【摘要】：背景胰島素樣生長(zhǎng)因子結(jié)合蛋白7(IGFBP7)被發(fā)現(xiàn)作為為數(shù)不多的角質(zhì)形成細(xì)胞特異基因中的一種。一項(xiàng)全基因表達(dá)研究顯示IGFBP7在銀屑病表皮中的表達(dá)下調(diào)。前期的研究發(fā)現(xiàn),IGFBP7下調(diào)對(duì)轉(zhuǎn)化生長(zhǎng)因子β(TGF-β)有影響,但是具體機(jī)制不明確。目的為了全面評(píng)估IGFBP7在銀屑病調(diào)節(jié)中的作用,以及其與TGF-β通路復(fù)雜效應(yīng)的分子機(jī)制,我們檢測(cè)了IGFBP7與轉(zhuǎn)化生長(zhǎng)因子β1(TGF-β1)和其下游p38 MAPK的活性之間的聯(lián)系。 方法HaCaT細(xì)胞是一種自主永生化的人類(lèi)角質(zhì)形成細(xì)胞系。我們通過(guò)在HaCaT細(xì)胞中使用IGFBP7特異性小干擾RNA (IGFBP7- siRNA)或重組IGFBP7 (rIGFBP7)來(lái)人為地下調(diào)或上調(diào)IGFBP7的水平。利用細(xì)胞免疫組化方法檢測(cè)IGFBP7和TGF-β1的表達(dá)水平。MTT法和annexin V/PI法用來(lái)檢測(cè)細(xì)胞活性和細(xì)胞凋亡的水平。蛋白印跡法用來(lái)檢測(cè)IGFBP7、TGF-β1和p38-MAPK的蛋白水平。 結(jié)果我們的研究發(fā)現(xiàn)IGFBP7下調(diào)顯著提高了HaCaT細(xì)胞的增殖能力,其與細(xì)胞的凋亡水平明顯降低有關(guān)。而重組IGFBP7可以逆轉(zhuǎn)上述效應(yīng)。IGFBP7沉默的細(xì)胞中,TGF-β1和磷酸化MAPK水平上調(diào),而IGFBP7過(guò)表達(dá)細(xì)胞中TGF-β1和磷酸化MAPK水平下調(diào)。 結(jié)論我們的結(jié)果提示IGFBP7在HaCaT細(xì)胞中對(duì)TGF-β1的調(diào)控可能通過(guò)MAPK通路,這為增厚性皮膚病的治療提供了線索。
[Abstract]:Background Insulin-like growth factor binding protein 7 (IGFBP7) has been identified as one of the few keratinocyte specific genes. A whole gene expression study showed that IGFBP7 expression was down-regulated in psoriatic epidermis. Previous studies have found that down-regulation of IGFBP7 has an effect on transforming growth factor 尾 (TGF- 尾), but the mechanism is unclear. Objective to evaluate the role of IGFBP7 in psoriasis regulation and the molecular mechanism of its complex effect on TGF- 尾 pathway, we examined the relationship between IGFBP7 and the activity of transforming growth factor 尾 1 (TGF- 尾 1) and its downstream p38 MAPK. Methods HaCaT cell line is an autonomous immortalized human keratinocyte cell line. We artificially down-regulated or upregulated IGFBP7 levels by using IGFBP7 specific small interfering RNA (IGFBP7- siRNA) or recombinant IGFBP7 (rIGFBP7) in HaCaT cells. The expression of IGFBP7 and TGF- 尾 1 was detected by immunohistochemistry, and the activity and apoptosis of cells were detected by MTT and annexin V/PI methods. Protein levels of IGFBP7,TGF- 尾 1 and p38-MAPK were detected by Western blot. Results in our study, down-regulation of IGFBP7 significantly increased the proliferation of HaCaT cells, which was related to the decrease of apoptosis level. The recombinant IGFBP7 could reverse the above effects. The levels of TGF- 尾 1 and phosphorylated MAPK were up-regulated in IGFBP7 silencing cells, while TGF- 尾 1 and phosphorylated MAPK levels were down-regulated in IGFBP7 overexpression cells. Conclusion our results suggest that the regulation of TGF- 尾 1 by IGFBP7 in HaCaT cells may be mediated by MAPK pathway, which may provide clues for the treatment of thickening dermatosis.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R329

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