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

【摘要】：目的研究外源性重組高遷移率組蛋白B1(HMGB1)對神經(jīng)干細胞增殖及分化的影響及其作用機制。方法在無血清的神經(jīng)干細胞培養(yǎng)基中培養(yǎng)SD鼠大腦皮層細胞,傳代擴增及純化神經(jīng)干細胞,免疫熒光檢測神經(jīng)干細胞標記物巢蛋白(nestin),分析神經(jīng)干細胞純度。CCK-8測定加入不同濃度的重組HMGB1對神經(jīng)干細胞增殖活性的影響,選擇重組HMGB1的最適濃度進行后續(xù)實驗;細胞免疫熒光檢測重組HMGB1對神經(jīng)干細胞分化的影響,Real-time PCR檢測晚期糖基化終末產(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)生長因子(NGF)mRNA的表達,Western blot檢測RAGE、TLRs、MMP-9、NGF蛋白的表達。結(jié)果大鼠大腦皮層細胞在培養(yǎng)至第3代時,nestin鑒定神經(jīng)干細胞純度可達99%及以上。在重組HMGB1 10ng/mL刺激下,神經(jīng)干細胞增殖活性最高。實驗組神經(jīng)Ⅲ類β-微管蛋白(TUJ1)表達高于對照組(P0.05),實驗組RAGE、TLRs、MMP-9、NGF mRNA及蛋白表達均高于對照組(P0.05)。結(jié)論外源性重組HMGB1或可通過RAGE、TLRs、MMP-9等信號通路促進神經(jīng)干細胞增殖及其向神經(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】：國家自然科學(xué)基金(No.81371283和No.81671146)資助
【分類號】：R743.3

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