Acquired semi-squamatization during chemotherapy suggests differentiation as a therapeutic strategy for bladder cancer

1. Download : Download high-res image (118KB)
2. Download : Download full-size image

     

Recent Advances in Pathology: the 2019 Annual Review Issue of The Journal of Pathology

In this Annual Review Issue of The Journal of Pathology, we present 15 invited reviews on topical aspects of pathology, ranging from the impacts of the microbiome in human disease through mechanisms of cell death and autophagy to recent advances in immunity and the uses of genomics for understanding, classifying and treating human cancers. Each of the reviews is authored by experts in their fields and our intention is to provide comprehensive updates in specific areas of pathology in which there has been considerable recent progress. Copyright © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

龙蛭汤对大鼠脑缺血再灌注损伤后Caspase-1和IL-18蛋白表达的影响

目的通过使用龙蛭汤对大脑中动脉闭塞(middle cerebral artery occlusion, MCAO)模型大鼠进行干预,观察脑组织中焦亡相关蛋白Caspase-1和IL-18表达情况,探讨其对脑缺血再灌注损伤(cerebral ischemia reperfusion injury, CIRI)的保护作用。方法将48只SD大鼠随机分为假手术组、模型组、龙蛭汤组、丁苯酞组,共4组,每组12只。除假手术组外,其余3组均采用Longa线栓法建立MCAO大鼠模型。造模成功后丁苯酞组、龙蛭汤组分别予丁苯酞[40 mg/(kg·d)]、龙蛭汤[13.6 g/(kg·d)]灌胃,假手术组、模型组均予等量生理盐水灌胃。术后第3天对各组大鼠行神经功能缺损评分;使用TTC染色法对新鲜脑片进行染色,同时行脑梗死体积测定;HE染色法观察脑皮质区病理形态变化;免疫组化法检测脑组织Caspase-1和IL-18蛋白表达情况。结果神经功能缺损评分结果显示,假手术组为0分;各造模组评分较假手术组升高,差异有统计学意义(P<0.05)。脑梗死体积百分比结果显示,假手术组无梗死灶;模型组最高,龙蛭汤组、丁苯酞组次之,均较模型组明显降低,差异有统计学意义(P<0.05)。脑组织HE染色结果显示,假手术组几乎未见受损细胞;模型组脑组织皮层神经细胞损伤严重,龙蛭汤组、丁苯酞组均较之有明显改善。免疫组化结果显示:假手术组仅见极少阳性细胞表达;与模型组比较,龙蛭汤组和丁苯酞组的Caspase-1和IL-18蛋白明显降低,差异有统计学意义(P<0.05)。结论龙蛭汤在改善大鼠CIRI后症状和保护大鼠损伤后神经细胞方面有显著的效果,其机制可能与下调Caspase-1和IL-18蛋白表达有关。     

扁塑藤素对LPS诱导的HUVEC细胞功能损伤和焦亡的作用

目的研究扁塑藤素对脂多糖(LPS)诱导人脐静脉血管内皮细胞(HUVEC)损伤的保护作用及可能机制。方法 建立LPS诱导HUVEC损伤模型,HUVEC细胞分为对照组、LPS组以及低、中、高剂量扁塑藤素组(0.1、0.2、0.4 μmol/L扁塑藤素)。CCK-8法测定细胞活力;试剂盒法检测乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量;ELISA法检测白细胞介素-1β(IL-1β)、IL-18蛋白水平;蛋白免疫印迹法、实时荧光定量PCR法检测焦亡相关分子NLRP3、Caspase-1、GSDMD蛋白和mRNA表达量。 结果与对照组比较,LPS组细胞活力和SOD含量显著下降(P＜0.05),LDH和MDA含量、NLRP3、Caspase-1、GSDMD的蛋白和mRNA表达量均显著升高(P＜0.05)。扁塑藤素呈剂量依赖性提高细胞活力和SOD含量,抑制LDH、MDA,降低NLRP3、Caspase-1、GSDMD的蛋白和mRNA表达水平(P＜0.05)。 结论扁塑藤素呈剂量依赖性抑制细胞焦亡和减轻氧化应激,从而改善LPS诱导的HUVEC功能损伤。     

