發(fā)布時(shí)間：2018-05-12 12:51

  本文選題：間充質(zhì)干細(xì)胞 + 白介素1受體拮抗劑��； 參考：《吉林大學(xué)》2008年博士論文

【摘要】： 白細(xì)胞介素1受體拮抗劑(Interleukin 1 receptor antagonist, IL-1Ra)是天然的IL-1拮抗物質(zhì),可作為基因治療的目的基因,通過合適的基因轉(zhuǎn)移途徑,在體內(nèi)發(fā)揮抗炎作用。間充質(zhì)干細(xì)胞(Mesenchymal stem cell, MSC)作為成體干細(xì)胞,不僅是目前組織工程和基因治療領(lǐng)域理想的種子細(xì)胞和基因載體,而且免疫原性低、并具有免疫調(diào)節(jié)作用。如將二者優(yōu)勢綜合,通過適當(dāng)途徑將外源性IL-1Ra基因轉(zhuǎn)入MSC,使其以旁分泌方式在體內(nèi)局部組織發(fā)揮抗炎和組織修復(fù)功能,將在類風(fēng)濕關(guān)節(jié)炎等免疫病理性炎癥損傷疾病的治療領(lǐng)域具有誘人的應(yīng)用前景,目前尚未見諸國內(nèi)外研究報(bào)道。 按照這一設(shè)想,本研究通過克隆人IL-1Ra基因,成功構(gòu)建AdhIL-1Ra重組腺病毒;在成功分離、培養(yǎng)和鑒定大鼠骨髓MSC后,首次將hIL-1Ra基因在腺病毒載體介導(dǎo)下轉(zhuǎn)入大鼠骨髓MSC,并證實(shí)hIL-1Ra基因在大鼠骨髓MSC中成功表達(dá);采用RT-PCR法,觀察外源性hIL-1Ra基因轉(zhuǎn)染對(duì)大鼠骨髓MSC免疫調(diào)節(jié)因子的表達(dá)調(diào)控。結(jié)果表明,正常培養(yǎng)的MSC可表達(dá)多種免疫調(diào)節(jié)因子,這些因子間的表達(dá)變化具有較高的相關(guān)性。LPS模擬的炎性刺激可使大鼠骨髓MSC的炎性因子表達(dá)水平升高。外源性hIL-1Ra基因修飾對(duì)大鼠骨髓MSC免疫調(diào)節(jié)因子基因表達(dá)具有明顯的調(diào)控作用,表現(xiàn)為增強(qiáng)抑炎性免疫調(diào)節(jié)因子表達(dá)及抑制LPS刺激下炎性因子表達(dá)水平的升高。另外,本研究發(fā)現(xiàn)hIL-1Ra基因修飾和LPS刺激均可影響MSC向軟骨細(xì)胞方向的分化。以上結(jié)果初步證明hIL-1Ra基因修飾MSC,發(fā)揮免疫調(diào)節(jié)作用的可行性,為今后進(jìn)一步開展相關(guān)機(jī)理研究和體內(nèi)研究打下實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:Interleukin-1 receptor antagonist (IL-1Ra) is a natural IL-1 antagonist, which can be used as the target gene of gene therapy and play an anti-inflammatory effect in vivo through the appropriate gene transfer pathway. Mesenchymal stem cell, MSC), as an adult stem cell, is not only the ideal seed cell and gene vector in tissue engineering and gene therapy, but also has low immunogenicity and immunomodulatory effect. If the two advantages are combined, the exogenous IL-1Ra gene can be transferred into MSCs by appropriate way, so that the exogenous IL-1Ra gene can play an anti-inflammatory and tissue repair function in local tissues in vivo by paracrine way. It will have an attractive application prospect in the treatment of immune pathological inflammatory injury diseases such as rheumatoid arthritis, which has not been reported at home and abroad. According to this assumption, the recombinant adenovirus of AdhIL-1Ra was successfully constructed by cloning human IL-1Ra gene, and the rat bone marrow MSC was successfully isolated, cultured and identified. HIL-1Ra gene was transferred into rat bone marrow by adenovirus vector for the first time, and the expression of hIL-1Ra gene in rat bone marrow MSC was confirmed successfully. The expression of MSC immunomodulatory factor in rat bone marrow was observed by RT-PCR method. The results showed that MSC in normal culture could express a variety of immunomodulatory factors, and the expression of these factors had a high correlation. LPS-mimic inflammatory stimulation could increase the expression of inflammatory factors in rat bone marrow MSC. Exogenous hIL-1Ra gene modification can obviously regulate the expression of MSC immunomodulator gene in bone marrow of rats, which is shown by enhancing the expression of anti-inflammatory immunomodulatory factor and inhibiting the increase of the expression level of inflammatory factor stimulated by LPS. In addition, it was found that both hIL-1Ra gene modification and LPS stimulation could affect the differentiation of MSC into chondrocytes. The above results preliminarily prove the feasibility of hIL-1Ra gene modification to play the role of immune regulation, and lay the experimental foundation for further research on related mechanism and in vivo study.
【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】：2008
【分類號(hào)】：R392

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本文編號(hào)：1878692