Jing Lei, Hu Baiwen, Song Qing Hua
Orthopedics Department, Ningbo First Hospital, Ningbo City, Zhejiang Province, China.
Plastic Surgery Center and Trauma Center, Ningbo First Hospital, Ningbo City, China.
Food Nutr Res. 2020 Dec 22;64. doi: 10.29219/fnr.v64.4208. eCollection 2020.
Obesity is closely related to osteoporosis. Lycium barbarum polysaccharides (LBPs) have anti-osteoporosis activity.
This study aimed to explore the role of LBPs in palmitic acid (PA)-induced osteoblast apoptosis.
The microarray data set 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 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.
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 . 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 group. 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 by miR-200b-3p inhibition were partly reversed by inhibition.
LBPs inhibit PA-induced MC3T3-E1 cell apoptosis by mainly decreasing miR-200b-3p to upregulate , but miR-200b-3p//PPARγ is not the only mechanism for LBPs protecting osteoblasts from PA.
肥胖与骨质疏松密切相关。枸杞多糖（LBPs）具有抗骨质疏松活性。
本研究旨在探讨枸杞多糖在棕榈酸（PA）诱导的成骨细胞凋亡中的作用。
从基因表达综合数据库（GEO）下载芯片数据集。基于STRING数据库，利用前300个差异表达基因（DEGs）构建蛋白质-蛋白质相互作用（PPI）网络，使用Cytoscape软件分析显著模块并筛选关键基因。COEXPEDIA数据库显示某基因与过氧化物酶体增殖物激活受体（PPARγ）之间存在共表达。用100 - 500μg/mL的棕榈酸处理MC3T3 - E1细胞。采用逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法检测mRNA和蛋白质水平。使用细胞计数试剂盒-8（CCK-8）法和流式细胞术检测细胞活力和细胞凋亡。
某基因是最显著模块中的关键基因，在用棕榈酸处理的MC3T3 - E1细胞中表达下调。在经棕榈酸处理的MC3T3 - E1细胞中，微小RNA miR-200b - 3p和PPARγ显著上调。荧光素酶报告基因检测结果表明miR-200b - 3p靶向某基因。过表达miR-200b - 3p可促进棕榈酸诱导的细胞凋亡并抑制细胞活力。在用棕榈酸和枸杞多糖预处理细胞后，MC3T3 - E1细胞凋亡率相对低于模拟物+棕榈酸组。某基因抑制部分逆转了miR-200b - 3p对经枸杞多糖和棕榈酸预处理的MC3T3 - E1细胞凋亡抑制的影响。miR-200b - 3p抑制导致的半胱天冬酶3（CASP3）、PPARγ降低和某基因增加，部分被某基因抑制所逆转。
枸杞多糖主要通过降低miR-200b - 3p来上调某基因，从而抑制棕榈酸诱导的MC3T3 - E1细胞凋亡，但miR-200b - 3p//PPARγ不是枸杞多糖保护成骨细胞免受棕榈酸损伤的唯一机制。
