本文關(guān)鍵詞：高糖對(duì)早期鵪鶉胚胎神經(jīng)系統(tǒng)發(fā)育的影響　出處：《暨南大學(xué)》2014年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 鵪鶉胚胎 高糖 糖尿病 神經(jīng)發(fā)育 活性氧自由基 維生素C

【摘要】：目的：孕期妊娠糖尿病的定義是葡萄糖耐受不良，表現(xiàn)為孕婦在懷孕期間血糖水平升高的現(xiàn)象。然而這種情況下對(duì)母體內(nèi)胎兒的發(fā)育有不利影響，但是它所涉及的機(jī)制仍不完全清楚。在這項(xiàng)研究中，我們以鵪鶉胚胎為模型，研究了高葡萄糖環(huán)境下對(duì)鵪鶉胚胎發(fā)育的影響，特別是對(duì)胚胎神經(jīng)系統(tǒng)發(fā)育的影響。 方法和結(jié)果：我們通過(guò)HE染色證實(shí)了高糖環(huán)境下可以損害中樞神經(jīng)系統(tǒng)，造成神經(jīng)管出現(xiàn)不閉合的缺陷。此外，我們通過(guò)NF免疫熒光和鍍銀染色發(fā)現(xiàn)高糖處理后胚胎的背根神經(jīng)節(jié)以及肢芽處的神經(jīng)元細(xì)胞發(fā)育都受到了明顯抑制的現(xiàn)象。出現(xiàn)了背根神經(jīng)節(jié)尺寸和數(shù)目的減少。我們了解到由于背根神經(jīng)節(jié)來(lái)源于一部分神經(jīng)嵴細(xì)胞的遷移和分化，所以我們探究高糖是否對(duì)從神經(jīng)管背側(cè)遷移出來(lái)的神經(jīng)嵴細(xì)胞的遷移量有影響。我們用HNK-1免疫熒光的方法在體內(nèi)和體外神經(jīng)嵴細(xì)胞的遷移實(shí)驗(yàn)證實(shí)高糖對(duì)神經(jīng)嵴細(xì)胞的遷移作用。結(jié)果顯示高糖的確對(duì)神經(jīng)嵴細(xì)胞的遷移有明顯的抑制作用。包括遷移量減少以及HNK-1陽(yáng)性表達(dá)變?nèi)�。最后，我們�?jīng)過(guò)添加高糖處理的培養(yǎng)基通過(guò)體外培養(yǎng)神經(jīng)元細(xì)胞的實(shí)驗(yàn)探究高糖對(duì)神經(jīng)元細(xì)胞的損害作用。研究結(jié)果表明與正常培養(yǎng)基中的神經(jīng)元細(xì)胞相比較，高糖培養(yǎng)基中神經(jīng)元的數(shù)目會(huì)變少，神經(jīng)元軸突的長(zhǎng)度變短以及培養(yǎng)的神經(jīng)元細(xì)胞的胞體折光度變?nèi)�。隨后我們檢測(cè)了神經(jīng)元細(xì)胞的活性氧自由基(ROS)的產(chǎn)生量，結(jié)果顯示ROS會(huì)隨著高糖的濃度增加而增加。于是我們假設(shè)，過(guò)量的活性氧自由基是神經(jīng)元的發(fā)育和分化異常的主要因素，如果除去神經(jīng)元細(xì)胞產(chǎn)生的過(guò)量的ROS，神經(jīng)元的生長(zhǎng)狀況會(huì)不會(huì)有所改善。因此我們往高糖培養(yǎng)基中加入一定量的維生素C(抗氧化劑)，再次測(cè)定神經(jīng)元的數(shù)目、軸突的長(zhǎng)度以及神經(jīng)元細(xì)胞的活性氧自由基(ROS)。我們的假說(shuō)得到了證實(shí)，結(jié)果表明通過(guò)添加維生素C可以一定程度的改善高葡萄糖對(duì)神經(jīng)元發(fā)育的損壞性影響。 結(jié)論：我們以上的研究結(jié)果表明高糖對(duì)神經(jīng)管，，背根神經(jīng)節(jié)和神經(jīng)在肢芽處的發(fā)育都有一定程度的損害作用。然而這些損害作用都是由于高糖過(guò)剩引起神經(jīng)元細(xì)胞內(nèi)的ROS的增加，繼而影響神經(jīng)嵴細(xì)胞的遷移以及分化而導(dǎo)致的。然而我們?cè)谂囵B(yǎng)的神經(jīng)元細(xì)胞的高糖培養(yǎng)基中添加了抗氧化劑維生素C后，維生素C可以減少ROS的產(chǎn)生，神經(jīng)元細(xì)胞的生長(zhǎng)狀況也得到了一定的改善。它意味著維生素C不是直接保護(hù)細(xì)胞發(fā)揮其作用而是通過(guò)抑制ROS的產(chǎn)生而對(duì)神經(jīng)元起到保護(hù)作用。
[Abstract]:Objective: gestational diabetes mellitus is defined as impaired glucose tolerance, which is characterized by increased blood glucose levels during pregnancy. However, this condition has adverse effects on maternal fetal development. However, the mechanism involved is still unclear. In this study, we used quail embryos as a model to study the effects of high glucose on the development of quail embryos. In particular, the effects on the development of the embryonic nervous system. Methods and results: we proved by HE staining that high glucose environment can damage the central nervous system, resulting in the defect of the neural tube is not closed. Through NF immunofluorescence and silver staining, we found that the development of neurons in dorsal root ganglion