乳杆菌微小膜蛋白对肠上皮细胞Caco-2的 细胞毒性研究

目的利用脂多糖(lipopolysaccharide,LPS)刺激模拟体外炎症环境,观察不同浓度下乳杆菌微小膜蛋白(micro integral membrane protein,MIMP)对肠上皮细胞Caco-2的生物学影响,评估其细胞毒性作用。方法首先通过CCK-8实验检测在LPS刺激后不同浓度MIMP(0.01、0.1和1ng/mL)对Caco-2细胞增殖活性的影响,并利用Toll样受体4(Toll-like receptor 4,TLR4)抑制剂作为阳性对照。其次利用流式细胞术检测不同浓度下MIMP对Caco-2细胞凋亡及细胞周期的影响。结果在12h的特定孵育时间内,单独应用不同浓度MIMP及TLR4抑制剂对Caco-2细胞增殖活性无显著影响(P0.05),但是MIMP可以拮抗LPS对Caco-2细胞的促增殖作用(P0.05)。不同浓度的MIMP对Caco-2细胞的周期和凋亡无明显影响(P0.05)。结论不同浓度MIMP对Caco-2细胞的增殖、凋亡及细胞周期无明显影响,并可以拮抗LPS的促细胞增殖作用,因此具有较高的安全性,有望用于炎症性肠病的治疗。

Cytotoxicity of micro integral membrane protein against intestinal epithelial Caco-2 cells

Abstract: Objective To evaluate the biological effects of micro integral membrane protein (MIMP) on intestinal epithelial cells Caco-2 and its cytotoxicity under different concentrations. Methods Firstly, Caco-2 cells were stimulated by using LPS, the proliferation of which was detected with CCK-8 assay under different concentrations of MIMP (0.01, 0.1 and 1 ng/mL), with toll-like receptor 4 (TLR4) inhibitors as the positive control. Secondly, flow cytometry was used to observe the impact of MIMP on the cell cycle and apoptosis of Caco-2 cells. Results Within specific incubation time of 12 hours, different concentrations of MIMP or TLR4 inhibitors alone had no significant effects on the proliferation of Caco-2 cells (P>0.05). However, MIMP antagonized the effect of LPS on the proliferation of Caco-2 cells (P<0.05). Different concentrations of MIMP had no significant influence on the cycle and apoptosis of Caco-2 cells, either (P>0.05). Conclusion MIMP can antagonize the effect of LPS on cell proliferation without obvious impact on the cell cycle, proliferation of Caco-2 cells. Therefore, MIMP may have a high safety in the clinical treatment of IBD.