【摘要】：膿毒癥的高致死率一直是臨床急需解決的難題。為了研究膿毒癥炎癥局部的免疫細(xì)胞應(yīng)答對(duì)疾病發(fā)生發(fā)展的影響,我們利用大腸桿菌腹腔注射的方法建立了小鼠膿毒癥性腹膜炎模型,并觀察了感染局部腹腔灌洗細(xì)胞(P LC)中的中性粒細(xì)胞(P MN)的數(shù)量、形態(tài)、亞群及活性的變化,發(fā)現(xiàn):1)腹膜炎小鼠PLC中PMN的數(shù)量明顯增多,出現(xiàn)巨大細(xì)胞,且與菌量正相關(guān);2)PMN的兩個(gè)亞群CD11b~+PMN和CD11b-PMN比例有差異,感染早期CD11b~+PMN降低而CD11b-PMN比例增高,隨著病程延長(zhǎng),兩群細(xì)胞比例均回歸正常;3)雖然在感染18h時(shí),C D11b~+PMN和CD11b-PMN中s TLR9+細(xì)胞比例均明顯增高,但在感染的更早時(shí)間(8h),s TLR9表達(dá)水平增高主要出現(xiàn)在CD11b~+PMN,而高菌量組的CD11b-PMN表達(dá)s TLR9水平降低;4)作為炎癥細(xì)胞因子的IL-17在感染早期(8h)的CD11b~+PMN和CD11b-PMN中的表達(dá)水平均明顯增高,但在感染18h時(shí)似乎只有C D11b~+PMN還在分泌IL-17;5)對(duì)于CD11b~+PMN,s TLR9和IL-17的表達(dá)不在同一群細(xì)胞內(nèi),但二者的變化規(guī)律是一致的;6)免疫抑制性O(shè)DN能提高膿毒癥腹膜炎小鼠的生存率,抑制P MN兩個(gè)亞群s TLR9的表達(dá)水平,也抑制C D11b~+PMN的IL-17表達(dá)水平及T細(xì)胞和巨噬細(xì)胞IL-17的表達(dá)水平。值得注意的是:在本研究中,我們通過(guò)流式細(xì)胞分選法證明細(xì)菌感染局部I L-17的產(chǎn)生不僅來(lái)源于C D3+T細(xì)胞,也來(lái)源于PMN。我們的研究提示:PMN在細(xì)菌感染中發(fā)揮重要的作用,但不同P MN亞群的功能有差異,s TLR9+PMN及IL17+PMN在感染后病情發(fā)展中有重要作用,可能成為膿毒癥干預(yù)治療或預(yù)后預(yù)測(cè)的關(guān)注對(duì)象。
[Abstract]:The high mortality rate of sepsis is an urgent problem in clinic. In order to study the effect of local immune cell response of sepsis inflammation on the occurrence and development of the disease, we established a mouse model of septic peritonitis by intraperitoneal injection of Escherichia coli. The number, morphology, subgroup and activity of neutrophil (P MN) in (P LC) of infected local peritoneal lavage cells were observed. The results showed that: 1) the number of PMN in PLC of peritonitis mice increased obviously, and giant cells appeared. And positive correlation with the amount of bacteria; 2) the proportion of CD11b~ PMN and CD11b-PMN in two subsets of PMN was different. The proportion of CD11b~ PMN decreased but CD11b-PMN increased in the early stage of infection. With the prolongation of the course of disease, the proportion of CD11b~ PMN and CD11b-PMN returned to normal. 3) although the proportion of s TLR9 cells in C D 11b ~ PMN and CD11b-PMN increased significantly at 18 h, the increase of), s TLR9 expression at the early stage of infection (8 h) was mainly found in CD11b~ PMN, while the CD11b-PMN expression s TLR9 level in high bacteria group was decreased. 4) the expression of IL-17, as an inflammatory cytokine, was significantly increased in CD11b~ PMN and CD11b-PMN at the early stage of infection (8 h), but only C D 11b ~ PMN seemed to secrete IL-17; at 18 h after infection. 5) the expression of CD11b~ PMN,s TLR9 and IL-17 was not in the same group of cells, but the changes of them were consistent. 6) immunosuppressive ODN could increase the survival rate of mice with septic peritonitis, inhibit the expression of s TLR9 in two subsets of P MN, IL-17 expression of C D11b- PMN and IL-17 expression of T cells and macrophages. It is worth noting that, in this study, we demonstrated by flow cytometry that the local production of L-17 in bacterial infection was derived not only from C D3 T cells, but also from PMN.. Our results suggest that PMN plays an important role in bacterial infection, but the function of different P MN subsets is different. S TLR9 PMN and IL17 PMN play an important role in the development of disease after infection. It may be the focus of sepsis intervention therapy or prognosis prediction.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R459.7

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