Role of astrocytic NLRP3 in lateral hypothalamus in anxiety-like behaviors after hemorrhagic shock and resuscitation in mice / 中华麻醉学杂志

ABSTRACT

Objective:To evaluate the role of astrocytic NOD-like receptor protein 3 (NLRP3) in the lateral hypothalamus (LHA) in anxiety-like behaviors after hemorrhagic shock and resuscitation in mice.Methods:Forty-eight clean-grade male C57BL/6 mice, aged 10 weeks, weighing 25-30 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (group C), hemorrhagic shock and resuscitation group (group H), hemorrhagic shock and resuscitation + adeno-associated virus group (group HI), and hemorrhagic shock and resuscitation + control virus group (group HIV). The model of hemorrhagic shock and resuscitation was developed by bleeding and re-transfusion through the femoral vein in H, HI and HIV groups. At 21 days before developing the model, AAV-GfaABC1D-EGFP-Cre was injected into bilateral LHA in group HI, and AAV-GfaABC1D-EGFP was administered as a control in group HIV. Anxiety-like behaviors were evaluated by EPM-maze and bead-burying tests at 14 days after resuscitation. Mice were immediately sacrificed at the end of behavioral tests, and LHA-containing brain tissues were obtained for determination of co-localization of NLRP3 with glial fibrillary acidic protein (GFAP), the fluorescence intensity of Wisteria floribunda agglutinin was measured using immunofluorescent staining to reflect the expression of extracellular matrix in the LHA, and the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells in total cells was calculated. Results:Compared with group C, the number of buried beads and percentage of time of staying at the open arm were significantly decreased, the expression of extracellular matrix in the LHA was down-regulated, and the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells was increased in H, HI and HIV groups, and the co-localization coefficient of NLRP3 and GFAP was significantly decreased in group HI ( P<0.01). Compared with group H, the number of buried particles and percentage of time of staying at the open arm were significantly decreased, the expression of extracellular matrix in the LHA was up-regulated, the co-location coefficient of NLRP3 and GFAP was decreased, the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells was decreased ( P<0.01), and no significant change was found in the parameters mentioned above in group HIV ( P>0.05). Conclusions:Anxiety-like behaviors after hemorrhagic shock and resuscitation is associated with astrocytic NLRP3-induced pyroptosis in the LHA and reduction of extracellular matrix in mice.