Fig. 8: The role of regulating ERK1/2 activity in GPER activation protecting 5-FU induced CIM. | Cell Death & Disease

Fig. 8: The role of regulating ERK1/2 activity in GPER activation protecting 5-FU induced CIM.

From: Activation of G protein coupled estrogen receptor prevents chemotherapy-induced intestinal mucositis by inhibiting the DNA damage in crypt cell in an extracellular signal-regulated kinase 1- and 2- dependent manner

Fig. 8

CIM model was induced by i.p. injection 5-FU (30 mg/kg/day) from day 0 to day 5. G-1 (0.03 mg/kg/day) and/or ERK1/2 inhibitor PD0325901 (5 mg/kg/day) were administrated intraperitoneally together with 5-FU. On the 5th day, the ileum was collected for H& E staining, immunofluorescence staining, immunohistochemical staining and western blot. Data were expressed as mean ± SEM (*P < 0.05, **P < 0.01, ***P < 0.001). a Representative immunohistochemical images of γH2AX staining in the ileum of the CIM model following G-1 administration or in combination with PD0325901 (Scale bars: 20 μm). b Statistical graph of γH2AX+ cells in ileal crypt within three subgroups to show the role of regulating ERK1/2 activity in GPER inhibiting DNA damage induced by CIM. 15 crypts were randomly calculated in each section, and the average value was obtained (n = 5). c Effect of ERK1/2 inhibitor PD0325901 on G-1 in reducing intestinal mucosal damage score and crypt damage score of CIM model (n = 9). d Effect of ERK1/2 inhibitor PD0325901 on G-1 inhibition of crypt depth reduction and villus shortening in the CIM model (n = 9). At least 20 villi were counted randomly for each sample, and the mean value was calculated for single sample. e Effect of ERK1/2 inhibitor PD0325901 on G-1 inhibition of weight loss in the CIM model (n = 9). f Effect of ERK1/2 inhibitor PD0325901on G-1 protected ileal mucosal permeability in the CIM model (n = 6). The mucosal permeability was evaluated by the concentration of Fluorescein isothiocyanate (FITC) -dextran in blood. g Representative immunohistochemical images of BrdU staining within four subgroups (scale bars: 20 μm) and statistical graph of BrdU+ cell to show the effect of ERK1/2 inhibitor PD0325901 on G-1 protecting proliferation of crypt cells in the CIM model. At least 20 crypts were counted randomly, and the mean value was calculated for single sample (n = 6). h Representative immunohistochemical images of Ki67 staining within four subgroups (scale bars: 20 μm) and statistical graph of Ki67+ cell to show the effect of ERK1/2 inhibitor PD0325901 on G-1 protecting proliferation of crypt cells in the CIM model. At least 20 crypts were counted randomly, and the mean value was calculated for single sample (n = 6). i Representative images of TUNEL staining within four subgroups (scale bars: 50 μm) and statistical graph of TUNEL + cell to show the effect of ERK1/2 inhibitor PD0325901 on G-1 inhibiting apoptosis of crypt cells in the CIM model. At least 20 crypts were counted randomly, and the mean value was calculated for single sample (n = 6). j Representative immunohistochemical images of Muc-2 staining within four subgroups (scale bars: 50 μm) and statistical graph of Muc-2+ cells to show the effect of ERK1/2 inhibitor PD0325901 on G-1 protecting the mucus barrier in the CIM model. At least 20 villi were counted randomly, and the mean value was calculated for single sample (n = 6).

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