CXC趋化因子配体14对高糖环境中脂肪细胞焦亡的影响

目的探讨CXC趋化因子配体14(CXCL14)对高糖暴露环境中脂肪细胞焦亡的影响。方法利用“鸡尾酒法”诱导3T3-L1细胞分化为成熟脂肪细胞,用5.5 mmol/L低糖(NG)或25 mmol/L高糖(HG)葡萄糖培养基培养脂肪细胞24 h;HG环境下用不同浓度CXCL14处理3T3-L1细胞不同时间。Western blot检测消化道皮肤素(GSDMD)、核苷酸结合寡聚化结构样受体蛋白3(NLRP3)、天冬氨酸蛋白水解酶-1(Caspase-1)蛋白、白细胞介素-6(IL-6)蛋白表达水平。实时荧光定量PCR检测GSDMD、NLRP3、白细胞介素-1β(IL-1β) mRNA转录水平。LDH测定试剂盒检测脂肪细胞上清液中乳酸脱氢酶(LDH)活力;Annexin V-FITC荧光检测细胞死亡情况;CCK-8法检测各组细胞增殖活力。结果高糖环境下脂肪细胞焦亡发生率升高,CXCL14处理可提高脂肪细胞增殖活力,但脂肪细胞焦亡相关指标却受到CXCL14浓度梯度的不同影响。50 nmol/L CXCL14处理可降低高糖环境下脂肪细胞GSDMD、NLRP3、IL-1β mRNA以及NLRP3蛋白的表达,下调脂肪细胞LDH活力,减少Annexin V-FITC荧光染色细胞死亡率,但25、100、200 nmol/L CXCL14对其焦亡的指标却呈反向趋势。并且50 nmol/L CXCL14干预后脂肪细胞NLRP3、Caspase-1蛋白随干预时间的延长呈先下降再上升趋势。结论CXCL14对高糖环境中脂肪细胞焦亡的影响与其浓度存在相关性。     

敲除S1PR3可缓解小鼠的急性肺损伤：基于抑制MAPK信号通路

目的 探讨敲除S1PR3是否通过抑制丝裂原活化蛋白激酶（MAPKs）途径改善脂多糖（LPS）诱导的小鼠急性肺损伤。方法 用LPS诱导小鼠急性肺损伤模型。雄性C57BL/6J和S1PR3敲除小鼠,随机分为4组：C57 NS组（C57,生理盐水处理）、C57 LPS组（C57,LPS诱导）、S1PR3-/- NS组（S1PR3敲除鼠,生理盐水处理）、S1PR3-/- LPS组（S1PR3敲除鼠,LPS诱导）,8只/组。RT-qPCR检测S1PR3,IL-β和IL-18表达,HE染色检测肺部组织损伤,流式细胞术检测细胞凋亡水平,Western blot法检测caspase-1,GSDMD,p-JNK,p-ERK p-p38 蛋白表达水平。同时,Ⅱ型肺泡上皮细胞（MLE-12 细胞）铺板后分为 4 组：PBS 组、LPS 组、CYM5541组（仅加入S1PR3激动剂）、CYM5541+LPS组（加入S1PR3激动剂+LPS）,检测各组细胞焦亡水平及MAPK信号通路分子的表达水平。结果 急性肺损伤小鼠肺组织S1PR3表达上调（P<0.001）;血清IL-1β和IL-18因急性肺损伤而明显升高（P<0.05）。急性肺损伤小鼠因敲除S1PR3肺出血、炎症渗出明显改善,肺湿干比重下降,细胞凋亡比例和焦亡相关蛋白如clv-caspase-1,GSDMD表达均降低（P<0.05）。MLE-12细胞因加入S1PR3激动剂,细胞焦亡相关蛋白表达均升高（P<0.05）。此外,MAPKs家族（JNK,ERK p38）的激活因S1PR3敲除后受到抑制,因加入S1PR3激动剂而表达明显升高（P<0.05）。结论 敲除S1PR3可通过抑制MAPK信号通路改善急性肺损伤。     

Role of AIM2 inflammasome in inflammatory diseases,cancer and infection

AIM2 is a cytosolic innate immune receptor which recognizes double‐stranded DNA (dsDNA) released during cellular perturbation and pathogenic assault. AIM2 recognition of dsDNA leads to the assembly of a large multiprotein oligomeric complex termed the inflammasome. This inflammasome assembly leads to the secretion of bioactive interleukin‐1β (IL‐1β) and IL‐18 and induction of an inflammatory form of cell death called pyroptosis. Sensing of dsDNA by AIM2 in the cytosol is crucial to mediate protection against the invading pathogens including bacteria, virus, fungi and parasites. AIM2 also responds to dsDNA released from damaged host cells, resulting in the secretion of the effector cytokines thereby driving the progression of sterile inflammatory diseases such as skin disease, neuronal disease, chronic kidney disease, cardiovascular disease and diabetes. Additionally, the protection mediated by AIM2 in the development of colorectal cancer depends on its ability to regulate epithelial cell proliferation and gut microbiota in maintaining intestinal homeostasis independently of the effector cytokines. In this review, we will highlight the recent progress on the role of the AIM2 inflammasome as a guardian of cellular integrity in modulating chronic inflammatory diseases, cancer and infection.     

