通过 ATP 释放和 P2Y 受体激活的自分泌调节伤口愈合。

Autocrine regulation of wound healing by ATP release and P2Y receptor activation.

机构信息

Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia; Molecular Horizons and School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia.

Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia; School of Medicine, University of Wollongong, Wollongong, NSW 2522, Australia.

出版信息

Life Sci. 2021 Oct 15;283:119850. doi: 10.1016/j.lfs.2021.119850. Epub 2021 Jul 24.

DOI:10.1016/j.lfs.2021.119850

PMID:34314735

Abstract

AIMS

Application of exogenous nucleotides can modulate wound healing via the activation of purinergic receptors. However, evidence for the release of endogenous nucleotides and the subsequent activation of purinergic receptors in this process has not been well defined. Therefore, the current study aimed to investigate wound-mediated nucleotide release and autocrine purinergic signalling during HaCaT keratinocyte wound closure following scratch injury.

MAIN METHODS

An in vitro scratch wound apparatus was employed to study wound healing over 24-h in the presence of modulators of ATP release, P2 receptors and pathways downstream of P2 receptor activation.

KEY FINDINGS

Adenosine 5'-triphosphate (ATP) was released from scratched cells. The ectonucleotidase apyrase and pharmacological inhibition of the nucleotide release hemichannel, pannexin-1, decreased wound closure over time. The non-selective P2Y receptor antagonist suramin and the selective P2Y receptor antagonist AR-C118925XX, but not other P2 antagonists, decreased wound closure. AR-C118925XX decreased wound closure in a concentration-dependent fashion. However, exogenous P2Y receptor agonists, ATP or uridine 5'-triphosphate, did not enhance wound closure. PCR and immunoblotting confirmed P2Y receptor expression in HaCaT cells. U73122, a phospholipase C antagonist, and 2-aminoethoxydiphenylborate, an inositol 1,4,5-trisphosphate receptor-sensitive Ca-release channel antagonist, decreased wound closure consistent with P2Y receptor activation. Absence of extracellular or intracellular Ca or inhibition of intracellular Ca-release also impaired wound closure.

SIGNIFICANCE

These data describe a novel autocrine signalling mechanism in which wound-mediated release of endogenous ATP in response to mechanical scratching of HaCaT cells activates P2Y receptors to facilitate wound closure.

摘要

目的

外源性核苷酸的应用可以通过嘌呤能受体的激活来调节伤口愈合。然而，在这个过程中，内源性核苷酸的释放及其随后的嘌呤能受体激活的证据尚未得到很好的定义。因此，本研究旨在探讨在划痕损伤后 HaCaT 角质形成细胞伤口闭合过程中，伤口介导的核苷酸释放和自分泌嘌呤能信号转导。

方法

采用体外划痕伤装置，在 ATP 释放调节剂、P2 受体及其下游途径激活的情况下，研究 24 小时内的伤口愈合。

主要发现

从划痕细胞中释放出腺苷 5'-三磷酸（ATP）。外核苷酸酶 apyrase 和核苷酸释放半通道，pannexin-1 的药理学抑制，随着时间的推移减少了伤口闭合。非选择性 P2Y 受体拮抗剂苏拉明和选择性 P2Y 受体拮抗剂 AR-C118925XX，但不是其他 P2 拮抗剂，减少了伤口闭合。AR-C118925XX 以浓度依赖的方式减少了伤口闭合。然而，外源性 P2Y 受体激动剂，ATP 或尿苷 5'-三磷酸，并没有增强伤口闭合。PCR 和免疫印迹证实 P2Y 受体在 HaCaT 细胞中的表达。PLC 拮抗剂 U73122 和肌醇 1,4,5-三磷酸受体敏感的 Ca 释放通道拮抗剂 2-氨基乙氧基二苯硼酸盐减少伤口闭合与 P2Y 受体激活一致。缺乏细胞外或细胞内 Ca 或抑制细胞内 Ca 释放也损害了伤口闭合。