Last modified: 2018-09-12
Abstract
Introduction: Amphotericin B acts as a classic antifungal agents. In current experiment, the main objective is investigating the effects of amphotericin B on specific miRNAs and the mechanism of specific miRNAs on meningoencephalitis.
Methods: Meningoencephalitis model is established through intervening with pathogenic or non-pathogenic C. neoformans, then intervened with/without amphotericin B, and bacterial culture in cerebrospinal fluid were acted at 72 hr. qRT-PCR was utilized to detect the levels of miR-294, -22-5p, 200c-5p, 218a-3p and 181a-5p in rat’s hippocampus and HBMECs, the expressions of HMGB1, RAGE and Cdc42 were measured though western blot. After miR-181a-5p mimics/inhibitors transfecting to HBMECs, HMGB1, RAGE and Cdc42 expressions were observed and the monolayer’s permeability were measured by ELISA.
Results: Bacterial colonies increase after C. neoformans infection, while amphotericin B markedly inhibited fungoid growth. C. neoformans up-regulated HMGB1, RAGE and Cdc42 levels and miR-294, -22-5p, 200c-5p, 218a-3p and 181a-5p expressions, while amphotericin B alleviated these affect. Meanwhile, OD values of HRP rised in vitro after C. neoformans infection. Overexpression of miR-181a-5p increased HMGB1, RAGE and Cdc42 expressions and accelerate OD values in HBMECs, while miR-181a-5p inhibition had the opposing effects.
Conclusions: Our study revealed the axis of HMGB1/RAGE and specific miRNAs are involved in the pathophysiology of meningoencephalitis. miR-181a-5p increased HMGB1 and RAGE expressions to deteriorate blood brain barrier permeability after C. neoformans infection, suggesting that miR-181a-5p might act as an essential positive regulator of meningoencephalitis.