Luteolin通過抑制Hsp90的活性來調節(jié)LPS誘導的RAW264.7細胞中炎癥因子的表達
發(fā)布時間：2019-01-26 21:54
【摘要】： 細菌嚴重感染引起的失控性炎癥反應能導致休克、全身炎癥反應綜合征,并進而誘發(fā)多器官功能障礙綜合征和多器官衰竭。LPS又叫內毒素,是革蘭氏陰性細菌細胞壁的主要成分,可激活哺乳動物的內皮細胞以及單核巨噬細胞。在關節(jié)炎,動脈粥樣硬化等疾病中單核巨噬細胞是介導炎癥反應的主要細胞,誘導致炎細胞因子、趨化因子、生長因子和其他多種因子如白細胞介素、腫瘤壞死因子等的合成和釋放。 木犀草素是廣泛存在于蔬菜水果和藥用植物中的一種黃酮類物質。木犀草素有兩個苯環(huán)和一個含兩個氧原子的環(huán),含有不飽和的鍵,有兩個碳雙鍵,在第四位有個羰基。研究表明Luteolin有抗癌、抗氧化、抗炎及保護心臟等作用。 我們本篇文章主要是對Luteolin抑制LPS誘導的RAW264.7細胞中炎癥調節(jié)因子的表達的機制進行研究。我們首次報道了Luteolin可能通過抑制Hsp90的活性來削弱LPS誘導的致炎因子的釋放。熱休克蛋白90是真核生物細胞中含量最為豐富的胞質蛋白之一,其重要的一項功能是作為分子伴侶參與新生肽鏈和環(huán)境壓力下變性的蛋白折疊成具有天然構象的功能蛋白,以及對客戶蛋白的穩(wěn)定性和活性進行調節(jié)。迄今為止,HSP90的客戶蛋白已發(fā)現(xiàn)有100多種,主要分為轉錄因子、蛋白激酶、聚合酶等幾大類。其中Akt、IRAK1、c-Jun和iNOS是已知的Hsp90的客戶蛋白。 我們的研究結果顯示:Luteolin能與Hsp90結合,并降低RAW264.7細胞中c-Jun,Akt,IRAK1的蛋白水平,但不影響它們的mRNA水平。說明Luteolin只是在蛋白水平而非轉錄水平下調Hsp90客戶蛋白的穩(wěn)定性。在過表達Hsp90后可以回升原先被Luteolin明顯下調的c-Jun,Akt的蛋白量;但對瞬轉Hsp90(1-401aa)的RAW264.7細胞中客戶蛋白的蛋白水平并沒有回復。并且Luteolin能把Akt的半衰期從12小時縮短到6小時,減少了其60%的表達量。我們由此猜測Luteolin可能是潛在的Hsp90抑制劑,抑制其伴侶活性,從而阻斷客戶蛋白的折疊或再折疊過程,最終影響細胞內轉錄調控和信號轉導等多種生命活動。 我們進一步研究發(fā)現(xiàn),對于MAPKs信號通路,Luteolin阻斷了MAPKs的激活,尤其是JNK的磷酸化,從而也抑制了c-Jun(Ser63)的活化。Jun-2熒光素酶報告基因分析實驗表明,Luteolin除了通過抑制JNK的激活影響c-Jun的磷酸化,也能夠下調c-Jun蛋白的穩(wěn)定性,最終有效地降低c-Jun總蛋白轉錄活性。c-Jun是序列特異性轉錄激活因子AP-1的成分之一,廣泛地參與細胞內轉錄調控和信號轉導。Luteolin下調活化轉錄因子和上游激酶蛋白水平,阻礙下游的炎癥調節(jié)因子iNOS,COX-2的表達和NO,TNF-α的產(chǎn)生。 綜上所述,Luteolin的抗炎效果是通過兩方面發(fā)揮的。一方面,Luteolin抑制了MAPKs的活化,削弱了炎癥因子的表達,從而對LPS引起的炎癥起到治療的效果。另一方面,Luteolin抑制了Hsp90的分子伴侶活性,致使炎癥相關通路重的關鍵調節(jié)因子蛋白水平降低。這為我們提供了Luteolin在治療炎癥方面的新靶標。
[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.
【學位授予單位】：南京師范大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R363
本文編號：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.
【學位授予單位】：南京師范大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R363
【參考文獻】
相關期刊論文 前1條
1 姜勇,趙克森,韓家淮;p38 MAPK 參與 LPS 誘導 RAW 細胞 TNF-α 基因表達的調控[J];中國病理生理雜志;1998年03期
，本文編號：2415948
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