【摘要】：目的:評(píng)價(jià)1.5%、3.0%以及4.5%的異氟醚后處理對(duì)大鼠海馬腦片氧糖剝奪損傷中的神經(jīng)保護(hù)作用;探討激活素A及其下游因子ERK1/2對(duì)異氟醚后處理對(duì)抗大鼠海馬腦片氧糖剝奪損傷中的保護(hù)作用。方法:新生SD大鼠80只,2周齡,體重50~70g,采用隨機(jī)數(shù)字表法,將其分為十組(n=8):正常對(duì)照組(Normal組),缺氧無(wú)糖損傷組(OGD組),異氟醚后處理組(1.5%、3.0%、4.5%ISPOC組)、激活素A抑制劑組(SB431542組)ERK1/2抑制劑組(U0126組)、異氟醚后處理+抑制劑組(ISPOC+SB431542或U0126組)、溶媒對(duì)照組(DMSO組)。抑制劑在不同異氟醚濃度組使用時(shí)間為OGD開(kāi)始之前。HE染色、TTC染色以及PI染色用于測(cè)定海馬腦片組織的活性。免疫熒光技術(shù)、Western blot技術(shù)以及實(shí)時(shí)熒光定量PCR技術(shù)用于檢測(cè)激活素A、Smad2/3、磷酸化Smad2/3(p-Smad2/3)、ERK1/2以及磷酸化ERK1/2(p-ERK1/2)蛋白以及m RNA的表達(dá)。結(jié)果:與Normal組相比較,OGD組壞死神經(jīng)元細(xì)胞的數(shù)量、PI染色的平均熒光強(qiáng)度增加,但甲瓚生成量降低、激活素A以及p-ER強(qiáng)度增加,但甲瓚產(chǎn)量、激活素A、p-Smad2/3以及p-ERK1/2蛋白表達(dá)水平以及激活素A m RNA水平有所降低。然而,4.5%ISPOC組中激活素A、p-Smad2/3以及p-ERK1/2蛋白表達(dá)水平以及激活素A m RNA水平要高于OGD組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05);其他實(shí)驗(yàn)結(jié)果在SB431542組/U0126組與3.0%ISPOC組之間相比較,PI染色的平均熒光強(qiáng)度增加,而激活素A、p-Smad2/3以及K1/2蛋白的表達(dá)水平、激活素A m RNA的水平有所降低(P0.05);與OGD組相比較,1.5%ISPOC組和3.0%ISPOC組壞死神經(jīng)元細(xì)胞的數(shù)量以及PI染色的平均熒光強(qiáng)度降低,但甲瓚產(chǎn)量、激活素A、p-Smad2/3以及p-ERK1/2蛋白表達(dá)水平以及激活素A m RNA水平顯著升高(P0.05);4.5%ISPOC組的實(shí)驗(yàn)室檢查結(jié)果與1.5%ISPOC、3.0%ISPOC組結(jié)果完全相反,該組中損傷神經(jīng)元細(xì)胞的數(shù)量以及平均熒光p-ERK1/2蛋白表達(dá)水平降低(P0.05);ERK1/2在所有組之間表達(dá)沒(méi)有統(tǒng)計(jì)學(xué)意義(p0.05)。結(jié)論:該研究證實(shí)與1.5%和4.5%異氟醚濃度相比,3.0%的異氟醚后處理能夠提供更好的神經(jīng)保護(hù)作用。激活素A、激活素A/Smads以及激活素A/ERK1/2信號(hào)調(diào)節(jié)通路參與了異氟醚后處理誘導(dǎo)的腦保護(hù)作用的機(jī)制。
[Abstract]:Objective: to evaluate the neuroprotective effect of 1.5% and 4.5% isoflurane on oxygen and glucose deprivation injury in rat hippocampal slices. To investigate the protective effect of activin A and its downstream factor ERK1/2 on the protective effect of isoflurane post-treatment on oxygen-glucose deprivation injury in rat hippocampal slices. Methods: 80 neonatal SD rats, aged 2 weeks and weighing 50 ~ 70g, were randomly divided into two groups. They were divided into 10 groups: normal control group (Normal group), hypoxic and glucose free injury group (OGD group), isoflurane post-treatment group (1.5%), activin A inhibitor group (SB431542 group), ERK1/2 inhibitor group (U0126 group), isoflurane post-treatment inhibitor group (ISPOC SB431542 or U0126 group). Solvent control group (DMSO group). In different isoflurane concentration groups, the activity of hippocampal slices was determined by HE staining and PI staining before the onset of OGD. Western blot and real-time fluorescent quantitative PCR were used to detect the expression of activin A Smad2 / 3, phosphorylated Smad2/3 (p-Smad2/3) / ERK1 / 2, phosphorylated ERK1/2 (p-ERK1/2) protein and mRNA. Results: compared with Normal group, the number of necrotic neurons in OGD group increased with the average fluorescence intensity of Pi staining, but the amount of Gan produced decreased, the intensity of activin A and p-ER increased, but the yield of Jia Zan increased. The expression of activin p-Smad2 / 3, p-ERK1/2 protein and activin A m RNA decreased. However, the levels of activin p-Smad2 / 3 and p-ERK1/2 protein and activin A m RNA in ISPOC group were significantly higher than those in OGD group (P0.05), and the average fluorescence intensity of Pi staining in SB431542 group / U0126 group was higher than that in 3.0%ISPOC group. The expression of activin p-Smad2 / 3 and K1 / 2 protein and the level of activin A m RNA were decreased (P0.05), and the number of necrotic neurons and the average fluorescence intensity of PI staining in OGD group and 3.0%ISPOC group were lower than those in OGD group, but the average fluorescence intensity of PI staining was decreased. The levels of activin p-Smad2 / 3 and p-ERK1/2 protein and activin A m RNA were significantly increased (P0.05). The results of laboratory examination in 4.5 ISPOC group were contrary to those in 1.5 ISPOC3.0 group. The number of injured neuron cells and the expression level of average fluorescent p-ERK1/2 protein decreased (P0.05) the expression of ERK1 / 2 in all groups was not statistically significant (p0.05). Conclusion: compared with 1.5% and 4.5% isoflurane concentrations, 3.0% isoflurane post treatment can provide better neuroprotective effect. Activin A, activin A/Smads and activin A/ERK1/2 signaling pathway are involved in the mechanism of brain protection induced by isoflurane postprocessing.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R614

