骨关节炎基于细胞焦亡机制的治疗手段研究进展

骨关节炎（OA）是一种与炎症、软骨磨损和衰老相关的疾病,目前的治疗手段仍以改善临床症状,延缓疾病进展为主。挖掘OA发病的分子学机制是寻找治疗方法的有效手段,近年的研究证实细胞焦亡参与OA的病理发展,缓解或者抑制细胞焦亡是OA治疗的新靶点。现基于OA细胞焦亡机制的相关研究已陆续展开,传统中药、现代医学治疗药物及某些非药物治疗方式均被证实可通过调控滑膜细胞焦亡或者软骨细胞焦亡而发挥作用,但具体的分子学机制及影响因素仍不明确。深入研究细胞焦亡可能是未来防治OA的重要方向,基于此,本文拟对目前已发表的治疗手段进行综述,以期为OA细胞焦亡研究提供参考思路。     

细胞焦亡在眼科相关疾病的研究进展

细胞焦亡是一种新的程序性细胞死亡方式,又称为细胞炎性坏死。炎性小体复合物在细胞内激活后,活化含半胱氨酸的天冬氨酸蛋白水解酶(Caspase),再通过Caspase介导Gasdermin家族蛋白D(GSDMD)的激活,使细胞膜完整性受到破坏,形成焦亡小体,最终诱发细胞焦亡。目前,很多研究认为细胞焦亡广泛参与了肿瘤、心血管以及自身免疫性疾病等的发生、发展过程,并且随着对细胞焦亡更深入的研究发现其也参与了多种眼科疾病的病变过程。本文就细胞焦亡的主要机制及其在糖尿病视网膜病变、年龄相关性黄斑变性、白内障等相关眼科学疾病中的最新研究进行综述。     

细胞焦亡在肺部疾病中的研究进展

细胞焦亡一直以来被看作是引起肺损害的潜在机理之一,特征为细胞不断膨胀以致胞膜破裂,从而造成胞内容物释放到外环境中,释放的炎性因子激活免疫途径刺激机体更强烈的炎症反应。焦亡过程中需要依赖特定半胱氨酸蛋白酶(Caspase)的活化和相应Gasdermin蛋白的激活,同时涵盖众多炎性因子的释放启动,将细胞焦亡与其他程序性死亡方式区分开来。本文通过对细胞焦亡在常见肺部疾病中的功能及机理作出了综述,有助于深入了解其与常见肺部疾病发病机制与转归关系,以期为肺部疾病的诊疗方法提供全新的临床思路。     

GSDMD介导的细胞焦亡在呼吸系统疾病中研究进展

细胞焦亡是一类新发现的程序性细胞死亡方式,由炎性半胱氨酸蛋白酶介导发生快速细胞膜破裂、细胞崩解,并释放细胞内容物和IL-18、IL-1β等大量炎性因子参与细胞死亡。GSDMD蛋白是细胞焦亡的最终执行蛋白,通过介导经典、非经典细胞焦亡途径发挥作用。在多种呼吸系统疾病中,均发现有GSDMD表达水平升高,提示GSDMD介导的细胞焦亡,可能成为呼吸系统疾病的潜在诊疗靶点,本文章就GSDMD介导的细胞焦亡在呼吸系统疾病中作用的研究进展进行综述。     

细胞焦亡与肺癌研究进展

细胞焦亡是由炎症小体引发的细胞炎症性坏死形式，其特点是半胱氨酸蛋白酶介导促炎因子和 Gasdermin 家族蛋白活化，导致膜孔形成、细胞肿胀、质膜裂解、染色质碎裂和细胞内促炎内容物释放。炎症小体存在于多种肿瘤细胞，可以通过介导细胞焦亡影响肿瘤的增殖、侵袭和转移。研究发现细胞焦亡与肺癌发病、进展、预后及耐药的发生密切相关，归纳总结细胞焦亡的分子机制及其在肺癌治疗中的研究进展，有利于为肺癌的诊疗和新药研发提供新的依据。     

