Lei Jing1, Baiwen Hu1, Qing Hua Song2. 1. Orthopedics Department, Ningbo First Hospital, Ningbo City, Zhejiang Province, China. 2. Plastic Surgery Center and Trauma Center, Ningbo First Hospital, Ningbo City, China.
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.
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.
Authors: Chanèle Cyr-Depauw; Jason J Northey; Sébastien Tabariès; Matthew G Annis; Zhifeng Dong; Sean Cory; Michael Hallett; Jonathan P Rennhack; Eran R Andrechek; Peter M Siegel Journal: Mol Cell Biol Date: 2016-05-02 Impact factor: 4.272
Authors: Ann-Kristin Picke; Graeme M Campbell; Matthias Blüher; Ute Krügel; Felix N Schmidt; Elena Tsourdi; Maria Winzer; Martina Rauner; Vladimir Vukicevic; Björn Busse; Juliane Salbach-Hirsch; Jan P Tuckermann; Jan C Simon; Ulf Anderegg; Lorenz C Hofbauer; Anja Saalbach Journal: Sci Transl Med Date: 2018-08-08 Impact factor: 17.956