cGAS-STING通路与老龄化疾病的研究进展

衰老是指随着年龄的增长生物体在分子、细胞、组织和系统水平上损伤的积累和功能的减退,逐渐趋向死亡的一种现象。衰老是导致年龄相关性疾病(aging-related diseases,ARDs)发生、发展和死亡的重要危险因素。衰老具有的特征有基因组不稳定、端粒损耗、表观遗传改变、蛋白质稳态丧失、大自噬失能、营养感应失调、线粒体功能障碍、细胞衰老、干细胞耗竭、细胞间通讯改变、慢性炎症和生态失调等。其中,细胞衰老主要由端粒缩短、DNA损伤、氧化应激以及细胞周期阻滞等因素所引起。DNA损伤累积导致促炎因子的过表达,包括各种细胞因子、趋化因子,统称为衰老相关分泌表型(senescence-associated secretory phenotype,SASP)。最近诸多科学研究表明,cGAS-STING通路会在衰老细胞中激活,从而进一步触发SASP表型。本文将对cGAS与细胞衰老的关系、SASP的调控以及相关老龄化疾病的影响进行综述。

cGAS-STING信号通路调节剂在免疫治疗中的研究进展

环鸟嘌呤-腺嘌呤核苷酸合成酶(cGAS)-干扰素基因刺激蛋白(STING)信号通路感知细胞质中的异常双链DNA后,诱导Ⅰ型干扰素(IFN-Ⅰ)和促炎细胞因子表达,从而激活宿主的免疫应答,增强机体抗肿瘤免疫反应和抗病原体感染。但是,cGAS-STING信号通路的持续激活会驱动自身免疫性疾病、衰老相关炎症和神经退行性病变等疾病。本文阐述了cGAS-STING信号通路参与调控多种免疫相关性疾病发生发展的机制,重点回顾了STING激动剂、cGAS抑制剂以及STING抑制剂的研发进展,为cGAS-STING调节剂的研发提供更多理论参考。

cGAS-STING通路在劳力性热射病所致心肌损伤中的作用研究

目的:探讨cGAS-STING通路在劳力性热射病(exertional heat stroke,EHS)所致心肌炎症中的作用。方法:将8周龄C57BL/6N雄性小鼠随机分为安静对照组(C组)、劳力性热射病发病即刻组(EHS0组)、劳力性热射病发病3天组(EHS3组)和劳力性热射病发病14天组(EHS14组)。发病组在温度为37.5℃±1℃、湿度为45%±5%的高温舱中进行力竭跑台运动,构建EHS小鼠模型。采用心动超声仪检测小鼠心功能指标,苏木精-伊红(HE)染色观察小鼠心肌组织病理变化,ELISA测定血清IL-6与cTnI水平,qPCR检测心肌组织cGAS、STING、IRF3、NF-κB、γH2AX、IL-6、TFAM、PGC-1αmRNA相对表达量,透射电镜观察小鼠心肌线粒体超微结构变化,Western blot检测心肌组织cGAS、p-NF-κB p65、γH2AX蛋白表达水平。结果:1)与C组比较,EHS组小鼠的心动超声指标(LVAWd、LVAWs、LVPWd、LVPWs、LVIDd和LVIDs)显著增加,LVEF和LVFS显著下降,心肌细胞出现肥大,肌纤维排列紊乱,伴有炎症细胞浸润,心肌线粒体固缩破裂,线粒体嵴溶解出现空泡改变,内部基质密度分布不均匀,Z线模糊,大量肌丝溶解断裂,且EHS发病后14天仍未完全恢复。2)EHS组小鼠的血清IL-6、cTnI水平和心肌组织NF-κB、PGC-1α、TFAM、cGAS、IRF3 mRNA表达量及cGAS、p-NF-κB p65、γH2AX蛋白表达水平均显著升高,EHS发病后14天基本恢复。结论:1)EHS发病后可引起小鼠心肌炎症反应,其机制可能与cGAS-STING信号通路的激活有关。2)EHS后心脏长期功能障碍与血清IL-6和cTnI的水平无关,而可能与心肌纤维紊乱、细胞肥大、心肌线粒体形态结构改变等有关。

