Fig. 5

CSO treatment relieved TLR4/NF-κB-mediated hyperactivation of astrocyte and microglia in the ischemic penumbra. A CSO treatment relieved astrocyte and microglia activation, respectively. A(a) Representative immunofluorescence images showing the morphologies of the astrocyte labeled with GFAP and microglia labeled with Iba1 in the ischemic penumbra 24 h after reperfusion. Scale bar = 20 μm. A(b) The average area of single microglial cells labeled by Iba1 and single astrocytic cells labeled by GFAP in the ischemic penumbra. ^##p < 0.01 vs. Sham-Con group, ^*p < 0.05 vs. MCAO-Con group. n = 5 per group. B CSO treatment relieved astrocyte and microglia co-activation. Representative immunofluorescence images showing the morphologies of the astrocyte labeled with GFAP and microglia labeled with Iba1 in the ischemic penumbra under normal conditions, and the activated microglia was close to the activated astrocyte in the ischemic penumbra 24 h after reperfusion, and CSO significantly inhibited co-activation of microglia and astrocyte. Scale bar = 20 μm. C CSO treatment reduced the expression of TLR4, P-NF-κB, GFAP, and Iba1 in the ischemic penumbra 24 h after reperfusion. C(a) Cropped gels and blots showing the protein expression of the TLR4, P-NF-κB, NF-κB, GFAP, and Iba1 in the ischemic penumbra 24 h after reperfusion. C(b) Graph of the protein expression of the TLR4, P-NF-κB, GFAP, and Iba1 in the ischemic penumbra 24 h after reperfusion. ^##p < 0.01, ^#p < 0.05 vs. Sham-Con group, ^*p < 0.05 vs. MCAO-Con group. n = 6 per group. D CSO treatment relieved P-NF-κB-mediated astrocyte activation and TLR4-mediated microglia activation, respectively. D(a) Representative immunofluorescence images showing the colocalization of P-NF-κB and GFAP in the ischemic penumbra 24 h after reperfusion. Scale bar = 5 μm. D(b) Representative immunofluorescence images showing the colocalization of TLR4 and Iba1 in the ischemic penumbra 24 h after reperfusion. Scale bar = 20 μm