Hyaluronidase inhibitor sHA2.75 alleviates ischemia-reperfusion-induced acute kidney injury.

CELL CYCLE 2024, VOL. 23, NO. 3, 248–261 https://doi.org/10.1080/15384101.2024.2309019 RESEARCH PAPER Hyaluronidase inhibitor sHA2.75 alleviates ischemia-reperfusion-induced acute kidney injury Yang Zhang, Huajiang Zhao, and Jing Zhang Department of Laboratory, Nanjing Jiangning Hospital of Traditional Chinese Medicine, Nanjing, Jiangsu, China ABSTRACT ARTICLE HISTORY Hyaluronidases (HAases) are enzymes that degrade hyaluronic acid (HA) in the animal king dom. The HAases-HA system is crucial for HA homeostasis and plays a significant role in biological processes and extracellular matrix (ECM)-related pathophysiological conditions. This study aims to explore the role of inhibiting the HAases-HA system in acute kidney injury (AKI). We selected the potent inhibitor “sHA2.75” to inhibit HAase activity through mixed inhibitory mechanisms. The ischemia-reperfusion mouse model was established using male BALB/c mice (7–9 weeks old), and animals were subjected to subcapsular injection with 50 mg/kg sHA2.75 twice a week to evaluate the effects of sHA2.75 on AKI on day 1, 5 and 14 after ischemiareperfusion or sham procedure. Blood and tissue samples were collected for immunohisto chemistry, biochemical, and quantitative analyses. sHA2.75 significantly reduced blood urea nitrogen (BUN) and serum creatinine levels in AKI mouse models. Expression of kidney injuryrelated genes such as Kidney injury molecule-1 (KIM-1), Neutrophil Gelatinase-Associated Lipocalin (NGAL), endothelial nitric oxide synthase (eNOS), type I collagen (Col1), type III collagen (Col3), alpha-smooth muscle actin (α-SMA) showed significant downregulation in mouse kidney tissues after sHA2.75 treatment. Moreover, sHA2.75 treatment led to decreased plasma levels of Interleukin-6 (IL-6) proteins and reduced mRNA levels in renal tissues of AKI mice. Inhibitor sHA2.75 administration in the AKI mouse model downregulated kidney injuryrelated biomarkers and immune-specific genes, thereby alleviating AKI in vivo. These findings suggest the potential use of HAase inhibitors for treating ischemic reperfusion-induced kidney injury. Received 12 May 2023 Revised 1 October 2023 Accepted 26 November 2023 1. Introduction Acute kidney injury (AKI) is a syndrome character ized by an abrupt decrease in kidney functions (including structural and loss of function impair ment) [1], a rapid increase in serum creatinine, and/or decline in urine output. This condition affects more than 1.6 million people per year in the United States alone [2,3]. Renal ischemia, inflammatory response, and acute renal tubular injury contribute to the rapid reduction in renal functions due to a variety of different rea sons [4]. Ischemia/reperfusion injury (IRI) is a common cause of AKI, which is featured by the injury of the tubular epithelial and vascular endothe lial cells and inflammatory responses such as leuko cyte infiltration and secretion of inflammatory factors in the kidney [5,6]. Currently, the underlying pathophysiological mechanisms of ischemia- KEYWORDS Acute kidney injury (AKI); hyaluronidase inhibitor; ischemic reperfusion injury (IRI) model reperfusion-induced AKI are not well known. In addition, there are no clinically recognized therapies available to attenuate AKI or expedite recovery. Therefore, it is of vital significance to explore the underlying mechanism of ischemia-reperfusioninduced AKI and develop efficient treatment strategies. Hyaluronic acid (HA), a non-sulfated glycosa minoglycan (GAG) existing in most vertebrates, is typically associated with the pathogenesis of inflammatory reactions, angiogenesis, and fibrosis [7,8]. Serum HA is reported to be at higher levels and correlated with proteinuria and serum albu min level in patients with chronic kidney disease (CKD) and is suggested as biomarker for differen tial diagnosis of AKI and chronic kidney disease [9]. The HA-based nanoparticles also show the potential in the drug delivery for the treatment of CONTACT Jing Zhang Department of Laboratory, Nanjing Jiangning Hospital of Traditional Chinese Medicine, No. 657, Tianyin Avenue, Nanjing, Jiangsu 211100, China Supplemental data for this article can be accessed online at https://doi.org/10.1080/15384101.2024.2309019. © 2024 Informa UK Limited, trading as Taylor & Francis Group CELL CYCLE AKI [10,11]. However, the direct effects of HA on kidney injury were rarely explored. HA is divided into a high molecular weight HA (HMW-HA) which exhibits anti-inflammatory and antifibrotic characteristics [12], and a low molecular weight HA (LMW-HA) which promotes inflam mation and fibrosis [13]. Subsequently to this datum, hyaluronidases’ paramount properties emerge due to being accountable for the cleavage of anti-fibrotic HMW-HA into pro-fibrotic LMWHA. Hyaluronidase enzymes that degrade hyaluro nic acid (HA), are involved in various pathophy siological processes. A previous study has reported that hyaluronidase deficiency promotes renal damage in the post-ischemic kidney [14]. The role of hyaluronidases in kidney repair spurs detailed research on their function in the cardio pulmonary system. For instance, a previous study has shown that HA shedding is mediated by hya luronidase 1 (HYAL1), correlated to the pathogen esis of a variety of diseases, including acute kidney injury (AKI), ischemia/reperfusion (I/R), and chronic kidney disease (CKD) [15]. Another study has disclosed that elevated hyaluronidase (HAase) levels are noteworthy in dialysis patients [16] and that HAase can serve as a feasible bio marker, and its inhibition is a prospect target to protect the HA composition of the endothelial glycocalyx [17]. Although HAases are renowned for their enzymatic activity, non-enzymatic HAases specifically thrive in basic environments detached from the CD44 receptor [18]. The nonenzymatic hyaluronidase 2 (HYAL2) was found to mediate Ras homolog family member A (RhoA), interact with the actin cytoskeleton, and promote inflammation and fibrosis [19]. Recently, hyaluro nidases inhibitors such as sodium aurothiomalate, high molecular mass poly (styrene-4-sulfonate) (PSS), glycyrrhizic acid, gossypol, fenoprofen, fatty acids, heparin, and O-sulfated HA (sHA) [20–31] are implicated in preventing a broad spec trum of disorders like pathogenic infections, inflammation, tissue damage, and venom-derived compounds [21,29,31–34]. Although various HAase inhibitors are exploited to inhibit specific HAases involved in many pathophysiologic disor ders, sHA2.75 is the most prominent hyaluroni dases inhibitor [35]. 249 Biomarkers of kidney injury are suggested as new tools for risk assessment, which also guide the progression of AKI therapy. Nitric oxide (NO) is engaged in the renal injury [36]. Blood urea nitrogen (BUN), creatinine, Kidney injury molecule-1 (KIM-1), and Neutrophil GelatinaseAssociated Lipocalin (NGAL) are the indicators of kidney function and are typically used to assess glomerular and tubular injury in AKI models [37– 39]. (...truncated)