cGAS-STING通路通过调节巨噬细胞参与骨折愈合的研究进展

骨折愈合是一个复杂缓慢的动态过程,巨噬细胞是最早到达骨折部位的细胞之一。长期以来人们一直认为巨噬细胞参与损伤部位的初始炎症并有助于清创,它们在正常骨稳态和骨折愈合过程中调节骨再生中的关键作用也越来越受到重视。近年来研究发现,cGAS-STING通路是先天免疫和病毒防御中重要的DNA传感机制,越来越多证据表明cGAS-STING通路与巨噬细胞密切相关,所以cGAS-STING通路可能通过调节巨噬细胞极化类型来参与骨折愈合。因此了解骨折修复过程中cGASSTING通路参与调节巨噬细胞的机制,将为促进骨折修复提供新的思路和策略。

青蒿琥酯调节cGAS-STING信号通路对抑郁症模型小鼠神经炎性反应的影响

目的探讨青蒿琥酯(ART)调节环磷酸鸟苷-腺苷酸合成酶(cGAS)-干扰素基因刺激因子(STING)通路对抑郁症小鼠神经炎性反应的影响。方法小鼠分为模型组、对照组、ART低剂量组、ART高剂量组、氟西汀组、ART高剂量+RocA(cGAS-STING通路激活剂)组。糖水消耗实验、强迫游泳实验评估小鼠的抑郁行为;HE染色检测海马组织病理变化;ELISA检测白细胞介素(IL)-6、肿瘤坏死因子-α(TNF-α)、五羟色胺(5-HT)、多巴胺(DA)水平;TUNEL染色检测神经元凋亡;Western blot检测Bcl-2相关X蛋白(Bax)、p53、cGAS、STING蛋白。结果与对照组相比,模型组小鼠表现出神经元性脓疱变性,糖水消耗率、5-HT、DA水平降低,强迫游泳静止时间延长,IL-6、TNF-α水平、神经元凋亡率及Bax、p53、cGAS、STING蛋白表达升高(P<0.05);与模型组相比,ART低剂量组、ART高剂量组、氟西汀组小鼠海马神经元损伤有所缓解,糖水消耗率、5-HT、DA水平升高,强迫游泳静止时间缩短,IL-6、TNF-α水平、神经元凋亡率及Bax、p53、cGAS、STING蛋白表达降低(P<0.05);RocA逆转了高剂量ART对小鼠抑郁症的改善作用。结论ART抑制抑郁症小鼠神经炎性反应及神经元凋亡,上调胺类神经递质水平的机制可能与阻断cGAS-STING通路有关。

人乳头瘤病毒16型E7基因靶向调控cGAS-STING信号通路抑制宫颈癌细胞免疫功能的研究

目的:探讨HPV16 E7靶向调控cGAS-STING信号通路对宫颈癌细胞免疫功能的影响。方法:体外构建HPV16 E7过表达和干扰质粒,转染至SiHa细胞,qPCR检测HPV16 E7 mRNA表达水平,流式细胞术检测CD95、MICA/B、CD73、CD39、CTLA-4和CD25表达。在此基础上分别转染cGAS-siRNA和STING-siRNA至SiHa细胞,CCK8检测细胞活力,流式细胞术检测细胞凋亡。qPCR和Western blot分别检测cGAS、STING、IFN-Ⅰ、HPV16 E7的mRNA及蛋白表达。流式细胞术检测细胞CD95、MICA/B、CD73、CD39、CTLA-4和CD25表达。结果:STING-siRNA+HPV16 E7过表达组CD95和MICA/B水平降低(P<0.05),CD73、CD39、CTLA-4和CD25水平升高(P<0.05)。cGAS、STING和IFN-Ⅰ蛋白表达水平降低(P<0.05)。STINGsiRNA+HPV16 E7干扰组结果与之相反。结论:HPV16 E7靶向调控cGAS-STING信号通路进而抑制宫颈癌细胞免疫功能。

