【摘要】：目的研究全反式維甲酸(ATRA)對(duì)成年小鼠神經(jīng)干細(xì)胞(NSCs)的調(diào)控、信號(hào)通路及對(duì)細(xì)胞色素P450(CYP450)家族的影響。方法分離并培養(yǎng)成年小鼠腦室下區(qū)NSCs,將細(xì)胞分為ATRA組和對(duì)照組,ATRA組加入10-6mol/L ATRA,對(duì)照組正常培養(yǎng)。采用流式細(xì)胞儀檢測(cè)兩組細(xì)胞表面標(biāo)記物,計(jì)算神經(jīng)元、星形膠質(zhì)細(xì)胞、少突膠質(zhì)細(xì)胞和NSCs占總細(xì)胞數(shù)的百分比;Real-time PCR方法檢測(cè)CYP450家族相關(guān)基因表達(dá)情況,篩選出有統(tǒng)計(jì)學(xué)意義的基因;ELISA法測(cè)定細(xì)胞色素P450還原酶(CPR)的活性;Western blotting法檢測(cè)P38 MAPK通路相關(guān)蛋白P38、p-P38蛋白的相對(duì)表達(dá)量。結(jié)果 ATRA組NSCs主要是向神經(jīng)元分化,也有一部分向星形膠質(zhì)細(xì)胞分化,少突膠質(zhì)細(xì)胞較少。與對(duì)照組比較,ATRA組P450家族基因中CYP26A1、CYP26B1、CYP26C1表達(dá)上調(diào)(P均0.05)。ATRA組CPR活性以及p-P38蛋白表達(dá)量均較對(duì)照組升高(P均0.05)。結(jié)論 ATRA促進(jìn)小鼠NSCs向神經(jīng)元分化,可能通過(guò)上調(diào)CYP450中的CYP26家族基因、增加CPR活性以及P38 MAPK通路發(fā)揮調(diào)控作用。
[Abstract]:Objective to study the regulation, signaling pathway and cytochrome P450 (CYP450) family of adult mouse neural stem cells (NSCs) by all-trans retinoic acid (ATRA). Methods NSCs in the subventricular region of adult mice were isolated and cultured. The cells were divided into ATRA group and control group and 10-6 mol / L ATRA was added into the control group. Cell surface markers were detected by flow cytometry, and the percentage of neurons, astrocytes, oligodendrocytes and NSCs in total cells were calculated. Real-time PCR was used to detect the expression of CYP450 family related genes. The activity of cytochrome P450 reductase (CPR) was detected by Elisa. Western blotting was used to detect the relative expression of P38 MAPK pathway related protein P38 p-P38. Results in ATRA group, NSCs were mainly differentiated into neurons, and some of them were differentiated into astrocytes, with less oligodendrocytes. Compared with the control group, the expression of CYP26A1C1-CYP26B1-CYP26C1 was up-regulated in ATRA group (P 0.05). The CPR activity and p-P38 protein expression in ATRA group were higher than those in control group (P 0.05). Conclusion ATRA can promote the differentiation of NSCs into neurons, which may play a regulatory role by up-regulating CYP26 family gene in CYP450, increasing CPR activity and P38 MAPK pathway.
【作者單位】： 蘇州大學(xué)附屬常州第一人民醫(yī)院;蘇州衛(wèi)生職業(yè)技術(shù)學(xué)院;徐州醫(yī)科大學(xué);
【基金】：江蘇省自然科學(xué)基金資助項(xiàng)目(BK20130219) 江蘇高�！扒嗨{(lán)工程”科研資助項(xiàng)目
【分類(lèi)號(hào)】：R741

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