Probenecid Pre-treatment Downregulates the Kidney Cl-/HCO3- Exchanger (Pendrin) and Potentiates Hydrochlorothiazide-Induced Diuresis

Abstract

Background: Probenecid is a uricosuric agent that in addition to exerting a positive ionotropic effect in the heart, blocks the ATP transporter Pannexin 1 and inhibits the Cl-/HCO3- exchanger, pendrin. In the kidney, pendrin blunts the loss of salt wasting secondary to the inhibition of the thiazide-sensitive Na+-Cl- co-transporter (NCC/SLC12A3).Hypothesis: Pre-treatment with probenecid down-regulates pendrin; therefore, leaving NCC as the main salt absorbing transporter in the distal nephron, and hence enhances the hydrochlorothiazide (HCTZ)-induced diuresis.Methods: Daily balance studies, blood and urine chemical analysis, immunofluorescence, as well as western and northern blot analyses were utilized to examine the effects of probenecid alone (at 250 mg/kg/day) or in combination with HCTZ (at 40 mg/kg/day) on kidney function and on salt and water transporters in the collecting duct.Results: Male Sprague Dawley rats were subjected to three different protocols: (1) HCTZ for 4 days, (2) probenecid for 10 days, and (3) primed with probenecid for 6 days followed by probenecid and HCTZ for 4 additional days. Treatment protocol 1 (HCTZ for 4 days) only mildly increased the urine volume (U Vol) from a baseline of 9.8–13.4 ml/day. In response to treatment protocol 2 (probenecid for 10 days), U Vol increased to 15.9 ml/24 h. Treatment protocol 3 (probenecid for 6 days followed by probenecid and HCTZ for 4 additional days) increased the U Vol to 42.9 ml/day on day 4 of co-treatment with HCTZ and probenecid (compared to probenecid p = 0.003, n = 5 or HCTZ alone p = 0.001, n = 5). Probenecid treatment at 250 mg/kg/day downregulated the expression of pendrin and led to a decrease in AQP2 expression. Enhanced diuresis by probenecid plus HCTZ was not associated with volume depletion.Conclusion: Probenecid pre-treatment downregulates pendrin and robustly enhances diuresis by HCTZ-mediated NCC inhibition in kidney.

Tuberous sclerosis complex exhibits a new renal cystogenic mechanism

“…Knock‐down of hamartin is known to reduce tuberin expression (Barone et al. ). (B) Section of mutant kidney cortex and medulla.…”

“…Single and double immunofluorescence labeling was performed using established protocols (Xu et al 2011;Zahedi et al 2013;Barone et al 2018;Varasteh Kia et al 2018). Briefly, animals were euthanized with an overdose of pentobarbital sodium, and kidneys were perfused with saline followed by 4% paraformaldehyde removed, cut in tissue blocks, and fixed in 4% paraformaldehyde solution overnight at 4°C.…”

Risk factors and management of hyperuricemia after renal transplantation

“…Multiple studies have reported that diuretics were directly related to a higher risk for HUA after renal transplantation ( 7 , 8 , 32 ). Among these, thiazide diuretics and loop diuretics can interact with renal OAT; they enter the proximal tubular cells from the blood by OAT1 and OAT3, which could probably compete with UA, causing the reduced secretion of UA ( 30 , 44 , 45 ). Moreover, diuretics also decrease UA excretion, possibly by causing blood volume depletion with a consequent increase in proximal tubular reabsorption of UA ( 38 ).…”

Solubility Measurement and Correlation of Probenecid in 12 Pure Organic Solvents and Thermodynamic Properties of Mixing of Solutions

“…57-66-9, Figure ) has been applied in the treatment of gout for decades. − Previous studies showed that probenecid can influence the kidney physiology in the human body by the inhibition of the ATP transporter, which is named Pannexin 1, in the collecting duct and proximal tubule. Recent studies showed that probenecid can also protect a mouse against transient global cerebral ischemia/reperfusion injury in mouse model experiments. , Probenecid has prime pharmaceutical value and broad application.…”

“…Recent studies showed that probenecid can also protect a mouse against transient global cerebral ischemia/reperfusion injury in mouse model experiments. 6,7 Probenecid has prime pharmaceutical value and broad application.…”