Studies have demonstrated that reactive oxygen species(ROS) are closely related to inflammatory disorders. Nicotinamide adenine dinucleotide phosphate oxidase(NOX), originally found in phagocytes, is the main source o...Studies have demonstrated that reactive oxygen species(ROS) are closely related to inflammatory disorders. Nicotinamide adenine dinucleotide phosphate oxidase(NOX), originally found in phagocytes, is the main source of ROS in nonphagocytic cells. Besides directly producing the detrimental highly reactive ROS to act on biomolecules(lipids, proteins, and nucleic acids), NOX can also activate multiple signal transduction pathways, which regulate cell growth, proliferation, differentiation and apoptosis by producing ROS. Recently, research on pancreatic NOX is no longer limited to inflammatory cells, but extends to the aspect of pancreatic acinar cells and pancreatic stellate cells, which are considered to be potentially associated with pancreatitis. In this review, we summarize the literature on NOX protein structure, activation, function and its role in the pathogenesis of pancreatitis.展开更多
背景胰腺腺泡细胞增殖及凋亡与急性胰腺炎(acute pancreatitis,AP)发生密切相关,miRNA可通过调控靶基因表达从而参与细胞增殖及凋亡过程,因而寻找与胰腺腺泡细胞增殖及凋亡相关的miRNA分子标志物对临床诊断及治疗AP具有重要意义.目的探...背景胰腺腺泡细胞增殖及凋亡与急性胰腺炎(acute pancreatitis,AP)发生密切相关,miRNA可通过调控靶基因表达从而参与细胞增殖及凋亡过程,因而寻找与胰腺腺泡细胞增殖及凋亡相关的miRNA分子标志物对临床诊断及治疗AP具有重要意义.目的探讨微小RNA-7a-5p(miR-7a-5p)对AP腺泡细胞增殖、凋亡的影响及机制.方法构建雨蛙肽诱导的AP模型并收集胰腺炎腺泡细胞AR42J(雨蛙肽组),采用qRT-PCR与Western blot分别检测未经雨蛙肽诱导的AR42J细胞(对照组)及雨蛙肽组AR42J细胞中miR-7a-5p相对表达量及活化信号转导和转录激活因子的蛋白抑制因子-1(proteininhibitor of activated signal transducer and activator oftranscription 1,PIAS1)表达.分别将anti-miR-7a-5p、pcDNA-PIAS1转染至雨蛙肽组AR42J细胞,采用MTT法检测AR42J细胞增殖能力,流式细胞术检测AR42J细胞凋亡.荧光素酶报告系统检测miR-7a-5p对PIAS1基因的靶向调控作用,并采用Western blot检测miR-7a-5p对PIAS1蛋白表达的调控作用.采用RNA干扰技术沉默PIAS1表达(si-PIAS1组),分别将si-PIAS1及其阴性对照转染至anti-miR-7a-5p组AR42J细胞,观察AR42J细胞增殖及凋亡能力.结果与对照组相比,雨蛙肽组AR42J细胞中miR-7a-5p表达水平显著升高(P<0.05),PIAS1蛋白表达显著降低(P<0.05);抑制miR-7a-5p表达可促进胰腺炎腺泡细胞AR42J增殖并抑制其凋亡;miR-7a-5p可负向调控靶基因PIAS1表达;PIAS1过表达可促进AR42J细胞增殖并抑制其凋亡;与anti-miR-7a-5p+si-NC组相比,anti-miR-7a-5p+si-PIAS1组AR42J细胞活性显著降低(P<0.05),细胞凋亡率显著升高(P<0.05).结论miR-7a-5p可通过抑制PIAS1表达进而促进AP腺泡细胞凋亡并降低细胞增殖能力.展开更多
