Huprine X Attenuates The Neurotoxicity Induced by Kainic Acid, Especially Brain Inflammation

Abstract

Huprine X (HX) is a synthetic anticholinesterasic compound that exerts a potent inhibitory action on acetylcholinesterase (AChE) activity, an agonist effect on cholinergic receptors, neuroprotective activity in different neurotoxicity models in vivo and in vitro and cognition enhancing effects in non-transgenic (C57BL/6) and transgenic (3xTg-AD, APPswe) mice. In this study, we assessed the ability of HX (0.8 mg/kg, 21 days) to prevent the damage induced by kainic acid (KA; 28 mg/kg) regarding apoptosis, glia reactivity and neurogenesis in mouse brain. KA administration significantly modified the levels of pAkt1, Bcl2, pGSK3b, p25/p35, increased the glial cell markers and reduced the neurogenesis process. We also observed that pre-treatment with HX significantly reduced the p25/p35 ratio and increased synaptophysin levels, which suggests a protective effect against apoptosis and an improvement of neuroplasticity. The increase in GFAP (88%) and Iba-1 (72%) induced by KA was totally prevented by HX pre-treatment, underlying a relevant anti-inflammatory action of the anticholinesterasic drug. Our findings highlight the potential of HX, in particular, and of AChEIs, in general, to treat a number of diseases that course with both cognitive deficits and chronic inflammatory processes.

Neuroprotective Effects of the Multitarget Agent AVCRI104P3 in Brain of Middle-Aged Mice

“…Neurogenesis decreases during neurodegenerative disease and the aging process as well. As it was observed in the case of huprine X [ 14 ], treatment with AVCRI104P3 did not change the expression of the neurogenesis marker DCX in the DG of the hippocampus under the present experimental conditions. This is in contrast to previous reports where it was observed that the AChEIs donepezil, rivastigmine, and galantamine were able to stimulate neurogenesis in different mouse models [ 50 ].…”

“…In this study, it was determined whether the anticholinesterasic AVCRI104P3 could modify the expression of cerebral inflammatory protein markers such as GFAP and Iba1 as it is known that activation of nicotinic receptor is involved in anti-inflammatory mechanisms [ 46 ]. Indeed, we previously observed an anti-inflammatory effect of huprine X in the kainic acid mouse model [ 14 ]. In this work, we have observed that AVCRI104P3 did not modify the astrogliosis marker (GFAP), even though it significantly reduced the expression of the activated microglia protein marker (Iba-1) (34%).…”

“…The procedure for the quantification of protein expression for immunoblotting was done as previously described [ 14 ]. In this study the following primary antibodies were used: Akt1 (1:1000; Sigma, St. Louis, MO, USA); PAkt1 (Ser473) (1:1000; Sigma); Bcl2 (1:1000; Bd Biosciences, New York, NY, USA); GSK3β (1:1000; Cell Signaling, Boston, MA, USA); P-GSK3β (Ser9) (1:1000; Cell Signaling); p35 (C-19) (1:1000; Santa Cruz, Santa Cruz, CA, USA); p25 (1:1000; Santa Cruz); synaptophysin (1:2000; Dako, Glostrup, Denmark); β-actin (1:10,000; Sigma).…”

“…Sagittal sections were obtained as previously described [ 14 ]. Brain sections were mounted on slides (6 cuts per slide).…”

“…Huprine X also improved cognition in six-month-old 3xTg-AD [ 12 ], and biochemical data, such as an increase in synaptophysin and a decrease in Aβ1-40, providing further evidence that this drug can modulate some of the fundamental processes that contribute to neurodegeneration [ 13 ]. Recently, we have also shown that HX treatment ameliorates the toxic effects in the kainic acid mouse model [ 14 ]. We have also demonstrated disease-modifying effects in several families of huprine-based hybrid compounds, which, apart from displaying very potent AChE inhibitory activity (IC 50 values towards human AChE in the single-digit nanomolar range), modulate several key factors involved in the underlying mechanisms of AD when administered to APP/PS1 mice, including amyloid burden, neuroinflammation and epileptogenic activity, and enhance cognition [ 15 , 16 , 17 ].…”