Hydrogen sulfide improves vessel formation of the ischemic adductor muscle and wound healing in diabetic db/db mice

Abstract

Objectives: It has been demonstrated that hydrogen sulfide plays a vital role in physiological and pathological processes such as regulating inflammation, oxidative stress, and vessel relaxation. The aim of the study was to explore the effect of hydrogen sulfide on angiogenesis in the ischemic adductor muscles of type 2 diabetic db/db mice and ischemic diabetic wound healing.

Materials and methods: The femoral arteries of diabetic db/db mice were isolated and ligated for preparation of ischemic hind limb model. Round incision was made on ischemic and non-ischemic limbs. The wounds were treated with sodium bisulfide (hydrogen sulfide donor). Real-time PCR and Western blotting were used to measure transcription of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), platelet derived growth factor (PDGF), hypoxia inducible factor-1α (HIF-1α) and endothelial nitric oxide synthase (eNOS) and protein expression of VEGF, VEGF receptor (VEGFR) and PDGF, PDGF receptor (PDGFR), respectively. Angiogenesis and morphological changes in adductor muscles were observed.

Results: Hydrogen sulfide significantly increased transcription of VEGF, EGF, PDGF, HIF-1α, eNOS and protein expression of VEGF, PDGF, and phosphorylated VEGFR and PDGFR. Treatment with hydrogen sulfide significantly improved ischemic wound healing and formation of granulation tissue, and increased the number of small vessels in the ischemic adductor muscles.

Conclusion: Our data suggested that hydrogen sulfide attenuated injury of ischemic adductor muscle, and promoted the ischemic diabetic wound healing via modulating angiogenesis in type 2 diabetic db/db mice.

Keywords: Adductor muscle Angiogenesis; Diabetes; Hydrogen sulfide; Ischemia; db/db mice.

Figure 1

Effect of hydrogen sulfide on ischemic diabetic wound healing mice

Figure 2

Histological analysis of ischemic adductor muscles

Figure 3

Serum levels of proangiogenic factors in ischemic diabetic and nonischemic mice

Figure 4

Gene expression of proangiogenic factors in ischemic diabetic and nonischemic adductor muscles

Figure 5

VEGF (A) and PDGF (B) signaling in ischemic diabetic and nonischemic adductor muscles. The relative density of protein expression levels of p-VEGFR (C), p-PDGFR (D), VEGF (E) and PDGF (F) in four studied groups analyzed by one-way ANOVA. Significant differences between groups are indicated by symbols (^**P-value<0.01 compared with the N-C group and ^$$P-value<0.01 compared with the I-C group)