焦亡在肿瘤治疗中作用的研究进展

细胞焦亡是一种伴随炎症反应的程序性细胞死亡,主要通过激活炎性半胱氨酸蛋白酶Caspase-1/4/5/11切割GSDMD或激活凋亡半胱氨酸蛋白酶Caspase-3切割GSDME两种途径被触发。细胞焦亡参与多种疾病的发生发展。近年来,焦亡在肿瘤治疗中的意义得到了广泛关注,积累了诸多新成果,形成了一些新见解。本文谨就此进行简要综述,以期为拓展细胞死亡方式与肿瘤治疗之间的研究提供参考。     

Streptococcus pneumoniae induces pyroptosis through the regulation of autophagy in murine microglia

Streptococcus pneumoniae is responsible for significant mortality and morbidity worldwide and causes invasive pneumococcal diseases including pneumococcal meningitis. Pyroptosis is caspase-1-dependent inflammatory cell death and is known to be induced by various microbial infections. In the present study, we investigated the molecular mechanisms that regulate pyroptosis induced by S. pneumoniae in microglia. Our results revealed that S. pneumoniae induced pyroptosis through caspase-1 activation and IL-1β production. We also found that the activation of caspase-1 and the maturation of IL-1β and IL-18 in the S. pneumoniae-triggered pyroptotic cell death process were mediated by NLRP3 inflammasome. In addition, pneumococcal infection increased the expression of autophagy-related genes and induced autophagosome formation. We also showed that the inhibition of autophagy promoted pneumococcus-induced pyroptosis. Furthermore, ROS was generated by pneumococcal infection and inhibited caspase-1 activation within 4 h of infection. However, in the late phase of infection, IL-1β secretion and caspase-1-dependent cell death were induced by ROS. These results suggest that autophagy induction transiently delay pyroptosis induced by S. pneumoniae in microglia. Our study also revealed that the activation of caspase-1 and the production of IL-1β were induced by pneumolysin and that pneumolysin triggered pyroptosis in microglial cells. Similar to the in vitro results, S. pneumoniae induced caspase-1 activation and caspase-1-dependent cytokine maturation in the mouse meningitis model. Thus, the present data demonstrate that S. pneumoniae induces pyroptosis in murine microglia and that NLRP3 inflammasome is critical for caspase-1 activation during the process. Furthermore, the induction of autophagy could transiently protect microglia from pyroptosis.     

The NLRP3 inflammasome contributes to host protection during Sporothrix schenckii infection

Sporotrichosis is a mycosis caused by fungi from the Sporothrix schenckii species complex, whose prototypical member is Sporothrix schenckii sensu stricto. Pattern recognition receptors (PRRs) recognize and respond to pathogen‐associated molecular patterns (PAMPs) and shape the following adaptive immune response. A family of PRRs most frequently associated with fungal recognition is the nucleotide‐binding oligomerization domain‐like receptor (NLR). After PAMP recognition, NLR family pyrin domain‐containing 3 (NLRP3) binds to apoptosis‐associated speck‐like protein containing a caspase recruitment domain (ASC) and caspase‐1 to form the NLRP3 inflammasome. When activated, this complex promotes the maturation of the pro‐inflammatory cytokines interleukin‐1β (IL‐1β) and IL‐18 and cell death through pyroptosis. In this study, we aimed to evaluate the importance of the NLRP3 inflammasome in the outcome of S. schenckii infection using the following three different knockout (KO) mice: NLRP3^−/−, ASC^−/− and caspase‐1^−/−. All KO mice were more susceptible to infection than the wild‐type, suggesting that NLRP3‐triggered responses contribute to host protection during S. schenckii infection. Furthermore, the NLRP3 inflammasome appeared to be critical for the ex vivo release of IL‐1β, IL‐18 and IL‐17 but not interferon‐γ. Additionally, a role for the inflammasome in shaping the adaptive immune response was suggested by the lower frequencies of type 17 helper T (Th17) cells and Th1/Th17 but not Th1 cells in S. schenckii‐infected KO mice. Overall, our results indicate that the NLRP3 inflammasome links the innate recognition of S. schenckii to the adaptive immune response, so contributing to protection against this infection.