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低氧條件下大鼠大腦皮質(zhì)星形膠質(zhì)細(xì)胞中IL-1β和CD44的表達(dá)及其意義

發(fā)布時(shí)間:2018-07-29 12:20
【摘要】:中樞神經(jīng)系統(tǒng)的損傷和大腦疾病產(chǎn)生是由于大腦內(nèi)細(xì)胞的損傷和死亡所造成的,而缺血后的腦損傷通常伴隨著炎癥反應(yīng)的發(fā)生并介導(dǎo)繼發(fā)性的腦損傷。缺血早期白細(xì)胞的侵潤(rùn)和大腦水腫的形成都是由炎癥反應(yīng)所誘導(dǎo)的。而且,大腦中的細(xì)胞包括星形膠質(zhì)細(xì)胞、小膠質(zhì)細(xì)胞以及內(nèi)皮細(xì)胞在缺血的刺激下被激活而成為反應(yīng)性的細(xì)胞。大多數(shù)的炎癥反應(yīng)都是由一些炎性細(xì)胞因子所介導(dǎo)的。IL-1β作為一類(lèi)重要的炎癥前細(xì)胞因子,在腦缺血的損傷中有著重要的作用。在早期的損傷過(guò)程中已經(jīng)發(fā)現(xiàn)IL-1β的表達(dá)對(duì)于腦梗塞的形成有著直接的關(guān)系。CD44是作為透明質(zhì)酸(HA)的受體在細(xì)胞增殖和腫瘤的形成及轉(zhuǎn)移有重要作用的一類(lèi)細(xì)胞粘附分子家族,除此之外,CD44和HA的相互作用還參與許多炎癥疾病的發(fā)生。最近在腦缺血模型中發(fā)現(xiàn),CD44表達(dá)的升高可能會(huì)通過(guò)介導(dǎo)炎癥而參與缺血后的損傷。星形膠質(zhì)細(xì)胞作為腦內(nèi)數(shù)量最多的一類(lèi)細(xì)胞,在神經(jīng)系統(tǒng)中發(fā)揮著重要的作用,而且在不同的病理?xiàng)l件下它的功能也是完全不同的。因此,我們想了解的是:它在缺氧狀態(tài)下對(duì)IL-1β和CD44的表達(dá)有沒(méi)有改變?細(xì)胞外基質(zhì)的代謝是否受到低氧條件的調(diào)控而介導(dǎo)炎癥的發(fā)生?如果有,這些改變和變化的意義是什么?為了探討這些問(wèn)題,本文對(duì)星形膠質(zhì)細(xì)胞進(jìn)行了體外培養(yǎng)的實(shí)驗(yàn)研究。 通過(guò)體外分離培養(yǎng)新生大鼠星形膠質(zhì)細(xì)胞,低氧培養(yǎng)20min,建立缺氧缺糖模型,缺氧后復(fù)氧1h,3h,6h,12h,24h,利用免疫熒光雙標(biāo)和RT-PCR及Western blot的方法觀察和檢測(cè)IL-1β和CD44,CD44v6及Ⅱ型透明質(zhì)酸合酶(HAS-2)在復(fù)氧不同時(shí)間的表達(dá),并以正常的星形膠質(zhì)細(xì)胞作為對(duì)照。結(jié)果顯示:同正常培養(yǎng)的細(xì)胞相比,在轉(zhuǎn)錄水平,低氧復(fù)氧后星形膠質(zhì)細(xì)胞的IL-1β,CD44,CD44v6及HAS-2 mRNA表達(dá)明顯升高;在蛋白水平,IL-1β在缺氧復(fù)氧后6h表達(dá)有明顯的升高,CD44在缺氧復(fù)氧后表達(dá)量的變化有時(shí)問(wèn)依賴(lài)性趨勢(shì)升高。我們的結(jié)果表明了在星形膠質(zhì)細(xì)胞缺氧缺糖后IL-1β和CD44及其配體的差異性表達(dá),暗示了在損傷過(guò)程中IL-1β可能會(huì)和CD44相互作用來(lái)介導(dǎo)缺血后損傷,此外細(xì)胞外基質(zhì)的改變也可能會(huì)促進(jìn)缺血損傷后炎癥的發(fā)生的重要因素。
[Abstract]:Central nervous system (CNS) injury and brain disease are caused by the injury and death of brain cells, and the ischemic brain damage is usually accompanied by inflammation and mediates the secondary brain damage. Leukocyte infiltration and cerebral edema are induced by inflammation at the early stage of ischemia. Moreover, cells in the brain include astrocytes, microglia and endothelial cells activated by ischemia to become reactive cells. Most inflammatory responses are mediated by some inflammatory cytokines. IL-1 尾, as an important preinflammatory cytokine, plays an important role in cerebral ischemia injury. It has been found that the expression of IL-1 尾 is directly related to the formation of cerebral infarction in the early stage of injury. CD44 is a kind of cell adhesion molecule family which plays an important role in cell proliferation and tumor formation and metastasis as a receptor of hyaluronic acid (HA). In addition, the interaction of CD 44 and HA is involved in many inflammatory diseases. Recently, it was found that the increased expression of CD44 may play a role in ischemic injury by mediating inflammation in cerebral ischemia model. Astrocytes, as the most abundant cells in the brain, play an important role in the nervous system, and their functions are completely different under different pathological conditions. So what we want to know is: does it change the expression of IL-1 尾 and CD44 in anoxic state? Is the metabolism of extracellular matrix mediated by inflammation due to hypoxia? If so, what is the meaning of these changes and changes? In order to investigate these problems, astrocytes were cultured in vitro. The astrocytes of newborn rats were isolated and cultured in vitro and cultured in hypoxia for 20 min. The model of hypoxia and glucose deficiency was established. The expression of IL-1 尾, CD44v6 and type 鈪,

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