LncRNA MALAT1对成骨细胞分化的作用机制研究

doi:10.3760/cma.j.cn441417-20241211-19013

摘要/Abstract

摘要：

目的　探讨LncRNA MALAT1对成骨细胞分化的作用机制。方法　本实验于2024年1月至6月在延安大学医学院生物实验中心开展，选取小鼠颅骨源性成骨前细胞MC3T3-E1诱导成骨细胞分化。将细胞分为对照组（未处理细胞）、LPS组（1 mg/L LPS处理细胞）和LPS+siMALAT1组（siMALAT1转染后LPS处理细胞）。采用MTT法测定MC3T3-E1细胞活力。通过测定线粒体三磷酸腺苷（ATP）水平、膜电位（MMP）和超氧化物，评估siMALAT1对MC3T3-E1细胞线粒体功能的影响。采用酶联免疫吸附试验（ELISA）和实时荧光定量PCR（RT-qPCR）检测成骨细胞分化相关标志物表达水平，包括碱性磷酸酶（ALP）、骨钙素（OCN）、骨桥蛋白（OPN）和RUNT相关转录因子2（RUNX2）。采用Western blot测定沉默信息调节因子-1（Sirt1）和过氧化物酶体增殖物激活受体γ辅激活物-1α（PCG-1α）表达水平。采用LSD检验和单因素方差分析进行统计学分析。结果　LPS可呈剂量依赖性降低MC3T3-E1细胞线粒体ATP水平及MMP，促进超氧化物产生（均P<0.05）。siMALAT1可改善LPS处理的MC3T3-E1细胞功能障碍，包括MC3T-E1细胞活力、线粒体ATP水平、MMP以及超氧化物的产生（均P<0.05）。siMALAT1可改善LPS对MC3T3-E1细胞成骨分化（ALP、OCN、OPN、RUNX2）的抑制作用（均P<0.05）。siMALAT1可提高LPS处理的MC3T3-E1细胞Sirt1和PCG-1α表达水平（0.62±0.06比0.30±0.03、0.75±0.07比0.42±0.04）（均P<0.05）。结论　siMALAT1可缓解LPS抑制的MC3T3-E1细胞成骨分化，减轻LPS诱导的MC3T3-E1细胞损伤，尤其是线粒体功能障碍。

关键词:

LncRNA MALAT1, 骨质疏松, 成骨细胞, 线粒体功能障碍

Abstract:

Objective　To explore the mechanism by which LncRNA MALAT1 affects the differentiation of osteoblast. Methods　This experiment was conducted at the Biological Experiment Center of Yan'an University Medical College from January to June 2024.The osteogenic precursor cells derived from mouse skull, MC3T3-E1, were induced to differentiate into osteoblasts. The cells were divided into the control group (untreated cells), the LPS group (1 mg/L LPS treatment of cells), and the LPS+siMALAT1 group (cells treated with LPS after siMALAT1 transfection). The viability of MC3T3-E1 cells was determined using the MTT assay. The effects of siMALAT1 on the mitochondrial function of MC3T3-E1 cells were evaluated by measuring the mitochondrial adenosine triphosphate (ATP) level, membrane potential (MMP), and superoxide. The expression levels of osteoblast differentiation-related markers, including alkaline phosphatase (ALP), osteocalcin (OCN), osteopontin (OPN), and RUNT-related transcription factor 2 (RUNX2), were detected by enzyme-linked immunosorbent assay (ELISA) and real-time fluorescence quantitative PCR (RT-qPCR). The expression levels of silent information regulator-1 (Sirt1) and peroxisome proliferator-activated receptor gamma coactivator-1α (PCG-1α) were determined by Western blot. LSD test and one-way analysis of variance were used for statistical analysis. Results　LPS can dose-dependently reduce the mitochondrial ATP level and MMP in MC3T3-E1 cells, and promote the production of superoxide (all P<0.05). siMALAT1 can improve the functional disorders of LPS-treated MC3T3-E1 cells, including the viability of MC3T3-E1 cells, mitochondrial ATP levels, MMP and the production of superoxide(all P<0.05). siMALAT1 can improve the inhibitory effect of LPS on the osteogenic differentiation of MC3T3-E1 cells (ALP, OCN, OPN, and RUNX2) (all P<0.05). siMALAT1 can increase the expression levels of Sirt1 and PCG-1α in LPS-treated MC3T3-E1 cells (0.62±0.06 vs. 0.30±0.03, 0.75±0.07 vs. 0.42±0.04) (both P<0.05). Conclusion　siMALAT1 can alleviate the osteogenic differentiation inhibition of LPS-suppressed MC3T3-E1 cells, and reduce the damage caused by LPS to MC3T3-E1 cells, especially the mitochondrial dysfunction.

Key words:

LncRNA MALAT1, Osteoporosis, Osteoblast, Mitochondrial dysfunction

张亮亮冯阳阳赵程锦周煜虎李楠楠曹强.

LncRNA MALAT1对成骨细胞分化的作用机制研究 [J]. 国际医药卫生导报, 2025, 31(19): 3231-3235.

Zhang Liangliang, Feng Yangyang, Zhao Chengjin, Zhou Yuhu, Li Nannan, Cao Qiang.

The study on the mechanism of LncRNA MALAT1 on osteoblast differentiation [J]. International Medicine and Health Guidance News, 2025, 31(19): 3231-3235.

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