SaRNA-mediated activation of TRPV5 reduces renal calcium oxalate deposition in rat via decreasing urinary calcium excretion

Abstract

Hypercalciuria is a main risk factor for kidney stone formation. TRPV5 is the gatekeeper protein for mediating calcium transport and reabsorption in the kidney. In the present study, we tested the effect of TRPV5 activation with small activating RNA (saRNA), which could induce gene expression by targeting the promoter of the gene, on ethylene glycol (EG)-induced calcium oxalate (CaOx) crystals formation in rat kidney. Five pairs of RNA activation sequences targeting the promoter of rat TRPV5 were designed and synthesized. The synthesized saRNA with the strongest activating effect was selected, and transcellular calcium transportation was tested by Fura-2 analysis. Subsequently, Sprague-Dawley rats were equally divided into three groups and fed with water, 1% EG for 28 days after injecting the negative control saRNA, 1% EG for 28 days after injecting the selected TRPV5-saRNA, respectively. The CaOx crystal formation and the 24-h urine components were assessed. In vitro study, saRNA ds-320 could significantly activate the expression of TRPV5 and transcellular calcium transportation. In vivo study, after 28 days treatment of EG, rats pre-infected with saRNA ds-320 had lower urinary calcium excretion and renal CaOx crystals formation as compared to that pre-infected with negative control saRNA. Activation of TRVP5 with saRNA ds-320 could inhibit EG-induced calcium oxalate crystals formation via promoting urine calcium reabsorption and decreasing urine calcium excretion in rats.

Activities of Ca2+-related ion channels during the formation of kidney stones in an infection-induced urolithiasis rat model

“…Wang et al (39) found that TRPV5 downregulation could decrease urinary reabsorption of Ca 2ϩ and result in severe hypercalciuria. Another study also (43) demonstrated that activation of the TRPV5 channel using a small activating RNA approach decreased CaOx crystal formation. Hence, it can be concluded that lack of TRPV5 could aggravate crystallization activity.…”

Mucin 4 Gene Silencing Reduces Oxidative Stress and Calcium Oxalate Crystal Formation in Renal Tubular Epithelial Cells Through the Extracellular Signal-Regulated Kinase Signaling Pathway in Nephrolithiasis Rat Model

“…Recent studies showed that the transportation of materials was closely relevant with the development of nephrolithiasis [16,17]. For example, TRPV5 overexpression transported by calcium can induce calcium oxalate crystallization in rats' kidney [18]. The above literature and findings demonstrated that MUC4, as transporter related protein, showed a strong correlation with the development of nephrolithiasis.…”

Confounding risk factors and preventative measures driving nephrolithiasis global makeup

“…Genes which potentially affect the predisposition only influences the risk factors of nephrolithiasis; such as comorbidities or phenotypical symptoms which may increase the risk of nephrolithiasis in the longer run. Several genes which code for TRPV5[ 72 ], SLC26A6 and NaDC-1[ 73 ] can play a physiological role in contributing to calcium stone formation because they play a major role in homeostasis in maintaining physiological conditions. Other genetic factors may have an indirect correlation to nephrolithiasis by contributing to common comorbidities or risk factors to nephrolithiasis.…”