本文選題：人臍帶間充質(zhì)干細(xì)胞 + 干擾素-γ�。� 參考：《青島大學(xué)》2017年碩士論文

【摘要】：目的:探討干擾素-γ預(yù)處理對人臍帶間充質(zhì)干細(xì)胞免疫調(diào)節(jié)活性的影響,及其對系統(tǒng)性紅斑狼瘡小鼠的治療作用。方法:hUC-MSC經(jīng)過IFN-γ(20ng/ml)預(yù)處理或者單獨培養(yǎng)24小時,實時熒光定量核酸擴增檢測系統(tǒng)和蛋白質(zhì)印跡法檢測細(xì)胞吲哚2,3雙加氧酶的表達水平。24只20周左右大的雌性Fas-/-敲除的SLE模型小鼠被隨機分為3組:PBS組、IFN-γ未處理h UC-MSC組、IFN-γ預(yù)處理h UC-MSC組。分別通過尾靜脈注射等劑量(0.2ml)的PBS,IFN-γ未處理h UC-MSC(1x106)懸液以及IFN-γ(20ng/ml)預(yù)處理24h的h UC-MSC(1x106)的細(xì)胞懸液。收集Fas-/-小鼠24h尿液,送檢驗科檢測尿蛋白水平。Fas-/-小鼠眼眶靜脈取血,利用Elisa試劑盒測定Fas-/-小鼠血清抗雙鏈DNA抗體(ds-DNA)水平以及血清肌酐酸(IMP)水平;免疫組織化學(xué)檢測腎臟是否有免疫復(fù)合物Ig G及C3沉積。采用流式細(xì)胞儀檢測小鼠血液中CD3+CD4+T細(xì)胞、CD3+CD8+T細(xì)胞、CD4+CD25+T細(xì)胞水平。結(jié)果:IFN-γ未處理h UC-MSC在RNA水平和蛋白水平均不表達IDO,而IFN-γ預(yù)處理h UC-MSC表達IDO。細(xì)胞移植后連續(xù)觀察8周,發(fā)現(xiàn)IFN-γ預(yù)處理h UC-MSC組和IFN-γ未處理h UC-MSC組Fas-/-小鼠尿蛋白、血清IMP和血清ds-DNA水平均明顯低于PBS組(P0.05或P0.01)。與移植治療之前比,PBS組Fas-/-小鼠尿蛋白、血清IMP和血清ds-DNA水平顯著提高(P0.05或P0.01);IFN-γ未處理h UC-MSC組Fas-/-小鼠尿蛋白、血清IMP和血清ds-DNA水平并沒有發(fā)生顯著性改變(P0.05);而IFN-γ預(yù)處理h UC-MSC組Fas-/-小鼠尿蛋白、血清IMP和血清ds-DNA水平則顯著下降的趨勢(P0.05或P0.01)。PBS組Fas-/-小鼠腎小球系膜區(qū)域呈現(xiàn)大量Ig G、C3免疫復(fù)合物,IFN-γ未處理h UC-MSC組Fas-/-小鼠腎小球系膜區(qū)域呈現(xiàn)少量Ig G、C3免疫復(fù)合物,IFN-γ預(yù)處理h UC-MSC組Fas-/-小鼠腎小球系膜區(qū)域沒有Ig G、C3免疫復(fù)合物。IFN-γ未處理h UC-MSC組Fas-/-小鼠CD3+CD4+T細(xì)胞、CD3+CD8+T細(xì)胞水平與PBS組Fas-/-小鼠相比沒有顯著性變化(P0.05),IFN-γ預(yù)處理h UC-MSC顯著地降低了Fas-/-小鼠CD3+CD4+T細(xì)胞、CD3+CD8+T細(xì)胞(P0.01)。與PBS組Fas-/-小鼠相比,IFN-γ未處理h UC-MSC升高了Fas-/-小鼠CD4+CD25+T水平(P0.01),同時IFN-γ預(yù)處理h UC-MSC組Fas-/-小鼠CD4+CD25+T水平明顯高于IFN-γ未處理hUC-MSC組(P0.01)。結(jié)論:IFN-γ預(yù)處理hUC-MSC可顯著提高h(yuǎn)UC-MSC的抗炎活性,及其對SLE小鼠的治療療效,對系統(tǒng)性紅斑狼瘡疾病的治療具有更好的前景。
[Abstract]:Aim: to investigate the effect of interferon-緯 preconditioning on the immunomodulatory activity of human umbilical cord mesenchymal stem cells and its therapeutic effect on systemic lupus erythematosus mice. Methods Twenty one HUC-MSC was pretreated with IFN- 緯 (20 ng / ml) or cultured alone for 24 hours. Real-time fluorescence quantitative nucleic acid amplification system and Western blotting were used to detect the expression of indocyanine 3-dioxygenase. Twenty-four female Fas-r-knockout SLE mice aged about 20 weeks were randomly divided into 3 groups: 1: 10% PBS group without treatment of IFN- 緯 for h UC-MSC group was pretreated with IFN- 緯 for h UC-MSC group. The cell suspensions of hUC-MSC1x106) were pretreated with the same dose of PBS- IFN- 緯 (0.2 ml) and IFN- 緯 (20ng / ml) for 24 h, respectively, and the cell suspensions of hUC-MSC1x106) were pretreated with IFN- 緯 20 ng / ml for 24 h. The urine samples of Fas-r / -mice were collected for 24 hours, and the urine protein