Tan Hui, Li Feiyan, Cui Yanchao, Li Ziwen, Yan Shiqiang, Deng Quanfeng
Department of Rehabilitation Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China.
School of Clinical Medicine, Chengdu Medical College, Chengdu, Sichuan, China.
Front Pharmacol. 2025 May 30;16:1598151. doi: 10.3389/fphar.2025.1598151. eCollection 2025.
P2X7 receptor (P2X7R) is reported involved in renal fibrosis and the activation of NOD-like receptor protein 3 (NLRP3) inflammasome. This study aimed to investigate the role of the P2X7R and NLRP3 in renal tubular epithelial-myofibroblast transdifferentiation (TEMT) and interstitial fibrosis using a rat unilateral ureteral obstruction (UUO) model.
Sprague‒Dawley rats were randomly divided into the following three groups: sham, UUO, and UUO + Brilliant Blue G (BBG). BBG (50 mg/kg/d)-an antagonist of the P2X7R-was injected intraperitoneally in UUO-treated rats. The adenosine 5'-triphosphate (ATP) concentration in kidney tissue was measured. Hematoxylin and eosin staining and Masson's trichrome staining were used to evaluate the renal injury and the deposition of the extracellular matrix. Collagen-I, collagen-III, α-smooth muscle actin (α-SMA), P2X7R and NLRP3 expression levels were measured via immunohistochemical staining. Furthermore, the mRNA levels of α-SMA, P2X7R and NLRP3 were investigated via a reverse transcription-quantitative polymerase chain reaction.
Significant histopathological damage, which involved tubular dilatation, interstitial inflammation, and collagen accumulation, was observed in UUO rats and was notably alleviated via BBG administration. In the UUO group, ATP concentration increased considerably in kidney tissues; however, this concentration did not decrease following BBG treatment. Collagen-Ⅰ and -III expression levels were upregulated in UUO rats and attenuated through the administration of BBG. Furthermore, BBG administration ameliorated the accumulation of myofibroblast. P2X7R and NLRP3 protein and mRNA expressions increased notably in obstructed kidneys, whereas the protein and mRNA expression of NLRP3 appeared to reduce significantly in the BBG group. However, the mRNA level of P2X7R did not change in response to BBG treatment.
The ATP/P2X7R/NLRP3 pathway is involved in renal TEMT and interstitial fibrosis. P2X7R antagonists attenuate renal interstitial fibrosis and may potentially be used as effective therapeutic agents.
据报道，P2X7受体（P2X7R）参与肾纤维化及NOD样受体蛋白3（NLRP3）炎性小体的激活。本研究旨在利用大鼠单侧输尿管梗阻（UUO）模型，探讨P2X7R和NLRP3在肾小管上皮细胞-肌成纤维细胞转分化（TEMT）及间质纤维化中的作用。
将Sprague-Dawley大鼠随机分为以下三组：假手术组、UUO组和UUO+亮蓝G（BBG）组。对接受UUO处理的大鼠腹腔注射P2X7R拮抗剂BBG（50 mg/kg/d）。检测肾组织中三磷酸腺苷（ATP）浓度。采用苏木精-伊红染色和Masson三色染色评估肾损伤及细胞外基质沉积。通过免疫组化染色检测Ⅰ型胶原、Ⅲ型胶原、α平滑肌肌动蛋白（α-SMA）、P2X7R和NLRP3的表达水平。此外，通过逆转录-定量聚合酶链反应研究α-SMA、P2X7R和NLRP3的mRNA水平。
在UUO大鼠中观察到明显的组织病理学损伤，包括肾小管扩张、间质炎症和胶原积累，而BBG给药可显著减轻这些损伤。在UUO组中，肾组织ATP浓度显著升高；然而，BBG治疗后该浓度并未降低。UUO大鼠中Ⅰ型和Ⅲ型胶原表达水平上调，而BBG给药可使其减弱。此外，BBG给药改善了肌成纤维细胞的积累。在梗阻肾脏中，P2X7R和NLRP3蛋白及mRNA表达显著增加，而在BBG组中，NLRP3的蛋白和mRNA表达似乎显著降低。然而，P2X7R的mRNA水平在BBG治疗后未发生变化。
ATP/P2X7R/NLRP3通路参与肾TEMT和间质纤维化。P2X7R拮抗剂可减轻肾间质纤维化，可能用作有效的治疗药物