细胞焦亡与肺动脉高压的关系及治疗肺动脉高压药物研究进展

细胞焦亡(pyroptosis)是指伴有炎性反应的一种细胞程序性死亡方式,其广泛参与了多种疾病的发生发展,例如感染性疾病、心血管疾病和神经性退行等。研究表明,细胞焦亡参与了心血管疾病中动脉性肺动脉高压(pulmonary arterial hypertension, PAH)的发病机制。PAH患者的肺部存在血管周围炎症浸润,肺血管病变存在于极度发炎的微环境里,细胞焦亡中的促炎因子驱动了PAH患者的肺部血管重塑。本文针对细胞焦亡在PAH发病机制中的作用及治疗PAH药物的相关研究进行综述,旨在为临床上治疗PAH提供新思路。     

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。     

细胞焦亡在原发性肝癌进展中的作用研究进展

细胞焦亡是近年来证实的一种新的程序性细胞死亡方式,由消化道皮肤素介导,表现为细胞不断胀大直至细胞膜破裂,导致细胞内容物的释放进而激活强烈的炎症反应,导致细胞程序性坏死。细胞焦亡的主要通路包含依赖半胱天冬氨酸蛋白酶（caspase）-1的经典通路和依赖caspase-4、5、11的非经典通路。细胞焦亡在原发性肝癌的癌前病变以及发展过程中起重要作用,归纳总结了细胞焦亡的分子机制及其在原发性肝癌治疗中的研究进展,以期为原发性肝癌的诊断、治疗以及新药研发提供新的依据。     

细胞焦亡的研究进展

摘要： 细胞焦亡（pyroptosis）是促炎形式的程序性细胞死亡, 并且依赖于半胱氨酸天冬氨酸特异性蛋白酶（caspases） 活性。由caspases通过切割gasermin D （GSDMD） 的氨基端和羧基端的连接体调控, 后者移位到膜上并穿孔, 诱导水分渗透, 细胞肿胀并释放炎性因子, 继而发生细胞焦亡。细胞焦亡的形态学特征、 发生及调控机制等均不同于凋亡、 坏死等其他细胞死亡方式, 其是机体一种重要的天然免疫反应, 在抗击感染以及疾病中均有重要作用。通过对细胞焦亡的更深入研究, 可以认知其在相关疾病中的作用, 为临床提供新的治疗思路。     

细胞焦亡与药物治疗及免疫治疗在胶质母细胞瘤中的研究进展

细胞焦亡是一种由焦孔素家族介导的炎性细胞死亡方式,能够促进炎性介质的释放从而活化肿瘤微环境(tumor microenvironment,TME)中的NK细胞、T细胞、巨噬细胞等免疫细胞群发挥调控免疫抗肿瘤作用。胶质母细胞瘤(glioblastoma,GBM)是最严重、最恶性的胶质瘤,被诊断GBM的患者中位生存期<2年,并且由于血脑屏障的存在导致药物难以抵送至脑部从而影响药物抗GBM的作用。因此探索新的措施及机制来治疗GBM是十分重要的。GBM具有复杂的TME,其TME中存在大量的免疫细胞群,然而这些免疫细胞群往往受到GBM的影响处于免疫抑制状态。细胞焦亡作为一种能够激活免疫的细胞死亡方式,具有活化机体的免疫帮助逆转TME免疫抑制的作用。本综述以细胞焦亡为出发点阐述细胞焦亡与免疫系统之间的关系、细胞焦亡如何影响GBM的TME的免疫细胞群以及细胞焦亡途径在GBM治疗中的药物研究新进展,为未来临床治疗GBM提供新的方向和策略。     

