Changes of ATP metabolism and purinergic receptors in inflammatory response stage of diabetic wound healing / 上海交通大学学报（医学版）

ABSTRACT

Objective:To investigate the changes of extracellular adenosine triphosphate (ATP) metabolism and its related indices of inflammatory response stage during diabetic wound healing. Methods:Ninety-six C57BL/6J mice were divided into 4 groups randomly, i.e. normal control group (NC, n=24), normal with ATP applied group (NA, n=24), diabetes control groups (DC, n=24), and diabetes with ATP applied group (DA, n=24). The mice in DA and DC group were induced by streptozocin multiple intraperitoneal injection. Full-thickness excisional wound was created on the dorsum of all mice. Distilled water was applied in NC and DC group. ATP was applied in NA and DA group. Hematoxylin and eosin staining (HE staining) was used to observe neutrophils infiltration. Myeloperoxidase deficiency (MPO) contents were detected by enzyme-linked immunosorbent assay (ELISA). The contents of ATP and its metabolites in tissues of wound margin were detected by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Immunohistochemistry was utilized to investigate the expression of Connexin43, Pannexin1, CD39, P2X7 and P2Y2. Healing speed were assessed by the calculation of healing rate on each point. Results:①During the inflammatory response stage of wound healing, the MPO contents, neutrophils infiltration numbers, the ATP contents, and the expressions of Connexin43, Pannexin1, CD39, P2X7 and P2Y2 of mice in DC group were all lower than those in NC group (all P<0.05). ② On the 14th day after injury, the wound healing rate of mice in DA group was significantly higher than that in DC group (P=0.000). Conclusion:Inflammatory response is insufficient on the early stage of diabetic wound healing, while ATP, ATP hydrolase and purinergic receptor also remain at low level at the same time. To a certain extent, exogenous application of ATP can accelerate diabetic wound healing rate.