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The Mechanism of MtDNA Cause of SIRS and on the Preliminary Study of Protection Effect to the the SIRS of Chloroquine and PDTC

Author: ZhouMeng
Tutor: SunTianSheng
School: Anhui Medical University,
Course: Surgery
Keywords: mtDNA SIRS chloroquine TLR9 NF-κB
CLC: R459.7
Type: Master's thesis
Year: 2012
Downloads: 38
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Objective The body is caused to a systemic inflammatory response syndrome, or evenmultiple organ failure by the trauma. The pathogenesis and prevention of the systemicinflammatory response syndrome has not yet completely understood. It is well knownthat the body can produce a large number of mtDNA after trauma, mtDNA can beactivated P38protein kinase pathway to produce inflammatory factors, leading to asystemic inflammatory syndrome, but the mtDNA whether through TLR-9activation ofNF-κB pathway and inflammation factor is still unknown. Using in vivo tests, the firstinjection of different concentrations of mtDNA, the level of detection in peripheralblood of various inflammatory factors change, observed lung inflammatory infiltration,to explore the mechanism of mtDNA lead to SIRS; blockers chloroquine and PDTC onthe basis of this preliminary understanding the protective effect of chloroquine andPDTC for mtDNA cause of SIRS lower of SIRS, prevention and treatment ofmulti-organ damage and complications related to new ideas for the treatment of severetrauma.Methods The first part of the rats were divided into8groups;①buffer group (n=2)the②nDNA and (10ug/ml) group (n=14)③of mtDNA (15ug/ml) group (n=14)(4)mitochondrial fragments (final concentration containingmtDNA1μg/ml, MTD1) group(n=14)⑤of mtDNA (5ug/ml) group (n=14)(6) mitochondrial fragments (finalconcentration is containing mtDNA5μg/ml, MTD5) group (n=14)⑦of mtDNA(15ug/ml) group(n=14), the⑧mitochondrial fragments (final concentrationcontaining mtDNA15μg/ml, MTD15) group (n=14). The second part of the rats were divided into four groups:①of mtDNA (15ug/ml)+chloroquine (10mg/kg) group (n=14)②of mtDNA (15ug/ml)+chloroquine (30mg/kg) group (n=14)③of mtDNA(15ug/ml)+of PDTC (30mg/kg) group (n=14)④of mtDNA (15ug/ml)+PDTC(120mg/kg) group (n=14). Two parts, respectively, in1hour,2hours,4hours,8hours,12hours,24hours and48hours observed by enzyme-linked immunosorbent assay(ELISA) in peripheral blood TNF-alpha, IL-10, IL-6, the expression level;Multi-channel physiological detector to check vital signs in rats;real-time quantitativePCR (Real-time PCR) detection of lung tissue NF-κB subunit p65mRNA, I-κBmRNA,TLR9mRNA expression; fluorescence quantitative PCR detection of peripheral bloodmtDNA levels; HE staining of liver changes in the pathology of the lung, kidney,intestine. The experimental data were presented as mean±standard error (x±s)represents. All data use SPSS17.0statistical software using single-factor analysis ofvariance (one way ANOVA) and regression and correlation analysis, P <0.05significantdifference.Results Between the groups in rat blood pressure and breathing did not changesignificantly in all experimental groups, significant changes in body temperature atdifferent points in time, but the temperature change between the groups was notstatistically significant.Buffer injection in rats and nDNA rats no significant changes ateach time point of peripheral blood levels of inflammatory factors; mitochondrialfragmentation and mtDNA groups began to rise in one hour levels of inflammatoryfactors, and eight hours to reach the peak, with the injection of mitochondrialfragmentation andmtDNA concentration, the higher the level of inflammation, andmitochondrial fragments containing the corresponding mtDNA concentration with highlevels of inflammatory factors than this concentration of mtDNA.p65mRNA, I-κBmRNA expression levels in lung tissue and inflammatory factors, TLR9expressionlevels increased linearly related.p65mRNA, I-κB mRNA expression levels in lung tissuenegatively correlated.Chloroquine and PDTC the higher the concentration, the lower the level of expression of inflammatory cytokines to block the better, but chloroquine blockthe effect of PDTC was no significant difference.Conclusions mtDNA can stimulate TLR9-induced NF-κB pathway leading to thebody’s systemic inflammatory response syndrome. The protective effect of chloroquineand PDTC can be caused by mtDNA SIRS.

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