Gasdermin D in pyroptosis

Pyroptosis is the process of inflammatory cell death. The primary function of pyroptosis is to induce strong inflammatory responses that defend the host against microbe infection. Excessive pyroptosis, however, leads to several inflammatory diseases, including sepsis and autoimmune disorders. Pyroptosis can be canonical or noncanonical. Upon microbe infection, the canonical pathway responds to pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs), while the noncanonical pathway responds to intracellular lipopolysaccharides (LPS) of Gram-negative bacteria. The last step of pyroptosis requires the cleavage of gasdermin D (GsdmD) at D275 (numbering after human GSDMD) into N- and C-termini by caspase 1 in the canonical pathway and caspase 4/5/11 (caspase 4/5 in humans, caspase 11 in mice) in the noncanonical pathway. Upon cleavage, the N-terminus of GsdmD (GsdmD-N) forms a transmembrane pore that releases cytokines such as IL-1β and IL-18 and disturbs the regulation of ions and water, eventually resulting in strong inflammation and cell death. Since GsdmD is the effector of pyroptosis, promising inhibitors of GsdmD have been developed for inflammatory diseases. This review will focus on the roles of GsdmD during pyroptosis and in diseases.     

Glycyrrhizin attenuates hepatic ischemia-reperfusion injury by suppressing HMGB1-dependent GSDMD-mediated kupffer cells pyroptosis

Gasdermin D (GSDMD), a genetic substrate for inflammatory caspases, plays a central role in pyroptosis of macrophages and release of interleukin‑1β (IL-1β), but was mainly referred to microbial infection. High mobility group box-1 (HMGB1), served as an alarm molecule during various pathological process, has been widely recognized to be involved in liver ischemia-reperfusion (I/R). Glycyrrhizin, a natural anti-inflammatory and antiviral triterpene in clinical use, was recently referred to have ability to prevent I/R induced liver injury by inhibiting HMGB1 expression and activity. However, the mechanisms responsible for damage amelioration subsequently to HMGB1 inhibition during liver I/R remain enigmatic. This study was designed to explore the functional role and molecular mechanism of glycyrrhizin in the regulation of I/R induced liver injury. We found that liver I/R promotes GSDMD-mediated pyroptotic cell death of Kupffer cells, which was inhibited by glycyrrhizin. Interestingly, endogenous HMGB1, not exogenous one, was involved in hypoxia-reoxygenation (H/R) induced pyroptosis. Moreover, GSDMD knockdown protects kupffer cells against H/R induced pyroptosis in vitro. Here, we report, for the first time, that glycyrrhizin attenuated tissue damage and kupffer cells pyroptosis during liver ischemia-reperfusion injury (LIRI) and identify a previously unrecognized HMGB1- dependent GSDMD- mediated signaling pathway in the mechanism of kupffer cells pyroptosis induced by H/R. Our findings provide the first demonstration of GSDMD-determined pyroptotic cell death responsible for I/R induced release of IL-1β and this would provide a mandate to better understand the unconventional mechanisms of cytokine release in the sterile innate immune system.     

Role of pyroptosis in liver diseases

Pyroptosis is known as a novel form of pro-inflammatory cell death program, which is exceptional from other types of cell death programs. Particularly, pyroptosis is characterized by Gasdermin family-mediated pore formation and subsequently cellular lysis, also release of several pro-inflammatory intracellular cytokines. In terms of mechanism, there are two signaling pathways involved in pyroptosis, including caspase-1, and caspase-4/5/11 mediated pathways. However, pyroptosis plays important roles in immune defense mechanisms. Recent studies have demonstrated that pyroptosis plays significant roles in the development of liver diseases. In our review, we have focused on the role of pyroptosis based on the molecular and pathophysiological mechanisms in the development of liver diseases. We have also highlighted targeting of pyroptosis for the therapeutic implications in liver diseases in the near future.     

The mechanism of Arhalofenate in alleviating hyperuricemia―Activating PPARγ thereby reducing caspase-1 activity

