【摘要】：LPS和Visfatin對細胞損傷方面的研究雖然有很多報道,但是有關兩者對心肌細胞炎癥共同作用的報道很少,所以有必要進行更加深入的研究。[研究背景]：Visfatin是最近幾年發(fā)現(xiàn)的由內臟脂肪分泌的脂肪因子,對細胞的炎性產生及發(fā)展有一定的促進作用。LPS是革蘭氏陰性細菌細胞壁的主要成分,是內毒素的特異性抗原,能夠引起機體的微循環(huán)障礙。本文主要是研究Visfatin在LPS誘導的心肌細胞炎癥作用方面的機制。[實驗原理]：心肌細胞在LPS炎性模型中加入Visfatin,細胞外刺激使得I-κB激酶復合體活化, I-κB磷酸化,使得以無活性形式存于細胞漿中的NF-κB和I-κB的復合體分離,NF-κB暴露核定位位點,游離的NF-κB迅速移位,進入到細胞核,與特異性KB序列結合,誘導相關基因轉錄,引起粘附因子ICAM-1和VCAM-1的表達,誘導了白細胞的聚集和粘附,導致了多種炎癥因子的表達,從而產生心肌損傷。[實驗方法]：MTT法檢測了不同濃度下的LPS和Visfatin對心肌的損傷程度,確定實驗濃度。利用RT-PCR檢測了粘附因子的mRNA表達情況后,并用Western blot法檢測了相應蛋白表達,最后通過Western blot法檢測了NF-κB p65的入核及I-κB的磷酸化情況。[實驗結果]：在LPS、Visfatin單獨處理的心肌細胞中發(fā)現(xiàn)ICAM-1和VCAM-1的表達量對比對照組均有所增加,二者共同作用時發(fā)現(xiàn)粘附因子的表達量明顯高于單獨處理組,而這個過程是通過I-KB的活化導致NF-κB入核,最終引起了下游粘附因子的表達途徑,從而引起了心肌細胞的損傷,活性下降。 結論：Visfatin對LPS誘導的原代心肌細胞炎性因子ICAM-1和VCAM-1的表達具有顯著的促進作用,而這一過程是通過I-κB的磷酸化降解釋放出NF-κB復合體,增加了NF-κB p65的入核,啟動了下游的粘附因子mRNA和蛋白的表達。
[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.
【學位授予單位】：東北師范大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R363

【參考文獻】

相關期刊論文 前1條

1 鄭煒,趙小燕;淺談脂多糖對機體的影響[J];甘肅中醫(yī)學院學報;1998年S1期

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本文編號：2477203