Visfatin對(duì)心肌細(xì)胞炎性因子ICAM-1，VCAM-1表達(dá)的影響
發(fā)布時(shí)間：2019-05-15 01:20
【摘要】：LPS和Visfatin對(duì)細(xì)胞損傷方面的研究雖然有很多報(bào)道,但是有關(guān)兩者對(duì)心肌細(xì)胞炎癥共同作用的報(bào)道很少,所以有必要進(jìn)行更加深入的研究。[研究背景]：Visfatin是最近幾年發(fā)現(xiàn)的由內(nèi)臟脂肪分泌的脂肪因子,對(duì)細(xì)胞的炎性產(chǎn)生及發(fā)展有一定的促進(jìn)作用。LPS是革蘭氏陰性細(xì)菌細(xì)胞壁的主要成分,是內(nèi)毒素的特異性抗原,能夠引起機(jī)體的微循環(huán)障礙。本文主要是研究Visfatin在LPS誘導(dǎo)的心肌細(xì)胞炎癥作用方面的機(jī)制。[實(shí)驗(yàn)原理]：心肌細(xì)胞在LPS炎性模型中加入Visfatin,細(xì)胞外刺激使得I-κB激酶復(fù)合體活化, I-κB磷酸化,使得以無(wú)活性形式存于細(xì)胞漿中的NF-κB和I-κB的復(fù)合體分離,NF-κB暴露核定位位點(diǎn),游離的NF-κB迅速移位,進(jìn)入到細(xì)胞核,與特異性KB序列結(jié)合,誘導(dǎo)相關(guān)基因轉(zhuǎn)錄,引起粘附因子ICAM-1和VCAM-1的表達(dá),誘導(dǎo)了白細(xì)胞的聚集和粘附,導(dǎo)致了多種炎癥因子的表達(dá),從而產(chǎn)生心肌損傷。[實(shí)驗(yàn)方法]：MTT法檢測(cè)了不同濃度下的LPS和Visfatin對(duì)心肌的損傷程度,確定實(shí)驗(yàn)濃度。利用RT-PCR檢測(cè)了粘附因子的mRNA表達(dá)情況后,并用Western blot法檢測(cè)了相應(yīng)蛋白表達(dá),最后通過(guò)Western blot法檢測(cè)了NF-κB p65的入核及I-κB的磷酸化情況。[實(shí)驗(yàn)結(jié)果]：在LPS、Visfatin單獨(dú)處理的心肌細(xì)胞中發(fā)現(xiàn)ICAM-1和VCAM-1的表達(dá)量對(duì)比對(duì)照組均有所增加,二者共同作用時(shí)發(fā)現(xiàn)粘附因子的表達(dá)量明顯高于單獨(dú)處理組,而這個(gè)過(guò)程是通過(guò)I-KB的活化導(dǎo)致NF-κB入核,最終引起了下游粘附因子的表達(dá)途徑,從而引起了心肌細(xì)胞的損傷,活性下降。 結(jié)論：Visfatin對(duì)LPS誘導(dǎo)的原代心肌細(xì)胞炎性因子ICAM-1和VCAM-1的表達(dá)具有顯著的促進(jìn)作用,而這一過(guò)程是通過(guò)I-κB的磷酸化降解釋放出NF-κB復(fù)合體,增加了NF-κB p65的入核,啟動(dòng)了下游的粘附因子mRNA和蛋白的表達(dá)。
[Abstract]:Although there are many reports on cell injury caused by LPS and Visfatin, there are few reports on the interaction between them on cardiomyocyte inflammation, so it is necessary to carry out more in-depth study. [background]: LPs is a fat factor secreted by visceral fat discovered in recent years, which can promote the inflammatory production and development of cells. LPs is the main component of Gram-negative bacteria cell wall. It is a specific antigen of lipopolysaccharide, which can cause microcirculatory disturbance of the body. The purpose of this study was to study the mechanism of Visfatin in cardiomyocyte inflammation induced by LPS. [principle]: the extracellular stimulation of Visfatin, into the LPS inflammatory model activated the I-kappa B kinase complex and the phosphorylation of I-kappa B, resulting in the separation of the complexes of NF- and I-魏 B, which were stored in the cytoplasm in the form of inactivity, and the extracellular stimulation of the cardiomyocytes resulted in the activation of the I-kappa B kinase complex and the phosphorylation of the I-kappa B complex in the cytoplasm. NF- kappa B exposed to nuclear localization sites, free NF- kappa B rapidly shifted into the nucleus, bound to the specific KB sequence, induced the transcription of related genes, resulting in the expression of adhesion factors ICAM-1 and VCAM-1. It induced the aggregation and adhesion of leukocytes and led to the expression of many inflammatory factors, resulting in myocardial injury. [method]: MTT method was used to detect the degree of myocardial injury induced by LPS and Visfatin at different concentrations, and the experimental concentration was determined. The mRNA expression of adhesion factor was detected by RT-PCR, and the corresponding protein expression was detected by Western blot method. Finally, the nucleation of NF- 魏 B p65 and the phosphorylation of I-魏 B were detected by Western blot method. [results]: the expression of ICAM-1 and VCAM-1 in cardiomyocytes treated with LPS,Visfatin alone was significantly higher than that in the control group, and the expression of adhesion factor was significantly higher than that in the control group. This process leads to the entry of NF- kappa B into the nucleus through the activation of I-KB, and finally leads to the expression of downstream adhesion factor, which leads to the injury of cardiomyocytes and the decrease of activity. Conclusion: Visfatin can significantly promote the expression of inflammatory factors ICAM-1 and VCAM-1 in primary cardiomyocytes induced by LPS, and this process releases the NF- kB complex through the phosphorylation and degradation of I-魏 B and increases the nucleation of NF- 魏 B p65. The expression of downstream adhesion factor mRNA and protein was initiated.