异常流体剪切力通过cGAS-STING信号通路参与髓核细胞的凋亡、炎症与自噬

目的探讨异常流体剪切力诱导髓核细胞(NPC)凋亡、炎症和自噬的作用及其可能的机制。方法对NPC加载24 dyn/cm^(2)流体剪切力,时间分别为0、60、90、120、150 min。使用细胞骨架染色研究流体剪切力对细胞骨架及细胞形态的影响;使用5-乙炔基-2'-脱氧尿苷染色验证NPC的增殖能力;用线粒体膜电位变化评估流体剪切力对线粒体的损伤效应;用赫斯特染色研究凋亡与流体剪切力的关系;用丹酰戊二胺染色研究自噬与流体剪切力的关系;探究细胞培养基酸碱度的变化,验证流体剪切力与炎症的关联;使用蛋白质印迹法研究cGAS-STING相关蛋白(cGAS、STING、TBK1)、炎症相关蛋白(肿瘤坏死因子-α、COX-2)、凋亡相关蛋白(Bcl-2、Bax)以及自噬蛋白(IL3-Ⅰ/Ⅱ、p62)的变化。结果作为一种细胞间的机械刺激,24 dyn/cm^(2)强度的流体剪切力可造成NPC形态由梭形转变为多边形,细胞骨架发生重组,细胞增殖能力下降,培养基酸性增强,线粒体JC-1多聚体减少、单体增加;同时,细胞内肿瘤坏死因子-α、COX-2、Bax蛋白表达量上调,LC3Ⅱ/LC3Ⅰ增加,p62、Bcl-2降解增加,出现凋亡、炎症、自噬以及线粒体损伤;加载24 dyn/cm^(2)的流体剪切力可以激活NPC内的cGAS-STING信号通路,且与时间呈正相关关系,120 min时强度最高。使用cGAS抑制剂RU.521可以减缓24 dyn/cm^(2)流体剪切力造成的NPC的凋亡、炎症、自噬以及线粒体损伤。结论流体剪切力诱导的NPC凋亡、炎症和自噬与cGAS-STING信号通路激活相关,抑制cGAS-STING信号通路的激活可以缓解异常流体剪切力造成的细胞损害。

曲美他嗪通过cGAS-STING通路对肺炎链球菌肺炎大鼠免疫功能的影响

目的:探究曲美他嗪(TMZ)通过环鸟苷酸-腺苷酸合成酶(cGAS)-干扰素基因刺激蛋白(STING)通路对肺炎链球菌(SP)肺炎大鼠免疫功能的影响。方法:大鼠随机分为模型组、TMZ组、DMXAA组(cGAS-STING信号通路激活剂),鼻腔内滴入SP构建SP肺炎大鼠模型,对照组大鼠滴入等量生理盐水。ELISA检测IL-1β、IL-10水平;流式细胞术检测CD4^(+)T、CD8^(+)T细胞水平;比较各组大鼠脾脏、胸腺指数;透射比浊法检测IgG、IgA水平;HE染色观察肺组织病理变化;Western blot检测cGAS-STING通路蛋白表达。结果:与对照组比较,模型组大鼠肺泡壁和肺间质增厚,伴有水肿出血、炎症细胞浸润,IL-1β、CD8^(+)T细胞水平、肺组织病理损伤评分、cGAS、STING表达升高,IL-10、CD4^(+)T细胞、CD4^(+)T/CD8^(+)T、脾脏、胸腺指数、IgG、IgA水平降低(P<0.05);与模型组比较,TMZ组大鼠肺泡壁增厚减轻,肺泡结构显著改善、炎症细胞浸润较少,IL-1β、CD8^(+)T水平、肺组织病理损伤评分、cGAS、STING表达降低,IL-10、CD4^(+)T、CD4^(+)T/CD8^(+)T、脾脏、胸腺指数、IgG、IgA水平升高(P<0.05);与TMZ组比较,DMXAA组IL-1β、CD8^(+)T细胞水平、肺组织病理损伤评分、cGAS、STING表达升高,IL-10、CD4^(+)T细胞、CD4^(+)T/CD8^(+)T、脾脏、胸腺指数、IgG、IgA水平降低(P<0.05)。结论:TMZ可能通过抑制cGAS-STING信号通路激活抑制炎症,增强SP肺炎大鼠免疫功能。

