ABSTRACT
Calcium oxalate (CaOx) crystals can activate autophagy, causing damage to renal tubular epithelial cells (TECs). Puerarin has been shown to have protective and therapeutic effects against a variety of diseases by inhibiting autophagy activation. However, the protective effect of puerarin against CaOx crystals and the underlying molecular mechanisms are unclear. Cell Counting Kit-8 (CCK-8) assays were used to evaluate the effects of puerarin on cell viability. Intracellular reactive oxygen species (ROS) levels were measured by the cell-permeable fluorogenic probe 2,7-dichlorofluorescein diacetate (DCFH-DA). Immunofluorescence, immunohistochemistry, and western blotting were used to examine the expression of SIRT1, Beclin1, p62, and LC3, and explore the underlying molecular mechanisms in vivo and in vitro. Puerarin treatment significantly attenuated CaOx crystal-induced autophagy of TECs and CaOx cytotoxicity to TECs by altering SIRT1 expression in vitro and in vivo, whereas the SIRT1-specific inhibitor EX527 exerted contrasting effects. In addition, we found that the protective effect of puerarin was related to the SIRT1/AKT/p38 signaling pathway. The findings suggest that puerarin regulates CaOx crystal-induced autophagy by activating the SIRT1-mediated signaling pathway, and they suggest a series of potential therapeutic targets and strategies for treating nephrolithiasis.
Subject(s)
Calcium Oxalate , Kidney Calculi , Autophagy , Calcium Oxalate/metabolism , Epithelial Cells/metabolism , Humans , Isoflavones , Kidney Calculi/metabolism , Oxidative Stress , Signal Transduction , Sirtuin 1/metabolism , Sirtuin 1/pharmacologyABSTRACT
OBJECTIVES: Calcium oxalate (CaOx) crystals can activate inflammatory cytokines by triggering inflammasomes, which cause damage to the adhered epithelium, a dysfunctional microenvironment and even renal failure. However, a comprehensive and in-depth understanding of the mechanisms underlying the effects of these crystals on damage and cytokine function in renal tubular epithelial cells (TECs) remains limited and to be explored. MATERIALS AND METHODS: We detected the pyroptosis of TECs induced after exposure to CaOx crystals and demonstrated the significance of cytokine activation in the subsequent inflammatory processes through a proteomic study. We then conducted animal and cell experiments to verify relevant mechanisms through morphological, protein, histological and biochemical approaches. Human serum samples were further tested to help explain the pathophysiological mechanism of H3 relaxin. RESULTS: We verified that crystal-induced extracellular adenosine triphosphate (ATP) upregulation via the membrane purinergic 2X7 receptor (P2X7 R) promotes ROS generation and thereby activates NLRP3 inflammasome-mediated interleukin-1ß/18 maturation and gasdermin D cleavage. Human recombinant relaxin-3 (H3 relaxin) can act on the transmembrane receptor RXFP1 to produce cAMP and subsequently improves crystal-derived damage via ATP consumption. Additionally, endogenous relaxin-3 was found to be elevated in patients with renal calculus and can thus serve as a biomarker. CONCLUSIONS: Our results provide previously unidentified mechanistic insights into CaOx crystal-induced inflammatory pyroptotic damage and H3 relaxin-mediated anti-inflammatory protection and thus suggest a series of potential therapeutic targets and methods for but not limited to nephrocalcinosis.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Calcium Oxalate/pharmacology , Pyroptosis/drug effects , Relaxin/pharmacology , Adenosine Triphosphate/metabolism , Animals , Calcium Oxalate/chemistry , Cell Line , Cyclic AMP/metabolism , Humans , Inflammasomes/metabolism , Interleukin-1beta/metabolism , Kidney/drug effects , Kidney/metabolism , Kidney/pathology , Kidney Tubules, Proximal/cytology , Kidney Tubules, Proximal/metabolism , Male , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Rats , Rats, Sprague-Dawley , Reactive Oxygen Species/metabolism , Receptors, G-Protein-Coupled/metabolism , Receptors, Peptide/metabolism , Receptors, Purinergic P2X7/metabolism , Relaxin/bloodABSTRACT
Premature ejaculation is a common male sexual dysfunction disorder, and there are many controversies over its definition. With deeper insights into the etiology and pathogenesis of premature ejaculation, more and more auxiliary examinations are used in its diagnosis, prognostic evaluation and treatment, such as transrectal ultrasonography of seminal vesicles, determination of serum 5-hydroxytryptamine (5-HT) concentration, serum hormone levels, penile sensitivity detection, brain function tests, and genetic sequencing. This review outlines the latest advances in the auxiliary examination of premature ejaculation and provides clinicians with some diagnostic indexes or methods of premature ejaculation for reference.