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 田安好;趙薇;張蕾;董佳亮;李偉偉;金輝;;卵泡抑素、激活素A與BMP-4在缺血缺氧性腦損傷大鼠腦組織中的表達(dá)[J];四川大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2014年05期

2 Hongliang Guo;Xiaoran Shen;Ye Xu;Junliang Yuan;Dongming Zhao;Wenli Hu;;Emodin prevents hypoxic-ischemic neuronal injury Involvement of the activin A pathway[J];Neural Regeneration Research;2013年15期

3 Guihua Xu;Jinting He;Hongliang Guo;Chunli Mei;Jiaoqi Wang;Zhongshu Li;Han Chen;Jing Mang;Hong Yang;Zhongxin Xu;;Activin A prevents neuron-like PC12 cell apoptosis after oxygen-glucose deprivation[J];Neural Regeneration Research;2013年11期

4 ;Dual Isoflurane-induced Preconditioning Improves Neuroprotection in Rat Brain In Vitro and the Role of Extracellular Signal-regulated Protein Kinase[J];Chinese Medical Sciences Journal;2011年01期

5 張令軍;黃鶴;潘平喜;劉小菁;吳文超;付華;;激活素A在心肌缺血后適應(yīng)中的表達(dá)變化及意義[J];四川大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2009年01期

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