AIM To elucidate the underlying mechanism that microRNA-22(miR-22) promotes the apoptosis of rat pancreatic acinar cells(AR42 J) and the elements that regulate the expression of miR-22.METHODS One hundred nanomoles pe...AIM To elucidate the underlying mechanism that microRNA-22(miR-22) promotes the apoptosis of rat pancreatic acinar cells(AR42 J) and the elements that regulate the expression of miR-22.METHODS One hundred nanomoles per liter of caerulein(Cae)was administrated to induce the apoptosis of AR42 J cells and the apoptosis rate was detected by flow cytometry analysis. An amylase assay kit was used to measure the amylase expression level in the supernatant. Quantitative real-time PCR(qRT-PCR)was adopted to measure miR-22 expression. We used online tools to predict the potential transcription promoter of miR-22 and the binding sites, which was further identified by using luciferase reporter analysis,chromatin immunoprecipitation(ChIP) and ChIPqP CR assays. Then, a mimic of miR-22, Nr3 c1 plasmid encoding the glucocorticoid receptor(GR), and siNr3 c1 were used to transfect AR42 J cells, respectively.The mRNA expression of miR-22, Nr3 c1, and Erb-b2 receptor tyrosine kinase 3(ErbB3) was confirmed by qRT-PCR and the apoptosis rate of AR42 J cells was detected by flow cytometry analysis. Western blot was used to detect the expression of ErbB3, GR, PI3 k, PI3 kp85α, Akt, p-Akt, Bad, Bax, Bcl-xl, Bcl-2, and cleaved caspase3.RESULTS After inducing apoptosis of AR42 J cells in vitro, the expression of miR-22 was significantly increased by2.20 ± 0.26 and 4.19 ± 0.54 times, respectively, at3 h and 6 h in comparison with the control group.As revealed by qRT-PCR assay, the expression of miR-22 was 78.25 ± 6.61 times higher in the miR-22 mimic group relative to the miRNA control group,accompanied with an obviously increased acinar cell apoptosis rate(32.53 ± 1.15 vs 18.07 ± 0.89, P =0.0006). The upregulation of miR-22 could suppress its target gene, ErbB3, and the phosphorylation of PI3 k and Akt. Furthermore, we predicted the potential transcription promoter of miR-22 and the binding sites using online tools. Luciferase reporter analysis and sitedirected mutagenesis indicated that the binding site(GACAGCCATGTACA) of the GR, whi展开更多
目的 观察Janus激酶2/信号转导与转录激活子3（JAK2/STAT3）信号通路在胰蛋白酶原激活肽（TAP）诱导大鼠胰腺腺泡细胞释放高迁移率族蛋白B1（HMGB1）中的作用。方法 提取大鼠胰腺腺泡细胞,随机分为正常对照组、TAP促进组（终质量浓度为3...目的 观察Janus激酶2/信号转导与转录激活子3（JAK2/STAT3）信号通路在胰蛋白酶原激活肽（TAP）诱导大鼠胰腺腺泡细胞释放高迁移率族蛋白B1（HMGB1）中的作用。方法 提取大鼠胰腺腺泡细胞,随机分为正常对照组、TAP促进组（终质量浓度为3 nmol/L）、JAK2抑制剂AG490预处理组（25μ.mol/L）、STAT3抑制剂雷帕霉素预处理组（40μg/L）,采用聚合酶链反应（PCR）及Western blot法检测3-24 h各组HMGB1 mRNA及蛋白表达水平。结果 与正常对照组HMGB1 mRNA表达水平（1.01±0.04）比较,TAP组6-24 h HMGB1 mRNA表达水平显著增高（2.87±0.08、32.85±1.48、38.43±1.60,P〈0.01）,与TAP组比较,AG490预处理组12-24 h HMGB1 mRNA表达均显著抑制（23.40±1.38、19.44±0.89,P〈0.01）,与TAP组比较,雷帕霉素预处理组12-24 h HMGB1 mRNA表达均显著抑制（21.14±1.32、17.76±0.57,P〈0.01）。与正常对照组HMGB1蛋白表达水平（0.39±0.02）比较,TAP组3-24 h HMGB1蛋白表达显著增高（0.46±0.02、0.67±0.03、0.72±0.02、0.90±0.03,P〈0.01）,与TAP组比较,AG490预处理组12-24 h HMGB1蛋白表达均显著抑制（0.60±0.02、0.54±0.01,P〈0.01）,与TAP组比较,雷帕霉素预处理组12-24 h HMGB1蛋白表达均显著抑制（0.59±0.02、0.58±0.01,P〈0.01）。结论 TAP可诱导大鼠胰腺腺泡细胞HMGB1的表达和释放,其机制可能与JAK2/STAT3信号通路有关。展开更多
