Fig 1.
Blood glucose, body weight, and uterine weight.
(A) Hourly blood glucose levels between 0 (initial) and 12 h after fasting are presented as line graphs. (B) Daily blood glucose levels after wounding are shown as line graphs. (C) Daily body weights after wounding are depicted as line graphs. (D) Uterine weights on day 14 after wounding are shown as dotted graphs. Values are expressed as means ± SEM, n = 3–6; ANOVA, Tukey’s honestly significant difference (HSD) tests, **p < 0.01: versus the db/db group, †p < 0.05 and ††p < 0.01: versus the db/db + estrogen group, and ¶p < 0.05, ¶¶p < 0.01 versus the db/db + vehicle group. (E) Macroscopic images of the uterus.
Fig 2.
(A) Wounds with a diameter of 4 mm were inflicted, and images were obtained to assess healing. Bar, 5 mm. (B) The wound area-to-the initial area ratios on day 0 are shown as line graphs based on each day. Values were expressed as mean ± SEM, n = 6–12; ANOVA, Tukey’s HSD test, *p < 0.05, **p < 0.01: versus the db/db group, †p < 0.05, ††p < 0.01: versus the db/db + estrogen group, and ¶p < 0.05 versus the db/db + vehicle group.
Fig 3.
Re-epithelialization and new blood vessels.
(A) H&E staining (bars, 200 μm and 20 μm for db/db [40×]) of granulation tissues on day 14 and (B) the ratio of re-epithelialization (%) are depicted as box graphs. (C) New blood vessels stained with an anti-CD31 antibody (bars, 200 μm) were observed in granulation tissues on day 14 and (D) the ratio of new blood vessels (%) are depicted as box graphs. Values are expressed as mean ± SEM, n = 5–10; ANOVA, Tukey’s HSD tests, *p < 0.05, **p < 0.01 versus the db/db group, ††p < 0.01: versus the db/db + estrogen group, and ¶¶p < 0.01: versus the db/db + vehicle group.
Fig 4.
Macrophages and ym1+ M2 macrophages.
(A) Immunofluorescence staining with Alexa Fluor 488 (green; anti-mac-3 antibody), Alexa Fluor 647 (pink; anti-ym1 antibody), and DAPI (blue) were observed on day 7. The panels labeled as a, b and c are negative controls without primary antibodies, and the panel labeled with an asterisk is an enlarged representative image of the db/db + estrogen group. Bars, 50 μm. (B) The number of macrophages per mm2 and (C) the number of ym1+ M2 macrophages per mm2 are depicted in box graphs. (D) The ratio of ym1+ M2 macrophages/macrophages (%) is shown in doted graphs. Values were expressed as mean ± SEM, n = 4–5; ANOVA, Tukey’s HSD test, *p < 0.05, **p < 0.01: versus the db/db group, ††p < 0.01: versus the db/db + estrogen group.
Fig 5.
(A) The scatter plots on days 7 and 14 after wounding are shown. The upregulated and downregulated genes are depicted in red and blue, respectively. (B) The volcano plots show DEG with a p- value <0.05 and an FC >2.0 on days 7 and 14 after wounding, depicted in red, in the db/db + estrogen or the db/db + vehicle group compared with the db/db group.
Table 1.
Top 10 upregulated genes in db/db mice.
Table 2.
Top 10 downregulated genes in db/db mice.
Fig 6.
(A) Gene set enrichment analysis of db/db + estrogen-specific upregulation on days 7 and 14 after wounding. (B) Gene set enrichment analysis of db/db + estrogen-specific downregulation on days 7 and 14 after wounding.
Table 3.
Top 10 genes upregulated specifically in db/db + estrogen mice.
Table 4.
Top 10 genes downregulated specifically in db/db + estrogen mice.