|
|
|
| Effect of CircRNA YAP1 on Angiogenesis of Femoral Head Microvascular Endothelial Cells |
| RIXIATI Paerhati, ZHAI Sheng, LV Qing |
| The Fifth Affiliated Hospital of Xinjiang Medical University, Xinjiang Urumqi 830000, China |
|
|
|
|
Abstract Objective: To explore the role and mechanism of circular RNA (circRNA) YAP1 in the regulation of microvascular endothelial cell (MVEC) angiogenesis in steroid-induced osteonecrosis of the femoral head (SONFH) rats. Methods: Adult male SD rats were injected with methylprednisolone (MPS, 20 mg/kg/d) for 3 weeks to induce the SONFH model (n=30). The control group (n=30) was injected with an equal amount of saline. Micro-CT scanning and HE staining were performed to evaluate whether the model was successfully established. MVECs were isolated from the femoral heads of the rats using CD31 as a marker and identified by immunofluorescence staining. The MVECs were divided into three groups: the pcDNA-YAP1 group (overexpression of circRNA YAP1), the pcDNA-NC group (negative control), and the pcDNA-YAP1+dickkopf-1 group (treated with Wnt1/β-catenin pathway inhibitor dickkopf-1). The expression of circRNA YAP1 was detected using qPCR, while the expressions of Wnt1, β-catenin, VEGF-A, and vimentin were detected using Western blotting. Cell proliferation was detected using the CCK-8 assay, while cell migration was detected using the Transwell assay. Tube formation assays were performed to detect the angiogenic ability of the cells. Results: The HE staining results showed that the control group was healthy, with no bone necrosis, while the SONFH group showed severe femoral head necrosis and deterioration. The results of micro-CT scanning showed that the control group had dense and regular bone trabeculae, with normal numbers and morphology of bone trabeculae. The SONFH group was accompanied by an empty bone pit, a decrease in the number of bone trabeculae, and shortened bone trabeculae. Compared with the control group (1.00±0.08; 1.00±0.12; 1.00±0.15), the expressions of circRNA YAP1 (0.27±0.04), Wnt1 (0.49±0.03), and β-catenin (0.33±0.03) in the SONFH group were significantly reduced (t=12.158, P=0.009; t=10.596, P=0.012; t=10.080, P=0.014). In addition, MVECs from the femoral heads of the Ctrl and SONFH groups were successfully isolated. Compared with the control group (1.00±0.00; 1.00±0.02; 1.00±0.01), the expressions of circRNA YAP1 (0.15±0.01), Wnt1 (0.28±0.05), and β-catenin (0.31±0.02) in the MVECs from the SONFH group were significantly reduced (t=17.625, P=0.004; t=16.154, P=0.005; t=13.596, P=0.007).In MVECs, the expressions of circRNA YAP1, Wnt1, β-catenin, VEGF-A and vimentin were up-regulated in pcDNA-YAP1 group compared with pcDNA-NC group. The cell proliferation rate, cell migration rate and tubule formation rate were significantly increased (all P<0.05). Compared with pcDNA-YAP1 group, the expression of circRNA YAP1 in pcDNA-YAP1+dickkopf-1 group had no significant change (P>0.05), while the expression of Wnt1, β-catenin, VEGF-A and vimentin were down-regulated. The cell proliferation rate, cell migration rate and tubule formation rate were significantly inhibited (all P<0.05). Conclusion: CircRNA YAP1 promotes the angiogenesis of bone endovascular endothelial cells in SONFH rats by activating Wnt1/β-catenin pathway.
|
|
|
|
|
|
[1] Yao T,Wang L,Ding Z F,et al.hsa_circ_0058122 knockdown prevents steroid-induced osteonecrosis of the femoral head by inhibiting human umbilical vein endothelial cells apoptosis via the miR-7974/IGFBP5 axis[J].Journal of Clinical Laboratory Analysis,2022,36(4):24134.
[2] Mao Z,Liu G,Xiao G Y,et al.CircCDR1as suppresses bone microvascular endothelial cell activity and angiogenesis through targeting miR-135b/FIH-1 axis[J].Orthopaedic surgery,2021,13(2):573-582.
[3] Sung I H,Son H J,Park J S,et al.Extraskeletal osteosarcoma misdiagnosed as heterotopic ossification after periprosthetic femoral fracture:A case report[J].Acta Orthopaedica Traumatologica Turcica,2020,54(1):117.
[4] Huang Y,Xiao D,Huang S,et al.Circular RNA YAP1 attenuates osteoporosis through up-regulation of YAP1 and activation of Wnt/β-catenin pathway[J].Biomedicine & Pharmacotherapy,2020,129:110365.
[5] Lee E,Ko J Y,Kim J,et al.Osteogenesis and angiogenesis are simultaneously enhanced in BMP2-/VEGF-transfected adipose stem cells through activation of the YAP/TAZ signaling pathway[J].Biomaterials science,2019,7(11):4588-4602.
[6] Jia L,Zhang Y,Ji Y,et al.YAP balances the osteogenic and adipogenic differentiation of hPDLSCs in vitro partly through the Wnt/β-catenin signaling pathway[J].Biochemical and Biophysical Research Communications,2019,518(1):154-160.
[7] Hsu HW,Rodriguez-Ortiz CJ,Zumkehr J,et al.Inflammatory cytokine IL-1β downregulates endothelial LRP1 via microRNA-mediated gene silencing[J].Neuroscience,2021,453:69-80.
[8] Wan F,Zhang Q,Wen P,et al.Effect of glucocorticoids on miRNA expression spectrum of rat femoral head microcirculation endothelial cells[J].Gene,2018,651:126-133.
[9] Yan M,Wang H,Gu Y,et al.Melatonin exerts protective effects on diabetic retinopathy via inhibition of Wnt/β-catenin pathway as revealed by quantitative proteomics[J].Experimental Eye Research,2021,205:108521.
[10] Zhong L,Pan Y,Shen J.FBXW7 inhibits invasion,migration and angiogenesis in ovarian cancer cells by suppressing VEGF expression through inactivation of β-catenin signaling[J].Exp Ther Med,2021,21(5):514-522.
[11] Li D,Yang Z,Luo Y,et al.Delivery of miR335-5p-pendant tetrahedron DNA nanostructures using an injectable heparin lithium hydrogel for challenging bone defects in steroid-associated osteonecrosis[J].Adv Health Mater,2022,11(1):2101412. |
|
|
|
2023, Vol. 29 
