发酵大麦分离蛋白对3T3-L1细胞棕色化的影响

Effect of Lactobacillus Plantarum Dy-1 Fermented Barley Protein on the Browning of 3T3-L1 Preadipocytes

摘要:

采用实验室自主分离的植物乳杆菌(Lactobacillus plantarumm dy-1)发酵大麦，研究其提取物分离蛋白（LFBE-P）对3T3-L1前脂肪细胞诱导分化为米色脂肪细胞的作用。采用盐析、G25凝胶柱层析和超滤等方法对发酵液中的蛋白质进行分离纯化和浓缩，再用适当浓度的LFBE-P对3T3-L1进行诱导试验，检测细胞葡萄糖消耗量和相关mRNA的相对含量。结果表明，与未发酵大麦分离蛋白（RBE-P）相比，LFBE-P可显著提高3T3-L1细胞葡萄糖消耗量和棕色化脂肪细胞特征基因的表达。对LFBE-P进一步进行纯化的研究结果显示，相对分子质量为3~20 ku的蛋白质是诱导细胞棕色化反应的主要成分，该范围的蛋白质可显著提高3T3-L1细胞中UCP1和PGC-1α等与细胞降脂产热直接关联基因的表达。综上，LFBE-P能够诱导3T3-L1细胞发生棕色化反应，促使白色脂肪向米色脂肪转变。

Abstract:

Lactobacillus plantarum dy-1 isolated from pickles was used for barley fermentation to obtain Lactobacillus-fermented barley extracted protein (LFBE-P), and the effect of LFBE-P on the browning of 3T3-L1 preadipocytes was investigated. Using salting out, gel filtration and ultrafiltration, the protein were purified and condensed. Glucose consumption and relative RNA expression of 3T3-L1 adipocytes were detected to support the browning effect of appropriate concentration of LFBE-P. Results showed that, compared with unfermented barley extracted protein(RBE-P), LFBE-P increased the glucose consumption and expression of the brown adipose tissue-specific genes including. Furthermore, LFBE-P was divided into different parts according to the molecular weight. It was found that LFBE-Ps with molecular weights between 3 ku and 20 ku were the key components inducing 3T3-L1 browning, which could improve the relative expression of uncoupling protein 1, peroxisome proliferator-activated receptor gamma coactivator 1α and PR domain-containing 16. These genes were essential for cells to generate heat and reduce lipid. In conclusion, LFBE-P significantly increased the relative expression of brown-specific genes and glucose consumption of 3T3-L1 adipocytes, inducing 3T3-L1 adipocytes into beige adipocytes.