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
OBJECTIVE: To investigate the efficacy of dihydromyricetin(DMY) on nasopharyngeal carcinoma(NPC) cell proliferation, apoptosis and to reveal the underlying mechanism in vitro experiments.METHODS: The CNE-2 cell line w...OBJECTIVE: To investigate the efficacy of dihydromyricetin(DMY) on nasopharyngeal carcinoma(NPC) cell proliferation, apoptosis and to reveal the underlying mechanism in vitro experiments.METHODS: The CNE-2 cell line was treated with different concentrations of DMY and the effects of DMY on cell viability and proliferation were evaluated using cell counting kit-8(CCK-8) assay and plate colony formation assay. Cellular apoptosis was detected by flow cytometry following Annexin V fluorescein isothiocyanate/propidine iodide staining.Nuclei morphology was observed under a fluorescence microscope following Hoechst 333258 staining. The expression of phosphorylated inhibitor of nuclear factor kappa-B kinase subunit beta(p-IKKβ), phosphorylated inhibitor of nuclear factor kappa-B kinase subunit alpha(p-IKKα), inhibitor of nuclear factor kappa-B alpha(IκB-α), nuclear factor kappa-B(NF-κB)/p65 was examined by Western blot analysis and the nuclear translocation of NF-κB/p65 was observed using a confocal laser scanning microscopy.RESULTS: DMY inhibited the proliferative capability and colony formation of NPC CNE-2 cells. Meanwhile, DMY induced apoptosis of CNE-2 cells in a dose and time-dependent manner via upregulating B-cell lymphoma-2 associated X, but downregulating B-cell lymphoma-2 and pro-caspase-3. Importantly, we found that DMY suppressed tumor necrosis factor alpha(TNF-α)-mediated NF-κB activation via inhibiting p-IKKβ, p-IKKα and blocking NF-κB subunit p65.CONCLUSION: Our experiments demonstrated that DMY had significant antiproliferative and apoptosisinducing effects on CNE-2 cells. Additionally, DMY promoted inactivation of p-IKKβ, p-IKKα,and blocked the nuclear translocation of NF-κB subunit p65. These results suggest that DMY may be an important therapeutic approach for NPC.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.
基金Supported by the National Natural Science Foundation of China (Regulation and Mechanism of Bone Marrow Mesenchymal Stem Cells on the Characteristics of Nasopharyngeal Carcinoma Stem Cells, No.81272434)the Medical Research Fund of Guangdong Province (Effect of Berberine on Growth of Transplanted Tumor of Nasopharyngeal Carcinoma in Mice Based on JAK/STAT3 Signaling Pathway, No.A2016431)。
文摘OBJECTIVE: To investigate the efficacy of dihydromyricetin(DMY) on nasopharyngeal carcinoma(NPC) cell proliferation, apoptosis and to reveal the underlying mechanism in vitro experiments.METHODS: The CNE-2 cell line was treated with different concentrations of DMY and the effects of DMY on cell viability and proliferation were evaluated using cell counting kit-8(CCK-8) assay and plate colony formation assay. Cellular apoptosis was detected by flow cytometry following Annexin V fluorescein isothiocyanate/propidine iodide staining.Nuclei morphology was observed under a fluorescence microscope following Hoechst 333258 staining. The expression of phosphorylated inhibitor of nuclear factor kappa-B kinase subunit beta(p-IKKβ), phosphorylated inhibitor of nuclear factor kappa-B kinase subunit alpha(p-IKKα), inhibitor of nuclear factor kappa-B alpha(IκB-α), nuclear factor kappa-B(NF-κB)/p65 was examined by Western blot analysis and the nuclear translocation of NF-κB/p65 was observed using a confocal laser scanning microscopy.RESULTS: DMY inhibited the proliferative capability and colony formation of NPC CNE-2 cells. Meanwhile, DMY induced apoptosis of CNE-2 cells in a dose and time-dependent manner via upregulating B-cell lymphoma-2 associated X, but downregulating B-cell lymphoma-2 and pro-caspase-3. Importantly, we found that DMY suppressed tumor necrosis factor alpha(TNF-α)-mediated NF-κB activation via inhibiting p-IKKβ, p-IKKα and blocking NF-κB subunit p65.CONCLUSION: Our experiments demonstrated that DMY had significant antiproliferative and apoptosisinducing effects on CNE-2 cells. Additionally, DMY promoted inactivation of p-IKKβ, p-IKKα,and blocked the nuclear translocation of NF-κB subunit p65. These results suggest that DMY may be an important therapeutic approach for NPC.
