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目的以干酪乳杆菌(Lactobacillus casei,L.casei)胞外多糖(exopolysaccharides,EPS)为研究对象,研究其在体外对Balb/c小鼠小肠上皮细胞(small intestinal epithelial cell,IECs)增殖及其分泌IL-8、IL-10和TGF-β的影响。方法利用嗜热菌蛋白酶消化联合差速贴壁以及免疫细胞化学鉴定的方法对Balb/c小鼠IECs进行分离和鉴定,获得实验用的IECs。用不同质量浓度的L.casei EPS与IECs共培养,分别观察0.5、1、2 h和4 h后IECs的增殖情况。当用不同质量浓度的L.casei EPS处理IECs 2 h后,采用双抗体夹心ELISA法检测细胞培养上清液中细胞因子IL-8、IL-10和TGF-β的质量浓度。结果嗜热菌蛋白酶消化联合差速贴壁的方法能够获得纯度高与活性好的IECs;L.casei EPS能够促进IECs的增殖,显著上调上清液中IL-8、IL-10和TGF-β的分泌水平(P<0.05),EPS质量浓度在0~150μg/ml范围内呈剂量依赖关系,并且在150μg/ml的质量浓度时达到最大值。结论 L.casei EPS能够促进IECs的增殖及IL-8、IL-10和TGF-β的分泌水平。
Objective To study the effects of exopolysaccharides (EPS) from Lactobacillus casei (L.casei) on the proliferation and secretion of Balb / c mouse small intestinal epithelial cells (IECs) IL-8, IL-10 and TGF-β. Methods The IECs of Balb / c mice were isolated and identified by thermophilic protease digestion combined with differential adherent and immunocytochemical identification to obtain experimental IECs. The LPSasei EPS with different mass concentrations was co-cultured with IECs to observe the proliferation of IECs after 0.5, 1, 2 h and 4 h respectively. When IECs were treated with different concentrations of L.casei EPS for 2 h, the concentrations of cytokines IL-8, IL-10 and TGF-β in the cell culture supernatants were determined by sandwich ELISA. Results The combination of thermophilic protease digestion and differential adherent method could obtain IECs with high purity and good activity. L.casei EPS could promote the proliferation of IECs and significantly upregulate the levels of IL-8, IL-10 and TGF-β (P <0.05). The EPS concentration was in a dose-dependent manner in the range of 0-150 μg / ml and reached a maximum at a concentration of 150 μg / ml. Conclusion L.casei EPS can promote the proliferation of IECs and the secretion of IL-8, IL-10 and TGF-β.