Function and mechanism of the pyrin inflammasome

Pyrin, encoded by the MEFV gene, is an intracellular pattern recognition receptor that assembles inflammasome complexes in response to pathogen infections. Mutations in the MEFV gene have been linked to autoinflammatory diseases such as familial Mediterranean fever (FMF) or pyrin‐associated autoinflammation with neutrophilic dermatosis (PAAND). Recent insights have now revealed how pyrin is activated during infection, providing a molecular basis for the understanding of such disease‐causing mutations in pyrin. Interestingly, pyrin does not directly recognize molecular patterns (pathogen‐ or host‐derived danger molecules), but rather responds to disturbances in cytoplasmic homeostasis caused by the infection. In the case of pyrin, these perturbations, recently defined as ‘homeostasis‐altering molecular processes’ (HAMPs), are processes leading to the inactivation of the RhoA GTPase. This review attempts to combine early observation and findings with the most recent discoveries on how pyrin detects inactivation of RhoA to shed light on the function and mechanism of pyrin activation.     

细胞焦亡在变应性鼻炎中的作用机制及研究进展

变应性鼻炎(AR)是由环境和遗传因素共同作用导致的鼻黏膜变应性炎症疾病,目前AR的发病率呈现每年不断升高的趋势,给人们的身心健康、生活等造成严重的影响,其主要的治疗手段包括避免接触过敏原、药物治疗和免疫治疗等,但是部分患者治疗效果并不理想,究其原因主要与其发病机制复杂多样有关。细胞焦亡是程序性细胞死亡的一种,近年来研究表明细胞焦亡通过多种途径参与了AR的发生发展,部分药物可以通过抑制细胞焦亡来治疗AR。因此,深入探索细胞焦亡对AR的调控机制可能为AR的治疗提供新思路。论文就细胞焦亡在AR中的作用机制及研究进展进行综述。     

AIM2 inflammasome activation may mediate high mobility group box 1 release in murine allergic rhinitis

IntroductionHigh mobility group box 1 protein participates in the pathogenesis of allergic rhinitis. Activation of the inflammasome can mediate the release of high mobility group box 1. The role of the absent in melanoma 2 inflammasome in allergic rhinitis remains unclear.ObjectiveThis study aimed to investigate the function of absent in melanoma 2 inflammasome in murine allergic rhinitis and the interaction between high mobility group box 1 and the absent in melanoma 2 inflammasome.MethodsA murine allergic rhinitis model was established using twenty Balb/c mice. Expression of the components of the absent in melanoma 2 inflammasome: absent in melanoma 2, apoptosis-associated speck-like protein containing a CARD (Asc), caspase-1 p20, and additional nod-like receptor family pyrin domain containing 3 (Nlrp3) were detected by western blotting during allergic rhinitis. Alterations of absent in melanoma 2, caspase-1, and high mobility group box 1 after ovalbumin challenge were demonstrated by immunohistochemistry. TdT-mediated dUTP Nick end labeling, TUNEL assay, and cleavage of caspase-3 and PARP-1 were used for the observation of pyroptosis.ResultsEosinophilia and goblet cell infiltration were observed in the nasal mucosa of mice in the allergic rhinitis group. Absent in melanoma 2, Asc, and caspase-1 p20 increased after ovalbumin exposure while Nlrp3 did not. High mobility group box 1 was released in the nasal mucosa of allergic rhinitis mice. TUNEL-positive cells increased in the epithelium and laminae propria, whereas cleavage of caspase-3 and PARP-1 was not observed.ConclusionsThe absent in melanoma 2 inflammasome was activated and pyroptosis may occur in the nasal mucosa after ovalbumin treatment. These may contribute to the translocation of high mobility group box 1 and the development of allergic rhinitis.     

细胞焦亡在肝病中的作用及中医药治疗的研究进展

细胞焦亡是一种与炎症相关的新型程序性细胞死亡,其发生与炎症小体密不可分,作为天然免疫系统的组成部分,是机体受到异常刺激后免疫系统清除有害因子进行组织修复而产生的防御反应。炎症反应可以提高机体清除损伤因子的能力,但过度活化则可能会对机体造成损害。随着研究逐步深入,细胞焦亡被认为与肝脏疾病的发生、发展密切相关。中医防治肝脏疾病由来已久,在临床中已经取得良好的疗效。研究显示,中药通过调控细胞焦亡相关通路蛋白与炎症因子,从而干预疾病的进展。现结合现代医学,对细胞焦亡在肝脏疾病中的作用机制进行归纳与总结,并对现阶段的中医治疗进展加以阐述。     

