Pravastatin Attenuates Acute Radiation-Induced Enteropathy and Improves Epithelial Cell Function

Figures

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  FIGURE 1. Pravastatin attenuates radiation-induced intestinal injury. (A) TUNEL assay of small intestine tissue from control (Con), irradiated (IR), and pravastatin-treated IR (IR + Prava) mice 12 h after application of 13.5 Gy of abdominal irradiation. Arrows indicate TUNEL-positive cells. Bar = 20 μm. (B) H&E-stained intestine tissues harvested from Con, IR, and IR + Prava mice 3 and 6 days after 13.5-Gy irradiation. Bar = 200 μm. (C) the number of TUNEL positive cells of small intestinal crypts from Con, IR, and IR + Prava mice 12 h after application of 13.5 Gy of abdominal irradiation. (D) Villi lengths in the small intestine from Con, IR, and IR + Prava mice at 3 and 6 days after irradiation. Data are presented as the mean ± standard error of the mean; n = 5 mice per group. ∗P < 0.05 compared to the control; #P < 0.05 compared to the IR group.

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  FIGURE 2. Pravastatin normalizes impaired epithelial cell differentiation in the irradiated intestine. Expression of (A) MUC2, (B) ChgA, and (C) Lyz1 in the intestine tissue of control (Con), irradiated (IR), and pravastatin-treated IR (IR + Prava) mice, as detected by immunohistochemistry. Bar = 100 μm. Data are presented as the mean ± standard error of the mean; n = 5 mice per group.

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  FIGURE 3. Pravastatin enhances epithelial cell proliferation after radiation-induced intestinal damage. (A) WST-1 assays and (B) SA β-gal staining of control (Con), irradiated (IR), and pravastatin-treated IR (IR + Prava) InEpC cells. n = 3 per group. (C) Counting of Ki-67-positive cell in the intestine of Con, IR, and IR + Prava mice. Bar = 200 μm. Data are presented as the mean ± standard error of the mean; n = 5 mice per group. ∗P < 0.05 compared to the control; #P < 0.05 compared to the IR group.

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  FIGURE 4. Pravastatin attenuates radiation-induced oxidative stress in intestinal epithelial cells. (A) MitoSox Red assay, (B) 2′,7′-dichlorofluorescein diacetate (DCF-DA) assay, and (C) glutathione (GSH) assay of control (Con), irradiated (IR), and pravastatin-treated IR (IR + Prava) InEpC cells. n = 3 per group. (D) Immunohistochemistry analysis of 8-OHdG in the small intestine of Con, IR, and IR + Prava mice. Arrows indicate 8-OHdG-positive cells. Bar = 50 μm. Data are presented as the mean ± standard error of the mean; n = 5 mice per group. ∗P < 0.05 compared to the control; #P < 0.05 compared to the IR group.

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  FIGURE 5. Pravastatin inhibits the inflammatory response during radiation-induced enteropathy. mRNA levels of (A) interleukin (IL)-6, (B) IL-1β, and (C) tumor necrosis factor (TNF)-α in control (Con), irradiated (IR), and pravastatin-treated IR (IR + Prava) InEpC cells, as determined by real-time RT-PCR. n = 3 per group. (D) Immunohistochemistry analysis of myeloperoxidase (MPO) and mRNA levels of (E) Il-1β, (F) Tnf-α, and (G) matrix metallopeptidase (Mmp)-9 in the intestine of Con, IR, and IR + Prava mice. Arrow indicate MPO-positive cells. Bar = 100 μm. Data are presented as the mean ± standard error of the mean; n = 5 mice for each group. ∗P < 0.05 compared to the control; #P < 0.05 compared to the IR group.

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  FIGURE 6. Pravastatin rescues impaired intestinal barrier function induced by radiation exposure. (A) The number of colonies from mLNs tissue and (B) FITC-dextran absorption assay of control (Con), irradiated (IR), and pravastatin-treated IR (IR + Prava) mice. (C) mRNA and (D) protein expression of claudin 3 (CLDN3) in the intestinal tissue of Con, IR, and IR + Prava mice. Bar = 100 μm. Data are presented as the mean ± standard error of the mean; n = 5 mice per group. ∗P < 0.05 compared to the control; #P < 0.05 compared to the IR group.