level. Fas-r-mouse orbital vein blood was collected. The serum levels of anti-ds-DNA and creatinine IMP were measured by Elisa kit. Immunoglobulin G and C 3 deposition were detected by immunohistochemistry. The levels of CD 3 CD 4 T cells, CD 3 CD 8 T cells and CD 4 CD 25 T cells in the blood of mice were detected by flow cytometry. Results the hUC-MSC did not express IDO at RNA level and protein level, but IFN- 緯 pretreated hUC-MSC expressed IDO. After 8 weeks of cell transplantation, it was found that the levels of urinary protein, serum IMP and serum ds-DNA were significantly lower in IFN- 緯 pretreated hUC-MSC group and untreated hUC-MSC group than in PBS group (P0.05 or P0.01). Compared with those before transplantation, the levels of urinary protein, serum IMP and serum ds-DNA of Fas-r-mice in PBS group were significantly higher than those in control group (P 0.05 or P 0.01) IFN- 緯 untreated Fas-r-mouse urine protein in HUC-MSC group. The levels of serum IMP and serum ds-DNA did not change significantly (P 0.05), but IFN- 緯 pretreated the urinary protein of Fas-r-mices in UC-MSC group. The level of serum IMP and serum ds-DNA decreased significantly (P0.05) or Fas-r-mil of P0.01U / PBS group showed a large number of immunoreactive immune complexes (IFN- 緯) in glomerular Mesangial region of mice. The glomerular Mesangial region of HUC-MSC group showed a small amount of Ig GfC 3 immune complex in untreated h UC-MSC group. No immunoglobulin G C 3 immune complex. IFN- 緯 untreated Fas-r mouse CD3 CD4 T cell CD3 CD8 T cell level in hUC-MSC group was not significantly changed compared with PBS group Fas-r-mouse group (P0.05 IFN- 緯 preconditioning h UC-MSC). CD3 CD 4 T cells and CD 3 CD 8 T cells in Fas-R-mice were significantly decreased (P 0. 01). Compared with PBS group, untreated hUC-MSC with IFN- 緯 increased CD4 CD25T level of Fas-r-mouse and P0.01T of IFN- 緯 pretreated h UC-MSC group, and the CD4 CD25T level of IFN- 緯 pretreated hour UC-MSC group was significantly higher than that of hUC-MSC group without IFN- 緯 treatment (P0.01). Conclusion the anti-inflammatory activity of hUC-MSC and its therapeutic effect on SLE mice can be significantly improved by preconditioning hUC-MSC with 10% IFN- 緯, and has a better prospect for the treatment of systemic lupus erythematosus.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R593.241

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相關(guān)期刊論文 前3條

1 王治國;劉學(xué)明;佟勝全;石哲群;饒莉;;骨髓間充質(zhì)干細(xì)胞移植治療系統(tǒng)性紅斑狼瘡[J];中國組織工程研究;2016年10期

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本文編號：2026509