本文選題：銀耳多糖　切入點(diǎn)：調(diào)節(jié)性T淋巴細(xì)胞　出處：《中國(guó)免疫學(xué)雜志》2016年03期 　論文類型：期刊論文

【摘要】：目的:探討銀耳多糖(TPS)對(duì)膿毒癥小鼠外周血調(diào)節(jié)性T細(xì)胞免疫調(diào)節(jié)的作用。方法:雄性BALB/c小鼠(20±1)g,采用燒傷后腹腔注射銅綠假單胞菌制作膿毒癥并予以TPS干預(yù),隨機(jī)分成5組,分別為正常對(duì)照組(未做任何處理),未刺激組(燒傷后銅綠假單胞菌感染未經(jīng)TPS處理組),TPS低劑量組(燒傷后銅綠假單胞菌感染+50 mg/kg治療組),TPS中劑量組(燒傷后銅綠假單胞菌感染+100 mg/kg治療組),TPS高劑量組(燒傷后銅綠假單胞菌感染+200 mg/kg治療組),從各組的外周血中分離調(diào)節(jié)性T細(xì)胞,并體外培養(yǎng)。自燒傷即刻(PBD0)起,至燒傷后4 d每天監(jiān)測(cè)培養(yǎng)上清液中IL-10、IFN-γ、IL-4的水平。磁珠孵育和磁性分離小鼠外周血的調(diào)節(jié)性T細(xì)胞( CD4~+CD25~(high)Tregs)和CD4~+T細(xì)胞,分別加入TPS后進(jìn)行培養(yǎng),以不加TPS的細(xì)胞作為對(duì)照,流式細(xì)胞儀分析 CD4~+CD25~(high)Tregs的表型,ELISA法檢測(cè)細(xì)胞因子的表達(dá)。結(jié)果:與對(duì)照組相比,TPS能顯著降低未刺激組小鼠的IL-10和IL-4的分泌,并顯著增加IFN-γ的分泌,并且IL-10的分泌水平與TPS濃度呈劑量效應(yīng)關(guān)系。體外培養(yǎng)試驗(yàn)中,與正常對(duì)照組相比,未刺激組的CD4~+T細(xì)胞增殖和IFN-γ水平顯著降低(P0.05),IL-4水平顯著升高(P0.05);與未刺激組相比,TPS刺激組的CD4~+T細(xì)胞增殖和IFN-γ水平顯著升高(P0.05),IL-4水平顯著降低(P0.05);與未刺激組相比,TPS刺激+抗體1組CD4~+T細(xì)胞增殖和IFN-γ水平顯著升高(P0.05),IL-4水平顯著降低(P0.05);與未刺激組相比,TPS刺激+抗體2組的CD4~+T細(xì)胞增殖和IFN-γ、IL-4水平無顯著性差異。結(jié)論:TPS可以通過降低IL-10的分泌抑制 CD4~+CD25~(high)Tregs對(duì)CD4~+T淋巴細(xì)胞增殖和極化的影響,并誘導(dǎo)CD4~+T淋巴細(xì)胞向Th1分型,從而使機(jī)體免疫活性增強(qiáng)。
[Abstract]:Objective: to investigate the effect of Tremella polysaccharide (TP) on the immune regulation of regulatory T cells in peripheral blood of septic mice. Methods: male BALB/c mice were injected intraperitoneally with Pseudomonas aeruginosa to make sepsis and treated with TPS after burn, and were randomly divided into 5 groups. They were normal control group (without any treatment) and unstimulated group (postburn Pseudomonas aeruginosa infection without TPS treatment group with low dose (50 mg/kg post-burn Pseudomonas aeruginosa infection treatment group). Pseudomonas aeruginosa infection for 100 mg/kg was used to isolate regulatory T cells from peripheral blood of patients with high dose of Pseudomonas aeruginosa infection (200 mg/kg after burn). The levels of IL-10 IFN- 緯 IL-4 in the supernatant were monitored daily from immediate burn to 4 days after burn. The regulatory T cells (CD _ 4 ~ CD _ (25) T _ (+)) and CD _ 4 ~ T cells were incubated with magnetic beads and separated from the peripheral blood of mice. The expression of cytokines was detected by flow cytometry in cells without TPS. Results: compared with the control group, TPS could significantly decrease the secretion of IL-10 and IL-4 and increase the secretion of IFN- 緯 in unstimulated mice. And the level of IL-10 secretion was dose-dependent with the concentration of TPS. In vitro culture test, compared with the normal control group, The proliferation of CD4 ~ T cells and the level of IFN- 緯 in unstimulated group were significantly lower than those in unstimulated group. The level of IL-4 in P0.05 was significantly higher than that in unstimulated group, while the proliferation of CD4 ~ T cell and the level of IFN- 緯 in TPS stimulated group were significantly higher than that in unstimulated group. The proliferation of CD4 ~ T cells and the level of IFN- 緯 increased significantly in group 1, and the level of IL-4 decreased significantly in group P0.05, but there was no significant difference in proliferation of CD4T cells and IL-4 level of IFN- 緯 IL-4 in TPS stimulated antibody group 2 compared with that in group 2. Conclusion the level of TPS can be reduced by decreasing the level of IL-4. The effect of IL-10 secretion on the proliferation and polarization of CD4 ~ T lymphocytes was inhibited by CD4 ~ CD25~(high)Tregs. CD4 ~ T lymphocytes were induced to type to Th1, and the immune activity was enhanced.
【作者單位】： 煤炭總醫(yī)院腎內(nèi)科;解放軍總醫(yī)院第一附屬醫(yī)院全軍燒傷研究所;
【基金】：國(guó)家自然科學(xué)基金(81000847,81272140) 中國(guó)博士后面上項(xiàng)目(201150M1530) 中國(guó)博士后第五批特別資助項(xiàng)目(2012T50863)
【分類號(hào)】：R459.7
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本文編號(hào)：1556560