乳杆菌在Caco-2细胞模型中对肠吸收短链脂肪酸的促进作用  

Enhancing intestinal short-chain fatty acid absorption by Lactobacillus in Caco-2 cell model

作　　者：陈大卫[1] 任晨瑜 范晨雨 瞿恒贤 陈春萌 张臣臣 关成冉 李启明 杨锁华 顾瑞霞[1] CHEN Dawei;REN Chenyu;FAN Chenyu;QU Hengxian;CHEN Chunmeng;ZHANG Chenchen;GUAN Chengran;LI Qiming;YANG Suohua;GU Ruixia(Key Laboratory of Dairy Biotechnology and Safety Control of Jiangsu Province,School of Food Science and Engineering,Yangzhou University,Yangzhou,Jiangsu 225127,China;New Hope Dairy Company Limited,Chengdu,Sichuan 610023,China;Synbiotech Biotechnology(Yangzhou)Company Limited,Yangzhou,Jiangsu 225100,China)

机构地区：[1]扬州大学食品科学与工程学院江苏省乳品生物技术与安全控制重点实验室,江苏扬州225127 [2]新希望乳业股份有限公司,四川成都610023 [3]生合生物科技(扬州)有限公司,江苏扬州225100

出　　处：《微生物学通报》2021年第5期1593-1603,共11页Microbiology China

基　　金：国家自然科学基金(31701627,31972094);江苏省高等学校自然科学研究项目(19KJA140004,17KJB550009);国家重点研发计划(2019YFF0217602);成都市重大科技应用示范项目(2019-YF09-00055-SN);扬州市社会发展项目(YZ2019069)。

摘　　要：【背景】短链脂肪酸(Short-Chain Fatty Acids,SCFAs)具有提供能量、调节营养物质代谢、抑制内源性胆固醇合成等广泛的生理活性和生物学效应。【目的】利用建立的Caco-2细胞吸收SCFAs模型研究乳杆菌对肠吸收SCFAs的影响。【方法】通过跨膜电阻值(Transepithelial Electrical Resistance,TEER)、细胞超微结构、苯酚红通透量及细胞增殖-毒性试验等来综合评价Caco-2细胞吸收SCFAs模型的完整性和稳定性,并利用气相色谱-质谱联用仪测定乳杆菌干预前后模型中Caco-2细胞内丙酸和丁酸的含量。【结果】Caco-2细胞单层培养至第11天时的TEER值为1290.73Ω·cm2,在第15天时为1319.31Ω·cm2,而且细胞间连接紧密并覆盖着一层垂直于细胞表面的微绒毛,苯酚红通透量小于1×10^(−6)cm/s,在1 mmol/L丙酸或丁酸分别作用3 h后,模型中Caco-2细胞的存活率较高,分别为99.03%和91.42%。植物乳杆菌(Lactobacillus plantarum)P54、P58、P67、P97、P123及P198干预后,模型中Caco-2细胞内丙酸含量均显著高于未接菌(P<0.05);而发酵乳杆菌(Lactobacillus fermentum)F146和植物乳杆菌(Lactobacillus plantarum)P1干预后,胞内丁酸含量均显著高于未接菌(P<0.05)。【结论】试验的乳杆菌在模型中能够促进Caco-2细胞对丙酸或丁酸的吸收。[Background] Short-chain fatty acids(SCFAs) have a wide range of physiological activities and biological effects, such as providing energy, regulating nutrient metabolism, inhibiting endogenous cholesterol synthesis, and so on. [Objective] The model of Caco-2 cell absorption SCFAs was established, which was used to research the effect of Lactobacillus on intestinal uptake of SCFAs. [Methods] The integrity and stability of Caco-2 cell model was evaluated by transepithelial electrical resistance(TEER), cell ultrastructure, apparent permeability coefficients of phenol red(Papp) and cell proliferation toxicity test. The content of propionic acid and butyric acid in the model was determined by gas chromatography mass spectrometry(GC-MS) before and after inoculating with Lactobacillus. [Results] The TEER of Caco-2 cell monolayer was 1 290.73 Ω·cm2 on the 11 th day, 1 319.31 Ω·cm2 on the 15 th day, and the Papp was less than 1×10-6 cm/s, the cells were closely connected and covered with a layer of microvilli perpendicular to the cell surface. The survival rates of Caco-2 cells were 99.03% and 91.42% after treating with 1 mmol/L propionic acid and 1 mmol/L butyric acid for 3 hours respectively. The propionic acid content of Caco-2 cells in the model inoculated with Lactobacillus plantarum P54, P58, P67, P97, P123 and P198 was higher than non-inoculate significantly(P<0.05), and the butyric acid content of Caco-2 cells in the model inoculated with Lactobacillus fermentum F146 and Lactobacillus plantarum P1 was higher than non-inoculate significantly(P<0.05). [Conclusion] The Lactobacillus strains used in this study can promote the absorption of propionic acid or butyric acid by Caco-2 cells in the model.

关 键 词：乳杆菌 CACO-2 细胞 促吸收 短链脂肪酸 

分 类 号：TS201.3[轻工技术与工程—食品科学]