MMP-9 inhibitors as a new drug that blocks the development of epilepsy

• Autorzy: Rejmak-Kozicka E. , Khomiak D. , Kaczmarek L.
• Języki publikacji: EN

Abstrakty

EN

INTRODUCTION: Matrix metalloproteinase-9 (MMP-9) is a member of matrix metalloproteinase family that remodels the extracellular matrix. Recently, cumulative evidence indicates that MMP-9 is upregulated in experimental epilepsy models. Increased MMP-9 is also implicated in clinical epilepsy studies. AIM(S): Thus, we hypothesize that MMP-9 may be a novel therapeutic target for epilepsy and some agents, such as OAT1 or OAT2, may be potential antiepileptogenic drugs. METHOD(S): First we estimated IC50 of selected inhibitors using recombinant MMP‑9 and DQ gelatin, fluorescent substrate of MMP-9. Next, we estimated the level of cleavage of MMP-9 substrate – Nectin-3. To determine whether Nectin-3 cleavage might be blocked by MMP-9 inhibitors, we added to hippocampal neurons different concentrations of inhibitors upon 50 μM glutamate stimulation. Extracts from whole cell lysates were analyzed on Western blots. We also adapted gel zymography, method for estimating the level of MMP-9. Due to the MMP-9 low brain expression level and its secretion on the synapse upon neuronal stimulation we decided to inject mice with PTZ (50 mg/kg, 15 min). Next we added inhibitors of MMP-9 directly to developing buffer. RESULTS: IC50 value for OAT1 is 0,5 nM and for OAT2 is 13 nM. Glutamate-dependent stimulation of Nectin-3 cleavage was abolished only in the presence of OAT1 in 5 μM concentration in culture. Further, the gel zymography analysis showed inhibition of MMP-9 activity in the gel with OAT1 in the buffer. The inhibitor’s specificity towards MMP-9 was supported by the absence of MMP-2 activity in this probe, which is another abundant in the brain metalloproteinase showing gelatinase activity. CONCLUSIONS: This results strongly confirm that OAT1 is specific towards MMP‑9 and could be used in animal models of epileptogenesis. FINANCIAL SUPPORT: PBS3/A8/36/2015 from The National Centre for Research and Development.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2017
• Tom: 77
• Numer: Suppl.1
• Opis fizyczny: p.56

Twórcy

autor

Rejmak-Kozicka E.

• Laboratory of Neurobiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Khomiak D.

• Laboratory of Neurobiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Kaczmarek L.

• Laboratory of Neurobiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland