Luteolin通过抑制Hsp90的活性来调节LPS诱导的RAW264.7细胞中炎症因子的表达
发布时间：2019-01-26 21:54
【摘要】： 细菌严重感染引起的失控性炎症反应能导致休克、全身炎症反应综合征,并进而诱发多器官功能障碍综合征和多器官衰竭。LPS又叫内毒素,是革兰氏阴性细菌细胞壁的主要成分,可激活哺乳动物的内皮细胞以及单核巨噬细胞。在关节炎,动脉粥样硬化等疾病中单核巨噬细胞是介导炎症反应的主要细胞,诱导致炎细胞因子、趋化因子、生长因子和其他多种因子如白细胞介素、肿瘤坏死因子等的合成和释放。 木犀草素是广泛存在于蔬菜水果和药用植物中的一种黄酮类物质。木犀草素有两个苯环和一个含两个氧原子的环,含有不饱和的键,有两个碳双键,在第四位有个羰基。研究表明Luteolin有抗癌、抗氧化、抗炎及保护心脏等作用。 我们本篇文章主要是对Luteolin抑制LPS诱导的RAW264.7细胞中炎症调节因子的表达的机制进行研究。我们首次报道了Luteolin可能通过抑制Hsp90的活性来削弱LPS诱导的致炎因子的释放。热休克蛋白90是真核生物细胞中含量最为丰富的胞质蛋白之一,其重要的一项功能是作为分子伴侣参与新生肽链和环境压力下变性的蛋白折叠成具有天然构象的功能蛋白,以及对客户蛋白的稳定性和活性进行调节。迄今为止,HSP90的客户蛋白已发现有100多种,主要分为转录因子、蛋白激酶、聚合酶等几大类。其中Akt、IRAK1、c-Jun和iNOS是已知的Hsp90的客户蛋白。 我们的研究结果显示:Luteolin能与Hsp90结合,并降低RAW264.7细胞中c-Jun,Akt,IRAK1的蛋白水平,但不影响它们的mRNA水平。说明Luteolin只是在蛋白水平而非转录水平下调Hsp90客户蛋白的稳定性。在过表达Hsp90后可以回升原先被Luteolin明显下调的c-Jun,Akt的蛋白量;但对瞬转Hsp90(1-401aa)的RAW264.7细胞中客户蛋白的蛋白水平并没有回复。并且Luteolin能把Akt的半衰期从12小时缩短到6小时,减少了其60%的表达量。我们由此猜测Luteolin可能是潜在的Hsp90抑制剂,抑制其伴侣活性,从而阻断客户蛋白的折叠或再折叠过程,最终影响细胞内转录调控和信号转导等多种生命活动。 我们进一步研究发现,对于MAPKs信号通路,Luteolin阻断了MAPKs的激活,尤其是JNK的磷酸化,从而也抑制了c-Jun(Ser63)的活化。Jun-2荧光素酶报告基因分析实验表明,Luteolin除了通过抑制JNK的激活影响c-Jun的磷酸化,也能够下调c-Jun蛋白的稳定性,最终有效地降低c-Jun总蛋白转录活性。c-Jun是序列特异性转录激活因子AP-1的成分之一,广泛地参与细胞内转录调控和信号转导。Luteolin下调活化转录因子和上游激酶蛋白水平,阻碍下游的炎症调节因子iNOS,COX-2的表达和NO,TNF-α的产生。 综上所述,Luteolin的抗炎效果是通过两方面发挥的。一方面,Luteolin抑制了MAPKs的活化,削弱了炎症因子的表达,从而对LPS引起的炎症起到治疗的效果。另一方面,Luteolin抑制了Hsp90的分子伴侣活性,致使炎症相关通路重的关键调节因子蛋白水平降低。这为我们提供了Luteolin在治疗炎症方面的新靶标。
[Abstract]:The uncontrolled inflammatory response caused by severe bacterial infection can lead to shock, systemic inflammatory response syndrome, and in turn induce multiple organ dysfunction syndrome and multiple organ failure. LPS, in turn, is an endotoxin, a major component of the cell wall of the gram-negative bacteria, and can activate the endothelial cells of the mammal as well as the mononuclear macrophages. In the case of arthritis, atherosclerosis, and other diseases, the mononuclear macrophages are the main cells that mediate the inflammatory response, inducing the synthesis and release of inflammatory cytokines, chemokines, growth factors and other factors such as interleukins, tumor necrosis factors, and the like. luteolin is a kind of flavonoids which is widely used in vegetable and fruit and medicinal plants. a substance. The luteus is known as two benzene rings and a ring containing two oxygen atoms, with unsaturated bonds, two carbon double bonds, and one in the fourth place. The study shows that Lteberry has anti-cancer, anti-oxidation, anti-inflammatory, and protective heart. The effect of this article is mainly on the expression of the inflammatory regulation factor in the cells of the RAW264.7 cells induced by Lteolin in the inhibition of LPS-induced RAW264.7 cells. The study was made. We first reported that the Ltesta could impair LPS-induced inflammation by inhibiting the activity of Hsp90. The heat shock protein 90 is one of the most abundant cytoplasmic proteins in the true nuclear biological cells, an important function of which is to fold the denatured protein as a molecular chaperone in the nascent peptide chain and the ambient pressure to have a natural conformation. Functional proteins, as well as the stability and activity of the customer protein To date, the customer protein of HSP90 has been found to be more than 100, mainly divided into transcription factor, protein kinase, polymerization, Enzymes and the like, where Akt, IRAK1, c-Jun, and iNOS are known Hsp90 The results of our study showed that LteI could bind to Hsp90 and reduce the protein level of c-Jun, Akt, and IRAK1 in RAW264.7 cells, but did not affect it. The mRNA level of the patients was described. It was shown that the Ltedin was only down-regulated at the level of the protein rather than the level of transcription. The stability of the client protein. After the overexpression of Hsp90, the protein amount of c-Jun, Akt which was apparently down-regulated by the Lutsche can be recovered; however, the egg of the client protein in the RAW264.7 cell of the transient Hsp90 (1-401aa) The white level did not respond, and the Ltetrich can shorten the half-life of Akt from 12 hours to 6 hours, reducing It's 60% of its expression. We thus guess that the LteI may be a potential Hsp90 inhibitor to inhibit its partner activity, thereby blocking the folding or re-folding of the customer's protein, ultimately affecting transcription regulation and signal rotation in the cell. We have further studied the discovery that, for MAPKs signaling pathways, the Lteolin blocks the activation of MAPKs, in particular the phosphorylation of JNK, thereby also inhibiting c-Ju. The activation of n (Ser63). The analysis of the Jun-2 luciferase reporter gene indicated that the addition of Ltedin in addition to the inhibition of the phosphorylation of c-Jun by inhibiting the activation of JNK, it was also possible to downregulate the stability of c-Jun protein and ultimately to reduce c-Jun is one of the components of the sequence-specific transcriptional activator, AP-1. Intracellular transcriptional regulation and signal transduction. The production of NO, TNF-1. In conclusion, the Luteolin The anti-inflammatory effect is achieved by two aspects. On the one hand, the Lteolin inhibits the activation of the MAPKs, impairing the expression of the inflammatory factor, and thus, on the other hand, the luteolin inhibits the activity of the molecular chaperone of hsp90 and leads to an inflammation-related pathway. The heavy key regulatory factor protein level is reduced. This provides Lteo for us.
【学位授予单位】：南京师范大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R363
本文编号：2415948
[Abstract]:The uncontrolled inflammatory response caused by severe bacterial infection can lead to shock, systemic inflammatory response syndrome, and in turn induce multiple organ dysfunction syndrome and multiple organ failure. LPS, in turn, is an endotoxin, a major component of the cell wall of the gram-negative bacteria, and can activate the endothelial cells of the mammal as well as the mononuclear macrophages. In the case of arthritis, atherosclerosis, and other diseases, the mononuclear macrophages are the main cells that mediate the inflammatory response, inducing the synthesis and release of inflammatory cytokines, chemokines, growth factors and other factors such as interleukins, tumor necrosis factors, and the like. luteolin is a kind of flavonoids which is widely used in vegetable and fruit and medicinal plants. a substance. The luteus is known as two benzene rings and a ring containing two oxygen atoms, with unsaturated bonds, two carbon double bonds, and one in the fourth place. The study shows that Lteberry has anti-cancer, anti-oxidation, anti-inflammatory, and protective heart. The effect of this article is mainly on the expression of the inflammatory regulation factor in the cells of the RAW264.7 cells induced by Lteolin in the inhibition of LPS-induced RAW264.7 cells. The study was made. We first reported that the Ltesta could impair LPS-induced inflammation by inhibiting the activity of Hsp90. The heat shock protein 90 is one of the most abundant cytoplasmic proteins in the true nuclear biological cells, an important function of which is to fold the denatured protein as a molecular chaperone in the nascent peptide chain and the ambient pressure to have a natural conformation. Functional proteins, as well as the stability and activity of the customer protein To date, the customer protein of HSP90 has been found to be more than 100, mainly divided into transcription factor, protein kinase, polymerization, Enzymes and the like, where Akt, IRAK1, c-Jun, and iNOS are known Hsp90 The results of our study showed that LteI could bind to Hsp90 and reduce the protein level of c-Jun, Akt, and IRAK1 in RAW264.7 cells, but did not affect it. The mRNA level of the patients was described. It was shown that the Ltedin was only down-regulated at the level of the protein rather than the level of transcription. The stability of the client protein. After the overexpression of Hsp90, the protein amount of c-Jun, Akt which was apparently down-regulated by the Lutsche can be recovered; however, the egg of the client protein in the RAW264.7 cell of the transient Hsp90 (1-401aa) The white level did not respond, and the Ltetrich can shorten the half-life of Akt from 12 hours to 6 hours, reducing It's 60% of its expression. We thus guess that the LteI may be a potential Hsp90 inhibitor to inhibit its partner activity, thereby blocking the folding or re-folding of the customer's protein, ultimately affecting transcription regulation and signal rotation in the cell. We have further studied the discovery that, for MAPKs signaling pathways, the Lteolin blocks the activation of MAPKs, in particular the phosphorylation of JNK, thereby also inhibiting c-Ju. The activation of n (Ser63). The analysis of the Jun-2 luciferase reporter gene indicated that the addition of Ltedin in addition to the inhibition of the phosphorylation of c-Jun by inhibiting the activation of JNK, it was also possible to downregulate the stability of c-Jun protein and ultimately to reduce c-Jun is one of the components of the sequence-specific transcriptional activator, AP-1. Intracellular transcriptional regulation and signal transduction. The production of NO, TNF-1. In conclusion, the Luteolin The anti-inflammatory effect is achieved by two aspects. On the one hand, the Lteolin inhibits the activation of the MAPKs, impairing the expression of the inflammatory factor, and thus, on the other hand, the luteolin inhibits the activity of the molecular chaperone of hsp90 and leads to an inflammation-related pathway. The heavy key regulatory factor protein level is reduced. This provides Lteo for us.
【学位授予单位】：南京师范大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R363
【参考文献】
相关期刊论文 前1条
1 姜勇,赵克森,韩家淮;p38 MAPK 参与 LPS 诱导 RAW 细胞 TNF-α 基因表达的调控[J];中国病理生理杂志;1998年03期
，本文编号：2415948
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