本文關(guān)鍵詞：誘導(dǎo)型多能干細(xì)胞通過修復(fù)ZO-1蛋白改善膿毒癥小鼠腸黏膜通透性和細(xì)菌移位　出處：《中山大學(xué)學(xué)報(bào)(醫(yī)學(xué)科學(xué)版)》2016年05期 　論文類型：期刊論文

  更多相關(guān)文章： 誘導(dǎo)型多能干細(xì)胞 膿毒癥 腸黏膜通透性 緊密連接 ZO-蛋白

【摘要】：【目的】探討誘導(dǎo)型多能干細(xì)胞(i PS-MSC)對(duì)膿毒癥小鼠腸黏膜通透性的改善作用和對(duì)腸道菌群移位的影響。【方法】采用尾靜脈注射脂多糖(LPS,10 mg/Kg)建立小鼠膿毒癥模型。常規(guī)培養(yǎng)及傳代i PS-MSC。在膿毒癥建立2 h和6 h經(jīng)小鼠尾靜脈注射進(jìn)行治療干預(yù)(1×106/只)。利用基因重組技術(shù)制備新質(zhì)粒p ET28b(+)-EGFP,轉(zhuǎn)染大腸桿菌,并在膿毒癥后連續(xù)2 d予小鼠進(jìn)行灌胃處理。在膿毒癥發(fā)生72 h后,取各組小鼠腹水及腸系膜淋巴結(jié)勻漿于Kan抗性的LB培養(yǎng)基培養(yǎng)鑒定,計(jì)算和比較菌群負(fù)荷。通過透射電鏡觀察小鼠小腸黏膜上皮細(xì)胞超微結(jié)構(gòu)的變化。Western blotting法檢測(cè)各組小鼠腸黏膜ZO-1蛋白的表達(dá)。熒光顯微鏡下觀察帶有GFP綠色熒光蛋白的ips-MSC在損傷腸黏膜的移行和歸巢情況。【結(jié)果】成功構(gòu)建表達(dá)綠色熒光的重組大腸桿菌p ET-28b(+)-EGFP作為實(shí)驗(yàn)用示蹤菌。灌胃處理后,ips-MSC治療的小鼠腹水和腸系膜淋巴結(jié)的示蹤菌的菌負(fù)荷較膿毒癥組明顯降低,其中ips-MSC 2 h治療組較6 h治療組降低的更明顯。透射電鏡觀察發(fā)現(xiàn)膿毒癥小鼠腸系膜上皮細(xì)胞間緊密連接結(jié)構(gòu)破壞,細(xì)胞間縫隙增寬,而經(jīng)i PS-MSC治療的小鼠細(xì)胞間緊密連接結(jié)構(gòu)破壞明顯改善,在2 h治療組表現(xiàn)更為明顯,伴隨緊密連接蛋白ZO-1的表達(dá)水平升高。【結(jié)論】靜脈注射i PS-MSC能夠向膿毒癥小鼠損傷的腸黏膜移行,通過提高連接蛋白ZO-1的表達(dá),修復(fù)緊密連接,改善膿毒癥小鼠腸黏膜的通透性,減少膿毒癥時(shí)腸道菌群的移位。
[Abstract]:[Objective] to investigate the induced pluripotent stem cells (I PS-MSC) to improve the role of sepsis in colonic mucosa of mice with sepsis and the influence on the permeability of bacterial translocation. [method] by intravenous injection of lipopolysaccharide (LPS, 10 mg/Kg) to establish a mouse model of sepsis. The conventional culture and subculture I PS-MSC. established in sepsis in 2 h and 6 h were injected into tail vein of mice were treated (1 x 106/). Preparation of plasmid P ET28b by gene recombination technology (+) -EGFP, transfected into Escherichia coli, and 2 consecutive d to mice in sepsis after gavage. In sepsis after 72 h from each group, the ascites and mesenteric lymph nodes of mice in homogenate Kan resistant LB medium identification, calculation and comparison of bacteria load. Transmission electron microscope was used to observe changes of.Western expression by blotting mice intestinal mucosal epithelial cell ultrastructure of intestinal mucosa of mice were detected ZO-1 protein fluorescence. Light microscope with GFP green fluorescent protein ips-MSC in intestinal mucosa injury of migration and homing. [results] the expression of green fluorescence in recombinant Escherichia coli P was successfully constructed ET-28b (+) -EGFP as tracer experiments with bacteria. After treatment by gavage, bacterial loads in ips-MSC treated mice ascites and mesenteric lymph node tracer bacteria than sepsis group significantly decreased, the ips-MSC 2 h treatment group compared with 6 h treatment group decreased more obvious. Transmission electron microscopy showed sepsis mouse mesenteric tight junctions between epithelial cells damage, cell gap widened, and the cell I of mice treated with PS-MSC significantly improved the close connection between the structural damage in 2, h in treatment group was more obvious, with increased expression of tight junction protein ZO-1. [Conclusion] the intravenous injection of I PS-MSC to septic mice intestinal mucosa injury by increasing the migration The expression of protein ZO-1, repair close connection, improve the permeability of intestinal mucosa in sepsis mice, and reduce the displacement of intestinal microflora in sepsis.

【作者單位】： 中山大學(xué)附屬第一醫(yī)院急診科;中山大學(xué)附屬第三醫(yī)院急診科;
【基金】：廣東省醫(yī)學(xué)科學(xué)技術(shù)研究基金(A2015486) 廣東省科技計(jì)劃項(xiàng)目(2012B031800366) 廣東省自然科學(xué)基金(S2013010015328)
【分類號(hào)】：R459.7
【正文快照】： 膿毒癥是常并發(fā)于創(chuàng)傷、手術(shù)、感染、休克等,是目前臨床常見的危急重癥之一,死亡率高,救治困難[1]。膿毒癥的應(yīng)激狀態(tài)下,患者的腸黏膜的結(jié)構(gòu)和功能極易受到損傷,表現(xiàn)為腸黏膜萎縮,腸道的通透性增高,腸腔內(nèi)細(xì)菌移位,從而誘發(fā)腸原性感染的二次打擊,進(jìn)一步誘發(fā)全身炎癥反應(yīng)綜合征

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