Lycium barbarum polysaccharide (LBP) inhibits palmitic acid (PA)-induced MC3T3-E1 cell apoptosis by regulating miR-200b-3p/Chrdl1/PPARγ

Abstract

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+PA200 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.