Related Experiment Video
Updated: Jul 14, 2026
LPS and ATP-induced Death of PMA-differentiated THP-1 Macrophages and its Validation
Published on: May 3, 2024
Pyrrolidine dithiocarbamate attenuate shock wave induced MDCK cells injury via inhibiting nuclear factor-kappa B
Xiang Li1, Dalin He, Linlin Zhang
1Department of Urology, No. 1 Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi Province 710061, China.
Abstract:
Shock wave lithotripsy (SWL)-induced renal damage appears to be multifactorial. Recent data indicated that the mechanism of renal tissue damage secondary to SWL is similar to that of ischemia reperfusion injury. Nuclear factor-kappa B (NFkappaB) and its target genes, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), have been demonstrated to play a very important role in a variety of cells or tissues ischemia reperfusion injuries. Thus in the present study, using an in vitro model MDCK cells, we investigated the role of NFkappaB and its target cytotoxic enzyme in shock wave-induced renal cellular damage. We also examined whether inhibition this pathway by pyrrolidine dithiocarbamate (PDTC) is contributed to alleviate SWL-caused cell damage. Suspensions of MDCK cells were placed in containers for shock wave exposure. Three groups of six containers each were examined: control group, no shock wave treatment and SWL group, which received 100 shocks at 18 kV; 3 SWL + PDTC group. PDTC were added to the suspensions before shock wave exposure. After shock wave 0, 2, 4, 6 and 8 h, respectively, the cell supernatants were detected for the level of MDA and release of LDH. At post-shock wave 8 h, cells were harvested to detect the nuclear translocation of NFkappaBp65 by immunofluorescence staining. Degradation of IkappaB-a (an inhibitor protein of NFkappaB) and expression of iNOS and COX-2 were also examined by western blotting. Our results indicated that shock wave initiated the apparent activation of NFkappaB, which in turn induced high expression of iNOS and COX-2. Blocking degradation of IkappaB-a by PDTC was contributed to decrease the expression of iNOS. And the level of MDA and the release of LDH were also significantly reduced by using PDTC. However, the degree of COX-2 expression does not differ significantly between SWL and SWL + PDTC groups. Activation of NFkappaB and subsequent expression of its target cytotoxic enzyme have been demonstrated to be a potential and crucial mechanism in SWL-induced renal cell damage. Blocking this pathway by PDTC is contributed to protect against cellular damage from shock wave.
Insights
Shock wave lithotripsy (SWL) causes kidney damage through Nuclear factor-kappa B (NFkappaB) activation, leading to harmful enzyme production. Inhibiting this pathway with pyrrolidine dithiocarbamate (PDTC) protects kidney cells from SWL-induced injury.
Area of Science:
- Nephrology
- Molecular Biology
- Cellular Biology
Background:
- Shock wave lithotripsy (SWL) can cause kidney damage, potentially through mechanisms similar to ischemia reperfusion injury.
- Nuclear factor-kappa B (NFkappaB) and its downstream targets, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), are implicated in cellular injury.
Purpose of the Study:
- To investigate the role of NFkappaB and its target enzymes in SWL-induced renal cellular damage using an in vitro model.
- To evaluate the protective effect of pyrrolidine dithiocarbamate (PDTC) in inhibiting this pathway and mitigating SWL-induced cell damage.
Main Methods:
- MDCK cells were exposed to SWL with or without PDTC treatment.
- Cell damage markers (MDA, LDH) and NFkappaB pathway activation (NFkappaBp65 translocation, IkappaB-a degradation, iNOS, COX-2 expression) were assessed at various time points post-SWL.
Main Results:
- SWL induced NFkappaB activation, increasing iNOS and COX-2 expression.
- PDTC treatment reduced iNOS expression, MDA levels, and LDH release.
- PDTC did not significantly alter COX-2 expression levels compared to SWL alone.
Conclusions:
- NFkappaB activation and subsequent cytotoxic enzyme expression are key mechanisms in SWL-induced renal cell damage.
- Inhibiting the NFkappaB pathway with PDTC offers protection against SWL-induced cellular damage, particularly by reducing iNOS levels.
