【摘要】： 目的研究CXC家族趨化因子I-TAC在內(nèi)皮細(xì)胞上的表達(dá)及其多種免疫學(xué)功能,同時(shí)用免疫抑制劑霉酚酸(MPA)對I-TAC的表達(dá)和移植免疫學(xué)功能進(jìn)行干預(yù),從而探討I-TAC在器官移植排斥反應(yīng)中新的作用以及MPA新的作用靶點(diǎn)。 方法采用RT-PCR半定量分析檢測人臍靜脈內(nèi)皮細(xì)胞株ECV304的I-TAC mRNA的表達(dá)。ELISA檢測同種異體外周血單個(gè)核細(xì)胞(PBMC)混合培養(yǎng)上清中IFN-γ和TNF-α濃度。通過體外趨化實(shí)驗(yàn)觀察I-TAC對PBMC的遷移作用。采用黏附實(shí)驗(yàn)檢測I-TAC促PBMC黏附功能。應(yīng)用MTT法測定I-TAC對PBMC、ECV304細(xì)胞增殖的影響。應(yīng)用流式細(xì)胞術(shù)檢測CXCR3在PBMC表面的表達(dá)。 結(jié)果ECV304細(xì)胞在IFN-γ作用8h后表達(dá)I-TAC mRNA,TNF-α對IFN-γ的這種作用具有協(xié)同效應(yīng),且呈劑量依賴性;同種異體PBMC的混合細(xì)胞培養(yǎng)上清中測定到高濃度的IFN-γ和TNF-α,且能誘導(dǎo)ECV304細(xì)胞I-TAC mRNA的表達(dá)。I-TAC對PBMC具有趨化作用,且在I-TAC濃度為200ng/ml時(shí)趨化功能最強(qiáng);同時(shí)I-TAC具有促PBMC增殖和黏附作用,且呈劑量依賴性,對內(nèi)皮細(xì)胞I-TAC具有抑制增殖作用。I-TAC的受體CXCR3在ECV304細(xì)胞上也有表達(dá)�；旌吓囵B(yǎng)后PBMC上的CXCR3表達(dá)與單獨(dú)培養(yǎng)相比明顯提高。 MPA對IFN-γ誘導(dǎo)ECV304細(xì)胞I-TAC mRNA表達(dá)以及I-TAC對PBMC趨化功能具有一定抑制作用,同時(shí)還發(fā)現(xiàn)MPA能夠降低CXCR3在PBMC上的表達(dá)。結(jié)論IFN-γ能夠誘導(dǎo)ECV304細(xì)胞表達(dá)I-TAC mRNA,TNF-α對IFN-γ的誘導(dǎo)作用具有協(xié)同效應(yīng)。I-TAC具有趨化、促增殖、促黏附等多重功能,它不僅是T細(xì)胞穿過內(nèi)皮細(xì)胞進(jìn)入移植物的關(guān)鍵因素,而且對淋巴細(xì)胞和內(nèi)皮細(xì)胞的功能具有調(diào)節(jié)作用。 免疫抑制劑MPA不僅對I-TAC的表達(dá)及其趨化功能具有抑制作用,而且對其受體CXCR3表達(dá)也有一定的影響。MPA的這些綜合活性可能是MMF發(fā)揮抗移植排斥作用的機(jī)制之一。
[Abstract]:Objective to study the expression of CXC family chemokine I-TAC in endothelial cells and its immunological functions, and to intervene the expression of I-TAC and the immunological function of transplantation with mycophenolate mofetil (MPA), an immunosuppressant. In order to explore the new role of I-TAC in organ transplantation rejection and the new target of MPA. Methods the expression of I-TAC mRNA in human umbilical vein endothelial cell line ECV304 was detected by RT-PCR semi-quantitative analysis, and the concentrations of IFN- 緯 and TNF- 偽 in (PBMC) mixed culture medium of allogenic peripheral blood mononuclear cells were detected by Elisa. The migration effect of I-TAC on PBMC was observed by chemotactic assay in vitro. The adhesion function of PBMC promoted by I-TAC was detected by adhesion test. The effect of I-TAC on the proliferation of PBMC,ECV304 cells was measured by MTT assay. The expression of CXCR3 on PBMC surface was detected by flow cytometry. Results the expression of I-TAC mRNA,TNF- 偽 in ECV304 cells for 8 h had synergistic effect on IFN- 緯 in a dose-dependent manner. High concentrations of IFN- 緯 and TNF- 偽 were detected in the mixed cell culture medium of allogenic PBMC, and I-TAC could induce the expression of I-TAC mRNA in ECV304 cells. I-TAC had chemotactic effect on PBMC, and the chemotactic function was the strongest when the concentration of I-TAC was 200ng/ml. At the same time, I-TAC can promote the proliferation and adhesion of PBMC in a dose-dependent manner, and inhibit the proliferation of endothelial cells I-TAC. I-TAC receptor CXCR3 is also expressed in ECV304 cells. The expression of CXCR3 on PBMC in mixed culture was significantly higher than that in culture alone. MPA could inhibit the expression of I-TAC mRNA in ECV304 cells induced by IFN- 緯 and the chemotactic function of PBMC induced by I-TAC. At the same time, it was also found that MPA could decrease the expression of CXCR3 on PBMC. Conclusion the expression of I-TAC mRNA,TNF- 偽 in ECV304 cells induced by IFN- 緯 has synergistic effect on the induction of IFN- 緯. I-TAC has many functions, such as chemotaxis, proliferation, adhesion and so on. It is not only a key factor for T cells to pass through endothelial cells into the graft, but also plays a regulatory role in the function of lymphocytes and endothelial cells. Immunosuppressive agent MPA has inhibitory effect not only on the expression of I-TAC and its chemotactic function, but also on the expression of its receptor CXCR3. These comprehensive activities of MPA may be one of the mechanisms of MMF against transplantation rejection.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 郭克泰;趨化因子及其受體的研究新進(jìn)展[J];國外醫(yī)學(xué)(免疫學(xué)分冊);2002年01期

2 張鋒;一種新的趨化因子—Fractalkine[J];國外醫(yī)學(xué)(免疫學(xué)分冊);2002年03期

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