Since ADSCs is imperative for wound healing, its pivotal to unravel the potential molecule-level causal links of increased apoptosis of ADSCs in patients with diabetes. To date, Circular RNAs have been associated with multiple mankind illnesses, such as diabetic diseases, heart and blood vessel illnesses, persistent inflammation illnesses and neurological disorders(24–27). There are few previous studies on hsa_circ_0008500. Only one study indicates that hsa_circ_0008500 is like a miR-1301-3p sponge to facilitate osteoblastic cell mineralisation via the upregulation of PADI4(28). Herein, hsa_circ_0008500 was focused on. Our results of qRT-PCR revealed that hsa_circ_0008500 was regulated downward in ADSCs at HG in vitro. In addition, our team overexpressed hsa_circ_0008500 in diabetic ADSCs, and it was discovered that the programmed cell death of ADSCs was remarkably decreased. The discoveries herein unveiled that hsa_circ_0008500 is able to suppress the programmed cell death of ADSCs and facilitate the healing of diabetic wounds.
Circular RNAs have diverse roles owing to diverse distributional status. To investigate the effects of hsa_circ_0008500 on the modulation of cell apoptosis in depth, our team conducted localization experiments. Hsa_circ_0008500 was mainly distributed in the cytoplasm, and cytoplasmic CircRNA primarily operates via ceRNA. Many Circular RNAs exert important biological functions by acting as protein suppressors (‘sponges’) or microRNA, via modulating protein functions or via their own translation (29). After that, by virtue of biological information forecast, our team discovered that miR-1273h-5p has a hsa_circ_0008500 binding spot. miRNAs interact with the AGO protein family to inhibit translation or decompose mRNA. After that, our team integrated the RIP and RNA pull-down assays to demonstrate that hsa_circ_0008500 is capable of binding to the AGO2 protein. The luciferase reporter assay confirmed as well that hsa_circ_0008500 was able to inhibit miR-1273h-5p expression. We performed qRT-PCR in the miR-1273h-5p-mimic group, demonstrating that hsa_circ_0008500 was decreased, which confirmed that hsa_circ_0008500 decoyed miR-1273h-5p as a sponge RNA.
CircRNAs can function as ceRNA of miRNAs by regulating apoptosis in mankind illnesses(30). For example, circERCC2 could ameliorate intervertebral disc degeneration (IVDD) via miR-182-5p/SIRT1 axis through stimulating mitochondrial autophagy and suppressing programmed cell death, and reveals that circERCC2 is an underlying valid treatment target of IVDD(31). There has been evidence that mmu_circ_0000250 inhibited programmed cell death by autophagy activation, and mmu_circ_0000250 reinforced the treatment potency of ADSCs-exosomes to facilitate wound repair in diabetic diseases via absorpting miR-128-3p and upregulating SIRT1(16). The present research highlighted the effects of hsa_circ_0008500 on regulating ADSCs apoptosis under high glucose, and demonstrated that high expressing of hsa_circ_0008500 inhibited apoptosis of ADSCs. The causal link of hsa_circ_0008500 in apoptosis of ADSCs is limited and needs more exploration.
MiRNAs participate in the modulation of post-transcription genetic expression in cytobiology. Coherent with the high expressing levels of hsa-miR-1273h-5p identified herein, the expression level of hsa-miR-1273h-5p was high under high glucose conditions relative to that under normal glucose conditions. In addition, the data herein revealed that the expressing of hsa_circ_0008500 was related to the expression of hsa-miR-1273h-5p in a negative way. At the same time, the outcomes in this research greatly reveal that hsa_circ_0008500 can be a ceRNA of miR-1273h-5p to realize the downregulation of its expression. A study reported miR-1273h-5p as a contributor to gastric cancer growth and metastasis(32). MiR-1273h-5p, which was remarkably elevated within the peripheral blood monocytes of SLE sufferers, might be a new biomarker for the diagnoses and assessment of SLE(33). Since hsa_circ_0008500 sponged miR-1273h-5p as a ceRNA, our team researched the effects of miR-1273h-5p on ADSC apoptosis and discovered that miR-1273h-5p accelerated apoptosis. In addition, biological information forecast and luciferase reporter assay unveiled that miR-1273h-5p was critical negative regulator of ELK1.
We sought to elucidate the effects of miR-1273h-5p on the modulation of ADSCs apoptosis in depth ; indeed, we found that ELK1 acted as the targeted protein of miR-1273h-5p and demonstrated that miR-1273h-5p suppressed ELK1 expression. ELK1, a TF of the ETS family, is a pivotal constituent of the mitogen modulation signal path activating the mitogen-stimulated protein kinase cascade(34). ELK1 primarily participated in the modulation of cellular proliferation, differentiation, and apoptosis(35). ELK1 is vital for miR-139-3p to suppress osteoblastic differentiation and facilitate osteoblastic programmed cell death(35). MiR-150 can modulate the expressing of ELK1, while ELK1 knockout can terminate the antiapoptotic effect of the suppressor miR-150 in endotheliocytes(36). Another study confirmed that Zuo Gui Jiang Tang Jie Yu Formula was capable of reversing the expression of ELK-1, JNK and c-Fos signals in hippocamp nerve cells in DD milieu and eserted antiapoptotic effects(37). In conclusion, we unveiled that hsa_circ_0008500 could modulate the expressing of ELK-1 via sponging miR-1273h-5p. We verified that hsa_circ_0008500 regulates ADSC apoptosis through miR-1273h-5p / ELK1, thus elucidating the mechanism of ADSC apoptosis under high-glucose stress.