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丁酸鈉對(duì)小鼠腦缺血再灌注損傷的保護(hù)作用

發(fā)布時(shí)間:2018-07-25 09:39
【摘要】:目的研究不同劑量丁酸鈉對(duì)腦缺血再灌注損傷急性期小鼠炎癥反應(yīng)、神經(jīng)元凋亡保護(hù)機(jī)制。方法選取由ICR小鼠124只,將小鼠隨機(jī)分為4組(每組31只):假手術(shù)組、模型組、低劑量實(shí)驗(yàn)組(丁酸鈉5 mg·kg~(-1))和高劑量試驗(yàn)組(丁酸鈉10 mg·kg~(-1));依據(jù)改良Zea-Longa線栓法制備大鼠大腦中動(dòng)脈栓塞模型,血流阻斷2 h后拔出線栓,形成再灌注。假手術(shù)組小鼠雙側(cè)的頸總動(dòng)脈暴露但血管不進(jìn)行結(jié)扎,表皮的切口正常的縫合,在24 h以后處死小鼠;模型組、低劑量試驗(yàn)組和高劑量試驗(yàn)組在假手術(shù)組基礎(chǔ)上,結(jié)扎雙側(cè)頸總動(dòng)脈,30 min以后解開結(jié)扎線,再灌注2 h后灌胃處理,24 h后處死小鼠。用WST-1檢測(cè)超氧化物歧化酶(SOD)活力,用硫代巴比妥酸檢測(cè)丙二醛(MDA)含量,用Western blot檢測(cè)B淋巴細(xì)胞癌-2(Bcl-2)、Bcl-2相關(guān)X蛋白(Bax)、絲氨酸/蘇氨酸激酶(Akt)和磷酸化Akt(p-Akt)蛋白表達(dá)量,用ELISA法檢測(cè)小鼠腦組織內(nèi)白細(xì)胞介素8(IL-8)、白細(xì)胞介素1β(IL~(-1)β)和腫瘤壞死因子-α(TNF-α)的含量。結(jié)果模型組、假手術(shù)組與高劑量試驗(yàn)組SOD為(115.46±25.07),(153.65±34.81),(146.88±15.63)U·mg~(-1),MDA為(2.13±0.27),(1.42±0.07),(1.43±0.08)nmol·mg~(-1),IL~(-1)β分別為(17.82±3.60),(7.64±1.14),(10.15±2.05)pg·mg~(-1);TNF-α分別為(16.03±1.67),(7.95±0.85),(11.08±0.83)pg·mg~(-1),IL-8分別為(15.03±2.66),(9.52±1.05),(10.28±1.17)pg·mg~(-1);Bax/Bcl-2比值分別為0.67±0.05,1.00±0.00,1.13±0.10;p-Akt/Akt比值分別為0.72±0.06,1.00±0.00,1.18±0.12,模型組分別與假手術(shù)組和高劑量試驗(yàn)組比較,差異均有統(tǒng)計(jì)學(xué)意義(均P0.05)。結(jié)論丁酸鈉能夠抑制炎性因子與氧化應(yīng)激,使Bcl-2蛋白的表達(dá)增加,Bax蛋白的表達(dá)減少,從而對(duì)神經(jīng)元的凋亡起到抑制作用。
[Abstract]:Aim to study the protective mechanism of different doses of sodium butyrate on inflammatory response and neuronal apoptosis in mice with acute cerebral ischemia reperfusion injury. Methods 124 ICR mice were randomly divided into 4 groups (31 mice in each group): sham operation group, model group, and model group. Low dose sodium butyrate (5 mg kg ~ (-1) and high dose group (10 mg kg ~ (-1) sodium butyrate) were used to establish the model of middle cerebral artery embolization according to modified Zea-Longa thread embolization method. In sham-operation group, the common carotid artery was exposed but the blood vessel was not ligated, the incision of epidermis was sutured normally, and the mice were killed after 24 hours, while the model group, low-dose test group and high-dose experimental group were on the basis of sham-operation group. After 30 min of ligation of bilateral common carotid artery, the ligation line was untied, and the mice were killed after 24 h of gastric perfusion after 2 h reperfusion. The activity of superoxide dismutase (SOD) was detected by WST-1, malondialdehyde (MDA) by thiobarbituric acid, and the expression of (Bax), serine / threonine kinase (Akt) and phosphorylated Akt (p-Akt) by Western blot. The contents of interleukin-8 (IL-8), interleukin-1 尾 (IL-1 尾) and tumor necrosis factor- 偽 (TNF- 偽) in brain tissue of mice were detected by ELISA method. 緇撴灉妯″瀷緇,

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