Hyperuricemia (HUA) is an important risk factor for renal diseases and contributes to gout. Arhalofenate (Arha) has been proved to have uricosuric activity as an inhibitor of URAT1, organic anion transporter 4 (OAT4) and OAT10. However, the effects of Arha on HUA remain unknown. The objective of this study was to investigate whether Arha could alleviate HUA and uncovered the underlying mechanism in vitro. HK-2 cells were exposed to uric acid (UA) to simulate HUA in vitro. Then cells were treated with Arha, caspase-1 inhibitor Belnacasan (Beln), caspase-11 inhibitor Wedelolactone (Wede) and PPARγ inhibitor Mifobate, respectively. The alteration of cell proliferation, inflammation, pyroptosis and expression of related proteins were detected. Results showed that UA exposure inhibited cell viability and increased IL-1β and IL-18 generation in a concentration dependent manner. Meanwhile, UA activated the cleavage of gasdermin D (GSDMD), enhanced the protein expression of URAT1, OAT4, TLR4, caspase-1, and caspase-11 and reduced PPARγ expression. While the presence of Arha or Beln enhanced cell viability and inhibited cleavage of GSDMD. Wede slightly increased cell viability but failed to prevent GSDMD cleavage. The expression of related proteins except caspase-11was also recovered by Arha. Beln and Wede partially rescued related proteins level except PPARγ compared with model group. Besides, the co-treatment of Mifobate blunted the effects of Arha on cell viability and expression of GSDMD, TLR4, and caspase-1. In conclusion, Arha inhibited UA transport as well as preventing inflammation and pyroptosis via activating PPARγ thereby blocking caspase-1 activation of HUA in vitro.     

Gasdermin D-mediated pyroptosis in myocardial ischemia and reperfusion injury: Cumulative evidence for future cardioprotective strategies

Cardiomyocyte death is one of the major mechanisms contributing to the development of myocardial infarction (MI) and myocardial ischemia/reperfusion (MI/R) injury. Due to the limited regenerative ability of cardiomyocytes, understanding the mechanisms of cardiomyocyte death is necessary. Pyroptosis, one of the regulated programmed cell death pathways, has recently been shown to play important roles in MI and MI/R injury. Pyroptosis is activated by damage-associated molecular patterns (DAMPs) that are released from damaged myocardial cells and activate the formation of an apoptosis-associated speck-like protein containing a CARD (ASC) interacting with NACHT, LRR, and PYD domains-containing protein 3 (NLRP3), resulting in caspase-1 cleavage which promotes the activation of Gasdermin D (GSDMD). This pathway is known as the canonical pathway. GSDMD has also been shown to be activated in a non-canonical pathway during MI and MI/R injury via caspase-4/5/11. Suppression of GSDMD has been shown to provide cardioprotection against MI and MI/R injury. Although the effects of MI or MI/R injury on pyroptosis have previously been discussed, knowledge concerning the roles of GSDMD in these settings remains limited. In this review, the evidence from in vitro, in vivo, and clinical studies focusing on cardiac GSDMD activation during MI and MI/R injury is comprehensively summarized and discussed. Implications from this review will help pave the way for a new therapeutic target in ischemic heart disease.     

灯盏花乙素通过调节线粒体融合-分裂平衡抑制氧糖剥夺/复糖复氧诱导的N2a细胞焦亡

目的: 探讨灯盏花乙素对氧糖剥夺/复氧复糖(OGD/R)诱导的N2a细胞焦亡的影响及作用机制。方法: 建立N2a细胞OGD/R损伤模型,设立对照组、模型组、灯盏花乙素给药组和线粒体分裂抑制剂Mdivi-1组。通过CCK-8检测N2a细胞增殖能力,检测各组细胞培养基中LDH和IL-1β水平,通过JC-1探针检测线粒体膜电位,Western blot检测线粒体融合分裂关键蛋白(Drp-1、Mfn-1、Mfn-2、OPA-1)和细胞焦亡标志蛋白(Caspase-1、NLRP-3和GSDMD)表达。结果: 与对照组相比,OGD/R可显著降低N2a细胞活力,灯盏花乙素可明显提高N2a细胞活力。与模型组相比,灯盏花乙素可减少LDH的释放,降低Caspase-1和IL-1β水平,提高线粒体膜势能,降低Drp-1的表达水平,上调Mfn-1、Mfn-2和OPA-1的表达,并减少细胞焦亡关键蛋白NLRP-3和GSDMD的表达。进一步给予Drp-1抑制剂Mdivi-1,与模型组相比,Mdivi-1可明显提高线粒体膜势能,减少Caspase-1、NLRP-3和GSDMD的表达。结论: 灯盏花乙素可通过抑制线粒体过度分裂,改善线粒体融合-分裂失衡与功能异常,进而缓解OGD/R所致N2a细胞焦亡。     

