤维化中的作用及其作为治疗靶点的展望

吸入晶体二氧化硅颗粒可诱导矽肺, 矽肺的发生与肺部炎症和肺纤维化的发生发展密切相关。NLRP3炎症小体已被确立为一种主要的促炎受体, 用于感知环境危险信号。肺巨噬细胞吞噬二氧化硅颗粒后活化NLRP3炎症小体可能是诱导肺部氧化应激和持续炎症反应的重要机制。本文总结了NLRP3炎症小体通路在矽肺炎性反应和肺纤维化中的作用, 并对其作为矽肺治疗的潜在靶点进行分析。     

microRNA-703调控NLRP3介导的焦亡和炎症反应抑制小鼠子宫内膜异位症研究

目的：探讨microRNA-703(miR-703)调控NLRP3介导的焦亡和炎症反应影响小鼠子宫内膜异位症(EM)进展机制。方法：采用小鼠-小鼠腹腔植入方法构建子宫内膜异位症BABL/C小鼠模型。将小鼠分为EM组和对照组。免疫印迹法和酶联免疫法检测两组焦亡标志蛋白(NLRP3、pro-caspase-1、caspase-1)和炎症因子表达(IL-1β、IL-18、TNF-α、IL-6)。实时荧光定量PCR检测两组miR-703含量。造模成功的小鼠分别腹腔注射miR-703 NC(EM+NC阴性组)、pre-miR-703(EM+miR-703组),检测两组小鼠子宫内膜异位组织大小,焦亡标志蛋白及炎性因子表达。结果：EM组小鼠NLRP3、caspase-1蛋白表达均高于对照组,炎症因子IL-1β、IL-18、TNF-α、IL-6水平(1.33±0.11ng/ml, 0.66±0.08ng/ml, 179.69±3.78pg/ml, 90.375±4.63pg/ml)均高于对照组(0.30±0.05ng/ml, 0.21±0.03ng/ml, 50.51±2.96pg/ml, 24.99...     

纳米二氧化硅诱导巨噬细胞焦亡对肺成纤维细胞的影响

目的 探讨纳米二氧化硅(SiNPs)诱导巨噬细胞焦亡对原代肺成纤维细胞基质金属蛋白酶及成纤维细胞向肌成纤维细胞转分化的影响。方法 构建SiNPs致巨噬细胞焦亡模型后,采用纳米高光谱定位巨噬细胞中SiNPs、光学显微镜观察巨噬细胞SiNPs暴露后形态、Western blot检测焦亡相关蛋白表达。通过胰酶和Ⅳ型胶原酶消化法提取小鼠原代肺成纤维细胞;以光学显微镜和免疫荧光技术表征并鉴定原代肺成纤维细胞;体外构建SiNPs诱导巨噬细胞焦亡与原代肺成纤维细胞共培养模型。以正常巨噬细胞上清干预原代肺成纤维细胞作为MF组,焦亡巨噬细胞上清干预原代肺成纤维细胞作为PF组,采用qRT-PCR、Western blot技术分别检测原代肺成纤维细胞中MMP2、MMP9的mRNA水平和蛋白表达,免疫荧光、免疫细胞化学染色检测肌成纤维细胞标志物α-SMA蛋白表达。结果 纳米高光谱、形态学观察和Western blot检测结果显示, SiNPs 成功诱导J774A.1 巨噬细胞发生焦亡,光学显微镜观察发现,与胰酶消化法相比,Ⅳ型胶原酶法提取小鼠原代肺成纤维细胞的纯度更高;免疫荧光结果提示Ⅳ型胶原酶法获得的成纤维细胞波形蛋白呈现典型的微丝结构,细胞骨架结构完整。qRT-PCR 结果显示,PF组MMP2 和MMP9 mRNA水平较MF组增加(P<0.05), Western blot结果显示,PF组MMP2和MMP9蛋白表达水平较MF组升高(P<0.05), 免疫荧光结果显示,与MF组相比,PF组α-SMA红色荧光强度增加,免疫细胞化学染色为强阳性。结论 SiNPs诱导巨噬细胞焦亡能够促进原代肺成纤维细胞MMP2和MMP9表达水平上调,并促进成纤维细胞向肌成纤维细胞转分化。     

P2X3受体介导的痛觉传递及其作用机制

目前,国际上认为疼痛是继呼吸、脉搏、血压、体温之后的第五个生命体征,为生命体征的重要指标。P2X3受体是ATP—门控性离子通道家族的一个亚型,在慢性疼痛的伤害性信息传递中起重要作用,由于P2X3受体的分布仅限于感觉神经元,对该通道的处理就可能提供一个镇痛作用既强又无副作用的慢性疼痛的治疗方法。     

