本文選題：CREG + 光損傷　； 參考：《內(nèi)蒙古民族大學(xué)》2017年碩士論文

【摘要】：目的:研究CREG蛋白對(duì)抗大鼠視網(wǎng)膜感光細(xì)胞光損害的作用,客觀評(píng)價(jià)其抗凋亡的能力,探索CREG在感光細(xì)胞光損傷中的抗凋亡機(jī)制。方法:1、造模:選取健康wistar大鼠30只,雌雄不限,將其隨機(jī)分成3組,對(duì)照組、光照1周組和光照2周組,每組10只。對(duì)3組大鼠鼠眼的冰凍切片行HE染色,確立模型為光照一周組。2、提取光照1周組和對(duì)照組大鼠視網(wǎng)膜蛋白,為檢測(cè)感光細(xì)胞凋亡的途徑,用Western blot檢測(cè)MAPK信號(hào)通路中關(guān)鍵蛋白ERK、P38、JNK及其磷酸化的表達(dá)量。3、選取30只wistar大鼠隨機(jī)分成3組,制備光損傷模型,制備完畢后將4μlCREG蛋白和4μlPBS分別注入大鼠的左右眼玻璃體腔。注射后第1天、3天、7天摘取鼠眼,行冰凍切片及HE染色,提取大鼠視網(wǎng)膜蛋白,Western blot檢測(cè)大鼠視網(wǎng)膜蛋白中凋亡因子casepase3、8、9的表達(dá)量。4、Western blot檢測(cè)MAPK凋亡通路及PI3K/AKT通路中關(guān)鍵蛋白P38、JNK、AKT及它們的磷酸化形式的表達(dá)量。將P38抑制劑SB203580,JNK抑制劑SP600125,AKT抑制劑LY2940002對(duì)3天組大鼠注射CREG蛋白眼進(jìn)行玻璃體腔注射,劑量為4μl,對(duì)照組注射同等劑量的PBS。Western blot檢測(cè)兩組中Caspase3的表達(dá)情況。結(jié)果:1、光照1周組視網(wǎng)膜外核層細(xì)胞較對(duì)照組變薄,光照2周視網(wǎng)膜外核層細(xì)胞消失。2、光照1周組的大鼠視網(wǎng)膜蛋白中p-P38、p-JNK的表達(dá)量較對(duì)照組的明顯增加(P0.001),P38、JNK、ERK、p-ERK的表達(dá)量較對(duì)照組無明顯變化。3、CREG蛋白注射3天組較對(duì)照組視網(wǎng)膜外核層細(xì)胞凋亡減少,而1天組和7天組無明顯變化。4、CREG蛋白注射3天組大鼠的p-JNK、p-P38、較對(duì)照組明顯減少(P0.001),p-AKT表達(dá)量明顯增加(P0.001)。而1天組和7天組較對(duì)照組無明顯變化。5、P38、JNK、AKT抑制劑注射組中的Caspase3的表達(dá)量較對(duì)照組明顯增加(P0.001)。結(jié)論:1、視網(wǎng)膜光損傷感光細(xì)胞的凋亡可以通過MAPK信號(hào)通路中關(guān)鍵蛋白P38和JNK介導(dǎo)。2、CREG蛋白注射大鼠玻璃體腔后3天組能夠抑制光損傷感光細(xì)胞的凋亡。3、CREG可以通過調(diào)節(jié)MAPK及PI3K/AKT信號(hào)通路,進(jìn)而抑制光損傷細(xì)胞的凋亡。
[Abstract]:Aim: to study the effect of CREG protein on photodamage of photoreceptor cells in rat retina, evaluate its ability of anti-apoptosis and explore the mechanism of anti-apoptosis of CREG in photoreceptor cells. Methods: 30 healthy wistar rats, male and female, were randomly divided into 3 groups: control group, 1 week illumination group and 2 week illumination group, 10 rats in each group. The frozen sections of the eyes of the three groups were stained with HE, and the model was established as one week light group. The retinal protein was extracted from the rats in the first week group and the control group, which was the way to detect the apoptosis of photoreceptor cells. Western blot was used to detect the expression of ERKN P38 Western blot and its phosphorylation in MAPK signal pathway. Thirty wistar rats were randomly divided into 3 groups to prepare the model of light injury. After the preparation, 4 渭 lCREG protein and 4 渭 lPBS were injected into the vitreous cavity of the rats. Mouse eyes were removed on the 1st day and 7th day after injection. Frozen sections and HE staining were performed. The expression of apoptotic factor casepase3m8k9 in rat retina protein was detected by Western blot. 4Western blot was used to detect the expression of the key protein P38 blot and its phosphorylated form in MAPK pathway and PI3K/AKT pathway. P38 inhibitor SB203580 JNK inhibitor SP600125AKT inhibitor LY2940002 was injected into the vitreous body cavity of the rats of the 3-day group, the dose was 4 渭 l. The expression of Caspase3 in the two groups was detected by injecting the same dose of PBS.Western blot into the control group. Results the cells in the outer nuclear layer of the retina were thinner in the 1 week group than in the control group. After 2 weeks of illumination, the cells of the outer nuclear layer of the retina disappeared. 2. The expression of p-P38 + p-JNK in the retinal protein of the rats exposed to light for 1 week was significantly higher than that in the control group. There was no significant change in the expression of p-P38-P3NK ERKPERKPERK in the control group after 3 days of injection of p0.001, and the expression of the p-JNK protein in the 3-day group was significantly higher than that in the control group. Apoptosis in the outer nuclear layer of omentum decreased, However, there was no significant change in the expression of p-JNKN p-P38 in the rats in the 1-day and 7-day groups. The expression of p-JNKK p-P38 in the 3-day group was significantly decreased compared with the control group, and the expression of p-AKT in the P0.001 group was significantly increased than that in the control group (P 0.001). There was no significant change in the expression of Caspase3 in the control group compared with the control group. The expression of Caspase3 in the control group was significantly higher than that in the control group. Conclusion the apoptosis of photoreceptor cells induced by retinal light injury can be inhibited by modulating the apoptosis of photoreceptor cells in the rat vitreous cavity 3 days after injection of the critical protein P38 and JNK in the MAPK signaling pathway. Node MAPK and PI3K/AKT signaling pathway, And then inhibit the apoptosis of the cells damaged by light.
【學(xué)位授予單位】：內(nèi)蒙古民族大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R774.1

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