基金Supported by The National Natural Science Foundation of China,No.81173393the Natural Science Foundation of Tianjin City,No.12YFJZJC00800+1 种基金the Scientific Research Foundation(No.WHM201222,FYM201114)the Innovation Team Program(No.WHTD201310)from Logistics University of the Chinese People’s Armed Police Forces
文摘Studies have demonstrated that reactive oxygen species(ROS) are closely related to inflammatory disorders. Nicotinamide adenine dinucleotide phosphate oxidase(NOX), originally found in phagocytes, is the main source of ROS in nonphagocytic cells. Besides directly producing the detrimental highly reactive ROS to act on biomolecules(lipids, proteins, and nucleic acids), NOX can also activate multiple signal transduction pathways, which regulate cell growth, proliferation, differentiation and apoptosis by producing ROS. Recently, research on pancreatic NOX is no longer limited to inflammatory cells, but extends to the aspect of pancreatic acinar cells and pancreatic stellate cells, which are considered to be potentially associated with pancreatitis. In this review, we summarize the literature on NOX protein structure, activation, function and its role in the pathogenesis of pancreatitis.
文摘背景胰腺腺泡细胞增殖及凋亡与急性胰腺炎(acute pancreatitis,AP)发生密切相关,miRNA可通过调控靶基因表达从而参与细胞增殖及凋亡过程,因而寻找与胰腺腺泡细胞增殖及凋亡相关的miRNA分子标志物对临床诊断及治疗AP具有重要意义.目的探讨微小RNA-7a-5p(miR-7a-5p)对AP腺泡细胞增殖、凋亡的影响及机制.方法构建雨蛙肽诱导的AP模型并收集胰腺炎腺泡细胞AR42J(雨蛙肽组),采用qRT-PCR与Western blot分别检测未经雨蛙肽诱导的AR42J细胞(对照组)及雨蛙肽组AR42J细胞中miR-7a-5p相对表达量及活化信号转导和转录激活因子的蛋白抑制因子-1(proteininhibitor of activated signal transducer and activator oftranscription 1,PIAS1)表达.分别将anti-miR-7a-5p、pcDNA-PIAS1转染至雨蛙肽组AR42J细胞,采用MTT法检测AR42J细胞增殖能力,流式细胞术检测AR42J细胞凋亡.荧光素酶报告系统检测miR-7a-5p对PIAS1基因的靶向调控作用,并采用Western blot检测miR-7a-5p对PIAS1蛋白表达的调控作用.采用RNA干扰技术沉默PIAS1表达(si-PIAS1组),分别将si-PIAS1及其阴性对照转染至anti-miR-7a-5p组AR42J细胞,观察AR42J细胞增殖及凋亡能力.结果与对照组相比,雨蛙肽组AR42J细胞中miR-7a-5p表达水平显著升高(P<0.05),PIAS1蛋白表达显著降低(P<0.05);抑制miR-7a-5p表达可促进胰腺炎腺泡细胞AR42J增殖并抑制其凋亡;miR-7a-5p可负向调控靶基因PIAS1表达;PIAS1过表达可促进AR42J细胞增殖并抑制其凋亡;与anti-miR-7a-5p+si-NC组相比,anti-miR-7a-5p+si-PIAS1组AR42J细胞活性显著降低(P<0.05),细胞凋亡率显著升高(P<0.05).结论miR-7a-5p可通过抑制PIAS1表达进而促进AP腺泡细胞凋亡并降低细胞增殖能力.
