本文選題：創(chuàng)傷性腦損傷 + miRNA　； 參考：《第三軍醫(yī)大學(xué)學(xué)報(bào)》2017年14期

【摘要】：目的觀察創(chuàng)傷性腦損傷(traumatic brain injury,TBI)后大鼠損傷皮質(zhì)區(qū)miRNA-9的表達(dá)變化規(guī)律,探討其對(duì)腦微血管內(nèi)皮細(xì)胞的保護(hù)作用。方法選取成年雄性SD大鼠60只制作控制性皮質(zhì)撞擊損傷模型(controlled cortical impact,CCI),分別應(yīng)用熒光定量PCR和Western blot檢測(cè)傷后6 h及1、3、7、14、28 d各時(shí)間點(diǎn)傷灶周?chē)M織中miRNA-9及CD31的表達(dá)情況。培養(yǎng)原代大鼠腦微血管內(nèi)皮細(xì)胞,將內(nèi)皮細(xì)胞分為正常組、損傷模型組[用依托泊苷(etoposide,ETO)損傷]、miRNA-9過(guò)表達(dá)損傷模型組、空轉(zhuǎn)染損傷模型組,應(yīng)用CCK-8檢測(cè)各組細(xì)胞活力;應(yīng)用Western blot檢測(cè)各組B細(xì)胞淋巴瘤/白血病-2蛋白(B-cell lymphoma-2,Bcl-2)、B細(xì)胞淋巴瘤/白血病-2相關(guān)X蛋白(Bcl-2 associated X,Bax)、活化半胱氨酸天冬氨酸特異性蛋白酶-3蛋白(cleaved cysteinyl aspartate specific proteinase 3,cl-caspase-3)表達(dá)水平。結(jié)果 (1)腦創(chuàng)傷后傷灶周?chē)鷧^(qū)域miRNA-9表達(dá)明顯增加,于傷后第14天達(dá)高峰(P0.01);內(nèi)皮細(xì)胞標(biāo)志物CD31蛋白表達(dá)水平從傷后第3~28天持續(xù)高于正常組(P0.05);(2)內(nèi)皮細(xì)胞建模轉(zhuǎn)染后,qPCR結(jié)果提示損傷模型組較正常組miRNA-9表達(dá)顯著降低(P0.01),但miRNA-9過(guò)表達(dá)損傷模型組miRNA-9表達(dá)顯著高于損傷模型組(P0.01);CCK-8結(jié)果同樣顯示miRNA-9過(guò)表達(dá)損傷模型組細(xì)胞活力明顯高于損傷模型組(P0.01);(3)相比于損傷模型組,miRNA-9過(guò)表達(dá)損傷模型組內(nèi)皮細(xì)胞Bcl-2蛋白表達(dá)增加(P0.01),Bcl-2/Bax值增加(P0.05),但Bax蛋白、cl-caspase-3蛋白表達(dá)降低(P0.05)。結(jié)論創(chuàng)傷性腦損傷后傷灶周?chē)鷧^(qū)域miRNA-9表達(dá)增多且過(guò)表達(dá)miRNA-9可提高依托泊苷誘導(dǎo)損傷的內(nèi)皮細(xì)胞活力,提示腦創(chuàng)傷后miRNA-9表達(dá)增加有助于腦血管重塑的發(fā)生。
[Abstract]:Objective to observe the expression of miRNA-9 in rat cortical area after traumatic brain injury (traumatic brain injura) and to explore its protective effect on cerebral microvascular endothelial cells (MECs). Methods 60 adult male Sprague-Dawley rats were selected to make the model of (controlled cortical impaction injury. The expression of miRNA-9 and CD31 in the tissues around the injured focus was detected by fluorescence quantitative PCR and Western blot. Primary rat brain microvascular endothelial cells were cultured. Endothelial cells were divided into normal group, injury model group [etoposideside ETO injury model group] miRNA-9 overexpression model group, empty transfection injury model group, and CCK-8 were used to detect the cell viability of each group. The expression of B cell lymphoma-2Bcl 2 protein and B cell lymphoma / leukemia associated X protein (Bcl-2 associated XnBax) and activated cysteine aspartate specific protease 3 (cleaved cysteinyl aspartate specific proteinase 3) were detected by Western blot. Results (1) the expression of miRNA-9 was significantly increased in the perifocal area after traumatic brain injury. The expression level of CD31 protein in endothelial cells was continuously higher than that in normal group (P0.05); (2) on the 14th day after injury (P0.01). The results showed that the expression of miRNA-9 in the model group was significantly lower than that in the normal group (P0.01), but the expression of CD31 protein in the model group was significantly lower than that in the normal group (P0.01). The expression of miRNA-9 in the injury model group was significantly higher than that in the injury model group (P0.01). The results also showed that the cell viability of the injury model group was significantly higher than that of the injury model group (P0.01); (3) compared with the injury model group. The expression of Bcl-2 protein increased (P0.01) and Bcl-2 / Bax increased (P0.05), but the expression of Bax protein cl-caspase-3 decreased (P0.05). Conclusion the increased expression of miRNA-9 and overexpression of miRNA-9 can increase the viability of endothelial cells induced by etoposide after traumatic brain injury, suggesting that the increased expression of miRNA-9 is helpful to the development of cerebral vascular remodeling after traumatic brain injury.
【作者單位】： 重慶醫(yī)科大學(xué)附屬第一醫(yī)院神經(jīng)外科;
【基金】：國(guó)家自然科學(xué)基金面上項(xiàng)目(81571159)~~
【分類(lèi)號(hào)】：R651.15

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