cGAS-STING信号通路在缺血性脑卒中作用机制的研究进展

缺血性脑卒中(IS)是世界上导致成人死亡和残疾的主要疾病之一,其复杂的病理机制至今尚未完全阐明。环鸟苷酸-腺苷酸合成酶(cGAS)-干扰素基因刺激因子(STING)信号通路是近年来新发现的一种广泛表达于哺乳动物细胞内的模式识别受体(PRR),在固有免疫应答过程中发挥着关键作用。研究发现cGAS-STING信号通路可通过调节神经炎症、细胞死亡、自噬等途径参与脑缺血的发生发展,有望成为治疗IS的新靶标。本文主要归纳总结了近年来cGAS-STING信号通路在IS中作用机制的研究成果,以期为脑缺血的治疗提供理论依据和新的研究思路。

cGAS-STING信号通路在系统性红斑狼疮中的研究进展

cGAS-STING信号通路在免疫反应调控中发挥着至关重要的作用,其通过感知细胞质内DNA的存在,激活下游一系列分子,导致Ⅰ型干扰素等炎症细胞因子的释放,最终参与和调控免疫反应的发生。SLE发病机制复杂,免疫功能紊乱及Ⅰ型干扰素等细胞因子参与SLE的发生发展。本文将就cGAS-STING信号通路在SLE中的研究进展进行综述,以期为SLE的诊断及靶向治疗提供新方向。

Identification of hub genes associated with Helicobacter pylori infection and type 2 diabetes mellitus:A pilot bioinformatics study

BACKGROUND Helicobacter pylori(H.pylori)infection is related to various extragastric diseases including type 2 diabetes mellitus(T2DM).However,the possible mechanisms connecting H.pylori infection and T2DM remain unknown.AIM To explore potential molecular connections between H.pylori infection and T2DM.METHODS We extracted gene expression arrays from three online datasets(GSE60427,GSE27411 and GSE115601).Differentially expressed genes(DEGs)commonly present in patients with H.pylori infection and T2DM were identified.Hub genes were validated using human gastric biopsy samples.Correlations between hub genes and immune cell infiltration,miRNAs,and transcription factors(TFs)were further analyzed.RESULTS A total of 67 DEGs were commonly presented in patients with H.pylori infection and T2DM.Five significantly upregulated hub genes,including TLR4,ITGAM,C5AR1,FCER1G,and FCGR2A,were finally identified,all of which are closely related to immune cell infiltration.The gene-miRNA analysis detected 13 miRNAs with at least two gene cross-links.TF-gene interaction networks showed that TLR4 was coregulated by 26 TFs,the largest number of TFs among the 5 hub genes.CONCLUSION We identified five hub genes that may have molecular connections between H.pylori infection and T2DM.This study provides new insights into the pathogenesis of H.pylori-induced onset of T2DM.

Mining elite loci and candidate genes for root morphology-related traits at the seedling stage by genome-wide association studies in upland cotton(Gossypium hirsutum L.)

