|
|
|
| The Role of Knockdown of CircCDR1as in Glucocorticoid-Induced Bone Microvascular Endothelial Cell Activity and Angiogenesis |
| Aikeremujiang Aerken, Rixiati Paerhati, ZHANG Zheng |
| The Fifth Affiliated Hospital of Xinjiang Medical University, Xinjiang Urumqi 830011, China |
|
|
|
|
Abstract Objective: To investigate the effect of knockdown of Circular RNAantisense to the cerebellar degeneration related protein 1 transcript (circCDR1as) on glucocorticoid-induced activity of human femoral head bone microvascular endothelial cells (BMECs) and angiogenesis. Methods: Femoral head tissue was collected from patients with glucocorticoid-induced osteonecrosis of the femoral head (ONFH), Histopathological features were observed by HE staining, the expression changes of CircCDR1as were detected by real-time quantitative polymerase chain reaction (qRT-PCR); the femoral heads of patients with total hip replacement were obtained, BMECs were isolated and cultured, and BMECs were identified by flow cytometry to detect the expression of cell surface markers CD31, CD34, CD45, CD54, CD144 and CD117, immunofluorescence staining was used to detect the expression of CD31 and vWF; the experimental groups included the control group, the GC group, the shNC+GC group and the shCDR1as+GC group, the glucocorticoid hydrocortisone-treated cells and the shCDR1as and shNC-transfected cells were treated accordingly, qRT-PCR to detect changes in the expression of CircCDR1as, the level of cell proliferation was detected by MTT method and EdU staining, in vitro tube formation experiments were performed to observe the formation of cell lumen, Western blot was used to determine the protein expressions of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR-2). Results: Bone marrow necrosis and empty bone cavity appeared in the femoral head tissue of ONFH patients, and the relative expression of CircCDR1as was significantly higher than that of the control group (P<0. 05). It was observed that the isolated cells grew in good condition, with positive expression of CD31, CD54 and CD144, negative expression of CD34, CD45 and CD117, and obvious immunofluorescence expression of CD31 and vWF in the cells, indicating that BMECs were successfully isolated. Compared with the control group, the relative expression of CircCDR1as in the GC group was up-regulated, the cell proliferation activity was decreased, the EdUpositive cell rate was decreased, the number of lumens formed by the cells was decreased, and the relative expressions of VEGF and VEGFR-2 proteins were down-regulated, the differences were statistically significant (P<0. 05); compared with the shNC+GC group, the relative expression of CircCDR1as in the shCDR1as+GC group was down-regulated, the cell proliferation activity and EdUpositive cell rate were increased, and the number of lumens formed by cells increased, at the same time, the relative expression of VEGF and VEGFR-2 proteins was up-regulated, the differences were all statistically significant (P<0. 05). Conclusion: CircCDR1as is highly expressed in the necrosis of femoral head tissue of patients. Knockdown of CircCDR1as can inhibit the activity of BMECs induced by glucocorticoids and promote angiogenesis in vitro, thereby protecting BMECs.
|
|
|
|
|
|
[1] Guggenbuhl P,Robin F,Cadiou S,et al.Etiology of avascular osteonecrosis of the femoral head[J].Morphologie,2021,105(349):80-84.
[2] Zhao D,Ma Z.Application of biomaterials for the repair and treatment of osteonecrosis of the femoral head[J].Regen Biomater,2020,7(1):1-8.
[3] 孙飞,彭昊.激素性股骨头坏死病理机制中细胞凋亡的研究进展[J].骨科,2022,13(1):86-91.
[4] 左伟,高福强,李配瑶,等.体外震波对骨微血管内皮细胞生理功能的影响[J].中国组织工程研究,2016,20(37):5504-5510.
[5] Guo Z,Cao Q,Zhao Z,et al.Biogenesis features functions and disease relationships of a specific Circular RNA:CDR1as[J].Aging Dis,2020,11(4):1009-1020.
[6] Shimba A,Ikuta K.Control of immunity by glucocorticoids in health and disease[J].Semin Immunopathol,2020,42(6):669-680.
[7] Hardy RS,Raza K,Cooper MS.Therapeutic glucocorticoids:mechanisms of actions in rheumatic diseases[J].Nat Rev Rheumatol,2020,16(3):133-144.
[8] Hachemi Y,Rapp AE,Picke AK,et al.Molecular mechanisms of glucocorticoids on skeleton and bone regeneration after fracture[J].Mol Endocrinol,2018,61(1):R75-R90.
[9] Yu QS,Guo WS,Cheng LM,et al.Glucocorticoids significantly influence the transcriptome of bone microvascular endothelial cells of human femoral head[J].Chin Med (Engl),2015,128(14):1956-1963.
[10] Yue J,Wan F,Zhang Q,et al.Effect of glucocorticoids on miRNA expression spectrum of rat femoral head microcirculation endothelial cells[J].Gene,2018,651:126-133.
[11] Yu H,Liu P,Zuo W,et al.Decreased angiogenic and increased apoptotic activities of bone microvascular endothelial cells in patients with glucocorticoid-induced osteonecrosis of the femoral head[J].BMC Musculoskelet Disord,2020,21(1):277.
[12] Aghajani Mir M,Dinmohammadi H,Moudi E,et al.Clinical values of expression signature of circCDR1AS and circHIAT1 in prostate cancer:Two circRNAs with regulatory function in androgen receptor (AR) and PI3K/AKT signaling pathways[J].Clin Lab Anal,2022,36(2):e24220.
[13] Jiang B,Zhu SH,Zeng JY,et al.Plasma and local expressions of CircRNA CDR1as are linked with disease severity in patients with non-traumatic osteonecrosis of femoral head[J].Orthop Surg Res,2020,15(1):592.
[14] Liu M,Zhao G,Wei BF.Attenuated serum vasoactive intestinal peptide concentrations are correlated with disease severity of non-traumatic osteonecrosis of femoral head[J].Orthop Surg Res,2021,16(1):325.
[15] Wenz A,Tjoeng I,Schneider I,et al.Improved vasculogenesis and bone matrix formation through coculture of endothelial cells and stem cells in tissue-specific methacryloyl gelatin-based hydrogels[J].Biotechnol Bioeng,2018,115(10):2643-2653.
[16] Liao H,Zhong Z,Liu Z,et al.Bone mesenchymal stem cells co-expressing VEGF and BMP-6 genes to combat avascular necrosis of the femoral head[J].Exp Ther Med,2018,15(1):954-962. |
|
|
|
2022, Vol. 28 