积雪草苷对矽肺大鼠NLRP3/IL-1β/IL-18/TGF-β1信号通路的干预作用

目的 拟从NLRP3/IL-1β/IL-18/TGF-β1通路探讨积雪草苷（asiaticoside，AS）对染矽尘大鼠肺纤维的干预作用，为临床治疗提供理论依据。方法 将32只SD雄性大鼠随机分为对照组、SiO2模型组以及积雪草苷低、高剂量组，每组8只。对照组大鼠经气管灌注1 ml生理盐水，其余各组灌注1 ml SiO2（50 mg/ml）混悬液。造模后的第2天开始灌胃，AS低、高剂量组分别给予40、60 mg/kg AS对照组和模型组给予等量的生理盐水。每天1次，连续给药28 d。于灌胃第28天处死大鼠，取大鼠肺组织行HE、Masson染色观察病理变化，检测大鼠肺组织中羟脯氨酸（HYP）、白介素（IL）-1β、IL-18的含量、I型胶原蛋白（Col-I）、转化生长因子（TGF）-β1和NLRP3、 caspase-1、ASC蛋白表达水平。结果 模型组大鼠肺组织中HYP、IL-18、IL-1β含量均高于对照组（P<0.05），Col-I、TGF-β1、NLRP3、caspase-1、ASC蛋白表达水平亦较对照组明显升高（P<0.05），积雪草苷干预后上述各项指标均较模型组显著降低（P<0.05）。结论 积雪草苷可减轻肺部炎性细胞浸润和肺泡结构的损伤，其机制可能通过NLRP3/IL-1β/IL-18/TGF-β1信号通路进行干预。     

血管紧张素Ⅱ激活NLRP3炎性小体通路促进子宫内膜上皮细胞的转分化作用

目的 分析NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)炎性小体在血管紧张素Ⅱ（angiotensinⅡ,AngⅡ）诱导的子宫内膜上皮细胞（endometrial epithelial cells,EECs）转化为间充质细胞中的作用及其机制。方法 2021年2月1日—8月26日选取正常的子宫内膜,经酶消化并分离出EECs。在细胞培养液中加入浓度不等的AngⅡ作用不同时间,噻唑蓝染色分析细胞数量的改变;酶联免疫技术（enzyme linked immunosorbent assay,ELISA）检测细胞释放入培养液里的Ⅰ型、Ⅲ型胶原蛋白的含量;Western blot评估细胞表型蛋白平滑肌肌动蛋白-α(smooth muscle actin,α-SMA)和钙黏蛋白E(E-cadherin,E-cad)的水平、细胞内NLRP3、半胱氨酸蛋白酶-1(Caspase-1)和白介素-1β(interleukin-1β,IL-1β)的含量。结果 与正常组相比,培养液中加入AngⅡ的量越多,作用时间越长,细胞的数量越多,以AngⅡ（10-6mol/L）培养4...     

Modulatory Properties of Food and Nutraceutical Components Targeting NLRP3 Inflammasome Activation

Inflammasomes are key intracellular multimeric proteins able to initiate the cellular inflammatory signaling pathway. NLRP3 inflammasome represents one of the main protein complexes involved in the development of inflammatory events, and its activity has been largely demonstrated to be connected with inflammatory or autoinflammatory disorders, including diabetes, gouty arthritis, liver fibrosis, Alzheimer’s disease, respiratory syndromes, atherosclerosis, and cancer initiation. In recent years, it has been demonstrated how dietary intake and nutritional status represent important environmental elements that can modulate metabolic inflammation, since food matrices are an important source of several bioactive compounds. In this review, an updated status of knowledge regarding food bioactive compounds as NLRP3 inflammasome modulators is discussed. Several chemical classes, namely polyphenols, organosulfurs, terpenes, fatty acids, proteins, amino acids, saponins, sterols, polysaccharides, carotenoids, vitamins, and probiotics, have been shown to possess NLRP3 inflammasome-modulating activity through in vitro and in vivo assays, mainly demonstrating an anti-NLRP3 inflammasome activity. Plant foods are particularly rich in important bioactive compounds, each of them can have different effects on the pathway of inflammatory response, confirming the importance of the nutritional pattern (food model) as a whole rather than any single nutrient or functional compound.     

