嗜酸乳杆菌在初始弱酸碱条件下对HT-29细胞免疫因子的调节作用

doi:10.7506/spkx1002-6630-20191110-108

食品科学 ›› 2020, Vol. 41 ›› Issue (19): 191-197.doi: 10.7506/spkx1002-6630-20191110-108

嗜酸乳杆菌在初始弱酸碱条件下对HT-29细胞免疫因子的调节作用

张秋月，王刚，陈坤朋，潘道东，曾小群，吴振，郭宇星

（1.宁波大学食品与药学学院，浙江省动物蛋白食品精深加工技术重点实验室，浙江?宁波 315211；2.南京师范大学食品与制药工程学院，江苏?南京 210097）

出版日期:2020-10-15 发布日期:2020-10-23
基金资助:
江苏省攻关项目（BE3018397）；江苏省农业科技自主创新资金项目（CX（18）3036）；国家自然科学基金面上项目（31972048）；浙江省教育厅一般科研项目（Y201839391）

Effect of Lactobacillus acidophilus on Regulating the Production of Immune Factors in HT-29 Cells under Initial Weak Acidic and Alkaline Conditions

ZHANG Qiuyue, WANG Gang, CHEN Kunpeng, PAN Daodong, ZENG Xiaoqun, WU Zhen, GUO Yuxing

(1. Key Laboratory of Animal Protein Food Processing Technology of Zhejiang Province, School of Food and Pharmaceutical Sciences, Ningbo University, Ningbo 315211, China; 2. School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing 210097, China)

Online:2020-10-15 Published:2020-10-23

摘要/Abstract

摘要： 为探究嗜酸乳杆菌对人体肠道细胞的免疫调节作用，将pH 5.5和pH 7.5条件下培养的具有不同黏附特性的嗜酸乳杆菌与人结肠癌细胞HT-29共同培养，测定其免疫因子的变化。通过反转录实时荧光定量聚合酶链式反应检测了热休克蛋白70（heat shock protein 70，Hsp70）、白细胞介素（interleukin，IL）-8、IL-10、核因子κB（nuclear factor κB，NF-κB）、Toll样受体4（Toll-like receptors 4，TLR-4）和肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）基因的相对表达量，采用酶联免疫吸附测定了免疫因子IL-8、IL-10和TNF-α蛋白的表达情况。结果表明，HT-29细胞与两种pH值条件下的嗜酸乳杆菌共同培养后，在转录水平上，较空白对照组均呈现出Hsp70和TNF-α基因表达的极显著上调，IL-8、NF-κB和TLR-4基因表达的极显著下调（P＜0.01）。pH 7.5组与pH 5.5组对比，Hsp70、IL-10和TNF-α的基因表达显著上调，NF-κB和TLR-4的基因表达显著下调（P＜0.05）；在两种pH值条件下，HT-29细胞的TNF-α质量浓度极显著高于空白对照组，而IL-8的质量浓度极显著降低，并且pH 7.5组的IL-10质量浓度极显著高于pH 5.5组（P＜0.01）。综上，弱酸、弱碱性肠道条件下的嗜酸乳杆菌能促进肠道细胞免疫，且pH 7.5的弱碱性肠道环境与pH 5.5的弱酸性肠道环境相比，更能显著增强嗜酸乳杆菌对肠道的免疫效应，维护肠道健康。

关键词: 嗜酸乳杆菌；HT-29细胞；免疫因子；反转录实时荧光定量聚合酶链式反应；酶联免疫吸附测定

Abstract: In order to investigate the immunomodulatory effects of Lactobacillus acidophilus on human intestinal cells, Lactobacillus acidophilus, showing different adhesion characteristics when cultured at pH 5.5 and 7.5, were co-cultured with human colon cancer HT-29 cells, and the changes of immune factors in HT-29 cells were determined. The relative gene expression levels of heat shock protein (Hsp) 70, interleukin (IL)-8, IL-10, nuclear factor κB (NF-κB), Toll-like receptor 4 (TLR-4) and tumor necrosis factor-α (TNF-α) were detected by reverse-transcription quantitative real-time polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA) was applied to determine the protein expression levels of IL-8, IL-10 and TNF-α. RT-qPCR results showed that after the co-cultivation of HT-29 cells and Lactobacillus acidophilus at either pH, at the transcriptional level, Hsp70 and TNF-α were significantly up-regulated, while IL-8, NF-κB and TLR-4 were significantly down-regulated when compared with the blank control cells without the presence of Lactobacillus acidophilus (P < 0.01). The gene expression levels of Hsp70, IL-10 and TNF-α were significantly up-regulated in the pH 7.5 group when compared with the pH 5.5 group; meanwhile, NF-κB and TLR-4 were significantly down-regulated (P < 0.05). The ELISA results showed that under both pH conditions, the TNF-α expression level in HT-29 cells was significantly higher than that in the blank control group, while the concentration of pro-inflammatory factor IL-8 was significantly decreased; the anti-inflammatory factor IL-10 in the pH 7.5 group was significantly higher than that in the pH 5.5 group (P < 0.01). In summary, Lactobacillus acidophilus under weak acidic and weak alkaline intestinal conditions can promote the immunity of intestinal cells, and the weak alkaline condition with pH 7.5 is more effective than the weak acidic intestinal environment with pH 5.5, which can significantly enhance the immunoregulatory effect of Lactobacillus acidophilus on the intestinal tract, thereby helping maintain intestinal health.

Key words: Lactobacillus acidophilus; HT-29 cells; immune factors; reverse transcription quantitative real-time polymerase chain reaction; enzyme-linked immunosorbent assay

中图分类号:

TS252.1

张秋月，王刚，陈坤朋，潘道东，曾小群，吴振，郭宇星. 嗜酸乳杆菌在初始弱酸碱条件下对HT-29细胞免疫因子的调节作用[J]. 食品科学, 2020, 41(19): 191-197.

ZHANG Qiuyue, WANG Gang, CHEN Kunpeng, PAN Daodong, ZENG Xiaoqun, WU Zhen, GUO Yuxing. Effect of Lactobacillus acidophilus on Regulating the Production of Immune Factors in HT-29 Cells under Initial Weak Acidic and Alkaline Conditions[J]. FOOD SCIENCE, 2020, 41(19): 191-197.

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嗜酸乳杆菌在初始弱酸碱条件下对HT-29细胞免疫因子的调节作用

Effect of Lactobacillus acidophilus on Regulating the Production of Immune Factors in HT-29 Cells under Initial Weak Acidic and Alkaline Conditions

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