Elucidating the role of the GC/GR/GLUT1 axis in steroid-induced osteonecrosis of the femoral head: A proteomic approach

AUTHORS

Hongbin Luo, Jie Wei, Songye Wu, Qunya Zheng, Xinhua Lin, Peng Chen

ABSTRACT

Background

Steroid-induced osteonecrosis of the femoral head (SONFH) is a prevalent and incapacitating condition that affects the hip joint. Unfortunately, early diagnostic and treatment measures are limited.

Methods

Our study employed Tandem Mass Tag (TMT) labeling mass spectrometry (MS)-based quantitative proteome to compare the proteins of femoral head tissues in patients with SONFH with those of patients who sustained femoral neck fracture (FNF). We investigated the level and effects of glucose transporter member 1 (GLUT1) in SONFH patients and MC3T3-E1 cells and examined the function and molecular mechanism of GLUT1 in the context of SONFH using in vivo and in vitro approaches.

Results

The SONFH group exhibited significant changes in protein expression levels compared to the fracture group. Specifically, we observed the up-regulation of 86 proteins and the down-regulation of 138 proteins in the SONFH group. Among the differentially expressed proteins, GLUT1 was down-regulated and associated with glucose metabolic processes in the SONFH group. Further analysis using Parallel Reaction Monitoring (PRM), WB, and PCR confirmed that the protein was significantly down-regulated in both femoral head tissue samples from SONFH patients and dexamethasone-treated MC3T3-E1 cells. Moreover, overexpression of GLUT1 effectively reduced glucocorticoid (GC)-induced apoptosis and the suppression of osteoblast proliferation and osteogenic differentiation in MC3T3-E1 cells, as well as GC-induced femoral head destruction in GC-induced ONFH rat models. Additionally, our research demonstrated that GC down-regulated GLUT1 transcription via glucocorticoid receptors in MC3T3-E1 cells.

Conclusions

GLUT1 was down-regulated in patients with SONFH; furthermore, down-regulated GLUT1 promoted apoptosis and inhibited osteoblast ossification in dexamethasone-induced MC3T3-E1 cells and contributed to GC-induced femoral head destruction in a SONFH rat model. Glucocorticoids inhibited the transcriptional activity of GLUT1, leading to a reduction in the amount and activity of GLUT1 in the cells and ultimately promoting apoptosis and inhibiting osteoblast ossification via the GC/GR/GLUT1 axis in SONFH.

Differences in the Microarchitectural and Histomorphologic Characteristics Between Glucocorticoid-induced Osteonecrosis of Femoral Head and Alcohol-induced Osteonecrosis of Femoral Head

AUTHORS

Yiwei Chen, Kexin Liu, Yu Miao, Bin Zhu, Feng Xue, Junhui Yin, Minghao Zheng, guangyi li, Changqing Zhang

ABSTRACT

Aims

To analyze microarchitecture and histomorphology characteristics of different regions in femoral heads from patients with glucocorticoid-induced osteonecrosis of femoral head (GIONFH) and alcohol-induced osteonecrosis of femoral head (AIONFH).

Methods

Patients diagnosed with GIONFH and AIONFH were recruited. Femoral heads were obtained after total hip replacement. Micro-CT was applied to evaluate the microstructure of 9 regions of interest (ROIs) in the femoral head. Along the supero-inferior orientation, the femoral head was divided into necrotic region, reactive interface, and normal region; along the medio-lateral orientation, the femoral head was divided into medial region, central region and lateral region. Decalcified and undecalcified bone histology were then performed to assess histopathological alterations and bone remodeling levels.

Results

42 GIONFH patients (50 hips) and 43 AIONFH patients (50 hips) anticipated in the study. In the necrotic region, most of the microarchitectural parameters did not differ significantly between GIONFH and AIONFH, whereas both the reactive interface and normal region illustrated significant differences in the microstructure and histomorphometry. The reactive interface and normal region exhibited a less sclerotic microarchitecture, but a higher bone remodeling level in GIONFH as compared with AIONFH. Despite similar necrotic pathological manifestations, subchondral trabecular microfracture in the necrotic region was more severe and vasculature of the reactive interface was more abundant in GIONFH.

Conclusions

Although these two subtypes of ONFH shared similar microarchitecture and pathological features in the necrotic region, GIONFH exhibited a less sclerotic microarchitecture and a more active bone metabolic status in both the reactive interface and normal region.