Recently,the roles of pyroptosis,a form of cell death induced by activated NODlike receptor protein 3(NLRP3)inflammasome,in the pathogenesis of diabetic cardiomyopathy(DCM)have been extensively investigated.However,mo...Recently,the roles of pyroptosis,a form of cell death induced by activated NODlike receptor protein 3(NLRP3)inflammasome,in the pathogenesis of diabetic cardiomyopathy(DCM)have been extensively investigated.However,most studies have focused mainly on whether diabetes increases the NLRP3 inflammasome and associated pyroptosis in the heart of type 1 or type 2 diabetic rodent models,and whether various medications and natural products prevent the development of DCM,associated with decreased levels of cardiac NLRP3 inflammasome and pyroptosis.The direct link of NLRP3 inflammasome and associated pyroptosis to the pathogenesis of DCM remains unclear based on the limited evidence derived from the available studies,with the approaches of NLRP3 gene silencing or pharmaceutical application of NLRP3 specific inhibitors.We thus emphasize the requirement for more systematic studies that are designed to provide direct evidence to support the link,given that several studies have provided both direct and indirect evidence under specific conditions.This editorial emphasizes that the current investigation should be circumspect in its conclusion,i.e.,not overemphasizing its role in the pathogenesis of DCM with the fact of only significantly increased expression or activation of NLRP3 inflammasome and pyroptosis in the heart of diabetic rodent models.Only clear-cut evidence-based causative roles of NLRP3 inflammasome and pyroptosis in the pathogenesis of DCM can help to develop effective and safe medications for the clinical management of DCM,targeting these biomarkers.展开更多
OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like g...OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like growth factor-binding protein-3(IGFBP-3)suppresses HCC cell proliferation in both IGF-dependent and independent manners.The present study is to investigate whether treatment with exogenous IGFBP-3 inhibits bF GF and PDGF production and the cell proliferation of HCC cells.METHODS Cell Counting Kit 8 assay were designed to detect HCC cell proliferation,transcription factor early growth response-1(EGR1)involving in IGFBP-3 regulation of b FGF and PDGF were detected by RT-PCR and Western blot assays.Western blot assay was adopted to detect the IGFBP-3 regulating insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS The present study demonstrates that IGFBP-3 suppressed IGF-1-induced b FGF and PDGF expression while it does not affect their expression in the absence of IGF-1.To delineate the underlying mechanism,Western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1(EGR1)is involved in IGFBP-3 regulation of b FGF and PDGF.IGFBP-3 inhibition of type 1 insulin-like growth factor receptor(IGF1R),ERK and AKT activation is IGF-1-dependent.Furthermore,transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1,b FGF and PDGF expression.CONCLUSION In conclusion,these findings suggest that IGFBP-3suppresses transcription of EGR1 and its target genes b FGF and PDGF through inhibiting IGF-1-dependent ERK and AKT activation.It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation,suggesting that IGFBP-3 could be a target for the treatment of HCC.展开更多
