發(fā)布時(shí)間：2018-03-23 21:36

  本文選題：孕酮　切入點(diǎn)：小膠質(zhì)細(xì)胞　出處：《蘭州大學(xué)》2014年碩士論文

【摘要】：孕酮是人體內(nèi)的一種類固醇激素類物質(zhì),其對(duì)于神經(jīng)系統(tǒng)類疾病的治療作用已經(jīng)為廣大的學(xué)者所認(rèn)可。近年來,孕酮已經(jīng)被應(yīng)用于臨床實(shí)驗(yàn)中對(duì)神經(jīng)元及神經(jīng)膠質(zhì)細(xì)胞的損傷進(jìn)行治療。但到目前為止,雖然發(fā)現(xiàn)孕酮對(duì)治療中風(fēng)類疾病具有一定的效果,但是其具體的作用機(jī)理還缺乏詳細(xì)的研究。由于孕酮的作用涉及的過程比較復(fù)雜,除了對(duì)神經(jīng)元具有保護(hù)作用外,還對(duì)神經(jīng)膠質(zhì)細(xì)胞起到一定的作用。但是,關(guān)于孕酮對(duì)小膠質(zhì)細(xì)胞進(jìn)行處理后所發(fā)揮的效果的研究還很有限。 實(shí)驗(yàn)采用小膠質(zhì)細(xì)胞帶有GFP熒光標(biāo)記的轉(zhuǎn)基因小鼠新生鼠作為實(shí)驗(yàn)材料,進(jìn)行原代小膠質(zhì)細(xì)胞培養(yǎng),同時(shí)采用BV-2小膠質(zhì)瘤細(xì)胞進(jìn)行傳代培養(yǎng)。對(duì)兩種細(xì)胞分別進(jìn)行氧糖剝奪模型(oxygen and glucose deprivation, OGD)的實(shí)驗(yàn)處理后,再使用適量的孕酮對(duì)其進(jìn)行處理,探索孕酮對(duì)小膠質(zhì)細(xì)胞的作用。本研究主要采用了Hoechst33258熒光染色,Iba-1和BrdU免疫細(xì)胞化學(xué)染色,MTT細(xì)胞活性檢測及流式細(xì)胞術(shù)等實(shí)驗(yàn)技術(shù)。 原代培養(yǎng)的小膠質(zhì)細(xì)胞在OGD處理24小時(shí)后,對(duì)其使用40μM的孕酮溶液處理后再進(jìn)行檢測,結(jié)果發(fā)現(xiàn)OGD模型處理可以使小膠質(zhì)細(xì)胞總數(shù)降低,凋亡增加。發(fā)現(xiàn)孕酮可以促進(jìn)經(jīng)過OGD處理的小膠質(zhì)細(xì)胞的增殖,同時(shí)還可以降低經(jīng)過OGD處理的小膠質(zhì)細(xì)胞的凋亡比率。對(duì)BV-2小膠質(zhì)瘤細(xì)胞進(jìn)行OGD處理6小時(shí)后,再用10μM的孕酮進(jìn)行處理。發(fā)現(xiàn)OGD模型處理可以使BV-2細(xì)胞細(xì)胞活力降低,細(xì)胞凋亡比例增加,細(xì)胞增殖降低。發(fā)現(xiàn)經(jīng)過OGD聯(lián)合孕酮處理后的BV-2細(xì)胞活性上升,細(xì)胞周期基本恢復(fù)正常,并且細(xì)胞的凋亡比例下降,細(xì)胞發(fā)生增殖。 綜覽上述結(jié)果,可以發(fā)現(xiàn)OGD模型分別處理原代培養(yǎng)的小膠質(zhì)細(xì)胞和傳代培養(yǎng)的BV-2細(xì)胞,都可以產(chǎn)生明顯的效果。之后再用適宜劑量的孕酮對(duì)經(jīng)過OGD處理后的原代培養(yǎng)的小膠質(zhì)細(xì)胞和BV-2細(xì)胞進(jìn)行處理,發(fā)現(xiàn)孕酮促進(jìn)了細(xì)胞活力的恢復(fù),同時(shí)促進(jìn)了細(xì)胞的增殖。因此,孕酮對(duì)于經(jīng)過OGD處理的小膠質(zhì)細(xì)胞的確會(huì)產(chǎn)生一定的保護(hù)作用,為臨床中孕酮的使用提供了一定的理論依據(jù)。
[Abstract]:Progesterone is a steroid hormone in human body, and its therapeutic effect on nervous system diseases has been recognized by many scholars in recent years. Progesterone has been used in clinical trials to treat injuries to neurons and glial cells. But so far, progesterone has been found to be effective in the treatment of apoplectic diseases. However, the specific mechanism of the action is still lack of detailed study. As the process involved in progesterone is relatively complex, in addition to the protection of neurons, it also plays a certain role in neuroglial cells. Studies on the effects of progesterone on microglia are limited. The primary microglia of transgenic mice with GFP fluorescent labeling were used as experimental materials. At the same time, BV-2 small glioma cells were subcultured. The two kinds of cells were treated with oxygen and glucose privacy, and then treated with appropriate amount of progesterone. In order to explore the effect of progesterone on microglia, Hoechst33258 fluorescence staining and BrdU immunocytochemical staining were used to detect the cell activity and flow cytometry. The primary cultured microglial cells were treated with OGD for 24 hours, then treated with 40 渭 M progesterone solution. The results showed that OGD model treatment could reduce the total number of microglia cells. Apoptosis increased. It was found that progesterone could promote the proliferation of microglia treated with OGD and decrease the apoptosis rate of microglia treated with OGD. Then treated with 10 渭 M progesterone, it was found that OGD model treatment could decrease the viability, increase the proportion of apoptosis and decrease the proliferation of BV-2 cells. The activity of BV-2 cells treated with OGD combined with progesterone was increased. The cell cycle returned to normal, and the proportion of apoptosis decreased and cell proliferation occurred. After reviewing the above results, we can find that the OGD model deals with the primary cultured microglia and the subcultured BV-2 cells, respectively. Then the primary cultured microglia and BV-2 cells treated with OGD were treated with the appropriate dose of progesterone, and it was found that progesterone promoted the recovery of cell viability. Therefore, progesterone does have a protective effect on microglia treated with OGD, which provides a theoretical basis for the use of progesterone in clinic.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.41

