Exploration and clinical validation of the repair mode of the sclerotic zone of steroid-induced osteonecrosis of the femoral head based on Tandem Mass Tags technology

doi:10.12307/2024.301

Abstract

Abstract: BACKGROUND: The sclerotic zone in the femoral head is an important imaging feature in the progression of steroid-induced femoral head necrosis, which is associated with disease prognosis. Peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) has been shown to possess biological activities such as osteogenesis, angiogenesis and anti-mitochondrial apoptosis, which may be closely related to bone repair of steroid-induced femoral head necrosis.
OBJECTIVE: To screen for the differential proteins in the sclerotic zone of steroid-induced osteonecrosis of the femoral head versus the normal zone, to screen for hub proteins in the sclerotic zone, and to verify the differential expression of hub proteins in the femoral head specimens following steroid-induced femoral head necrosis, and to to explore the repair pattern of the sclerotic zone following steroid-induced femoral head necrosis.
METHODS: Femoral head samples were collected from patients with steroid-induced osteonecrosis of the femoral head receiving total hip arthroplasty. The differentially expressed genes in the sclerotic zone and the normal zone were screened by Tandem Mass Tags and analyzed by GO and KEGG signaling pathways to construct a protein-protein interaction network and screen hub genes. In addition, the expression of hub genes in the sclerotic zone was verified by immunohistochemistry and western blot.
RESULTS AND CONCLUSION: Quantitative protein profiling by Tandem Mass Tags revealed that 609 proteins were significantly differentially expressed (Log2FC > 1.20, Log2FC < 0.84 and P < 0.05) in the sclerotic zone of the femoral head compared with the normal zone, of which 290 proteins were upregulated and 319 proteins were downregulated. The GO and KEGG pathway enrichment analyses revealed that among the top 10 enriched pathways, Wnt signaling pathway and life-cycle regulatory pathway were closely related to bone repair; in the life-cycle regulatory pathway, PGC-1α was one of the important proteins. In addition, western blot results verified the low expression of PGC-1α and NRF1 in the sclerotic zone and high expression of Cleaved Caspase-3 in the sclerotic zone compared with the normal zone of steroid-induced femoral head necrosis specimens. Light microscopic immunohistochemical results showed the distribution of PGC-1α, NRF1 and Cleaved Caspase-3 positive expression in the sclerotic and normal zones in the femoral head tissue specimens, indicating the presence of their expression in bone trabeculae, osteoblasts and bone marrow. In contrast, the brown area of the sclerotic zone of femoral head necrosis stained darker and showed more obvious expression of Cleaved Caspase-3. To conclude, in the sclerotic zone of steroid-induced femoral head necrosis, biological behaviors including activation of osteogenesis-related pathways such as Wnt and oxidative apoptosis characterized by low expression of PGC-1 are observed. Low expression of PGC-1α in the sclerotic zone of steroid-induced femoral head necrosis may be associated with the activation of oxidative apoptosis.

Key words: Tandem Mass Tags, steroid-induced osteonecrosis of the femoral head, peroxisome proliferative activated receptor γ coactivator 1α, mitochondrial oxidative apoptosis, bone repair

CLC Number:

Zhuang Zhikun, He Mincong, Lin Tianye, Wu Rongkai, Guo Jinhua, Wu Zhaoke, Wei Qiushi. Exploration and clinical validation of the repair mode of the sclerotic zone of steroid-induced osteonecrosis of the femoral head based on Tandem Mass Tags technology[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(14): 2191-2196.

