摘要/Abstract
摘要： 目的·探讨胞外腺嘌呤核苷三磷酸（adenosine triphosphate，ATP）代谢及其相关指标在糖尿病创面损伤修复炎症反应阶段的变化。方法·将96只C57BL/6J小鼠随机分成4组，每组24只，包括正常空白对照组（NC组）、正常ATP干预组（NA组）、糖尿病空白对照组（DC组）和糖尿病ATP干预组（DA组），其中采用多次小剂量链脲佐菌素腹腔注射诱导DC组和DA组小鼠建立糖尿病模型。所有小鼠于背部制造全层皮肤缺损创面模型，而后NC组和DC组小鼠的创面局部外用双蒸水处理，NA组和DA组小鼠的创面局部外用ATP溶液处理。通过苏木精 - 伊红染色观察创面中性粒细胞浸润。采用酶联免疫吸附测定检测髓过氧化物酶（myeloperoxidase，MPO）含量，超高效液相色谱 - 串联质谱法检测ATP及其代谢产物含量。利用免疫组织化学染色法观察Connexin43、Pannexin1、CD39、P2X7及P2Y2的阳性表达。通过计算创面愈合速率评估创面愈合速度。结果·①创面愈合炎症反应阶段，DC组小鼠创缘组织MPO含量，中性粒细胞浸润数量，ATP含量，Connexin43、Pannexin1、CD39、P2X7及P2Y2的表达量均低于NC组（均PP0.000）。结论·在糖尿病创面愈合炎症反应阶段，炎症反应存在一定的不足，ATP、ATP水解酶及嘌呤信号受体均呈低水平状态，外源性补充ATP可在一定程度上加快糖尿病创面愈合速度。
关键词: 糖尿病, ATP, 炎症反应, 创面愈合
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, n24), normal with ATP applied group (NA, n24), diabetes control groups (DC, n24), and diabetes with ATP applied group (DA, n24). The mice in DA and DC group were inducedstreptozocin 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 detectedenzyme-linked immunosorbent assay (ELISA). The contents of ATP and its metabolites in tissues of wound margin were detectedultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Immunohistochemistry was utilized to investigate the of Connexin43, Pannexin1, CD39, P2X7 and P2Y2. Healing speed were assessedthe 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 s of Connexin43, Pannexin1, CD39, P2X7 and P2Y2 of mice in DC group were all lower than those in NC group (all PP0.000). 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.
Key words: diabetes mellitus, adenosine triphosphate (ATP), inflammation, wound healing
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