罗汉果醇对脂多糖诱导急性肺损伤的作用及机制研究北大核心CSCD

目的通过脂多糖刺激小鼠以及人单核细胞THP-1诱导的急性炎症模型, 探索罗汉果醇促进AMPK磷酸化后靶向调控炎症通路的作用以及相关机制, 为罗汉果醇(MO)能否对急性肺损伤的临床治疗中得到应用提供研究证据。方法动物实验:取用24只6~8周龄清洁级C57BL/6雄性小鼠, 采用随机(随机数字法)法将小鼠分为对照组、罗汉果醇(MO)组、脂多糖组、脂多糖+MO组, 每组各分配6只。对照组小鼠腹腔注射生理盐水(30 mL/kg),MO组小鼠腹腔注射MO(30 mg/kg),脂多糖组小鼠腹腔注射脂多糖(10 mg/kg), 脂多糖+MO组小鼠腹腔注射MO(30 mg/kg), 30 min后另一侧腹腔注射脂多糖(10 mg/kg)。经过12 h后, 处死小鼠取材, 并进行病理学、分子生物学检测。细胞实验:取用状态良好的THP-1细胞用含10%胎牛血清的RPMI1640培养基中培养24 h后, 用100 ng/mL PMA诱导分化成巨噬细胞, 设置对照组、罗汉果醇(MO)组、脂多糖组、脂多糖+MO组。药物刺激结束后收集各组细胞悬液, 所得细胞及培养基上清液用于后续检测。结果与对照组相比, 脂多糖组的损伤程度明显, 组织切片的H&E染色结果中可见肺泡腔结构破坏, 炎症细胞浸润增多, 且有出血以及肺泡腔间隔明显增厚;在加入MO干预下, 小鼠肺组织的损伤程度大幅度改善, MPO、肺湿/干重比等也有了显著的降低。肺组织中炎症因子IL-1β、IL-6、TNF-α的mRNA水平也同样在MO的干预下显著下调[(2.96±0.10)vs. (5.53±0.14), (8.62±0.17)vs. (12.31±0.09), (3.01±0.09)vs.(4.85±0.36)]。在脂多糖组小鼠肺组织中炎性小体的主要成分蛋白NLRP3、caspase-1 p20、GSDMD-N、ASC表达量显著高于对照组,而脂多糖+MO组中AMPK磷酸化水平提高, 焦亡相关蛋白的表达均得到有效地抑制[(0.58±0.09)vs.(0.89±0.15)、(0.19±0.08)vs. (0.93±0.16)、(0.65±0.09)vs. (0.86±0.14)、(0.30±0.12)vs. (0.47±0.10), 均P<0.05];同时通过ELISA法测得在组织中焦亡的主要标志物炎症因子IL-1β、IL-18的分泌同样可以被MO缓解。而细胞实验中MO同样促进了AMPK的磷酸化, 抑制NLRP3炎症小体相关成分蛋白表达, 同时明显提高了细胞活力。结论 MO通过促进AMPK的磷酸化抑制NLRP3介导的细胞焦亡, 进而缓解脂多糖诱导的急性肺损伤。     

氧糖剥夺/复氧所致细胞焦亡在肺微血管内皮细胞损伤中的作用

目的:探讨氧糖剥夺/复氧(OGD/R)是否导致人肺微血管内皮细胞(HPMVECs)焦亡及其在细胞损伤中的作用.方法:采用HPMVECs复制OGD/R细胞模型,模拟体外循环中HPMVECs缺血/再灌注过程.将细胞分为对照(control)组(正常培养)、OGD/R组(OGD 8 h+恢复12 h)及VX-765(casp...     

清肺通络方通过胱天蛋白酶-1/Gasdermin D蛋白细胞焦亡途径对支原体肺炎小鼠的干预机制

目的:探讨清肺通络方通过胱天蛋白酶-1(Caspase-1)/Gasdermin D(GSDMD)蛋白细胞焦亡途径对支原体肺炎小鼠干预机制。方法:将30只BALB/c小鼠随机分为空白对照组、模型组、清肺通络方组,每组10只。采用肺炎支原体菌株滴鼻法建立支原体肺炎小鼠模型。造模同时予灌胃治疗,连续治疗3 d,实验第4天处死各组小鼠。PCR检测小鼠肺组织MP-DNA含量,HE染色观察小鼠肺组织病理改变,Western blot检测胱天蛋白酶-1前体(pro-Caspase-1)、Caspase-1、GSDMD、Gasdermin D氮端活性结构域多肽(GSDMD-N)表达,ELISA检测肺泡灌洗液中白介素-1β(IL-1β)、白介素-18(IL-18)含量,生化法检测肺组织细胞乳酸脱氢酶(LDH)和Na⁺-K⁺-ATP酶活性。结果:镜下观察模型组小鼠肺间质出现炎性细胞浸润、渗出明显。与空白对照组比较,模型组小鼠pro-Caspase-1、Caspase-1、GSDMD、GSDMD-N表达及IL-1β、IL-18含量均升高,LDH活性上升,Na     

