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目的探讨干酪乳酸杆菌对新生小鼠坏死性小肠结肠炎(NEC)模型TH17细胞、白细胞介素(IL)-17A和IL-6的影响及其免疫调节的可能机制。方法将出生24h内的60只新生小鼠随机分为NEC组、NEC+益生菌组、空白对照组及单纯益生菌组,每组各15只。益生菌组出生后即给予干酪乳酸杆菌活菌悬液,共3周,在第3周末NEC组予缺氧+冷刺激,每天2次,共3天,最后一天予腹腔注射脂多糖(LPS)造模。所有小鼠在NEC组造模后6h进行心脏取血0.4ml,用流式细胞术检测新生小鼠血TH17细胞,ELISA检测肠道组织IL-17A、IL-6含量,HE染色病理切片观察新生鼠肠道病理变化。结果 (1)NEC组TH17细胞(%)、IL-17A含量(pg/ml)、IL-6含量(pg/ml)及肠组织病理评分(分)均高于NEC+益生菌组和空白对照组[TH17细胞:(2.50±0.63)比(0.24±0.09)、(0.14±0.06);IL-17A:(197.8±101.3)比(20.3±16.6)、(3.1±2.9);IL-6:(609.1±93.2)比(116.0±90.3)、(16.9±8.9);病理评分:(3.8±0.4)比(1.9±0.6)、(0.2±0.4),P均<0.001]。空白对照组与单纯益生菌组差异无统计学意义(P>0.05)。(2)肠组织病理评分与TH17细胞、IL-17A、IL-6水平成正相关(r值分别为0.808、0.856、0.754,P均<0.01)。结论 TH17细胞及其细胞因子可能参与NEC的发病过程,干酪乳酸杆菌可能通过降低TH17细胞、IL-17A、IL-6含量改善肠道及机体免疫功能,从而减轻NEC严重程度。
Objective To investigate the effect of Lactobacillus casei on TH17 cells, interleukin (IL) -17A and IL-6 in neonatal mouse model of necrotizing enterocolitis (NEC) and the possible mechanism of immune regulation. Methods Sixty newborn mice were randomly divided into NEC group, NEC + probiotics group, blank control group and probiotics group, 15 in each group. The probiotic group was given live Lactobacillus casei suspension after birth for 3 weeks. On the 3rd weekend, the NEC group was given hypoxia + cold stimulation twice a day for 3 days, and the last day was injected intraperitoneally with lipopolysaccharide (LPS) Modeling. All mice were sacrificed at 4h after NEC injection. Blood samples were collected from neonatal mice for blood flow cytometry. TH17 cells were detected by flow cytometry. The contents of IL-17A and IL-6 in gut were detected by ELISA. Rat intestinal pathological changes. Results (1) The expression of TH17 cells (%), IL-17A (pg / ml), IL-6 content (pg / ml) and intestinal histopathological score in NEC group were higher than those in NEC + probiotics group and blank control group IL-17A: (197.8 ± 101.3) vs. (20.3 ± 16.6), (3.1 ± 2.9); IL-6: (609.1 ± 0.63); (2.50 ± 0.63) ± 93.2), (116.0 ± 90.3) and (16.9 ± 8.9) respectively. The pathological score was (3.8 ± 0.4) vs (1.9 ± 0.6) and (0.2 ± 0.4) respectively, P <0.001. There was no significant difference between blank control group and simple probiotics group (P> 0.05). (2) The histopathological score of intestinal tissue was positively correlated with the level of TH17 cells, IL-17A and IL-6 (r = 0.808,0.856,0.754, P <0.01 respectively). Conclusion TH17 cells and their cytokines may be involved in the pathogenesis of NEC. Lactobacillus casei may reduce the severity of NEC by decreasing the content of TH17 cells, IL-17A and IL-6, and improving the immune function of the intestine and the body.