Humanin (HN), a mitochondrial derived peptide, plays cyto-protective role under various stress. In this study, we aimed to investigate the effects of HNGF6A, an analogue of HN, on osteoblast apoptosis and differentiation and the underlying mechanisms. Cell proliferation of murine osteoblastic cell line MC3TC-E1 was examined by CCK8 assay and Edu staining. Cell apoptosis was detected by Annexin V assay under H₂O₂ treatment. The differentiation of osteoblast was determined by Alizarin red S staining. We also tested the expression of osteoblast phenotype related protein by real-time PCR and Western blot. The interaction between Circ_0001843 and miR-214, miR-214 and TAFA5 was examined by luciferase report assay. Circ_0001843 was inhibited by siRNA and miR-214 was suppressed by miR-214 inhibitor to determine the effects of Circ_0001843 and miR-214 on cell proliferation, apoptosis, and differentiation. HNGF6A, an analogue of HN, exerted cyto-protection and osteogenesis-promotion in MC3T3-E1 cells. The expression of osteoblast phenotype related protein was significantly induced by HNGF6A. Additionally, HNGF6A treatment decreased Circ_0001843 and increased miR-214 levels, as well as inhibited the phosphorylation of p38 and JNK. We further found that Circ_0001843 directly bound with miR-214, which in turn inhibited the phosphorylation of p38 and JNK. Furthermore, both Circ_0001843 overexpression and miR-214 knockdown significantly decreased the cyto-protection and osteogenic promotion of HNGF6A. In summary, our data showed that HNGF6A protected osteoblasts from oxidative stress-induced apoptosis and osteoblast phenotype inhibition by targeting Circ_0001843/miR-214 pathway and the downstream kinases, p38 and JNK.

线粒体衍生的肽Humanin（HN）在各种压力下均具有细胞保护作用。在这项研究中，我们旨在研究HN类似物HNGF6A对成骨细胞凋亡和分化及其潜在机制的影响。通过CCK8测定和Edu染色检查鼠成骨细胞系MC3TC-E1的细胞增殖。在H ₂ O ₂下通过Annexin V法检测细胞凋亡。治疗。通过茜素红S染色确定成骨细胞的分化。我们还通过实时PCR和Western印迹测试了成骨细胞表型相关蛋白的表达。通过荧光素酶报告测定法检查了Circ_0001843与miR-214，miR-214和TAFA5之间的相互作用。Circ_0001843被siRNA抑制，miR-214被miR-214抑制剂抑制，以确定Circ_0001843和miR-214对细胞增殖，凋亡和分化的影响。HNGF6A是HN的类似物，可在MC3T3-E1细胞中发挥细胞保护作用和促进成骨作用。HNGF6A明显诱导成骨细胞表型相关蛋白的表达。此外，HNGF6A处理可降低Circ_0001843并增加miR-214的水平，并抑制p38和JNK的磷酸化。我们进一步发现Circ_0001843直接与miR-214结合，进而抑制p38和JNK的磷酸化。此外，Circ_0001843的过表达和miR-214的抑制均显着降低了HNGF6A的细胞保护和成骨促进作用。总而言之，我们的数据表明，HNGF6A通过靶向Circ_0001843 / miR-214途径以及下游激酶p38和JNK，保护成骨细胞免受氧化应激诱导的细胞凋亡和成骨细胞表型的抑制。