Figures/Tables 9

References

[1] MONT MA, SALEM HS, PIUZZI NS, et al. Nontraumatic Osteonecrosis of the Femoral Head: Where Do We Stand Today?: A 5-Year Update. J Bone Joint Surg Am. 2020;102(12):1084-1099.
[2] LIN T, CHEN W, YANG P, et al. Bioinformatics analysis and identification of genes and molecular pathways in steroid-induced osteonecrosis of the femoral head. J Orthop Surg Res. 2021;16(1):327.
[3] 魏秋实，何伟，张庆文，等. 岭南袁氏中医药防治股骨头坏死传承文化研究[J]. 新中医,2022,54(23):216-220.
[4] 何伟，刘予豪，周驰，等. 非手术保髋治疗非创伤性股骨头坏死的临床研究[J]. 中国中西医结合杂志,2020,40(2):176-181.
[5] CHEN WH, GUO WX, LI JX, et al. Application of protective weight-bearing in osteonecrosis of the femoral head: A systematic review and meta-analysis of randomized controlled trials and observational studies. Front Surg. 2022;9: 1000073.
[6] CHEN W, LI J, GUO W, et al. Outcomes of surgical hip dislocation combined with bone graft for adolescents and younger adults with osteonecrosis of the femoral head: a case series and literature review. BMC Musculoskelet Disord. 2022;23(1):499.
[7] 魏秋实,何晓铭,何伟,等. 非手术保髋治疗ARCO Ⅱ期股骨头坏死的临床疗效及影响因素分析[J].中华骨与关节外科杂志,2022,15(6):424-430.
[8] 石少辉. 股骨头坏死硬化带的观察[D]. 北京:中国协和医科大学,2009.
[9] 陈玥，刘敏，铁位有. 基于CT影像的形态学对双侧股骨头坏死塌陷的危险因素分析[J].中国CT和MRI杂志,2020,18(7):158-161.
[10] 牟永莹,顾培明,马博,等.基于质谱的定量蛋白质组学技术发展现状[J].生物技术通报,2017,33(9):73-84.
[11] PAGEL O, KOLLIPARA L, SICKMANN A. Quantitative Proteome Data Analysis of Tandem Mass Tags Labeled Samples. Methods Mol Biol. 2021;2228:409-417.
[12] PAPPIREDDI N, MARTIN L, WÜHR M. A Review on Quantitative Multiplexed Proteomics. Chembiochem. 2019;20(10):1210-1224.
[13] YOON BH, MONT MA, KOO KH, et al. The 2019 Revised Version of Association Research Circulation Osseous Staging System of Osteonecrosis of the Femoral Head. J Arthroplasty. 2020;35(4):933-940.
[14] 王慧婷,张岩晨,徐梦怡,等. 能量代谢关键调控因子PGC-1α的研究进展[J].生理学报,2020,72(6):804-816.
[15] 魏秋实,方斌,陈镇秋,等. 股骨头前外侧骨质状态在股骨头坏死塌陷进展中的作用[J]. 中国组织工程研究,2019,23(16):2516-2522.
[16] TINGART M, BECKMANN J, OPOLKA A, et al. Influence of factors regulating bone formation and remodeling on bone quality in osteonecrosis of the femoral head. Calcif Tissue Int. 2008;82(4):300-308.
[17] 于潼，谢利民. 硬化带在股骨头坏死塌陷作用中的研究进展[J]. 医学综述, 2015,21(1):84-86.
[18] 曾平，陈金龙，李金溢，等. 创伤性股骨头坏死血清差异蛋白质组学的研究[J]. 中国矫形外科杂志,2019,27(5):453-458.
[19] 胡兆林,常峰. 激素性股骨头坏死发病机制及相关信号通路研究进展[J].医学综述,2022,28(3):466-470.
[20] CHEN XJ, SHEN YS, HE MC, et al. Polydatin promotes the osteogenic differentiation of human bone mesenchymal stem cells by activating the BMP2-Wnt/β-catenin signaling pathway. Biomed Pharmacother. 2019;112:108746.
[21] CHENG CF, KU HC, LIN H. PGC-1α as a Pivotal Factor in Lipid and Metabolic Regulation. Int J Mol Sci. 2018;19(11):3447.
[22] HALLING JF, PILEGAARD H. PGC-1α-mediated regulation of mitochondrial function and physiological implications. Appl Physiol Nutr Metab. 2020;45(9):927-936.
[23] KONG S, CAI B, NIE Q. PGC-1α affects skeletal muscle and adipose tissue development by regulating mitochondrial biogenesis. Mol Genet Genomics. 2022;297(3):621-633.
[24] 陈黎,张世昌,张国英. 基因过氧化物酶体增殖物激活受体γ共激因子-1α/雌激素相关受体-α共修饰间充质干细胞对血管化的影响[J].中华实验外科杂志,2018,35(7):1203-1205.
[25] 闫春雷,黄欣,苏乐群. PGC-1α的转录调节和翻译后修饰[J].中国生物化学与分子生物学报,2015,31(1):12-19.
[26] XIA SR, WEN XY, FAN XL, et al. Wnt2 overexpression protects against PINK1 mutant‑induced mitochondrial dysfunction and oxidative stress. Mol Med Rep. 2020;21(6):2633-2641.
[27] 耿慧霞.李莺歌.石贞玉.等. PGC-1α过表达逆转OGD/R诱导的神经元线粒体功能降低和凋亡[J]. 中国病理生理杂志,2017,33(11):2078-2083.
[28] WALDMAN M, BELLNER L, VANELLA L, et al. Epoxyeicosatrienoic Acids Regulate Adipocyte Differentiation of Mouse 3T3 Cells, Via PGC-1α Activation, Which Is Required for HO-1 Expression and Increased Mitochondrial Function. Stem Cells Dev. 2016;25(14):1084-1094.
[29] HE Q, WANG T, NI H, et al. Endoplasmic reticulum stress promoting caspase signaling pathway-dependent apoptosis contributes to bone cancer pain in the spinal dorsal horn. Mol Pain. 2019;15:1744806919876150.
[30] 温宇华,刘培培,宋利格. PGC-1α在骨代谢中的作用[J].同济大学学报(医学版),2021,42(2):266-270.