Background:The nucleotide-binding and oligomerization domain-like receptor protein 3 (NLRP3) inflammasome composed of NLRP3,apoptosis-associated speck-like protein containing CARD (ASC),and caspase-1 is engaged in the...Background:The nucleotide-binding and oligomerization domain-like receptor protein 3 (NLRP3) inflammasome composed of NLRP3,apoptosis-associated speck-like protein containing CARD (ASC),and caspase-1 is engaged in the inflammatory response of many kidney diseases and can be activated by purinergic 2X7 receptor (P2X7R).This study was conducted to explore whether P2X7R plays a pathogenic role in the podocyte damage of obesity-related glomerulopathy (ORG) and whether this role is mediated by the activation ofNLRP3 inflammasome.Methods:A mouse model of ORG was established by high-fat diet feeding.The conditionally immortalized mouse podocytes were cultured with leptin or with leptin and P2X7R antagonist (KN-62 or A438079).The mRNA and protein expression of the P2X7R and NLRP3 inflammasome components including NLRP3,ASC,and caspase-1,as well as the podocyte-associated molecules including nephrin,podocin,and desmin in mouse renal cortex or cultured mouse podocytes were tested by real-time-polymerase chain reaction and Westem blot analysis,respectively.Results:The significantly upregulated expression of P2X7R and NLRP3 inflammasome components and the NLRP3 inflammasome activation were observed in the renal cortex (in fact their location in podocytes was proved by confocal microscopy) of ORG mice in vivo,which were accompanied with the morphological changes of podocyte damage and the expression changes of podocyte-associated molecules.Similar changes in the expression of P2X7R and NLRP3 inflammasome components as well as in the expression ofpodocyte-associated molecules were also observed in the cultured podocyte studies treated by leptin in vitro,and all of the above changes were significantly attenuated by the P2X7R antagonist KN-62 or A438079.Conclusions:P2X7R could trigger the activation ofNLRP3 inflammasome,and the activated P2X7R/NLRP3 inflammasome in podocytes might be involved in the podocyte damage of ORG.展开更多
AIM To validate the effects of receptor interacting protein kinase-3(RIP3) deletion in non-alcoholic fatty liver disease(NAFLD) and to clarify the mechanism of action.METHODS Wild-type(WT) and RIP3 knockout(KO) mice w...AIM To validate the effects of receptor interacting protein kinase-3(RIP3) deletion in non-alcoholic fatty liver disease(NAFLD) and to clarify the mechanism of action.METHODS Wild-type(WT) and RIP3 knockout(KO) mice werefed normal chow and high fat(HF) diets for 12 wk. The body weight was assessed once weekly. After 12 wk, the liver and serum samples were extracted. The liver tissue expression levels of RIP3, microsomal triglyceride transfer protein, protein disulfide isomerase, apolipoprotein-B, X-box binding protein-1, sterol regulatory element-binding protein-1c, fatty acid synthase, cluster of differentiation-36, diglyceride acyltransferase, peroxisome proliferator-activated