膿毒癥患者外周血DC成熟與分泌的變化及OXPAPC的影響
發(fā)布時(shí)間：2018-11-27 08:08
【摘要】：背景：膿毒癥患者樹突狀細(xì)胞(DC)的成熟度與分泌功能受到抑制,而DC活性的恢復(fù)有助于膿毒癥的好轉(zhuǎn),持續(xù)LPS刺激是體外模擬膿毒癥感染的常用且有效方式,且持續(xù)LPS對(duì)DC的效應(yīng)亦為抑制。因此探討能減輕DC成熟與分泌的抑制效應(yīng)的因素,對(duì)膿毒癥的發(fā)展與調(diào)控有重要意義。在此基礎(chǔ)上,尋找減輕持續(xù)LPS對(duì)DC成熟與分泌功能抑制作用的藥物,能為膿毒癥的治療提供理論依據(jù)。 目的：觀察膿毒癥患者外周血DC成熟度與分泌功能的變化及不同濃度OXPAPC對(duì)其的影響,初步探討OXPAPC對(duì)膿毒癥作用的機(jī)制。 方法：①選擇健康志愿者和膿毒癥患者無菌條件下獲得外周血中單個(gè)核細(xì)胞(包括淋巴細(xì)胞和單核細(xì)胞)。體外使用GM-CSF、IL-4和TNF-α誘導(dǎo)其定向DC分化成熟。在倒置顯微鏡和透射電鏡下觀察細(xì)胞形態(tài),流式細(xì)胞儀檢測(cè)CDla、HLA-DR和CD86的表達(dá)水平差異,ELISA法檢測(cè)培養(yǎng)上清液IL-12p70的變化。 ②采用持續(xù)性LPS刺激單個(gè)核細(xì)胞模擬體內(nèi)膿毒癥中DC反應(yīng)。無菌條件下分離健康志愿者外周血單個(gè)核細(xì)胞,使用GM-CSF、IL-4和TNF-α誘導(dǎo)單個(gè)核細(xì)胞定向DC分化成熟,加入持續(xù)性LPS和不同濃度OXPAPC進(jìn)行干預(yù)。在倒置顯微鏡和透射電鏡下觀察細(xì)胞形態(tài),流式細(xì)胞儀檢測(cè)CDla、HLA-DR和CD86的表達(dá)水平差異,ELISA法檢測(cè)培養(yǎng)上清液IL-12p70的變化。 結(jié)果：①與健康志愿者相比,膿毒癥患者組HLA-DR、CD86的表達(dá)水平及IL-12p70的分泌水平均較低,均P0.05。 ②持續(xù)LPS刺激干預(yù)組HLA-DR、CD86的表達(dá)水平及IL-12p70的水平均較對(duì)照組降低,均P0.05、OXPAPC成濃度依賴性地升高持續(xù)LPS刺激下HLA-DR、CD86和IL-12p70的水平,與持續(xù)LPS刺激組比較,差異均有統(tǒng)計(jì)學(xué)意義(均P0.05)。 結(jié)論：1.膿毒癥患者外周血DC的成熟度與分泌功能是受抑制的；2.OXPAPC成濃度依賴性減輕持續(xù)LPS刺激對(duì)DC成熟度和分泌功能的抑制效應(yīng)。
[Abstract]:Background: the maturity and secretory function of dendritic cells (DC) in septic patients are inhibited, and the recovery of DC activity contributes to the improvement of sepsis. Continuous LPS stimulation is a common and effective way to simulate sepsis infection in vitro. The effect of continuous LPS on DC was also inhibited. Therefore, it is important for the development and regulation of sepsis to explore the factors that can reduce the inhibitory effect of DC maturation and secretion. On this basis, to find a drug to reduce the inhibitory effect of LPS on DC maturation and secretory function, can provide theoretical basis for the treatment of sepsis. Aim: to observe the changes of DC maturity and secretory function in peripheral blood of septic patients and the effect of different concentrations of OXPAPC on it, and to explore the mechanism of OXPAPC on sepsis. Methods: 1 Mononuclear cells (including lymphocytes and monocytes) in peripheral blood were obtained from healthy volunteers and septic patients. GM-CSF,IL-4 and TNF- 偽 were used to induce DC differentiation and maturation in vitro. The cell morphology was observed under inverted microscope and transmission electron microscope. The expression levels of CDla,HLA-DR and CD86 were detected by flow cytometry and the changes of IL-12p70 in culture supernatant were detected by ELISA method. 2 continuous LPS was used to stimulate mononuclear cells to mimic the DC response in sepsis in vivo. The peripheral blood mononuclear cells (PBMC) of healthy volunteers were isolated under aseptic conditions. GM-CSF,IL-4 and TNF- 偽 were used to induce the differentiation and maturation of mononuclear cells (MNCs), and continuous LPS and different concentrations of OXPAPC were added to the mononuclear cells. The cell morphology was observed under inverted microscope and transmission electron microscope. The expression levels of CDla,HLA-DR and CD86 were detected by flow cytometry and the changes of IL-12p70 in culture supernatant were detected by ELISA method. Results: 1 the expression of HLA-DR,CD86 and the secretion of IL-12p70 in sepsis patients were lower than those in healthy volunteers (P 0.05). 