Recombinant human erythropoietin protects long-term cultured ageing primary nerve cells by upregulating the PI3K/Akt pathway

Abstract

Objective Previous studies have found that recombinant human erythropoietin (rhEPO) protects long-term cultured ageing primary nerve cells by enhancing the endogenous antioxidant capacity of cells; however, its signalling pathways are not clear. This study aimed to explore the relationship between the rhEPO and PI3K/Akt pathways in the protection of senescent nerve cells at the cellular level. Methods Primary nerve cells were cultured for 22 days to mimic the natural ageing process of nerve cells. rhEPO and LY294002 were administered as an intervention on the 11th day of culture. Western blot, immunochemistry, 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide, immunofluorescence double-labelling staining, Annexin V-FITC/PI double-labelling flow cytometry, and SA-β-gal staining experiments were used to observe the expression levels of erythropoietin receptor (EPOR) and phosphorylated Akt (p-Akt) protein and the related indices of nerve cell senescence. Results Western blot experiments showed that in ageing long-term cultured primary neurons, the EPOR and p-Akt decreased and rhEPO upregulated the expression levels of EPOR and p-Akt protein. The rest showed that the PI3K/Akt pathway blockade reduced the antioxidation capacity, cell viability, cell morphology, and ratio of apoptotic cells and senescent cells of rhEPO on ageing long-term cultured primary nerve cells. Conclusions This study explored the relationship between the rhEPO and PI3K/Akt pathways in the protection of ageing nerve cells at the cellular level and found that rhEPO protects long-term cultured ageing primary nerve cells by upregulating the PI3K/Akt pathway. These findings provide a theoretical basis and experimental evidence for the antiaeging mechanism of EPO in the nervous system.

Environmentally friendly strategy for discovering the toxicity and mechanisms of nerve injury induced by acrylamide via network toxicology combined with molecular dynamics simulation

“…Studies have shown that nerve growth factor (NGF) promotes the formation of lamentous pseudopods and branches of axons through the PI3K-Akt signaling pathway [39] . The PI3K-Akt signaling pathway has been shown to be associated with ND [40][41][42] . Considering the regulatory effect of the PI3K-Akt signaling pathway and key targets such as TP53, PI3KCA, PIK3R1, RTK2, and GRB2 on nerve injury and combined with the ndings of previous studies, acrylamide was found to regulate the PI3K-Akt signaling pathway [43][44] .…”

Recombinant human erythropoietin protects against immature brain damage induced by hypoxic/ischemia insult

“…It has the same amino acid sequence and similar biological activity as natural EPO, also can cross the blood-brain barrier [10]. Recent years, the protective effect of rhEPO has been proved in many studies in vivo [11] and in vitro [12], however, the role and mechanism of rhEPO on premature immature brain injury induced by HI insult still remain unclear. Thus, in the present study, we investigated the neuroprotective effects of rhEPO as well as its potential mechanism in immature brain injury induced by HI insult.…”