Role of pyroptosis in cardiovascular diseases

Pyroptosis is a form of programmed necrosis, and is morphologically and mechanistically unique form of programmed cell death compared to others, such as apoptosis and autophagic cell death. More specifically, pyroptosis features gasdermin family-mediated membrane pore formation and subsequent cell lysis, as well as release of pro-inflammatory intracellular contents including IL-1β, IL-18 and HMGB1. Mechanistically, pyroptosis is driven by two main signaling pathways - one mediated by caspase-1 and the other by caspase-4/5/11. Recent studies show that pyroptosis is implicated in several cardiovascular diseases. In this review, we summarize recent scientific discoveries of pyroptosis's involvement in atherosclerosis, myocardial infarction, diabetic cardiomyopathy, reperfusion injury and myocarditis. We also organized new and emerging evidence suggesting that pyroptosis signaling pathways may be potential therapeutic targets in cardiovascular diseases.     

Melatonin prevents endothelial cell pyroptosis via regulation of long noncoding RNA MEG3/miR-223/NLRP3 axis

OBJECTIVE Atherosclerosis(AS) is an inflammatory disease linked to endothelial dysfunction.Melatonin is reported to possess substantial anti-inflammatory properties, which has proven to be effective in AS. Emerging literature suggests that pyroptosis plays a critical role during AS progression. However, whether pyroptosis contributes to endothelial dysfunction and the underlying molecular mechanisms remained unexploited.This study was designed to investigate the antipyroptotic effects of melatonin in atherosclerotic endothelium and to elucidate the potential mechanisms. METHODS ApoE-/-mice were fed a high-fat diet to establish an atherosclerotic model, then divided into normal diet(ND) group, normal diet+melatonin(ND+melatonin) group, high fat diet(HFD)group and high fat diet+melatonin(HFD+melatonin) group.After 12 weeks, HE and oil Red O staining were used to detect the formation of atherosclerosis; qRT-RCR and Western blotting were used to detect the expression of NLRP3, ASC, IL-1β, IL-18, GSDMD, NF-κB, miR-223 and MEG3 in aortic endothelium; The luciferase assay was used to detect the binding of mi R-223 to MEG3. Human aortic endothelial cells(HAECs) were pretreated with ox-LDL. After melatonin treatment, qRT-RCR was used to detect the expression of mi R-223 and MEG3. Western blotting was used to detect NLRP3, ASC, c-caspase1,p-caspase1, GSDMD expression. In addition, after overexpressing MEG3 and knocking out mi R-223, the pyroptosis of HAECs was also detected. RESULTS We found intragastric administration of melatonin for 12 weeks markedly reduced the atherosclerotic plaque in aorta.Meanwhile, melatonin also attenuated the expression of pyroptosis-related genes, including NLRP3, ASC,cleaved caspase1, NF-κB/GSDMD, GSDMD N-termini,IL-1β, and IL-18 in aortic endothelium of melatonin-treated animals. Consistent antipyroptotic effects were also observed in ox-LDL-treated human aortic endothelial cells(HAECs). We found that lnc RNA MEG3 enhanced pyroptosis in HAECs. Moreover, MEG3 acted as an endogenous sponge by sequence complementarity to suppress the function of mi R-223 and to increase NLRP3 expression and enhance endothelial cel pyroptosis. Furthermore, knockdown of mi R-223 blocked the antipyroptotic actions of melatonin in ox-LDL-treated HAECs. CONCLUSION Our results suggest that melatonin prevents endothelial cell pyroptosis via MEG3/mi R-223/NLRP3 axis in atherosclerosis, and therefore, melatonin replacement might be considered a new strategy for protecting endothelium against pyroptosis, thereby for the treatment of atherosclerosis associated with pyroptosis.