Open AccessLycium barbarum polysaccharide (LBP) inhibits palmitic acid (PA)-induced MC3T3-E1 cell apoptosis by regulating miR-200b-3p/Chrdl1/PPARγ
Author(s) -
Jing Lei,
Baiwen Hu,
Song Qu
Publication year - 2020
Publication title -
food and nutrition research/food and nutrition research. supplement
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.041
H-Index - 37
eISSN - 1654-6628
pISSN - 1654-661X
DOI - 10.29219/fnr.v64.4208
Subject(s) - viability assay , apoptosis , downregulation and upregulation , peroxisome proliferator activated receptor , flow cytometry , cell growth , microrna , cell , microbiology and biotechnology , luciferase , chemistry , transfection , biology , receptor , gene , biochemistry
Background Obesity is closely related to osteoporosis. Lycium barbarum polysaccharides (LBPs) have anti-osteoporosis activity. Objective This study aimed to explore the role of LBPs in palmitic acid (PA)-induced osteoblast apoptosis. Methods The microarray data set GSE37676 was downloaded from Gene Expression Ominibus (GEO) database. Top 300 differentially expressed genes (DEGs) were used to construct a protein–protein interaction (PPI) network based on STRING database, and significant modules were analyzed and their key genes were screened by using Cytoscape software. COEXPEDIA database showed that there was co-expression between Chrdl1 and peroxisome proliferator-activated receptor (PPARγ). MC3T3-E1 cells were treated with 100–500 μg/mL of PA. Reverse transcription polymerase chain reaction (RT-PCR) and western blot assays were used to detect mRNA and protein levels. Cell Counting Kit-8 (CCK-8) assay and flow cytometry were used to detect cell viability and cell apoptosis. Results Chrdl1 was the key gene from the most significant module and downregulation in MC3T3-E1 cells treated with PA. MicroRNA miR-200b-3p and PPARγ were significantly upregulated among PA-treated MC3T3-E1 cells. The results of luciferase reporter gene assay showed that miR-200b-3p targeted Chrdl1 3’-UTR . Over-expressing miR-200b-3p promoted PA-induced cell apoptosis and inhibited cell viability. After pre-treating cells with PA and LBP, MC3T3-E1 cell apoptosis rate was relatively lower than that of mimics+PA 200 group. Chrdl1 inhibition partly reversed miR-200b-3p effect on inhibiting apoptosis among MC3T3-E1 cells pre-treated with LBP and PA. Decreased C CASP3, PPARγ and increased Chrdl1 by miR-200b-3p inhibition were partly reversed by Chrdl1 inhibition. Conclusions LBPs inhibit PA-induced MC3T3-E1 cell apoptosis by mainly decreasing miR-200b-3p to upregulate Chrdl1 , but miR-200b-3p/ Chrdl1 /PPARγ is not the only mechanism for LBPs protecting osteoblasts from PA.