receptor alpha, tumor necrosis factor-alpha(TNF-α), and interleukin-6 were assessed. Oleic acid treated primary hepatocytes from WT and RIP3 KO mice were stained with Nile red. The expression of inflammatory cytokines, including chemokine(C-X-C motif) ligand(CXCL) 1, CXCL2, and TNF-α, in monocytes was evaluated.RESULTS RIP3 KO HF diet fed mice showed a significant gain in body weight, and liver weight, liver to body weight ratio, and liver triglycerides were increased in HF diet fed RIP3 KO mice compared to HF diet fed WT mice. RIP3 KO primary hepatocytes also had increased intracellular fat droplets compared to WT primary hepatocytes after oleic acid treatment. RIP3 overexpression decreased hepatic fat content. Quantitative real-time polymerase chain reaction analysis showed that the expression of very-low-density lipoproteins secretion markers(microsomal triglyceride transfer protein, protein disulfide isomerase, and apolipoprotein-B) was significantly suppressed in RIP3 KO mice. The overall NAFLD Activity Score was the same between WT and RIP3 KO mice; however, RIP3 KO mice had increased fatty change and decreased lobular inflammation compared to WT mice. Inflammatory signals(CXCL1/2, TNF-α, and interleukin-6) increased after lipopolysaccharide and pancaspase inhibitor(necroptotic condition) treatment in monocytes. Neutrophil chemokines(CXCL1, and CXCL2) were decreased, and TNF-α was increased after RIP3 inhibitor treatment in monocytes.CONCLUSION RIP3 deletion exacerbates steatosis, and partially inhibits inflammation in the HF diet induced NAFLD model.展开更多
目的研究骆驼刺提取物(Alhagi pseudalhagi(M.B.)Desv.Extract,APE)对脂多糖诱导的大鼠小肠隐窝上皮细胞(Intestinal epithelial cell,IEC-6)损伤模型NLRP3炎症小体及相关细胞因子的影响。方法培养IEC-6细胞,将其分为空白组、模型组、AP...目的研究骆驼刺提取物(Alhagi pseudalhagi(M.B.)Desv.Extract,APE)对脂多糖诱导的大鼠小肠隐窝上皮细胞(Intestinal epithelial cell,IEC-6)损伤模型NLRP3炎症小体及相关细胞因子的影响。方法培养IEC-6细胞,将其分为空白组、模型组、APE低、中、高浓度组,用1.0μg/mL的脂多糖(Lipopolysaccharide,LPS)诱导建立细胞炎症损伤模型,APE(低、中、高浓度:15、25、35μg/mL)干预后采用CCK-8法检测细胞的存活率,通过ELISA试剂盒检测炎症因子IL-1β、IL-18、TNF-α的分泌水平。蛋白质印迹法(WB)检测核苷酸结合寡聚化结构域样受体蛋白3(Nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)炎症小体信号通路5个关键蛋白:NLRP3、半胱氨酸天冬氨酸蛋白酶1(Cystein-asparate protease-1,Caspase-l)、凋亡相关斑点样蛋白(Apoptosis-associated speck-like protein containing a CARD,ASC)及抗凋亡蛋白Bcl-2(Anti-apoptosis Protein Bcl-2)和Bcl-xl(Anti-apoptosis Protein Bcl-xl)表达。结果与空白组比较,模型组IEC-6细胞的存活率降低,NLRP3、Caspase-1、ASC蛋白表达水平升高,抗凋亡蛋白Bcl-2、Bcl-xl的表达水平降低,促炎因子IL-1β、IL-18和TNF-α的分泌水平升高,差异有统计学意义(P<0.05)。与模型组比较,APE低、中、高浓度组细胞存活率升高,35μg/mL APE组IEC-6细胞的NLRP3、Caspase-1、ASC蛋白相对表达水平降低,抗凋亡蛋白Bcl-2、Bcl-xl的表达水平升高,差异有统计学意义(P<0.05)。中、高浓度的APE能够抑制炎症因子分泌,25μg/mL APE对IL-1β、IL-18、TNF-α炎症因子分泌水平抑制率分别为31.60%、31.19%和31.09%(P<0.05)。结论骆驼刺提取物通过提高抗凋亡蛋白Bcl-2、Bcl-xl的表达水平,下调NLRP3炎症小体组成成分以及促炎因子IL-1β、IL-18和TNF-α分泌,从而抑制NLRP3炎症小体组装和激活,实现缓解LPS对IEC-6细胞的损伤。展开更多
目的探究GATA结合蛋白3(GATA binding protein 3,GATA3)对乳腺癌细胞迁移能力的影响。方法在MCF7细胞中利用慢病毒载体介导的基因干涉技术敲低GATA3基因,使用实时定量荧光PCR(qRT-PCR)和蛋白质印迹检测GATA3和LIFR的mRNA和蛋白表达水平,...目的探究GATA结合蛋白3(GATA binding protein 3,GATA3)对乳腺癌细胞迁移能力的影响。方法在MCF7细胞中利用慢病毒载体介导的基因干涉技术敲低GATA3基因,使用实时定量荧光PCR(qRT-PCR)和蛋白质印迹检测GATA3和LIFR的mRNA和蛋白表达水平,Transwell实验检测MCF7细胞的迁移能力。在MCF7和T47D细胞中用染色质免疫沉淀(ChIP-qPCR)实验检测GATA3在LIFR的启动子区的结合位点。在敲低GATA3基因的MCF7细胞中回补LIFR,通过细胞划痕实验和Transwell实验检测MCF7细胞的迁移能力。结果与对照组相比,敲低GATA3基因的MCF7细胞的迁移能力增强(P_(均)<0.05)。与对照组相比,敲低GATA3基因的MCF7细胞的LIFR表达水平降低(P_(均)<0.05)。乳腺癌细胞MCF7与T47D中GATA3在LIFR的启动子区有结合(P_(均)<0.05)。在敲低GATA3基因的MCF7细胞中稳定过表达LIFR可以部分挽救GATA3基因敲低引起的细胞迁移能力的增强(P_(均)<0.05)。结论GATA3通过转录激活LIFR抑制乳腺癌细胞MCF7的迁移。展开更多
