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[摘要]
目的 研究氧化苦参碱(OM)对脂多糖(LPS)诱导的大鼠胰腺星状细胞株(PSCs)NOD样受体蛋白3(NLRP3)活化的影响。方法 通过设置不同剂量(0、2.5、5、10、20、40 μg/mL)的LPS并设置相同剂量(10 μg/mL)的LPS刺激LTC14细胞不同时间(0、1、3、6、9、12 h),利用Western blotting检测技术,考察LPS引起炎性小体活化的量效关系;使用OM、Janus蛋白酪氨酸激酶2(JAK2)抑制剂AG490干预LPS诱导的LTC14细胞株后,通过Westernblotting技术检测Janus蛋白酪氨酸激酶2/信号转导子和转录激活子3(JAK2/STAT3)信号通路中相关分子以及NLRP3炎性小体相关蛋白表达的变化。结果 与对照组比较,LTC14细胞接受不同浓度的LPS刺激后JAK2、STAT3、NLRP3、caspase1、白细胞介素-1β(IL-1β)蛋白表达呈显著增高趋势;与对照组比较,LTC14细胞接受相同浓度LPS刺激不同时间后JAK2、STAT3、NLRP3、caspase1、IL-1β蛋白表达呈显著增高趋势;与LPS组对比,OM和LPS共同处理组、AG490和LPS共同处理组JAK2、STAT3、NLRP3、caspase1及IL-1β的蛋白表达显著降低。结论 LPS呈一定的时间-剂量关系激活LTC14细胞JAK2/STAT3信号通路及NLRP3炎性小体;OM可能通过抑制JAK2/STAT3信号通路起到抑制NLRP3炎性小体活化的作用。
[Key word]
[Abstract]
Objective To study the effects of oxymatrine (OM) on the activation of NLRP3 in pancreatic stellate cells (PSCs) induced by lipopolysaccharide (LPS).Methods By setting different doses of LPS (0, 2.5, 5, 10, 20, 40 μg/mL) and setting the same dosage of LPS (10 μg/mL) to stimulate LTC14 cells at different time (0, 1, 3, 6, 9, 12 h), the dose-effect relationship of LPS-induced inflammatory corpuscles activation was investigated by Western blotting. After using OM and JAK2 inhibitor AG490 to intervene LPS-induced LTC14 cell lines, the expression of related molecules in JAK2/STAT3 signaling pathway and NLRP3 inflammatory corpuscles -related protein were detected by Western blotting.Results Compared with the control group, the expression of JAK2, STAT3, NLRP3, caspase 1 and IL-1β protein in LTC14 cells were increased significantly after LPS stimulation at different concentrations. Compared with the control group, the expression of JAK2, STAT3, NLRP3, caspase 1 and IL-1β protein in LTC14 cells were increased significantly after LPS stimulation at the same concentration for different time. The protein expressions of JAK2, STAT3, NLRP3, caspase 1 and IL-1β were significantly decreased in LPS-treated group. Conclusion LPS activates JAK2/STAT3 signaling pathway and NLRP3 inflammatory corpuscles in LTC14 cells in a time-dose relationship. OM may inhibit NLRP3 inflammatory body activation by inhibiting JAK2/STAT3 signaling pathway.
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[基金项目]
国家自然科学基金资助项目(81500489);天津市自然科学基金联合资助青年项目(15JCQNJC45600);天津市西青医院科研基金(XQYYLX201603)