Root system architecture plays an essential role in water and nutrient acquisition in plants,and it is significantly involved in plant adaptations to various environmental stresses.In this study,a panel of 242 cotton accessions was collected to investigate six root morphological traits at the seedling stage,including main root length(MRL),root fresh weight(RFW),total root length(TRL),root surface area(RSA),root volume(RV),and root average diameter(AvgD).The correlation analysis of the six root morphological traits revealed strong positive correlations of TRL with RSA,as well as RV with RSA and AvgD,whereas a significant negative correlation was found between TRL and AvgD.Subsequently,a genome-wide association study(GWAS)was performed using the root phenotypic and genotypic data reported previously for the 242 accessions using 56,010 single nucleotide polymorphisms(SNPs)from the CottonSNP80K array.A total of 41 quantitative trait loci(QTLs)were identified,including nine for MRL,six for RFW,nine for TRL,12 for RSA,12 for RV and two for AvgD.Among them,eight QTLs were repeatedly detected in two or more traits.Integrating these results with a transcriptome analysis,we identified 17 candidate genes with high transcript values of transcripts per million(TPM)≥30 in the roots.Furthermore,we functionally verified the candidate gene GH_D05G2106,which encodes a WPP domain protein 2in root development.A virus-induced gene silencing(VIGS)assay showed that knocking down GH_D05G2106significantly inhibited root development in cotton,indicating its positive role in root system architecture formation.Collectively,these results provide a theoretical basis and candidate genes for future studies on cotton root developmental biology and root-related cotton breeding.

cGAS蛋白促进PCV-2茎环结构诱导cGAS-STING信号通路天然免疫反应研究

为了探索猪圆环病毒2型(PCV-2)DNA茎环结构是否可以激活环鸟苷酸腺苷酸合成酶(cGAS),以及是否诱导cGAS-STING信号通路天然免疫反应,试验将含有PCV-2茎环结构的单链DNA(SL-DNA)转染至PK15细胞中,通过Western-blot检测cGAS蛋白是否在PK15细胞中表达;将SL-DNA转染至过表达及敲除cGAS蛋白的PK15细胞中,通过实时荧光定量PCR检测细胞中cGAS、干扰素刺激基因(STING)、TANK结合激酶1(TBK1)、干扰素调节因子3(IRF3)、干扰素-β(IFN-β)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)基因的表达水平。结果表明:SL-DNA转染后,PK15细胞中cGAS蛋白表达量上升;过表达cGAS蛋白的PK15细胞与PK15细胞相比,SL-DNA转染后cGAS、STING、TBK1、IRF3、IFN-β、IL-1β、IL-6、TNF-α基因的相对表达量均极显著升高(P<0.01);敲除cGAS蛋白的PK15细胞与PK15细胞相比,SL-DNA转染后cGAS、STING、TBK1、IRF3、IFN-β、IL-1β、IL-6、TNF-α基因的相对表达量均极显著降低(P<0.01)。说明cGAS蛋白可以识别PCV-2的茎环结构,并且促进其诱导的cGAS-STING信号通路天然免疫反应。

Knockdown of the atypical protein kinase genes GhABC1K2-A05 and GhABC1K12-A07 make cotton more sensitive to salt and PEG stress