Interplay between Gut Microbiota and NLRP3 Inflammasome in Intracerebral Hemorrhage

The pathophysiological process of intracerebral hemorrhage (ICH) is very complex, involving various mechanisms such as apoptosis, oxidative stress and inflammation. As one of the key factors, the inflammatory response is responsible for the pathological process of acute brain injury and is associated with the prognosis of patients. Abnormal or dysregulated inflammatory responses after ICH can aggravate cell damage in the injured brain tissue. The NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome is a multiprotein complex distributed in the cytosol, which can be triggered by multiple signals. The NLRP3 inflammasome is activated after ICH, thus promoting neuroinflammation and aggravating brain edema. In addition, there is evidence that the gut microbiota is crucial in the activation of the NLRP3 inflammasome. The gut microbiota plays a key role in a variety of CNS disorders. Changes in the diversity and species of the gut microbiota affect neuroinflammation through the activation of the NLRP3 inflammasome and the release of inflammatory cytokines. In turn, the gut microbiota composition can be influenced by the activation of the NLRP3 inflammasome. Thereby, the regulation of the microbe–gut–brain axis via the NLRP3 inflammasome may serve as a novel idea for protecting against secondary brain injury (SBI) in ICH patients. Here, we review the recent evidence on the functions of the NLRP3 inflammasome and the gut microbiota in ICH, as well as their interactions, during the pathological process of ICH.     

The effect of P2X7 receptor 1513 polymorphism on susceptibility to tuberculosis: A meta-analysis

Studies of the association between the purinergic receptor P2X, ligand-gated ion channel 7 (P2X7 receptor) gene 1513A/C polymorphism and susceptibility to tuberculosis have yielded inconsistent results. We performed this meta-analysis to help clarify these inconsistencies. After systematically searching PUBMED, MEDLINE, EMBASE and ISI Web of knowledge, two of the authors independently extracted relevant data and a meta-analysis was performed by using STATA11.0 software. A total number of nine studies involving 2195 cases and 2036 controls were identified. The results indicated that P2X7 receptor gene 1513C allele (OR 1.389, 95% CI 1.161–1.660, p < 0.001) and CC genotype (1.582, 95% CI 1.129–2.217, p = 0. 012) were significantly associated with increased susceptibility to tuberculosis. Subgroup analysis indicated that this SNP greatly contributed to susceptibility to tuberculosis in Asians. The C allele of P2X7 receptor gene 1513A/C polymorphism was also associated with increased susceptibility to pulmonary tuberculosis in Asians (C vs. A: OR 1.420, 95% CI 1.163–1.733, p = 0.001; (CC + AC) vs. AA: OR 1.522, 95% CI 1.186–1.953, p = 0.001). Greater association between P2X7 receptor gene 1513A/C polymorphism and susceptibility to extra-pulmonary tuberculosis with bigger ORs were found (C vs. A, OR 2.035, 95% CI 1.236–3.352, p = 0.005; CC vs. AA, OR 3.788, 95% CI 1.434–10.009, p = 0.007; AC vs. AA, OR 2.148, 95% CI 1.252–3.684, p = 0.005; (CC + AC) vs. AA, OR 2.386, 95% CI 1.302–4.374, p = 0.005; CC vs. (AC + AA), OR 2.692, 95% CI 1.242–5.836, p = 0. 012). This meta-analysis indicates that the C allele of P2X7 receptor gene 1513A/C polymorphism is a risk factor for pulmonary tuberculosis in Asians, while not in Africans or Latinos and a risk for extra-pulmonary tuberculosis. Further well-designed, large scale studies are required to confirm this conclusion.     

Krill Oil Inhibits NLRP3 Inflammasome Activation in the Prevention of the Pathological Injuries of Diabetic Cardiomyopathy

Diabetic cardiomyopathy (DCM) is a common complication of diabetes mellitus (DM), resulting in high mortality. Myocardial fibrosis, cardiomyocyte apoptosis and inflammatory cell infiltration are hallmarks of DCM, leading to cardiac dysfunction. To date, few effective approaches have been developed for the intervention of DCM. In the present study, we investigate the effect of krill oil (KO) on the prevention of DCM using a mouse model of DM induced by streptozotocin and a high-fat diet. The diabetic mice developed pathological features, including cardiac fibrosis, apoptosis and inflammatory cell infiltration, the effects of which were remarkably prevented by KO. Mechanistically, KO reversed the DM-induced cardiac expression of profibrotic and proinflammatory genes and attenuated DM-enhanced cardiac oxidative stress. Notably, KO exhibited a potent inhibitory effect on NLR family pyrin domain containing 3 (NLRP3) inflammasome that plays an important role in DCM. Further investigation showed that KO significantly upregulated the expression of Sirtuin 3 (SIRT3) and peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α), which are negative regulators of NLRP3. The present study reports for the first time the preventive effect of KO on the pathological injuries of DCM, providing SIRT3, PGC-1α and NLRP3 as molecular targets of KO. This work suggests that KO supplementation may be a viable approach in clinical prevention of DCM.     

