Journal: Frontiers in Immunology
Article Title: Gut Microbial Metabolite Pravastatin Attenuates Intestinal Ischemia/Reperfusion Injury Through Promoting IL-13 Release From Type II Innate Lymphoid Cells via IL−33/ST2 Signaling
doi: 10.3389/fimmu.2021.704836
Figure Lengend Snippet: PA protected against intestinal I/R injury via an IL-33/ST2 signal. (A) IL-33 immunohistochemical staining in the ileum from sham, I/R and I/R + PA mice, scale bar is 100 μm (n = 8). (B, C) Relative mRNA levels of IL-33 and IL-33 receptor (ST2) (n = 8). (D) HE staining, ZO-1, Occludin and Ki67 immunofluorescent staining and IL-33 immunohistochemical staining in WT mice and IL-33 -/- mice, scale bar is 100 μm (n = 8). (E) Pathological damage score in the ileum. (F–H) The relative fluorescence intensity quantification analysis of ZO-1, Occludin and Ki67 in the ileum. (I) The relative mRNA level of Lgr5 in the ileum was measured by quantitative PCR (n = 8). (J) The relative IL-33 intensity quantification analysis in the ileum. (K, L) Intestinal lamina propria cells were analyzed by flow cytometry for the ratio of ILC2/ILCs and IL-13 + ILC2/ILC2 in WT mice (n = 3-4). (M, N) IL-13 immunohistochemical staining and intensity quantification analysis in the ileum, scale bar is 100 μm (n = 8). The results are expressed as the mean ± SEM (B, C, E–J, N) . * p < 0.05, ** p < 0.01, *** p < 0.001, NS means No statistically significant difference by one-way ANOVA (Tukey’s test). PA, pravastatin; I/R, ischemia/reperfusion; ILC2, type II innate lymphoid cells.
Article Snippet: The cell surface was stained using APC-Cyanine7-conjugated CD45 Antibody (Cat# A15395, eBioscience), FITC-conjugated mouse Hematopoietic Lineage Antibody Cocktail (Cat# 22-7770-72, eBioscience), and BV421-conjugated Rat Anti-Mouse IL-33R (ST2) antibody (Cat# 566309, BD Biosciences, San Jose, CA, USA).
Techniques: Immunohistochemical staining, Staining, Fluorescence, Real-time Polymerase Chain Reaction, Flow Cytometry