基金National Natural Science Foundation of China,No.31671440
文摘AIM To elucidate the underlying mechanism that microRNA-22(miR-22) promotes the apoptosis of rat pancreatic acinar cells(AR42 J) and the elements that regulate the expression of miR-22.METHODS One hundred nanomoles per liter of caerulein(Cae)was administrated to induce the apoptosis of AR42 J cells and the apoptosis rate was detected by flow cytometry analysis. An amylase assay kit was used to measure the amylase expression level in the supernatant. Quantitative real-time PCR(qRT-PCR)was adopted to measure miR-22 expression. We used online tools to predict the potential transcription promoter of miR-22 and the binding sites, which was further identified by using luciferase reporter analysis,chromatin immunoprecipitation(ChIP) and ChIPqP CR assays. Then, a mimic of miR-22, Nr3 c1 plasmid encoding the glucocorticoid receptor(GR), and siNr3 c1 were used to transfect AR42 J cells, respectively.The mRNA expression of miR-22, Nr3 c1, and Erb-b2 receptor tyrosine kinase 3(ErbB3) was confirmed by qRT-PCR and the apoptosis rate of AR42 J cells was detected by flow cytometry analysis. Western blot was used to detect the expression of ErbB3, GR, PI3 k, PI3 kp85α, Akt, p-Akt, Bad, Bax, Bcl-xl, Bcl-2, and cleaved caspase3.RESULTS After inducing apoptosis of AR42 J cells in vitro, the expression of miR-22 was significantly increased by2.20 ± 0.26 and 4.19 ± 0.54 times, respectively, at3 h and 6 h in comparison with the control group.As revealed by qRT-PCR assay, the expression of miR-22 was 78.25 ± 6.61 times higher in the miR-22 mimic group relative to the miRNA control group,accompanied with an obviously increased acinar cell apoptosis rate(32.53 ± 1.15 vs 18.07 ± 0.89, P =0.0006). The upregulation of miR-22 could suppress its target gene, ErbB3, and the phosphorylation of PI3 k and Akt. Furthermore, we predicted the potential transcription promoter of miR-22 and the binding sites using online tools. Luciferase reporter analysis and sitedirected mutagenesis indicated that the binding site(GACAGCCATGTACA) of the GR, whi
文摘目的 观察Janus激酶2/信号转导与转录激活子3（JAK2/STAT3）信号通路在胰蛋白酶原激活肽（TAP）诱导大鼠胰腺腺泡细胞释放高迁移率族蛋白B1（HMGB1）中的作用。方法 提取大鼠胰腺腺泡细胞,随机分为正常对照组、TAP促进组（终质量浓度为3 nmol/L）、JAK2抑制剂AG490预处理组（25μ.mol/L）、STAT3抑制剂雷帕霉素预处理组（40μg/L）,采用聚合酶链反应（PCR）及Western blot法检测3-24 h各组HMGB1 mRNA及蛋白表达水平。结果 与正常对照组HMGB1 mRNA表达水平（1.01±0.04）比较,TAP组6-24 h HMGB1 mRNA表达水平显著增高（2.87±0.08、32.85±1.48、38.43±1.60,P〈0.01）,与TAP组比较,AG490预处理组12-24 h HMGB1 mRNA表达均显著抑制（23.40±1.38、19.44±0.89,P〈0.01）,与TAP组比较,雷帕霉素预处理组12-24 h HMGB1 mRNA表达均显著抑制（21.14±1.32、17.76±0.57,P〈0.01）。与正常对照组HMGB1蛋白表达水平（0.39±0.02）比较,TAP组3-24 h HMGB1蛋白表达显著增高（0.46±0.02、0.67±0.03、0.72±0.02、0.90±0.03,P〈0.01）,与TAP组比较,AG490预处理组12-24 h HMGB1蛋白表达均显著抑制（0.60±0.02、0.54±0.01,P〈0.01）,与TAP组比较,雷帕霉素预处理组12-24 h HMGB1蛋白表达均显著抑制（0.59±0.02、0.58±0.01,P〈0.01）。结论 TAP可诱导大鼠胰腺腺泡细胞HMGB1的表达和释放,其机制可能与JAK2/STAT3信号通路有关。