中医药抑制细胞焦亡治疗脓毒症的研究进展与探析

脓毒症（sepsis）是一种与炎症相关的高死亡率疾病，目前临床的治疗手段多围绕抗感染、液体复苏及使用血管活性药物进行，但疗效有限，仍需进行机制研究以开发新疗法。细胞焦亡（pyroptosis）是一种受多靶点调控的细胞炎性程序性死亡模式，与机体炎症的调控密切相关。大量证据表明抑制细胞焦亡可以显著控制脓毒症发展、改善脓毒症造成的器官功能损伤。中医药具有多靶点、多通路治疗疾病的优势，近些年的研究显示中医药在通过抑制细胞焦亡多种途径的相关靶点治疗脓毒症方面可以发挥较好作用。本文旨在对近几年中医药抑制细胞焦亡多种途径治疗脓毒症的研究成果进行综述与探析，以期为未来探索中医药治疗脓毒症新型治疗策略的研究开展提供理论依据。     

Nanoemulsion-based mucosal adjuvant induces apoptosis in human epithelial cells

Nanoemulsions (NEs) are adjuvants that enhance antigen penetration of the nasal mucosa, increase cellular uptake of antigens by both epithelial and dendritic cells, and promote the migration of antigen-loaded dendritic cells to regional lymph nodes within 24-h of vaccine administration. The objective of this study was to elucidate cell death caused by W₈₀5EC NE and identify caspases and genes associated with death pathways. Consistent with this aim, we show that exposure of human epithelial cells (EC), both RPMI 2650 and FaDu, to NE results in the activation of caspases (1, 3/7, 6, 8, and 9) and the expression of genes involved in apoptotic as well as authophagy and necrosis pathways. Interestingly, the NE activates caspase 8 which promotes “immunogenic apoptosis”. The rescue assay was employed to investigate the fate of RPMI 2650 cells treated with W₈₀5EC NE. After four-hour treatment with as little as 0.03% of NE no cells were rescued at 72 h. Remarkably, immediately after four-hour treatment, the cells morphologically resembled untreated cells and most of the cells were alive. Altogether, these results suggest that NE induces death of human ECs through multiple pathways. Epithelial cell death caused by W₈₀5EC may have further implications on antigen uptake, processing, and presentation by DC's.     

煤焦沥青烟提取物诱导人支气管上皮细胞发生炎性凋亡的研究

目的 探讨煤焦沥青烟提取物(coal tar pitch smoke extract,CTP)刺激人支气管上皮细胞(BEAS-2B)是否发生炎性凋亡(pyroptosis).方法 用l、3μg/ml的CTP分别染毒BEAS-2B细胞8、24 h,以二甲基亚砜(DMSO)为对照,用乳酸脱氢酶(lactic dehydrogenase,LDH)活力测定试剂盒和白细胞介素-1β(Interleukin-1 beta,IL-1β) ELISA试剂盒分别检测细胞培养上清中的LDH活力和IL-1β含量,用半胱氨酸天冬氨酸特异性蛋白酶-1(caspase-1)活力测定试剂盒检测细胞caspase-1活力.结果 染毒8h,1、3 μg/ml CTP染毒组细胞中caspase-1的活力分别为(9.29±0.30) μmol/ml和(8.67±0.59) μmol/ml,均明显高于DMSO对照组[(7.42±0.59) μmol/ml],差异有统计学意义(P＜0.05);1、3μg/mlCTP染毒组LDH活力和IL-1β含量与DMSO对照组的差异无统计学意义(P＞0.05).染毒24 h时,1、3μg/ml染毒组与DMSO对照组细胞中caspase-1活力差异无统计学意义(P＞0.05);1、3μg/ml染毒组培养液上清中LDH活力[(1323.03±28.53)、(1148.45±16.42) U/dl]和IL-1β含量[(125.37±25.00)、(92.04±19.09) pg/m]均高于对照组[LDH:(1091.93±26.64) U/dl,IL-1β:(46.20±14.43) pg/ml],差异有统计学意义(P＜0.05).结论 煤焦沥青烟提取物刺激BEAS-2B细胞早期caspase-1活力升高,继之LDH酶活力及IL-1β含量均增高,可能发生pyroptosis.