Tiwari Rajiv Lochan, Singh Vishal, Singh Ankita, Rana Minakshi, Verma Anupam, Kothari Nikhil, Kohli Monica, Bogra Jaishri, Dikshit Madhu, Barthwal Manoj Kumar
Pharmacology Division, Council of Scientific and Industrial Research-Central Drug Research Institute, Lucknow, India.
Department of Transfusion Medicine, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, India.
J Lipid Res. 2014 Jul;55(7):1226-44. doi: 10.1194/jlr.M045658. Epub 2014 May 2.
This study examined the role of interleukin (IL)-1 receptor-associated kinase (IRAK) and protein kinase C (PKC) in oxidized LDL (Ox-LDL)-induced monocyte IL-1β production. In THP1 cells, Ox-LDL induced time-dependent secretory IL-1β and IRAK1 activity; IRAK4, IRAK3, and CD36 protein expression; PKCδ-JNK1 phosphorylation; and AP-1 activation. IRAK1/4 siRNA and inhibitor (INH)-attenuated Ox-LDL induced secreted IL-1β and pro-IL-1β mRNA and pro-IL-1β and mature IL-1β protein expression, respectively. Diphenyleneiodonium chloride (NADPH oxidase INH) and N-acetylcysteine (free radical scavenger) attenuated Ox-LDL-induced reactive oxygen species generation, caspase-1 activity, and pro-IL-1β and mature IL-1β expression. Ox-LDL-induced secretory IL-1β production was abrogated in the presence of JNK INH II, Tanshinone IIa, Ro-31-8220, Go6976, Rottlerin, and PKCδ siRNA. PKCδ siRNA attenuated the Ox-LDL-induced increase in IRAK1 kinase activity, JNK1 phosphorylation, and AP-1 activation. In THP1 macrophages, CD36, toll-like receptor (TLR)2, TLR4, TLR6, and PKCδ siRNA prevented Ox-LDL-induced PKCδ and IRAK1 activation and IL-1β production. Enhanced Ox-LDL and IL-1β in systemic inflammatory response syndrome (SIRS) patient plasma demonstrated positive correlation with each other and with disease severity scores. Ox-LDL-containing plasma induced PKCδ and IRAK1 phosphorylation and IL-1β production in a CD36-, TLR2-, TLR4-, and TLR6-dependent manner in primary human monocytes. Results suggest involvement of CD36, TLR2, TLR4, TLR6, and the PKCδ-IRAK1-JNK1-AP-1 axis in Ox-LDL-induced IL-1β production.
本研究检测了白细胞介素(IL)-1受体相关激酶(IRAK)和蛋白激酶C(PKC)在氧化型低密度脂蛋白(Ox-LDL)诱导单核细胞产生IL-1β中的作用。在THP1细胞中,Ox-LDL诱导IL-1β分泌和IRAK1活性呈时间依赖性;诱导IRAK4、IRAK3和CD36蛋白表达;诱导PKCδ-JNK1磷酸化以及AP-1激活。IRAK1/4小干扰RNA(siRNA)和抑制剂(INH)分别减弱了Ox-LDL诱导的IL-1β分泌以及前IL-1β mRNA、前IL-1β和成熟IL-1β蛋白表达。二苯碘鎓氯化物(NADPH氧化酶INH)和N-乙酰半胱氨酸(自由基清除剂)减弱了Ox-LDL诱导的活性氧生成、半胱天冬酶-1活性以及前IL-1β和成熟IL-1β表达。在存在JNK INH II、丹参酮IIa、Ro-31-8220、Go6976、rottlerin和PKCδ siRNA的情况下,Ox-LDL诱导的IL-1β分泌产生被消除。PKCδ siRNA减弱了Ox-LDL诱导的IRAK1激酶活性增加、JNK1磷酸化以及AP-1激活。在THP1巨噬细胞中,CD36、Toll样受体(TLR)2、TLR4、TLR6和PKCδ siRNA可阻止Ox-LDL诱导的PKCδ和IRAK1激活以及IL-1β产生。全身炎症反应综合征(SIRS)患者血浆中升高的Ox-LDL和IL-1β彼此之间以及与疾病严重程度评分呈正相关。含Ox-LDL的血浆以CD36、TLR2、TLR4和TLR6依赖性方式诱导原代人单核细胞中的PKCδ和IRAK1磷酸化以及IL-1β产生。结果提示CD36、TLR2、TLR4、TLR6以及PKCδ-IRAK1-JNK1-AP-1轴参与了Ox-LDL诱导的IL-1β产生。