本文選題：mtDNA + 肺微血管內(nèi)皮細(xì)胞�。� 參考：《中國人民解放軍醫(yī)學(xué)院》2013年博士論文

【摘要】：目的研究線粒體DNA(mitochondrial DNA, mtDNA)對大鼠肺微血管內(nèi)皮細(xì)胞活化和通透性的影響及其分子生物學(xué)機制,探索創(chuàng)傷后組織細(xì)胞破裂釋放的線粒體DNA導(dǎo)致系統(tǒng)性炎癥及急性肺損傷的機制,為創(chuàng)傷后SIRS及急性肺損傷的救治提供實驗依據(jù)。 方法本研究采用分子生物學(xué)、流式細(xì)胞儀等技術(shù),探索mtDNA作為一利,源頭性損傷相關(guān)分子模式直接激活內(nèi)皮細(xì)胞,參與啟動全身炎癥反應(yīng)綜合征(systemic inflammatory response syndrome,SIRS)的分子機制,探索線粒體DNA啟動、轉(zhuǎn)導(dǎo)、調(diào)控失控性炎癥反應(yīng)機制。通過大鼠尾靜脈注入臨床濃度的mtDNA導(dǎo)致急性肺損傷,檢測外周血炎癥因子TNF-α、IL-6及血管性血友病因子(von Willebrand factor, vWF)、sE-selectin和sICAM-1濃度,檢測肺血管通透性；以臨床濃度mtDNA刺激體外培養(yǎng)大鼠肺微血管內(nèi)皮細(xì)胞,檢測其通透性變化及vWF、sE-selectin和sICAM-1表達(dá)水平以及TLR-9mRNA、NF-κB亞單位p65mRNA、I-κB mRNA表達(dá)水平；將大鼠肺微血管內(nèi)皮細(xì)胞與中性粒細(xì)胞共培養(yǎng),予mtDNA刺激后觀察微血管內(nèi)皮細(xì)胞存活率、中性粒細(xì)胞粘附情況。 結(jié)果10ug/ml mtDNA體內(nèi)注射導(dǎo)致大鼠急性肺損傷；mtDNA刺激大鼠微血管內(nèi)皮細(xì)胞中的TLR-9mRNA、NF-κB亞單位p65mRNA表達(dá)增加,I-κ BmRNA表達(dá)降低：mtDNA能直接激活大鼠微血管內(nèi)皮細(xì)胞,顯著增加vwF、sE-selectin和sICAM-1表達(dá),使大鼠微血管內(nèi)皮細(xì)胞通透性增加,顯著促進(jìn)大鼠微血管內(nèi)皮細(xì)胞與中性粒細(xì)胞的粘附。 結(jié)論mtDNA能夠通過TLR-9激活大鼠微血管內(nèi)皮細(xì)胞NF-κB通路,刺激大鼠微血管內(nèi)皮細(xì)胞產(chǎn)生vwF、sE-selectin和sICAM-1,增加內(nèi)皮細(xì)胞通透性,顯著促進(jìn)與中性粒細(xì)胞的粘附。mtDNA激活內(nèi)皮細(xì)胞是創(chuàng)傷后SIRS及ALI的重要啟動機制。
[Abstract]:Objective to investigate the effect of mitochondrial DNA(mitochondrial DNA (mtDNA) on the activation and permeability of rat pulmonary microvascular endothelial cells and its molecular biological mechanism, and to explore the mechanism of systemic inflammation and acute lung injury induced by mitochondrial DNA released by tissue cell rupture after trauma. To provide experimental evidence for the treatment of SIRS and acute lung injury after trauma. Methods in this study, molecular biology, flow cytometry and other techniques were used to explore the molecular mechanism of mtDNA, as a beneficial molecular model associated with source injury, directly activating endothelial cells and initiating systemic inflammatory response syndromes (SIRs). To explore the mechanism of mitochondrial DNA activation, transduction and control of uncontrolled inflammatory response. Acute lung injury was induced by injecting clinical concentration of mtDNA into the tail vein of rats. The levels of TNF- 偽 IL-6, von Willebrand factor, vWFU E-selectin and sICAM-1 in peripheral blood were measured to determine the pulmonary vascular permeability. Pulmonary microvascular endothelial cells were cultured in vitro with clinical concentration of mtDNA. The permeability, the expression of vWFN E-selectin and sICAM-1 and the expression of TLR-9mRNA-NF- 魏 B mRNA were detected, and the p65mRNA-kappa B subunit p65mRNA- 魏 B mRNA were co-cultured with neutrophils. The survival rate of microvascular endothelial cells and the adhesion of neutrophils were observed after mtDNA stimulation. Results 10ug/ml mtDNA injection in vivo stimulated the expression of TLR-9 mRNA-NF- 魏 B subunit p65mRNA in rat microvascular endothelial cells (MECs) and increased the expression of TLR-9mRNA-kappa B subunit p65mRNA. The expression of TLR-9mRNA-kappa B subunit in rat microvascular endothelial cells (MECs) and the expression of VWFNs E-selectin and sICAM-1 were directly activated by 10ug/ml mtDNA. The permeability of rat microvascular endothelial cells was increased, and the adhesion of rat microvascular endothelial cells to neutrophils was significantly promoted. Conclusion mtDNA can activate the NF- 魏 B pathway of rat microvascular endothelial cells through TLR-9, stimulate the production of vwFN E-selectin and sICAM-1 by rat microvascular endothelial cells, and increase the permeability of endothelial cells. Promoting the adhesion with neutrophil. Mt DNA activated endothelial cells is an important mechanism of SIRS and ALI after trauma.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2013
【分類號】：R563

【共引文獻(xiàn)】

相關(guān)期刊論文 前9條

1 蔡文利;苗書齋;王沛育;曲青山;;雜合式腎臟替代與持續(xù)床旁血液濾過在移植腎功能延遲的應(yīng)用比較[J];廣東醫(yī)學(xué);2011年16期

2 王俊琳;嚴(yán)玉澄;;老年人急性腎損傷的特點及研究進(jìn)展[J];中國中西醫(yī)結(jié)合腎病雜志;2012年03期

3 李慧妍;吳學(xué)敏;魏濵;;急性腎衰竭的腎替代療法研究進(jìn)展[J];臨床薈萃;2009年16期

4 李謙平;張軍;羅衛(wèi)民;原野;程棟梁;曾敏;;重癥心臟瓣膜病術(shù)后腎功能不全患者連續(xù)性腎替代治療體會[J];嶺南心血管病雜志;2011年01期

5 彭家清;;老年急性腎損傷的臨床特點及死亡危險因素分析[J];中國中西醫(yī)結(jié)合腎病雜志;2013年08期

6 樊永亮;袁忠祥;;心臟術(shù)后急性腎損傷的研究進(jìn)展[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2009年20期

7 王芳;何強;洪大情;蒲蕾;張月;王莉;;持續(xù)靜靜脈血液濾過和持續(xù)低效每日透析治療重癥急性腎損傷的臨床研究[J];中國血液凈化;2011年06期

8 王莉;蒲蕾;;連續(xù)性腎臟替代治療在危重癥患者合并急性腎損傷中的應(yīng)用[J];中國血液凈化;2011年07期

9 曹長琦;李西慧;;心臟外科手術(shù)后急性腎功能衰竭的防治進(jìn)展[J];中國胸心血管外科臨床雜志;2008年06期

相關(guān)碩士學(xué)位論文 前1條

1 束偉斌;急性腎損傷后腎功能遠(yuǎn)期追蹤的初步研究[D];山東大學(xué);2011年

，

本文編號：1899969