目的 探讨急性脑梗死(Acute cerebral infarct,ACI)患者静脉溶栓前后核苷酸结合寡聚化结构域样受体蛋白3(Nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)炎性小体信号通路因子变化,分析NLRP3炎性小体信号...目的 探讨急性脑梗死(Acute cerebral infarct,ACI)患者静脉溶栓前后核苷酸结合寡聚化结构域样受体蛋白3(Nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)炎性小体信号通路因子变化,分析NLRP3炎性小体信号通路因子与功能结局的关系。方法 选取2022年6月-2023年6月北京市大兴区人民医院收治的180例进行溶栓治疗的ACI患者,检测患者溶栓前后NLRP3炎性小体信号通路因子[NLRP3、半胱氨酰天冬氨酸特异性蛋白酶1(Caspase-1)]水平。根据日常生活能力量表(Activity of daily living scale,ADL)评分将ACI患者分为功能结局良好组(ADL评分>60分)及功能结局不良组(ADL评分≤60分)。收集两组患者一般资料,分析两组ACI患者溶栓前后外周血NLRP3和Caspase-1水平。Logistic回归模型分析ACI功能结局的影响因素,根据受试者工作特征(Receiver operating characteristic curve,ROC)曲线分析NLRP3炎性小体信号通路因子对ACI功能结局的预测价值。结果 与溶栓前比较,溶栓1周后,ACI患者外周血NLRP3[(1.81±0.35)vs(1.26±0.22)]及Caspase-1[(1.89±0.31)vs(1.33±0.19)] mRNA水平均降低(P<0.05);180例ACI患者中45例功能结局不良,功能结局不良发生率为25%(45/180);功能结局不良组溶栓前及溶栓1周后的NLRP3及Caspase-1水平均高于功能结局良好组(P<0.05);功能结局不良组美国国立卫生院卒中量表(National institute of health stroke scale,NIHSS)评分、空腹血糖(Fasting blood glucose,FBG)水平均高于功能结局良好组(P<0.05);多因素Logistic回归分析结果显示,NIHSS评分高、溶栓1周后的NLRP3及Caspase-1 mRNA高水平是影响ACI功能结局的独立危险因素(P<0.05);ROC分析结果显示:溶栓1周后NLRP3、Caspase-1mRNA单独及联合预测ACI功能结局的曲线下面积(Area under curve,AUC)分别为0.738(95%CI:0.667~0.800)、0.684(95%CI:0.684~0.815)、0.893(95%CI:0.838~0.934),溶栓1周后NLRP3、Caspase-1 mRNA联合的预测效能高于单独检测(Z=3.093、2.634,P均<0.05)。结论 ACI患者溶栓后外周血NLRP3、Caspase-1mRNA水平降低,外周血NLRP3、Caspase-1mRNA联合检测用于预测ACI功能结局具有较高的参考价值。展开更多
文摘Recently,the roles of pyroptosis,a form of cell death induced by activated NODlike receptor protein 3(NLRP3)inflammasome,in the pathogenesis of diabetic cardiomyopathy(DCM)have been extensively investigated.However,most studies have focused mainly on whether diabetes increases the NLRP3 inflammasome and associated pyroptosis in the heart of type 1 or type 2 diabetic rodent models,and whether various medications and natural products prevent the development of DCM,associated with decreased levels of cardiac NLRP3 inflammasome and pyroptosis.The direct link of NLRP3 inflammasome and associated pyroptosis to the pathogenesis of DCM remains unclear based on the limited evidence derived from the available studies,with the approaches of NLRP3 gene silencing or pharmaceutical application of NLRP3 specific inhibitors.We thus emphasize the requirement for more systematic studies that are designed to provide direct evidence to support the link,given that several studies have provided both direct and indirect evidence under specific conditions.This editorial emphasizes that the current investigation should be circumspect in its conclusion,i.e.,not overemphasizing its role in the pathogenesis of DCM with the fact of only significantly increased expression or activation of NLRP3 inflammasome and pyroptosis in the heart of diabetic rodent models.Only clear-cut evidence-based causative roles of NLRP3 inflammasome and pyroptosis in the pathogenesis of DCM can help to develop effective and safe medications for the clinical management of DCM,targeting these biomarkers.
