本文選題：神經(jīng)干細(xì)胞　切入點(diǎn)：重組高遷移率組蛋白B　出處：《四川大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年03期 　論文類(lèi)型：期刊論文

【摘要】：目的研究外源性重組高遷移率組蛋白B1(HMGB1)對(duì)神經(jīng)干細(xì)胞增殖及分化的影響及其作用機(jī)制。方法在無(wú)血清的神經(jīng)干細(xì)胞培養(yǎng)基中培養(yǎng)SD鼠大腦皮層細(xì)胞,傳代擴(kuò)增及純化神經(jīng)干細(xì)胞,免疫熒光檢測(cè)神經(jīng)干細(xì)胞標(biāo)記物巢蛋白(nestin),分析神經(jīng)干細(xì)胞純度。CCK-8測(cè)定加入不同濃度的重組HMGB1對(duì)神經(jīng)干細(xì)胞增殖活性的影響,選擇重組HMGB1的最適濃度進(jìn)行后續(xù)實(shí)驗(yàn);細(xì)胞免疫熒光檢測(cè)重組HMGB1對(duì)神經(jīng)干細(xì)胞分化的影響,Real-time PCR檢測(cè)晚期糖基化終末產(chǎn)物受體(RAGE)mRNA、Toll樣受體(TLRs)mRNA、基質(zhì)金屬蛋白酶9(MMP-9)mRNA、神經(jīng)生長(zhǎng)因子(NGF)mRNA的表達(dá),Western blot檢測(cè)RAGE、TLRs、MMP-9、NGF蛋白的表達(dá)。結(jié)果大鼠大腦皮層細(xì)胞在培養(yǎng)至第3代時(shí),nestin鑒定神經(jīng)干細(xì)胞純度可達(dá)99%及以上。在重組HMGB1 10ng/mL刺激下,神經(jīng)干細(xì)胞增殖活性最高。實(shí)驗(yàn)組神經(jīng)Ⅲ類(lèi)β-微管蛋白(TUJ1)表達(dá)高于對(duì)照組(P0.05),實(shí)驗(yàn)組RAGE、TLRs、MMP-9、NGF mRNA及蛋白表達(dá)均高于對(duì)照組(P0.05)。結(jié)論外源性重組HMGB1或可通過(guò)RAGE、TLRs、MMP-9等信號(hào)通路促進(jìn)神經(jīng)干細(xì)胞增殖及其向神經(jīng)元方向分化。
[Abstract]:Objective to study the effect and mechanism of exogenous recombinant histone B1 HMGB1 on the proliferation and differentiation of neural stem cells. Methods SD rat cerebral cortex cells were cultured in serum-free neural stem cell culture medium. The neural stem cells were amplified and purified by subculture and purified. The nestin of neural stem cells was detected by immunofluorescence. The purity of neural stem cells was analyzed. CCK-8 was used to determine the effect of different concentrations of recombinant HMGB1 on the proliferation of neural stem cells. The optimal concentration of recombinant HMGB1 was selected for follow-up experiment. Effects of Recombinant HMGB1 on differentiation of Neural Stem cells by Cellular Immunofluorescence Assay the expression of advanced glycosylation end product receptor RAGEN mRNA-Toll-like receptor TLRsmRNAs, matrix metalloproteinase 9 mRNA-9 mRNAs, nerve growth factor nongfen mRNA expression and the expression of RAGEGE-TLsMMP-9NGF protein by real-time PCR. Results the purity of neural stem cells was 99% or more at the third passage of cultured rat cerebral cortex cells, and stimulated by recombinant HMGB1 10 ng / mL. The expression of 尾 -tubulin TUJ1 in the experimental group was higher than that in the control group (P 0.05), and the expression of NGF mRNA and protein in the experimental group was higher than that in the control group (P 0.05). Conclusion exogenous recombinant HMGB1 may be promoted by signal pathway such as RAGEG-TLRsMMP-9. Neural stem cells proliferate and differentiate into neurons.
【作者單位】： 四川大學(xué)華西醫(yī)院神經(jīng)內(nèi)科;
【基金】：國(guó)家自然科學(xué)基金(No.81371283和No.81671146)資助
【分類(lèi)號(hào)】：R743.3

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