Effects of bifidobacterium on mRNA expression of tumor necrosis factor receptor-associated factor 6, glycogen synthase kinase-3β and miRNA-146a in intestinal epithelial cells induced by lipopolysaccharide in rats
10.3760/cma.j.issn.2095-428X.2015.02.009
- VernacularTitle:双歧杆菌对脂多糖刺激的大鼠肠上皮细胞肿瘤坏死因子受体相关因子6、糖原合成激酶-3β和miRNA-146a mRNA表达的影响
- Author:
Yuan YUAN
;
Wei ZHOU
;
Jing LI
;
Weiming YUAN
;
Longguang HUANG
;
Shaowei ZHENG
- Publication Type:Journal Article
- Keywords:
Intestinal epithelial cell;
Rat;
Bifidobacterium;
Tumor necrosis factor receptor-associated factor 6;
Glycogen synthase kinase-3β;
miRNA-146a
- From:
Chinese Journal of Applied Clinical Pediatrics
2015;30(2):110-113
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect the effects of bifidobacterium or bifidobacterium cultured supernatant on the mRNA expression of tumor necrosis factor receptor-associated factor 6 (TRAF6),glycogen synthase kinase-3β (GSK-3β) and the miRNA-146a in rat small intestinal epithelial cell(IEC-6) induced by lipopolysaccharide (LPS).Methods IEC-6 in logarithmic phase were randomly divided into LPS group,cultured supernatant group and inactivated bacteria group.All the 3 groups were exposed to 5 mg/L LPS for 5 hours,and then 1 mL sterile saline was added in LPS group and culturing continued for 24 hours ; 1 mL bifidobacterium cultured supernatant was added in cultured supernatant group and culturing continued for 24 hours;1 mL inactivated bifidobacterium 1 x 1010 CFU/L added in inactivated bacteria group and continued culturing for 24 hours.The mRNA expressions of TRAF6,GSK-3 β and miRNA-146a were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results The level of TRAF6,GSK-3 β of culture supematant group (0.72 ± 0.05,0.46 ± 0.14) were all lower than LPS group (1.01 ± 0.14,1.02 ± 0.25),but the level of miRNA-146a(3.05 ± 0.40) was higher than that in LPS group(1.01 ± 0.12),and there were significant differences between them (t =5.278,6.316,13.218,P =0.000).The level of GSK-3 β of inactivated bacteria group(0.59 ±0.13) was significantly lower than that in LPS group(t =4.837,P =0.000).The levels of TRAF6 and miRNA-146a of inactivated bacteria group(1.05 ±0.11,0.78 ±0.22) had no significant differences with LPS group (t =0.732,1.463,P ＞ 0.05).The level of TRAF6 of cultured supernatant group was lower than that in inactivated bacteria group,and the level of miRNA-146a was higher than that in inactivated bacteria group,and there were significant differences between 2 groups (t =6.009,14.687,P =0.000).Conclusions Bifidobacterium cultured supernatant and inactivated bacteria both have certain protective effect on the IEC-6 induced by LPS.One of the protective mechanisms of bifidobacterium cultured supernatant may be achieved by elevating the expression of miRNA-146a,and decreasing the levels of inflammation related factor TRAF6 and damage related factor GSK-3β.The protective effects of inactivated bifidobacterium may be achieved by decreasing the level of damage related factor GSK-3β.