【參考文獻(xiàn)】

相關(guān)期刊論文 前6條

1 周麗華;;中風(fēng)患者的康復(fù)護(hù)理[J];中外醫(yī)療;2012年07期

2 劉鋒,朱長庚;小膠質(zhì)細(xì)胞激活的分子機(jī)制[J];解剖科學(xué)進(jìn)展;2003年02期

3 李麗霞;神經(jīng)膠質(zhì)細(xì)胞的研究現(xiàn)狀[J];陜西師范大學(xué)繼續(xù)教育學(xué)報(bào);2005年03期

4 張光運(yùn),曹玉紅,許燕,吳中亮,萬琪;光化學(xué)誘導(dǎo)局灶性腦梗死大鼠小膠質(zhì)細(xì)胞形態(tài)變化及環(huán)磷酰胺預(yù)治療的作用[J];中國臨床康復(fù);2005年29期

5 周杰;章翔;蔣曉帆;高大寬;常洪波;梁景文;宋蕾;;大鼠腦創(chuàng)傷后小膠質(zhì)細(xì)胞激活的時(shí)程及形態(tài)變化研究[J];中國臨床神經(jīng)外科雜志;2006年04期

6 雷萬龍,袁群芳,張懷波,姚志彬;腦缺血性半影區(qū)膠質(zhì)細(xì)胞和神經(jīng)元重組的形態(tài)學(xué)觀察[J];中山醫(yī)科大學(xué)學(xué)報(bào);2002年01期

，

本文編號(hào)：1655254