基金supported by National Natural Science Foundation of China(81502123 and81330081)Natural Science Foundation of Anhui Province(1308085QH130)Anhui Province Nature Science Foundation in University(KJ2014A119)
文摘OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like growth factor-binding protein-3(IGFBP-3)suppresses HCC cell proliferation in both IGF-dependent and independent manners.The present study is to investigate whether treatment with exogenous IGFBP-3 inhibits bF GF and PDGF production and the cell proliferation of HCC cells.METHODS Cell Counting Kit 8 assay were designed to detect HCC cell proliferation,transcription factor early growth response-1(EGR1)involving in IGFBP-3 regulation of b FGF and PDGF were detected by RT-PCR and Western blot assays.Western blot assay was adopted to detect the IGFBP-3 regulating insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS The present study demonstrates that IGFBP-3 suppressed IGF-1-induced b FGF and PDGF expression while it does not affect their expression in the absence of IGF-1.To delineate the underlying mechanism,Western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1(EGR1)is involved in IGFBP-3 regulation of b FGF and PDGF.IGFBP-3 inhibition of type 1 insulin-like growth factor receptor(IGF1R),ERK and AKT activation is IGF-1-dependent.Furthermore,transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1,b FGF and PDGF expression.CONCLUSION In conclusion,these findings suggest that IGFBP-3suppresses transcription of EGR1 and its target genes b FGF and PDGF through inhibiting IGF-1-dependent ERK and AKT activation.It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation,suggesting that IGFBP-3 could be a target for the treatment of HCC.
基金grants from the National Natural Science Foundation of China (No.81573745and No.8160140274)Beijing Municipal Natural Science Foundation (No.7172066) Beijing Development Foundation of Traditional Chinese Medicine (QN2016-23).
文摘Background:The nucleotide-binding and oligomerization domain-like receptor protein 3 (NLRP3) inflammasome composed of NLRP3,apoptosis-associated speck-like protein containing CARD (ASC),and caspase-1 is engaged in the inflammatory response of many kidney diseases and can be activated by purinergic 2X7 receptor (P2X7R).This study was conducted to explore whether P2X7R plays a pathogenic role in the podocyte damage of obesity-related glomerulopathy (ORG) and whether this role is mediated by the activation ofNLRP3 inflammasome.Methods:A mouse model of ORG was established by high-fat diet feeding.The conditionally immortalized mouse podocytes were cultured with leptin or with leptin and P2X7R antagonist (KN-62 or A438079).The mRNA and protein expression of the P2X7R and NLRP3 inflammasome components including NLRP3,ASC,and caspase-1,as well as the podocyte-associated molecules including nephrin,podocin,and desmin in mouse renal cortex or cultured mouse podocytes were tested by real-time-polymerase chain reaction and Westem blot analysis,respectively.Results:The significantly upregulated expression of P2X7R and NLRP3 inflammasome components and the NLRP3 inflammasome activation were observed in the renal cortex (in fact their location in podocytes was proved by confocal microscopy) of ORG mice in vivo,which were accompanied with the morphological changes of podocyte damage and the expression changes of podocyte-associated molecules.Similar changes in the expression of P2X7R and NLRP3 inflammasome components as well as in the expression ofpodocyte-associated molecules were also observed in the cultured podocyte studies treated by leptin in vitro,and all of the above changes were significantly attenuated by the P2X7R antagonist KN-62 or A438079.Conclusions:P2X7R could trigger the activation ofNLRP3 inflammasome,and the activated P2X7R/NLRP3 inflammasome in podocytes might be involved in the podocyte damage of ORG.