Activity of bc1 complex kinase(ABC1K)is an atypical protein kinase(aPK)that plays a crucial role in plant mitochondrial and plastid stress responses,but little is known about the responses of ABC1Ks to stress in cotton(Gossypium spp.).Here,we identified 40 ABC1Ks in upland cotton(Gossypium hirsutum L.)and found that the Gh ABC1Ks were unevenly distributed across 17 chromosomes.The GhABC1K family members included 35 paralogous gene pairs and were expanded by segmental duplication.The GhABC1K promoter sequences contained diverse cis-acting regulatory elements relevant to hormone or stress responses.The qRT-PCR results revealed that most Gh ABC1Ks were upregulated by exposure to different stresses.Gh ABC1K2-A05 and Gh ABC1K12-A07 expression levels were upregulated by at least three stress treatments.These genes were further functionally characterized by virus-induced gene silencing(VIGS).Compared with the controls,the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced cotton lines exhibited higher malondialdehyde(MDA)contents,lower catalase(CAT),peroxidase(POD)and superoxide dismutase(SOD)activities and reduced chlorophyll and soluble sugar contents under NaCl and PEG stress.In addition,the expression levels of six stress marker genes(Gh DREB2A,Gh SOS1,Gh CIPK6,Gh SOS2,Gh WRKY33,and Gh RD29A)were significantly downregulated after stress in the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced lines.The results indicate that knockdown of Gh ABC1K2-A05 and Gh ABC1K12-A07 make cotton more sensitive to salt and PEG stress.These findings can provide valuable information for intensive studies of Gh ABC1Ks in the responses and resistance of cotton to abiotic stresses.

Predictive model using four ferroptosis-related genes accurately predicts gastric cancer prognosis

BACKGROUND Gastric cancer(GC)is a common malignancy of the digestive system.According to global 2018 cancer data,GC has the fifth-highest incidence and the thirdhighest fatality rate among malignant tumors.More than 60%of GC are linked to infection with Helicobacter pylori(H.pylori),a gram-negative,active,microaerophilic,and helical bacterium.This parasite induces GC by producing toxic factors,such as cytotoxin-related gene A,vacuolar cytotoxin A,and outer membrane proteins.Ferroptosis,or iron-dependent programmed cell death,has been linked to GC,although there has been little research on the link between H.pylori infection-related GC and ferroptosis.AIM To identify coregulated differentially expressed genes among ferroptosis-related genes(FRGs)in GC patients and develop a ferroptosis-related prognostic model with discrimination ability.METHODS Gene expression profiles of GC patients and those with H.pylori-associated GC were obtained from The Cancer Genome Atlas and Gene Expression Omnibus(GEO)databases.The FRGs were acquired from the FerrDb database.A ferroptosis-related gene prognostic index(FRGPI)was created using least absolute shrinkage and selection operator–Cox regression.The predictive ability of the FRGPI was validated in the GEO cohort.Finally,we verified the expression of the hub genes and the activity of the ferroptosis inducer FIN56 in GC cell lines and tissues.RESULTS Four hub genes were identified(NOX4,MTCH1,GABARAPL2,and SLC2A3)and shown to accurately predict GC and H.pylori-associated GC.The FRGPI based on the hub genes could independently predict GC patient survival;GC patients in the high-risk group had considerably worse overall survival than did those in the low-risk group.The FRGPI was a significant predictor of GC prognosis and was strongly correlated with disease progression.Moreover,the gene expression levels of common immune checkpoint proteins dramatically increased in the highrisk subgroup of the FRGPI cohort.The hub genes were also confirmed to be highly overexpressed in GC cell lines and tissues and were found to be primarily localized at the cell membrane.The ferroptosis inducer FIN56 inhibited GC cell proliferation in a dose-dependent manner.CONCLUSION In this study,we developed a predictive model based on four FRGs that can accurately predict the prognosis of GC patients and the efficacy of immunotherapy in this population.