IL-17、TLR4、P2X7基因多态与慢性阻塞性肺疾病的关联研究

目的 探讨炎症相关基因白细胞介素17（interleukin-17,IL-17）、Toll样受体4（toll-like receptors 4,TLR4）、嘌呤受体（purinergic receptor P2X 7,P2X7）遗传多态与慢性阻塞性肺疾病（chronic obstructive pulmonary disease,COPD）易感性的关系。方法 采用病例对照研究设计,病例组为2015年6月~2016年5月南通市第三人民医院收集的COPD确诊患者,共152例。对照组来自参加健康体检的居民,按年龄和性别与病例进行频数匹配,共201例。基因分型采用TaqMan分型技术,关联强度采用OR值及95%CI值表示。结果 采用Bonferroni校正后,IL-17基因rs2275913、rs763780;TLR4基因rs10759932、rs2737190;P2X7基因rs1718119遗传多态与COPD的易感性关联均有统计学意义（均有P<0.05）。rs2275913A等位基因（OR=0.62,95%CI：0.46~0.86,P=0.003）;rs763780C等位基因（OR=1.96,95%CI：1.29~2.98,P=0.001）;rs10759932C等位基因（OR=0.49,95%CI：0.34~0.73,P<0.001）;rs2737190G等位基因（OR=0.51,95%CI：0.37~0.71,P<0.001）。结论 IL-17、TLR4、P2X7基因多态影响COPD遗传易感性。     

Caffeine Has Different Immunomodulatory Effect on the Cytokine Expression and NLRP3 Inflammasome Function in Various Human Macrophage Subpopulations

Besides its well-known psychoactive effects, caffeine has a broad range of actions. It regulates several physiological mechanisms as well as modulates both native and adaptive immune responses by various ways. Although caffeine is assumed to be a negative regulator of inflammation, the effect on the secretion of pro- and anti-inflammatory cytokines is highly controversial. Macrophages are major mediators of inflammatory responses; however, the various subpopulations develop different effects ranging from the initiation to the resolution of inflammation. Here we report a comparative analysis of the effect of caffeine on two subpopulations of human monocyte-derived macrophages differentiated in the presence of macrophage colony-stimulating factor (M-CSF) or granulocyte-macrophage colony-stimulating factor (GM-CSF), resulting in M-MΦs and GM-MΦs, respectively. We showed that although TNF-α secretion was downregulated in both LPS-activated MΦ subtypes by caffeine, the secretion of IL-8, IL-6, and IL-1β as well as the expression of Nod-like receptors was enhanced in M-MΦs, while it did not change in GM-MΦs. We showed that caffeine (1) altered adenosine receptor expression, (2) changed Akt/AMPK/mTOR signaling pathways, and (3) inhibited STAT1/IL-10 signaling axis in M-MΦs. We hypothesized that these alterations play an important modulatory role in the upregulation of NLRP3 inflammasome-mediated IL-1β secretion in LPS-activated M-MΦs following caffeine treatment.     

Cyanidin-3-O-glucoside and Peonidin-3-O-glucoside-Rich Fraction of Black Rice Germ and Bran Suppresses Inflammatory Responses from SARS-CoV-2 Spike Glycoprotein S1-Induction In Vitro in A549 Lung Cells and THP-1 Macrophages via Inhibition of the NLRP3 Inflammasome Pathway

Black rice is a functional food that is high in anthocyanin content, primarily C3G and P3G. It possesses nutraceutical properties that exhibit a range of beneficial effects on human health. Currently, the spike glycoprotein S1 subunit of SARS-CoV-2 (SP) has been reported for its contribution to pathological inflammatory responses in targeting lung tissue and innate immune cells during COVID-19 infection and in the long-COVID phenomenon. Our objectives focused on the health benefits of the C3G and P3G-rich fraction of black rice germ and bran (BR extract) on the inhibition of inflammatory responses induced by SP, as well as the inhibition of NF-kB activation and the NLRP3 inflammasome pathway in an in vitro model. In this study, BR extract was identified for its active anthocyanins, C3G and P3G, using the HPLC technique. A549-lung cells and differentiated THP-1 macrophages were treated with BR extract, C3G, or P3G prior to exposure to 100 ng/mL of SP. Their anti-inflammatory properties were then determined. BR extract at concentrations of 12.5–100 μg/mL exhibited anti-inflammation activity for both A549 and THP-1 cells through the significant suppression of NLRP3, IL-1β, and IL-18 inflammatory gene expressions and IL-6, IL-1β, and IL-18 cytokine secretions in a dose-dependent manner (p < 0.05). It was determined that both cell lines, C3G and P3G (at 1.25–10 μg/mL), were compatibly responsible for the significant inhibition of SP-induced inflammatory responses for both gene and protein levels (p < 0.05). With regard to the anti-inflammation mechanism, BR extract, C3G, and P3G could attenuate SP-induced inflammation via counteraction with NF-kB activation and downregulation of the inflammasome-dependent inflammatory pathway proteins (NLRP3, ASC, and capase-1). Overall, the protective effects of anthocyanins obtained from black rice germ and bran can be employed in potentially preventive strategies that use pigmented rice against the long-term sequelae of COVID-19 infection.