In vitro effect of antiretroviral drugs on cultured primary astrocytes: analysis of neurotoxicity and matrix metalloproteinase inhibition

Abstract

There is little information available on the possible toxic effects that antiretroviral (ARV) drugs used for the treatment of human immunodeficiency virus (HIV)-infected subjects, may have on the central nervous system (CNS) resident cells. Moreover, it remains unclear whether the efficacy of the ARV drugs may also be due to their ability to exert extravirological effects on factors responsible for the development of HIV brain injury, e.g., matrix metalloproteinases (MMPs). This study investigates the toxicity of three different ARV drugs and on their ability to modulate levels and expression of gelatinases A (MMP-2) and B (MMP-9) in astrocytes. Primary cultures of rat astrocytes were activated by exposure to lipopolysaccaride (LPS) and simultaneously treated with darunavir, maraviroc, or raltegravir, used alone or in combination. Among the tested drugs, maraviroc was the less toxic for astrocytes. At toxic concentration (TC ), the studied drugs induced the production of reactive oxygen species (ROS), suggesting that the oxidative stress may represent a mechanism of ARV toxicity. As assessed by gelatin zymography and RT-PCR, the single antiretroviral drugs reduced levels and expression of both MMP-2 and MMP-9 through the inhibition of the signaling transduction pathway of extracellular signal-regulated kinase1/2, which is involved in the regulation of MMP-9 gene. A synergistic inhibition of MMP-2 and MMP-9 was observed with combinations of the studied ARV drugs. The present results indicate that maraviroc, darunavir, and raltegravir, through their ability to inhibit MMP-2 and MMP-9 at doses non-toxic for astrocytes, might have a great potential for the management of HIV-associated neurological complications.

The Glutamate System as a Crucial Regulator of CNS Toxicity and Survival of HIV Reservoirs

“…For example, neurotoxicity has been described for several viral proteins including gp120 and the transactivator of transcription (Tat) ( Table 1 ), as well as for host factors such as TNF-α, IL-1β, and IL-6 released from latently HIV-1-infected or -activated cells (Koller et al, 2001 ; Zhou et al, 2017 ). In addition, several ART drugs have been demonstrated to generate toxicity on their own (Brier et al, 2015 ; Underwood et al, 2015 ; Latronico et al, 2018 ). Thus, the combination of HIV-1-infection, low-level secretion of viral proteins, and ART toxicity probably contribute to CNS dysfunction.…”

Neuroprotective potential of isothiocyanates in an in vitro model of neuroinflammation

“…For this reason, growing interest has been focused in the last decade on the potential of drugs and natural compounds to inhibit MMPs. Numerous in vitro studies have evidenced that anti-inflammatory and antioxidant action of several drugs and bioactive compounds present in vegetables and marine organisms are associated with their abilities to inhibit the expression of MMPs (Latronico et al 2016(Latronico et al , 2018Di Bari et al 2015). In this study we demonstrated that among the tested ITCs, SFN and PEITC were the most effective in inhibiting MMP-2 and MMP-9 levels and expression in LPS-activated astrocytes.…”

“…The animals were euthanized by exposure to carbon dioxide (CO 2 ) and killed by rapid decapitation. The neocortical tissues were dissected and used for purification of primary glial cell cultures following the method reported by Latronico et al (2018). Briefly, after the dissection, rat brains were depleted of meninges and blood vessels, mechanical minced and digested with 0.25% trypsin in presence of 0.01% DNase.…”

“…ERK 1/2 were detected by immunoblot analysis as reported in Latronico et al (2018). Briefly, confluent primary astrocytes plated in 6-well plates, made quiescent in serumfree medium for 24 h, were pretreated for 1 h with AITC (400 μM), PEITC (10 μM) or SFN (25 μM) in phenol red-free DMEM.…”

Cerebral Vascular Toxicity of Antiretroviral Therapy

“…A recent study demonstrated that exposure of astrocytes to Maraviroc, Raltegravir and Darunavir at high concentrations can lead to a reduction in MMP synthesis and activation. This effect was also shown to synergize when drugs were combined (Latronico et al 2018). Indinavir was demonstrated to inhibit the conversion of ProMMP2 into active MMP2 and decrease angiogenesis (Barillari et al 2014).…”