松萝酸调节cGAS-STING信号通路对大鼠心肌缺血再灌注损伤的保护作用研究

目的 探究松萝酸调节环磷酸鸟苷-磷酸腺苷合成酶(cGAS)-干扰素基因刺激因子(STING)信号通路对大鼠心肌缺血再灌注损伤的保护作用。方法 选取50只SD健康大鼠,分为假手术组、模型组、松萝酸低剂量组(25 mg/kg)、松萝酸高剂量组(100 mg/kg)和RU.521组(cGAS抑制剂,5 mg/kg),每组10只通过结扎冠状动脉左前降支再灌注方法建立大鼠心肌缺血再灌注模型。HE染色观察大鼠心肌组织病理学变化;TUNEL染色检测心肌细胞凋亡情况;NBT染色检测心肌梗死面积百分比;商品化试剂盒检测心肌组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和丙二醛(MDA)水平;ELISA法检测大鼠血清肌酸激酶同工酶(CKMB)、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)水平;RT-PCR法检测心肌组织cGAS mRNA、STING mRNA表达;Western blot检测大鼠心肌组织中Bax、Bcl-2、Caspase-3、cGAS、STING蛋白表达。结果 假手术组大鼠心肌组织结构形态正常,凋亡率较低;与假手术组比较,模型组大鼠心肌细胞受损严重、凋亡率、心肌梗死面积百分比、血清CKMB、TNF-α、IL-6水平、MDA水平、cGAS mRNA、STING mRNA表达水平、Bax、Caspase-3、cGAS、STING蛋白表达水平显著升高,SOD、CAT、Bcl-2蛋白表达水平显著降低(P<0.05);与模型组相比,松萝酸低、高剂量组和RU.521组大鼠心肌细胞形态结构明显改善,炎性细胞浸润显著减少、细胞凋亡率、心肌梗死面积百分比、血清CKMB、TNF-α、IL-6水平、MDA、cGAS mRNA、STING mRNA表达水平、Bax、Caspase-3、cGAS、STING蛋白表达水平显著降低,大鼠心肌组织SOD、CAT、Bcl-2蛋白表达水平显著升高(P<0.05);RU.521组各项指标与松萝酸高剂量组几乎相当。结论 松萝酸对心肌缺血再灌注大鼠具有保护作用,可能是通过抑制cGAS-STING信号通路实现的。

Key genes and regulatory networks for diabetic retinopathy based on hypoxia-related genes:a bioinformatics analysis

AIM:To prevent neovascularization in diabetic retinopathy(DR)patients and partially control disease progression.METHODS:Hypoxia-related differentially expressed genes(DEGs)were identified from the GSE60436 and GSE102485 datasets,followed by gene ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Potential candidate drugs were screened using the CMap database.Subsequently,a protein-protein interaction(PPI)network was constructed to identify hypoxia-related hub genes.A nomogram was generated using the rms R package,and the correlation of hub genes was analyzed using the Hmisc R package.The clinical significance of hub genes was validated by comparing their expression levels between disease and normal groups and constructing receiver operating characteristic curve(ROC)curves.Finally,a hypoxia-related miRNA-transcription factor(TF)-Hub gene network was constructed using the NetworkAnalyst online tool.RESULTS:Totally 48 hypoxia-related DEGs and screened 10 potential candidate drugs with interaction relationships to upregulated hypoxia-related genes were identified,such as ruxolitinib,meprylcaine,and deferiprone.In addition,8 hub genes were also identified:glycogen phosphorylase muscle associated(PYGM),glyceraldehyde-3-phosphate dehydrogenase spermatogenic(GAPDHS),enolase 3(ENO3),aldolase fructose-bisphosphate C(ALDOC),phosphoglucomutase 2(PGM2),enolase 2(ENO2),phosphoglycerate mutase 2(PGAM2),and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(PFKFB3).Based on hub gene predictions,the miRNA-TF-Hub gene network revealed complex interactions between 163 miRNAs,77 TFs,and hub genes.The results of ROC showed that the except for GAPDHS,the area under curve(AUC)values of the other 7 hub genes were greater than 0.758,indicating their favorable diagnostic performance.CONCLUSION:PYGM,GAPDHS,ENO3,ALDOC,PGM2,ENO2,PGAM2,and PFKFB3 are hub genes in DR,and hypoxia-related hub genes exhibited favorable diagnostic performance.

