Fig. 5: ITE conferred retinal protection against microglia derived toxins induced by LPS in retinal IR injury.

A Experimental protocols. LPS was added for 20 h with or without ITE pre-treatment for 4 h. BV2 microglia were then washed with PBS and medium was changed. After 24 hours, BV-2 conditioned medium was collected and concentrated as LPS induced BV2-CCM or ITE + LPS induced BV2-CCM. Mice were subjected to IR one hour before intravitreal injection with relative medium. Mice were euthanatized at day 3 after IR injury. B H&E staining of frozen tissue sections showed retinas in Control or IR groups receiving different conditioned medium. GCL ganglion cell layer, IPL inner plexiform layer, INL inner nuclear layer, ONL outer nuclear layer. Scale bar = 50 µm. C Quantitative analysis of the thickness of inner plexiform layer and retinal nerve fiber layer and cell numbers in ganglion cell layer in each group. D Apoptosis of the retina from Control and IR groups were detected by TUNEL (green) staining. Cell nuclei were counterstained with DAPI (blue). Scale bar = 50 µm. All data are presented as mean ± SEM, and are representative of 4-8 independent experiments. *P < 0.05, **P < 0.01. Compared with Control group: ^#P < 0.05, ^##P < 0.01, ^###P < 0.001. P value was obtained either by Student’s t test or by one way ANOVA followed by multiple comparisons.