目的:研究核因子κB抑制蛋白(IκBα)在急性胰腺炎(AP)大鼠胰腺和肝脏组织中的表达及中药新清胰汤Ⅱ号[Q ing Y i TangⅡ(QYT)]的影响。方法:70只SD大鼠随机被分为正常对照组(n=10)、AP+QYT组(n=30)、AP+生理盐水(NS)组(n=30)。经胰胆...目的:研究核因子κB抑制蛋白(IκBα)在急性胰腺炎(AP)大鼠胰腺和肝脏组织中的表达及中药新清胰汤Ⅱ号[Q ing Y i TangⅡ(QYT)]的影响。方法:70只SD大鼠随机被分为正常对照组(n=10)、AP+QYT组(n=30)、AP+生理盐水(NS)组(n=30)。经胰胆管逆行注射4%去氧胆酸钠复制AP模型;给予QYT(AP+QYT组)或NS(AP+NS组)灌胃,每5 h重复1次。造模后1 h、4 h和10 h分批处死动物。荧光定量RT-PCR、W estern b lot-ting分别检测胰腺和肝脏组织IκBα在mRNA水平和蛋白水平的表达,后者包括磷酸化形式和非磷酸化形式;ELISA法检测血清白三烯C4(LTC4)浓度;HE染色观察胰腺和肺组织病理学变化。结果:AP大鼠肝脏组织中IκBαmRNA表达在各个时点均显著高于正常对照组(P<0.01,P<0.05);AP+QYT组显著低于AP+NS组(P<0.05)。与IκBα的mRNA表达相一致,肝脏和胰腺组织中IκBα蛋白表达(无论其磷酸化形式和非磷酸化形式)在AP大鼠均高于正常对照组(P<0.05);AP+NS组IκBα蛋白磷酸化形式表达在观测时间内增高;QYT可抑制该形式的表达(P<0.05)。AP大鼠血清LTC4浓度明显高于正常对照组,呈时间依赖性;AP+QYT组血清LTC4浓度明显低于AP组(P<0.05)。病理学检查见AP大鼠胰腺组织水肿、出血、肺间质水肿、出血,炎症细胞大量浸润,随AP的病程而加重;QYT治疗组的病理变化较轻。结论:QYT可通过抑制IκBα的磷酸化等机制减轻AP时局部和全身的炎症反应。展开更多
AIM: To identify whether JTE-522 can induce apoptosis in AGS cells and ROS also involved in the process, and to investigate the changes in NF-kB, p53, bcl-2 and caspase in the apoptosis process. METHODS: Cell culture,...AIM: To identify whether JTE-522 can induce apoptosis in AGS cells and ROS also involved in the process, and to investigate the changes in NF-kB, p53, bcl-2 and caspase in the apoptosis process. METHODS: Cell culture, MTT, Electromicroscopy, agarose gel electrophoresis, lucigenin, Western blot and electrophoretic mobility shift assay (EMSA) analysis were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanisms. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Lucigenin assay showed the generation of ROS in cells under incubation with JTE-522. The increased ROS generation might contribute to the induction of AGS cells to apoptosis. EMSA and Western blot revealed that NF-kB activity was almost completely inhibited by preventing the degradation of IkBalpha. Additionally, by using Western blot we confirmed that the level of bcl-2 was decreased, whereas p53 showed a great increase following JTE-522 treatment. Their changes were in a dose-dependent manner. CONCLUSION: These findings suggest that reactive oxygen species, NF-kB, p53, bcl-2 and caspase-3 may play an important role in the induction of apoptosis in AGS cells after treatment with JTE-522.展开更多
文摘目的:研究核因子κB抑制蛋白(IκBα)在急性胰腺炎(AP)大鼠胰腺和肝脏组织中的表达及中药新清胰汤Ⅱ号[Q ing Y i TangⅡ(QYT)]的影响。方法:70只SD大鼠随机被分为正常对照组(n=10)、AP+QYT组(n=30)、AP+生理盐水(NS)组(n=30)。经胰胆管逆行注射4%去氧胆酸钠复制AP模型;给予QYT(AP+QYT组)或NS(AP+NS组)灌胃,每5 h重复1次。造模后1 h、4 h和10 h分批处死动物。荧光定量RT-PCR、W estern b lot-ting分别检测胰腺和肝脏组织IκBα在mRNA水平和蛋白水平的表达,后者包括磷酸化形式和非磷酸化形式;ELISA法检测血清白三烯C4(LTC4)浓度;HE染色观察胰腺和肺组织病理学变化。结果:AP大鼠肝脏组织中IκBαmRNA表达在各个时点均显著高于正常对照组(P<0.01,P<0.05);AP+QYT组显著低于AP+NS组(P<0.05)。与IκBα的mRNA表达相一致,肝脏和胰腺组织中IκBα蛋白表达(无论其磷酸化形式和非磷酸化形式)在AP大鼠均高于正常对照组(P<0.05);AP+NS组IκBα蛋白磷酸化形式表达在观测时间内增高;QYT可抑制该形式的表达(P<0.05)。AP大鼠血清LTC4浓度明显高于正常对照组,呈时间依赖性;AP+QYT组血清LTC4浓度明显低于AP组(P<0.05)。病理学检查见AP大鼠胰腺组织水肿、出血、肺间质水肿、出血,炎症细胞大量浸润,随AP的病程而加重;QYT治疗组的病理变化较轻。结论:QYT可通过抑制IκBα的磷酸化等机制减轻AP时局部和全身的炎症反应。
基金National Natural Science Foundation of China,No.39770300,30070873the Overseas Chinese Affairs Office of the State Council Foundation,No.98-33
文摘AIM: To identify whether JTE-522 can induce apoptosis in AGS cells and ROS also involved in the process, and to investigate the changes in NF-kB, p53, bcl-2 and caspase in the apoptosis process. METHODS: Cell culture, MTT, Electromicroscopy, agarose gel electrophoresis, lucigenin, Western blot and electrophoretic mobility shift assay (EMSA) analysis were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanisms. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Lucigenin assay showed the generation of ROS in cells under incubation with JTE-522. The increased ROS generation might contribute to the induction of AGS cells to apoptosis. EMSA and Western blot revealed that NF-kB activity was almost completely inhibited by preventing the degradation of IkBalpha. Additionally, by using Western blot we confirmed that the level of bcl-2 was decreased, whereas p53 showed a great increase following JTE-522 treatment. Their changes were in a dose-dependent manner. CONCLUSION: These findings suggest that reactive oxygen species, NF-kB, p53, bcl-2 and caspase-3 may play an important role in the induction of apoptosis in AGS cells after treatment with JTE-522.
