植物乳杆菌KLDS 1.0318产酸、耐酸、耐胆盐能力及其免疫特性研究

Study on the Acid Producing Ability,Acid and Bile Salt Tolerance of Lactobacillus plantarum KLDS 1.0318 and Its Immunologic Properties

目的:通过体外实验,研究植物乳杆菌KLDS 1.0318的生长曲线、产酸能力、耐酸耐胆盐性能,在此基础上对其可能的免疫调节活性进行研究。方法:采用平板计数法和紫外-可见分光光度法测定菌株生长曲线,pH计测定其产酸能力,平板计数法测定该菌株在酸性及高胆盐环境作用后的活菌数及存活率。采用CCK-8法观察不同剂量的植物乳杆菌KLDS 1.0318对小鼠脾淋巴细胞增殖能力以及对小鼠腹腔巨噬细胞能量代谢水平的影响,采用巨噬细胞吞噬中性红法评估不同剂量的植物乳杆菌KLDS 1.0318对巨噬细胞吞噬作用的影响。结果:植物乳杆菌KLDS 1.0318在4 h后进入对数期,12 h后达到稳定期。其产酸能力较强,pH在12 h后降至4.16。菌株在pH为2.5、3.5条件下培养至3 h时,其存活率分别为91.20%、97.50%,在胆盐质量浓度分别为0.3、0.5 g/100 mL的培养基中作用4 h后,其存活率分别为95.40%、87.76%。一定剂量的植物乳杆菌KLDS 1.0318能够促进脾淋巴细胞和巨噬细胞活性,并呈现剂量依赖关系。结论:植物乳杆菌KLDS 1.0318生长较快,具有较强的产酸、耐酸、耐胆盐能力,并具有潜在免疫调节功能。

Objective ： The growth curve, acid production rate, acid and bile salt resistance of LactobaciUus plantarum KLDS 1.0318 and its possible immunomodulation function in vitro were studied. Methods： The growth curve of the strain was determined by plate count method and UV visible spectrophotometry, and its acid producing capacity was determined by pH meter, the viabiblity of the strain during exposure to acid and bile salt MRS broth measured by plate count method. CCK-8 method was used to observe the effects of different doses of the strain on the proliferation of mouse spleen lymphocytes and the level of energy metabolism, and its effect on phagocytosis of macrophages were evaluated by the method of macrophage phagoeytosis of neutral red. Results： The strain reached a logarithmic phase after 4 h and a stable period after 12 h. It had a stronger ability of acid production,the pH dropped to 4.16 at 12 h.The survival rates of the strain were 91.20% at pH2.5 and 97.50% at pH3.5 after 3 h of culture, respectively. After being cultured for 4 h at a bile salt concentration of 0.3 and 0.5 g/100 mL,the survival rates of the strain were 95.40% and 87.76% ,respectively.A certain dose of Lactobacillus plantarum KLDS 1.0318 could promote the activity of splenic lymphocytes and macrophages and the promoting effect was dose- dependent.Conclusion：LactobaciUus plantarum KLDS 1.0318 grew fast, had strong acid producing ability, acid resistance, bile salt tolerance, and potential immunomodulatory function.