基金Supported by National Research Foundation of Korea(NRF)funded by the South Korean Government,No.NRF-2017M3A9C8028794
文摘AIM To validate the effects of receptor interacting protein kinase-3(RIP3) deletion in non-alcoholic fatty liver disease(NAFLD) and to clarify the mechanism of action.METHODS Wild-type(WT) and RIP3 knockout(KO) mice werefed normal chow and high fat(HF) diets for 12 wk. The body weight was assessed once weekly. After 12 wk, the liver and serum samples were extracted. The liver tissue expression levels of RIP3, microsomal triglyceride transfer protein, protein disulfide isomerase, apolipoprotein-B, X-box binding protein-1, sterol regulatory element-binding protein-1c, fatty acid synthase, cluster of differentiation-36, diglyceride acyltransferase, peroxisome proliferator-activated receptor alpha, tumor necrosis factor-alpha(TNF-α), and interleukin-6 were assessed. Oleic acid treated primary hepatocytes from WT and RIP3 KO mice were stained with Nile red. The expression of inflammatory cytokines, including chemokine(C-X-C motif) ligand(CXCL) 1, CXCL2, and TNF-α, in monocytes was evaluated.RESULTS RIP3 KO HF diet fed mice showed a significant gain in body weight, and liver weight, liver to body weight ratio, and liver triglycerides were increased in HF diet fed RIP3 KO mice compared to HF diet fed WT mice. RIP3 KO primary hepatocytes also had increased intracellular fat droplets compared to WT primary hepatocytes after oleic acid treatment. RIP3 overexpression decreased hepatic fat content. Quantitative real-time polymerase chain reaction analysis showed that the expression of very-low-density lipoproteins secretion markers(microsomal triglyceride transfer protein, protein disulfide isomerase, and apolipoprotein-B) was significantly suppressed in RIP3 KO mice. The overall NAFLD Activity Score was the same between WT and RIP3 KO mice; however, RIP3 KO mice had increased fatty change and decreased lobular inflammation compared to WT mice. Inflammatory signals(CXCL1/2, TNF-α, and interleukin-6) increased after lipopolysaccharide and pancaspase inhibitor(necroptotic condition) treatment in monocytes. Neutrophil chemokines(CXCL1, and CXCL2) were decreased, and TNF-α was increased after RIP3 inhibitor treatment in monocytes.CONCLUSION RIP3 deletion exacerbates steatosis, and partially inhibits inflammation in the HF diet induced NAFLD model.