【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R363
本文編號(hào)：2477203
[Abstract]:Although there are many reports on cell injury caused by LPS and Visfatin, there are few reports on the interaction between them on cardiomyocyte inflammation, so it is necessary to carry out more in-depth study. [background]: LPs is a fat factor secreted by visceral fat discovered in recent years, which can promote the inflammatory production and development of cells. LPs is the main component of Gram-negative bacteria cell wall. It is a specific antigen of lipopolysaccharide, which can cause microcirculatory disturbance of the body. The purpose of this study was to study the mechanism of Visfatin in cardiomyocyte inflammation induced by LPS. [principle]: the extracellular stimulation of Visfatin, into the LPS inflammatory model activated the I-kappa B kinase complex and the phosphorylation of I-kappa B, resulting in the separation of the complexes of NF- and I-魏 B, which were stored in the cytoplasm in the form of inactivity, and the extracellular stimulation of the cardiomyocytes resulted in the activation of the I-kappa B kinase complex and the phosphorylation of the I-kappa B complex in the cytoplasm. NF- kappa B exposed to nuclear localization sites, free NF- kappa B rapidly shifted into the nucleus, bound to the specific KB sequence, induced the transcription of related genes, resulting in the expression of adhesion factors ICAM-1 and VCAM-1. It induced the aggregation and adhesion of leukocytes and led to the expression of many inflammatory factors, resulting in myocardial injury. [method]: MTT method was used to detect the degree of myocardial injury induced by LPS and Visfatin at different concentrations, and the experimental concentration was determined. The mRNA expression of adhesion factor was detected by RT-PCR, and the corresponding protein expression was detected by Western blot method. Finally, the nucleation of NF- 魏 B p65 and the phosphorylation of I-魏 B were detected by Western blot method. [results]: the expression of ICAM-1 and VCAM-1 in cardiomyocytes treated with LPS,Visfatin alone was significantly higher than that in the control group, and the expression of adhesion factor was significantly higher than that in the control group. This process leads to the entry of NF- kappa B into the nucleus through the activation of I-KB, and finally leads to the expression of downstream adhesion factor, which leads to the injury of cardiomyocytes and the decrease of activity. Conclusion: Visfatin can significantly promote the expression of inflammatory factors ICAM-1 and VCAM-1 in primary cardiomyocytes induced by LPS, and this process releases the NF- kB complex through the phosphorylation and degradation of I-魏 B and increases the nucleation of NF- 魏 B p65. The expression of downstream adhesion factor mRNA and protein was initiated.
【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R363
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 鄭煒,趙小燕;淺談脂多糖對(duì)機(jī)體的影響[J];甘肅中醫(yī)學(xué)院學(xué)報(bào);1998年S1期
，本文編號(hào)：2477203
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