2 the expression of HLA-DR,CD86 and the level of IL-12p70 in the intervention group of continuous LPS stimulation were lower than those in the control group, and the levels of HLA-DR,CD86 and IL-12p70 were increased in a concentration-dependent manner by P0.05OXPAPC, and the levels of HLA-DR,CD86 and IL-12p70 were increased in a concentration-dependent manner. Compared with the continuous LPS stimulation group, the difference was statistically significant (P0.05). Conclusion: 1. The maturity and secretory function of DC in peripheral blood of septic patients were inhibited, and the inhibitory effect of continuous LPS stimulation on DC maturity and secretory function was alleviated in a concentration-dependent manner by 2.OXPAPC.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R459.7
本文編號(hào)：2359994
[Abstract]:Background: the maturity and secretory function of dendritic cells (DC) in septic patients are inhibited, and the recovery of DC activity contributes to the improvement of sepsis. Continuous LPS stimulation is a common and effective way to simulate sepsis infection in vitro. The effect of continuous LPS on DC was also inhibited. Therefore, it is important for the development and regulation of sepsis to explore the factors that can reduce the inhibitory effect of DC maturation and secretion. On this basis, to find a drug to reduce the inhibitory effect of LPS on DC maturation and secretory function, can provide theoretical basis for the treatment of sepsis. Aim: to observe the changes of DC maturity and secretory function in peripheral blood of septic patients and the effect of different concentrations of OXPAPC on it, and to explore the mechanism of OXPAPC on sepsis. Methods: 1 Mononuclear cells (including lymphocytes and monocytes) in peripheral blood were obtained from healthy volunteers and septic patients. GM-CSF,IL-4 and TNF- 偽 were used to induce DC differentiation and maturation in vitro. The cell morphology was observed under inverted microscope and transmission electron microscope. The expression levels of CDla,HLA-DR and CD86 were detected by flow cytometry and the changes of IL-12p70 in culture supernatant were detected by ELISA method. 2 continuous LPS was used to stimulate mononuclear cells to mimic the DC response in sepsis in vivo. The peripheral blood mononuclear cells (PBMC) of healthy volunteers were isolated under aseptic conditions. GM-CSF,IL-4 and TNF- 偽 were used to induce the differentiation and maturation of mononuclear cells (MNCs), and continuous LPS and different concentrations of OXPAPC were added to the mononuclear cells. The cell morphology was observed under inverted microscope and transmission electron microscope. The expression levels of CDla,HLA-DR and CD86 were detected by flow cytometry and the changes of IL-12p70 in culture supernatant were detected by ELISA method. Results: 1 the expression of HLA-DR,CD86 and the secretion of IL-12p70 in sepsis patients were lower than those in healthy volunteers (P 0.05). 2 the expression of HLA-DR,CD86 and the level of IL-12p70 in the intervention group of continuous LPS stimulation were lower than those in the control group, and the levels of HLA-DR,CD86 and IL-12p70 were increased in a concentration-dependent manner by P0.05OXPAPC, and the levels of HLA-DR,CD86 and IL-12p70 were increased in a concentration-dependent manner. Compared with the continuous LPS stimulation group, the difference was statistically significant (P0.05). Conclusion: 1. The maturity and secretory function of DC in peripheral blood of septic patients were inhibited, and the inhibitory effect of continuous LPS stimulation on DC maturity and secretory function was alleviated in a concentration-dependent manner by 2.OXPAPC.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R459.7
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