文摘目的研究骆驼刺提取物(Alhagi pseudalhagi(M.B.)Desv.Extract,APE)对脂多糖诱导的大鼠小肠隐窝上皮细胞(Intestinal epithelial cell,IEC-6)损伤模型NLRP3炎症小体及相关细胞因子的影响。方法培养IEC-6细胞,将其分为空白组、模型组、APE低、中、高浓度组,用1.0μg/mL的脂多糖(Lipopolysaccharide,LPS)诱导建立细胞炎症损伤模型,APE(低、中、高浓度:15、25、35μg/mL)干预后采用CCK-8法检测细胞的存活率,通过ELISA试剂盒检测炎症因子IL-1β、IL-18、TNF-α的分泌水平。蛋白质印迹法(WB)检测核苷酸结合寡聚化结构域样受体蛋白3(Nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)炎症小体信号通路5个关键蛋白:NLRP3、半胱氨酸天冬氨酸蛋白酶1(Cystein-asparate protease-1,Caspase-l)、凋亡相关斑点样蛋白(Apoptosis-associated speck-like protein containing a CARD,ASC)及抗凋亡蛋白Bcl-2(Anti-apoptosis Protein Bcl-2)和Bcl-xl(Anti-apoptosis Protein Bcl-xl)表达。结果与空白组比较,模型组IEC-6细胞的存活率降低,NLRP3、Caspase-1、ASC蛋白表达水平升高,抗凋亡蛋白Bcl-2、Bcl-xl的表达水平降低,促炎因子IL-1β、IL-18和TNF-α的分泌水平升高,差异有统计学意义(P<0.05)。与模型组比较,APE低、中、高浓度组细胞存活率升高,35μg/mL APE组IEC-6细胞的NLRP3、Caspase-1、ASC蛋白相对表达水平降低,抗凋亡蛋白Bcl-2、Bcl-xl的表达水平升高,差异有统计学意义(P<0.05)。中、高浓度的APE能够抑制炎症因子分泌,25μg/mL APE对IL-1β、IL-18、TNF-α炎症因子分泌水平抑制率分别为31.60%、31.19%和31.09%(P<0.05)。结论骆驼刺提取物通过提高抗凋亡蛋白Bcl-2、Bcl-xl的表达水平,下调NLRP3炎症小体组成成分以及促炎因子IL-1β、IL-18和TNF-α分泌,从而抑制NLRP3炎症小体组装和激活,实现缓解LPS对IEC-6细胞的损伤。
文摘目的探究GATA结合蛋白3(GATA binding protein 3,GATA3)对乳腺癌细胞迁移能力的影响。方法在MCF7细胞中利用慢病毒载体介导的基因干涉技术敲低GATA3基因,使用实时定量荧光PCR(qRT-PCR)和蛋白质印迹检测GATA3和LIFR的mRNA和蛋白表达水平,Transwell实验检测MCF7细胞的迁移能力。在MCF7和T47D细胞中用染色质免疫沉淀(ChIP-qPCR)实验检测GATA3在LIFR的启动子区的结合位点。在敲低GATA3基因的MCF7细胞中回补LIFR,通过细胞划痕实验和Transwell实验检测MCF7细胞的迁移能力。结果与对照组相比,敲低GATA3基因的MCF7细胞的迁移能力增强(P_(均)<0.05)。与对照组相比,敲低GATA3基因的MCF7细胞的LIFR表达水平降低(P_(均)<0.05)。乳腺癌细胞MCF7与T47D中GATA3在LIFR的启动子区有结合(P_(均)<0.05)。在敲低GATA3基因的MCF7细胞中稳定过表达LIFR可以部分挽救GATA3基因敲低引起的细胞迁移能力的增强(P_(均)<0.05)。结论GATA3通过转录激活LIFR抑制乳腺癌细胞MCF7的迁移。
文摘目的 探讨急性脑梗死(Acute cerebral infarct,ACI)患者静脉溶栓前后核苷酸结合寡聚化结构域样受体蛋白3(Nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)炎性小体信号通路因子变化,分析NLRP3炎性小体信号通路因子与功能结局的关系。方法 选取2022年6月-2023年6月北京市大兴区人民医院收治的180例进行溶栓治疗的ACI患者,检测患者溶栓前后NLRP3炎性小体信号通路因子[NLRP3、半胱氨酰天冬氨酸特异性蛋白酶1(Caspase-1)]水平。根据日常生活能力量表(Activity of daily living scale,ADL)评分将ACI患者分为功能结局良好组(ADL评分>60分)及功能结局不良组(ADL评分≤60分)。收集两组患者一般资料,分析两组ACI患者溶栓前后外周血NLRP3和Caspase-1水平。Logistic回归模型分析ACI功能结局的影响因素,根据受试者工作特征(Receiver operating characteristic curve,ROC)曲线分析NLRP3炎性小体信号通路因子对ACI功能结局的预测价值。结果 与溶栓前比较,溶栓1周后,ACI患者外周血NLRP3[(1.81±0.35)vs(1.26±0.22)]及Caspase-1[(1.89±0.31)vs(1.33±0.19)] mRNA水平均降低(P<0.05);180例ACI患者中45例功能结局不良,功能结局不良发生率为25%(45/180);功能结局不良组溶栓前及溶栓1周后的NLRP3及Caspase-1水平均高于功能结局良好组(P<0.05);功能结局不良组美国国立卫生院卒中量表(National institute of health stroke scale,NIHSS)评分、空腹血糖(Fasting blood glucose,FBG)水平均高于功能结局良好组(P<0.05);多因素Logistic回归分析结果显示,NIHSS评分高、溶栓1周后的NLRP3及Caspase-1 mRNA高水平是影响ACI功能结局的独立危险因素(P<0.05);ROC分析结果显示:溶栓1周后NLRP3、Caspase-1mRNA单独及联合预测ACI功能结局的曲线下面积(Area under curve,AUC)分别为0.738(95%CI:0.667~0.800)、0.684(95%CI:0.684~0.815)、0.893(95%CI:0.838~0.934),溶栓1周后NLRP3、Caspase-1 mRNA联合的预测效能高于单独检测(Z=3.093、2.634,P均<0.05)。结论 ACI患者溶栓后外周血NLRP3、Caspase-1mRNA水平降低,外周血NLRP3、Caspase-1mRNA联合检测用于预测ACI功能结局具有较高的参考价值。
