植物乳杆菌KLDS 1.0344体外抗氧化特性及抑制脂肪沉积的研究

Antioxidant properties and inhibition of fat deposition of Lactobacillus plantarum KLDS 1.0344 in vitro

目的:研究了植物乳杆菌(Lactobacillus plantarum)KLDS 1.0344体外抗氧化能力,及其在3T3-L1前体脂肪细胞培养中,所产生抑制脂肪沉积的作用。方法:首先,将植物乳杆菌(Lactobacillus plantarum)KLDS 1.0344与鼠李糖乳杆菌(Lactobacillus rhamnosus)GG商业菌株进行抗氧化能力比较,以DPPH清除率、羟基清除率、脂质抗氧化能力、超氧阴离子清除能力、还原能力5项指标进行检测与评定;其次,探究其能否对3T3-L1细胞成脂过程产生直接影响,通过油红O染色进行观察测定;最后,利用实时荧光定量聚合酶链式反应(q-RT PCR)测定其对细胞内氧化应激通路Keap1-Nrf2、脂肪合成相关通路Wnt-GSK3β-βcatenin中基因表达的变化。结果:KLDS1.0344菌悬液的抗氧化能力最佳;通过油红O染色,与空白对照组相比,KLDS 1.0344组能明显减少3T3-L1脂肪细胞中的脂肪沉积量(p<0.05);同时,植物乳杆菌KLDS1.0344通过调节细胞内氧化应激通路,改善细胞内氧化还原水平;并通过调节脂肪合成通路,对脂肪的沉积起到了抑制作用。结论:获得了一株在体外具有抑制脂肪沉积能力的植物乳杆菌KLDS 1.0344,该菌株可以作为抑制肥胖发生的潜在益生菌制剂。

Objective： The in vitro antioxidative capacity of Lactobacillus plantarum KLDS 1.0344 was studied, and its effect on fat deposition inhibition in 3 T3-L1 precursor adipocyte culture was studied.Methods： First, this experiment compared the antioxidant capacity of the Lactobacillus plantarum KLDS1.0344 against the Lactobacillus rhamnosus GG reference strain, DPPH clearance, hydroxyl clearance,lipid antioxidant capacity, superoxide anion scavenging ability and reducing ability were tested andevaluated. Secondly, to investigate whether it can directly affect the adipogenic process of 3 T3-L1 cells,observe and determine by oil red O staining. Finally, the use of real-time fluorescence quantitative polymerase chain reaction（q-RT PCR） to determine the intracellular oxidation Keap1-Nrf 2, a gene related to fat synthesis pathway, Wnt-GSK3β-βcatenin gene expression changes. Results： The antioxidation capacity of KLDS 1.0344 suspension was the best. By oil red O staining, KLDS 1.0344 group could significantly reduce the fat deposition in 3 T3-L1 adipocytes compared with the blank control group（P〈0.05）.At the same time, Lactobacillus plantarum KLDS 1.0344 improves intracellular redox levels by regulating intracellular oxidative stress pathways, and inhibits lipid deposition by regulating fat synthesis pathways.Conclusion： This experiment obtained a strain of Lactobacillus plantarum KLDS 1.0344 with the ability to inhibit fat deposition in vitro. This strain can be used as a potential probiotic to inhibit obesity.