非洲猪瘟病毒拮抗宿主cGAS-STING信号通路的研究进展

非洲猪瘟病毒(African swine fever virus,ASFV)拥有多种逃逸宿主免疫应答的策略,造成病毒难以被宿主清除。cGAS-STING信号通路介导的天然免疫在抗ASFV感染中发挥了重要作用,然而病毒编码的多个蛋白靶向该通路中的不同分子以拮抗宿主的I型干扰素应答。利用基因编辑技术敲除这些病毒基因后,ASFV对宿主的致病性降低,成为基因缺失疫苗的研制潜在靶点。本文对目前已知参与调控宿主cGAS-STING信号通路的病毒蛋白进行总结,旨在阐明这些蛋白免疫逃逸cGAS-STING信号通路的分子机制,加深对ASFV免疫逃逸策略的理解,以期为ASFV致病机制研究与疫苗创制提供参考。

RPLP0/TBP are the most stable reference genes for human dental pulp stem cells under osteogenic differentiation

BACKGROUND Validation of the reference gene(RG)stability during experimental analyses is essential for correct quantitative real-time polymerase chain reaction(RT-qPCR)data normalisation.Commonly,in an unreliable way,several studies use genes involved in essential cellular functions[glyceraldehyde-3-phosphate dehydro-genase(GAPDH),18S rRNA,andβ-actin]without paying attention to whether they are suitable for such experimental conditions or the reason for choosing such genes.Furthermore,such studies use only one gene when Minimum Information for Publication of Quantitative Real-Time PCR Experiments guidelines recom-mend two or more genes.It impacts the credibility of these studies and causes dis-tortions in the gene expression findings.For tissue engineering,the accuracy of gene expression drives the best experimental or therapeutical approaches.We cultivated DPSCs under two conditions:Undifferentiated and osteogenic dif-ferentiation,both for 35 d.We evaluated the gene expression of 10 candidates for RGs[ribosomal protein,large,P0(RPLP0),TATA-binding protein(TBP),GAPDH,actin beta(ACTB),tubulin(TUB),aminolevulinic acid synthase 1(ALAS1),tyro-sine 3-monooxygenase/tryptophan 5-monooxygenase activation protein,zeta(YWHAZ),eukaryotic translational elongation factor 1 alpha(EF1a),succinate dehydrogenase complex,subunit A,flavoprotein(SDHA),and beta-2-micro-globulin(B2M)]every 7 d(1,7,14,21,28,and 35 d)by RT-qPCR.The data were analysed by the four main algorithms,ΔCt method,geNorm,NormFinder,and BestKeeper and ranked by the RefFinder method.We subdivided the samples into eight subgroups.RESULTS All of the data sets from clonogenic and osteogenic samples were analysed using the RefFinder algorithm.The final ranking showed RPLP0/TBP as the two most stable RGs and TUB/B2M as the two least stable RGs.Either theΔCt method or NormFinder analysis showed TBP/RPLP0 as the two most stable genes.However,geNorm analysis showed RPLP0/EF1αin the first place.These algorithms’two least stable RGs were B2M/GAPDH.For BestKeeper,ALAS1 was ranked as the most stable RG,and SDHA as the least stable RG.The pair RPLP0/TBP was detected in most subgroups as the most stable RGs,following the RefFinfer ranking.CONCLUSION For the first time,we show that RPLP0/TBP are the most stable RGs,whereas TUB/B2M are unstable RGs for long-term osteogenic differentiation of human DPSCs in traditional monolayers.

cGAS-STING信号通路在骨质疏松症中的作用机制

cGAS-STING信号通路是一种新型的、重要的先天免疫信号通路,其被核酸物质激活,并与其他免疫应答相互作用,在炎症、感染、癌症等多种病理过程中发挥重要作用。近期研究发现cGAS-STING参与了各种与年龄相关的肌肉骨骼疾病的发生和发展,并对骨代谢产生影响。然而,在骨质疏松症(osteoporosis,OP)中确切的作用机制尚不清楚。因此,本文综述了cGAS-STING及其下游在OP中的作用机制,为开发OP新的治疗策略提供思路。