and limb bud of embryos treated with high glucose was obviously inhibited. The size and number of dorsal root ganglion decreased. We understand that the dorsal root ganglion is derived from the migration and differentiation of some neural crest cells. So we explored whether high glucose had an effect on the migration of neural crest cells from the dorsal side of the neural tube. We used HNK-1 immunofluorescence assay in vivo and in vitro to confirm the migration of neural crest cells. The results showed that high glucose did inhibit the migration of neural crest cells, including the decrease of migration amount and the weakening of HNK-1 positive expression. The effects of high glucose on neuronal cells were investigated by the experiment of cultured neurons in vitro. The results showed that the effects of high glucose on neuronal cells were compared with those in normal culture medium. The number of neurons in high glucose medium will decrease. The length of neuronal axons became shorter and the somatic diopter of cultured neuron cells weakened. Then we measured the production of reactive oxygen species (Ros) of neuron cells. The results showed that ROS increased with the increase of high glucose concentration, so we hypothesized that excessive Ros was the main factor of abnormal development and differentiation of neurons. If the excess ROSs produced by neuron cells were removed, the growth of neurons would be improved, so we added a certain amount of vitamin C (antioxidants) to high glucose medium. The number of neurons, the length of axons and the reactive oxygen species (Ros) of neuronal cells were determined again. Our hypothesis has been confirmed. The results showed that vitamin C could improve the damage effect of high glucose on neuron development to some extent. Conclusion: the results of our above studies show that high glucose on the neural tube. The development of dorsal root ganglion (DRG) and nerve in limb bud are damaged to some extent, however, these damages are due to the increase of ROS in neuronal cells caused by high glucose excess. However, vitamin C can reduce the production of ROS by adding vitamin C to the high sugar medium of cultured neuron cells, which affects the migration and differentiation of neural crest cells. The growth of neuronal cells has also been improved, which means that vitamin C not directly protects cells but protects neurons by inhibiting the production of ROS.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R714.256

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 李力;妊娠高血壓綜合征研究進(jìn)展[J];重慶醫(yī)學(xué);2002年09期

2 翁光明;雷桂蘭;;妊娠糖尿病的新進(jìn)展[J];現(xiàn)代中西醫(yī)結(jié)合雜志;2008年02期

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