目的探讨金盏花苷E对脂多糖(LPS)诱导炎症反应的抑制作用及可能的分子机制。方法 CCK-8实验检测不同浓度(0、2、4、6、8、10、20、25、30μg/mL)的金盏花苷E对RAW264.7细胞活力的影响;不同浓度的金盏花苷E(0、6、8、10μg/mL)预处理RAW2...目的探讨金盏花苷E对脂多糖(LPS)诱导炎症反应的抑制作用及可能的分子机制。方法 CCK-8实验检测不同浓度(0、2、4、6、8、10、20、25、30μg/mL)的金盏花苷E对RAW264.7细胞活力的影响;不同浓度的金盏花苷E(0、6、8、10μg/mL)预处理RAW264.7细胞2 h,然后用LPS(100 ng/mL)刺激细胞特定的时间,ELISA检测炎症因子TNF-α、IL-1β释放;Western blotting检测iNOS、COX-2的表达水平及JAK-stats、MAPKs及NF-кB信号途径的磷酸化;ROS检测试剂盒检测RAW264.7细胞内ROS含量;激光共聚焦实验检测转录因子stat3的核转位。结果 CCK-8结果显示,金盏花苷E浓度在低于20μg/mL时对RAW264.7细胞无明显毒性作用;金盏花苷E浓度依赖性地下调LPS诱导的iNOS和COX-2的表达(P<0.01 vs LPS组);抑制LPS诱导的促炎细胞因子TNF-α及IL-1β的释放,且1 0μg/mL组抑制作用尤为显著(P<0.01 vs LPS组);抑制LPS诱导的JAK1-stat3信号途径激活及stat3的核转位;降低LPS诱发的ROS产生(P<0.01 vs LPS组)。结论金盏花苷E通过抑制ROS介导的JAK1-stat3信号途径,抑制LPS诱导的炎症反应。展开更多
Cervical cancer(CC)is recognized as the most common neoplasm in the female reproductive system worldwide.The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipa...Cervical cancer(CC)is recognized as the most common neoplasm in the female reproductive system worldwide.The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipated prognosis of patients.Aloperine(ALO)is a natural quinolizidine alkaloid with marked anti-cancer effects on multiple malignancies as well as favorable activity in relieving inflammation,allergies and infection.However,its therapeutic efficacy and underlying mechanism in CC are still unclear.In the current study,MTT assay was employed to evaluate the viability of HeLa cells exposed to ALO to preliminarily estimate the effectiveness of ALO in CC.Then,the effects of ALO on the proliferation and apoptosis of HeLa cells were further investigated by plate colony formation and flow cytometry,respectively,while the migration and invasion of ALO-treated HeLa cells were evaluated using Transwell assay.Moreover,nude mice were subcutaneously inoculated with HeLa cells to demonstrate the anti-CC properties of ALO in vivo.The molecular mechanisms underlying these effects of ALO were evaluated by Western blot and immunohistochemical analysis.This study experimentally demonstrated that ALO inhibited the proliferation of HeLa cells via G2 phase cell cycle arrest.Simultaneously,ALO promoted an increase in the percentage of apoptotic HeLa cells by increasing the Bax/Bcl-2 ratio.Additionally,the migration and invasion of HeLa cells were attenuated by ALO treatment,which was considered to result from inhibition of epithelial-to-mesenchymal transition.For molecular mechanisms,the expression and activation of the IL-6-JAK1-STAT3 feedback loop were markedly suppressed by ALO treatment.This study indicated that ALO markedly suppresses the proliferation,migration and invasion and enhances the apoptosis of HeLa cells.In addition,these prominent anti-CC properties of ALO are associated with repression of the IL-6-JAK1-STAT3 feedback loop.展开更多
文摘目的探讨金盏花苷E对脂多糖(LPS)诱导炎症反应的抑制作用及可能的分子机制。方法 CCK-8实验检测不同浓度(0、2、4、6、8、10、20、25、30μg/mL)的金盏花苷E对RAW264.7细胞活力的影响;不同浓度的金盏花苷E(0、6、8、10μg/mL)预处理RAW264.7细胞2 h,然后用LPS(100 ng/mL)刺激细胞特定的时间,ELISA检测炎症因子TNF-α、IL-1β释放;Western blotting检测iNOS、COX-2的表达水平及JAK-stats、MAPKs及NF-кB信号途径的磷酸化;ROS检测试剂盒检测RAW264.7细胞内ROS含量;激光共聚焦实验检测转录因子stat3的核转位。结果 CCK-8结果显示,金盏花苷E浓度在低于20μg/mL时对RAW264.7细胞无明显毒性作用;金盏花苷E浓度依赖性地下调LPS诱导的iNOS和COX-2的表达(P<0.01 vs LPS组);抑制LPS诱导的促炎细胞因子TNF-α及IL-1β的释放,且1 0μg/mL组抑制作用尤为显著(P<0.01 vs LPS组);抑制LPS诱导的JAK1-stat3信号途径激活及stat3的核转位;降低LPS诱发的ROS产生(P<0.01 vs LPS组)。结论金盏花苷E通过抑制ROS介导的JAK1-stat3信号途径,抑制LPS诱导的炎症反应。
基金This work was supported by the National Natural Science Foundation of China(No.82001850)Shanxi Basic Application Research(No.201901D211491)+2 种基金the Scientific Research Project of Shanxi Health Commission(No.2019038)the Doctoral Research Project of Shanxi Medical University(No.XD1901)the Doctoral Research Project of Shanxi Province(No.SD1901).
文摘Cervical cancer(CC)is recognized as the most common neoplasm in the female reproductive system worldwide.The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipated prognosis of patients.Aloperine(ALO)is a natural quinolizidine alkaloid with marked anti-cancer effects on multiple malignancies as well as favorable activity in relieving inflammation,allergies and infection.However,its therapeutic efficacy and underlying mechanism in CC are still unclear.In the current study,MTT assay was employed to evaluate the viability of HeLa cells exposed to ALO to preliminarily estimate the effectiveness of ALO in CC.Then,the effects of ALO on the proliferation and apoptosis of HeLa cells were further investigated by plate colony formation and flow cytometry,respectively,while the migration and invasion of ALO-treated HeLa cells were evaluated using Transwell assay.Moreover,nude mice were subcutaneously inoculated with HeLa cells to demonstrate the anti-CC properties of ALO in vivo.The molecular mechanisms underlying these effects of ALO were evaluated by Western blot and immunohistochemical analysis.This study experimentally demonstrated that ALO inhibited the proliferation of HeLa cells via G2 phase cell cycle arrest.Simultaneously,ALO promoted an increase in the percentage of apoptotic HeLa cells by increasing the Bax/Bcl-2 ratio.Additionally,the migration and invasion of HeLa cells were attenuated by ALO treatment,which was considered to result from inhibition of epithelial-to-mesenchymal transition.For molecular mechanisms,the expression and activation of the IL-6-JAK1-STAT3 feedback loop were markedly suppressed by ALO treatment.This study indicated that ALO markedly suppresses the proliferation,migration and invasion and enhances the apoptosis of HeLa cells.In addition,these prominent anti-CC properties of ALO are associated with repression of the IL-6-JAK1-STAT3 feedback loop.
