Transforming growth factor-1 functions as a competitive endogenous RNA that ameliorates intracranial hemorrhage injury by sponging microRNA-93-5p

Intracerebral hemorrhage (ICH) has the highest mortality rate of all stroke subtypes but an effective treatment has yet to be clinically implemented. Transforming growth factor-beta 1 (TGF-beta 1) has been reported to modulate microglia-mediated neuroinflammation after ICH and promote functional recovery; however, the underlying mechanisms remain unclear. Non-coding RNAs such as microRNAs (miRNAs) and competitive endogenous RNAs (ceRNAs) have surfaced as critical regulators in human disease. A known miR-93 target, nuclear factor erythroid 2-related factor 2 (Nrf2), has been shown to be neuroprotective after ICH. It was hypothesized that TGF-beta 1 functions as a ceRNA that sponges miR-93-5p and thereby ameliorates ICH injury in the brain. Short interfering RNA (siRNA) was used to knock down TGF-beta 1 and miR-93 expression was also pharmacologically manipulated to elucidate the mechanistic association between miR-93-5p, Nrf2, and TGF-beta 1 in an in vitro model of ICH (thrombin-treated human microglial HMO6 cells). Bioinformatics predictive analyses showed that miR-93-5p could bind to both TGF-beta 1 and Nrf2. It was found that neuronal miR-93-5p was dramatically decreased in these HMO6 cells, and similar changes were observed in fresh brain tissue from patients with ICH. Most importantly, luciferase reporter assays were used to demonstrate that miR-93-5p directly targeted Nrf2 to inhibit its expression and the addition of the TGF-beta 1 untranslated region restored the levels of Nrf2. Moreover, an miR-93-5p inhibitor increased the expression of TGF-beta 1 and Nrf2 and decreased apoptosis. Collectively, these results identified a novel function of TGF-beta 1 as a ceRNA that sponges miR-93-5p to increase the expression of neuroprotective Nrf2 and decrease cell death after ICH. The present findings provided evidence to support miR-93-5p as a potential therapeutic target for the treatment of ICH.