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Updated: Mar 18, 2026
Author Spotlight: Developing a Rat Model for Weight-Bearing Intervention to Investigate Osteonecrosis of the Femoral Head
Published on: September 27, 2024
Differential expression of miR-672-5p and miR-146a-5p in osteoblasts in rats after steroid intervention
Pengfei Li1, Nan Sun1, Jianchun Zeng2
1Guangzhou University of Chinese Medicine, Guangzhou, China.
Background:
Apoptosis of osteoblasts and osteocytes is one cause of steroid-induced osteonecrosis of the femoral head; however, the molecular mechanism of steroid affecting osteoblasts at the genetic level is unclear. The aim of the present work is to examine differential expression of osteoblasts in rats after steroid intervention and to verify expression by real-time polymerase chain reaction (RT-PCR).
Methods:
Primary culture, passaging and identification of osteoblasts of SD neonatal rats were conducted; osteoblasts were divided into two groups, the control group, and the steroid group. Total RNA was extracted separately, and quality control was performed; by means of RNA labeling and microarray hybridization, data were collected and then standardized to ascertain differences in miRNA expression between the two groups. The gene expression spectrum was analyzed. Obvious differential expression of miR-672-5p and miR-146a-5p was verified by RT-PCR. Miranda, microcosm and mirdb bioinformatics software were used to predict target genes.
Results:
Compared with the control group, morphologically, the osteoblasts in the steroid group were more irregular and showed various shapes. The number of miRNAs (fold change >2) in the steroid group was six. Four miRNAs were upregulated and two miRNAs were downregulated. In particular, upregulated miR-672-5p expression and downregulated miR-146a-5p expression were significant. RT-PCR results showed that the 2(-△△) CT value of miR-672-5p in the steroid group was 3.743-fold of that in the control group, and the 2(-△△) CT value of miR-146a-5p in the steroid group was 0.322-fold of that in the control group. Angptl4, Ccdc51, Ssbp3 and RGD1306991 were predicted as the target gene of miR-672-5p, while Hrp12 was that of miR-146a-5p.
Conclusion:
Expression profiles of miR-672-5p and miR-146a-5p had the most significant changes in the osteoblasts of rats with steroid intervention, which may provide a new viewpoint to pathogenesis of osteonecrosis of the femoral head.
Insights
Steroid intervention significantly altered microRNA expression in rat osteoblasts, with notable changes in miR-672-5p and miR-146a-5p. These findings offer new insights into the genetic mechanisms underlying steroid-induced osteonecrosis of the femoral head.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Steroid-induced osteonecrosis of the femoral head involves osteoblast apoptosis.
- The precise genetic mechanisms by which steroids affect osteoblasts remain largely unknown.
- Understanding these mechanisms is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate differential gene expression in rat osteoblasts following steroid intervention.
- To identify specific microRNAs (miRNAs) involved in the steroid-induced changes.
- To validate expression changes using real-time polymerase chain reaction (RT-PCR).
Main Methods:
- Primary osteoblast cultures from neonatal rats were established and divided into control and steroid-treated groups.
- Microarray analysis was employed to identify differentially expressed miRNAs.
- RT-PCR was used to confirm the expression levels of selected miRNAs (miR-672-5p and miR-146a-5p).
- Bioinformatics tools predicted target genes for the identified miRNAs.
Main Results:
- Steroid treatment led to morphological changes in osteoblasts.
- Six miRNAs exhibited differential expression (fold change >2), with four upregulated and two downregulated.
- miR-672-5p showed a significant upregulation (3.743-fold), while miR-146a-5p was significantly downregulated (0.322-fold).
- Potential target genes for miR-672-5p and miR-146a-5p were predicted.
Conclusions:
- Expression profiles of miR-672-5p and miR-146a-5p were significantly altered in osteoblasts after steroid intervention.
- These miRNA changes may represent a key factor in the pathogenesis of steroid-induced osteonecrosis of the femoral head.
- This study provides a novel perspective on the genetic underpinnings of this condition.
