Citrate and vitamin E blunt the shock wave-induced free radical surge in an in vitro cell culture model

Fernando C. Delvecchio, Ricardo M. Brizuela, Saeed R. Khan, Karen Byer, Zaiquan Li, Pei Zhong, Glenn M. Preminger

Research output: Contribution to journalArticlepeer-review

Abstract

Free radical formation plays a major role in shock wave lithotripsy induced renal damage. Moreover, previous studies suggest that free radicals may also promote de novo calcium oxalate crystallization of previously damaged urothelium. Citrate is a known inhibitor of renal stone formation and has also been used as a free radical scavenger. Using an in vitro model with Madin-Darby canine kidney (MDCK) cells, we investigated the influence of two free radical scavengers, citrate and vitamin E, on the prevention of the shock wave-induced free radical surge. Suspensions of MDCK cells were placed in containers for shock wave exposure. Six groups of six containers each were examined: (a) no scavengers 0 shocks, (b) no scavengers 100 shocks, (c) citrate 0 shocks, (d) citrate 100 shocks, (e) vitamin E 0 shocks, (f) vitamin E 100 shocks. An unmodified HM3 was used to deliver 100 shocks at 24 kV. The cell groups that were not shocked acted as the control group and were handled identically, except for the lack of shock wave exposure. After shock wave administration, the containers were emptied and cell suspensions were immediately centrifuged. The supernatant was examined for lactate dehydrogenase (LDH) and 8-isoprostane (8-IP), markers of cellular injury and free radical formation, respectively. Intracellular LDH uniformly increased in all groups exposed to shock wave energy. Similarly, 8-IP increased in all shocked groups. However, the 8-IP increase was significantly reduced when the free radical scavengers were employed. As citrate is a well-known inhibitor of calcium nephrolithiasis, its mechanism of action may be further enhanced, based on its ability to reduce free radical formation, by a protective effect on the urothelium. These data further support the use of citrate based medications during the peri-operative period of shock wave lithotripsy, not only to inhibit stone formation and facilitate fragment passage, but also to reduce the incidence of shock wave induced renal damage. Further studies are warranted to clinically test this hypothesis.

Original languageEnglish (US)
Pages (from-to)448-452
Number of pages5
JournalUrological Research
Volume33
Issue number6
DOIs
StatePublished - Dec 2005
Externally publishedYes

Keywords

  • Cellular damage
  • Citrate
  • Free radical scavengers
  • Free radicals
  • MDCK cells
  • Shock wave lithotripsy cell culture
  • SWL
  • Vitamin E

ASJC Scopus subject areas

  • Urology

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