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双歧杆菌对脂多糖刺激的大鼠肠上皮细胞中TRAF6、GSK - 3β和微小RNA - 146a mRNA表达水平的影响

Effect of Bifidobacterium on the mRNA expression levels of TRAF6, GSK-3β, and microRNA-146a in LPS-stimulated rat intestinal epithelial cells.

作者信息

Zhou W, Yuan Y, Li J, Yuan W M, Huang L G, Zheng S W

机构信息

Department of Neonatology, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, Guangdong Province, China.

出版信息

Genet Mol Res. 2015 Aug 21;14(3):10050-6. doi: 10.4238/2015.August.21.11.

Abstract

We investigated the effect of inactivated Bifidobacterium on the mRNA expression of TRAF6, GSK-3β, and microRNA-146a in lipopolysaccharide (LPS)-stimulated rat small intestinal epithelial cells (IEC-6s). IEC-6s were randomly divided into an LPS group, a culture supernatant group, and an inactivated bacteria group. After stimulation with LPS for 5 h, the three groups were treated as follows: the LPS group was cultured for 24 h with sterile saline; the culture supernatant group was cultured with Bifidobacterium (infantis strain) culture supernatant for 24 h; and the inactivated bacteria group was cultured with inactivated infantis Bifidobacterium for 24 h. Reverse transcription polymerase chain reaction was used to determine mRNA expression levels. The mRNA expression levels of TRAF-6 and GSK-3β in the culture supernatant group were lower, and microRNA-146a expression was higher, compared with the LPS group (t = 5.278, P = 0.000; t = 6.316, P = 0.000; t = 13.218, P = 0.000, respectively). GSK-3β mRNA expression in the inactivated bacteria group was lower than in the LPS group (t = 4.837, P = 0.000). There was no difference in the mRNA expression levels of TRAF-6 and microRNA-146a between the two groups (t = 0.732, P = 0.472 and t = 1.463, P = 0.164). Both the culture supernatant and the inactivated Bifidobacterium had a protective effect on LPS-stimulated IEC-6s. The protective effect of Bifidobacterium may be achieved through increased microRNA-146a by reducing levels of TRAF6 and GSK-3β; the protective effect of inactivated Bifidobacterium may be achieved by reducing levels of GSK-3β.

摘要

我们研究了灭活双歧杆菌对脂多糖(LPS)刺激的大鼠小肠上皮细胞(IEC-6s)中TRAF6、GSK-3β和微小RNA-146a的mRNA表达的影响。IEC-6s被随机分为LPS组、培养上清液组和灭活细菌组。用LPS刺激5小时后,三组进行如下处理:LPS组用无菌盐水培养24小时;培养上清液组用双歧杆菌(婴儿株)培养上清液培养24小时;灭活细菌组用灭活的婴儿双歧杆菌培养24小时。采用逆转录聚合酶链反应测定mRNA表达水平。与LPS组相比,培养上清液组中TRAF-6和GSK-3β的mRNA表达水平较低,而微小RNA-146a表达较高(t分别为5.278,P = 0.000;t = 6.316,P = 0.000;t = 13.218,P = 0.000)。灭活细菌组中GSK-3β mRNA表达低于LPS组(t = 4.837,P = 0.000)。两组之间TRAF-6和微小RNA-146a的mRNA表达水平无差异(t = 0.732,P = 0.472和t = 1.463,P = 0.164)。培养上清液和灭活双歧杆菌对LPS刺激的IEC-6s均有保护作用。双歧杆菌的保护作用可能是通过降低TRAF6和GSK-3β水平增加微小RNA-146a来实现的;灭活双歧杆菌的保护作用可能是通过降低GSK-3β水平来实现的。

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