L-Carnitine inhibits eryptosis induced by uremic serum and the related mechanisms

Abstract

Objective: To investigate whether L-carnitine (LC) inhibits eryptosis induced by uremic serum and the related mechanism. Methods: One percent erythrocyte suspension was cultured by three kinds of mediums in vitro, which was included in the control group (Group C, phosphate buffered saline [PBS]), the uremic serum group (Group U, 30% uremic serum + 70% PBS) and the LC group (Group L, 30% uremic serum + 70% PBS + 200 umol/L LC), respectively. Erythrocytes were collected at 24 and 48 h, respectively. Phosphatidylserine (PS) was estimated from Annexin-V-binding and reactive oxygen species (ROS) by flow cytometry, glutathione (GSH) was estimated from Enzyme linked immunosorbent assays (ELISA) by Microplate reader. Results: Eryptosis in Group C increased as the incubating time extended (3.43 ± 0.37 at 24 h, 4.21 ± 0.44 at 48 h). Eryptosis increased in Group U compared with Group C (6.5 1 ± 0.71 at 24 h, p < 0.01; 8.55 ± 0.76 at 48 h, p < 0.01), while decreased in Group L compared with Group U (5.80 ± 0.69 at 24 h, p < 0.05; 7.87 ± 0.76 at 48 h, p < 0.05). Meanwhile, ROS of erythrocytes increased in Group U compared with Group C (33.12 ± 1.61 versus 14.83 ± 2.22 at 24 h, p < 0.01; 42.06 ± 1.81 versus 20.94 ± 1.78 at 48 h, p < 0.01), and GSH decreased in Group U compared with Group C (25.66 ± 0.32 versus 31.27 ± 0.38 at 24 h, p < 0.01; 8.53 ± 0.59 versus 17.29 ± 0.54 at 48 h, p < 0.01). ROS of erythrocytes decreased in Group L compared with Group C (26.29 ± 1.69 at 24 h, p < 0.01; 36.21 ± 2.00 at 48 h, p < 0.01). GSH increased in Group L compared with Group U (27.54 ± 0.60 at 24 h, p < 0.01; 15.18 ± 0.42 at 48 h, p < 0.01). Conclusions: LC inhibits eryptosis induced by uremic serum, which possibly relates to oxidative stress in part.

• Eryptosis
• glutathione
• L-carnitine
• oxidative stress
• reactive oxygen species
• uremia

Acknowledgments

We thank research assistant Junli Liu (Department of Central Laboratory, the Second Hospital of Shandong University, Jinan, Shandong Province, China) for her elaborately measure in the trial.

Declaration of interest

The authors declare no conflicts of interest. This work was supported by Natural Science Foundation of Shandong Province (ZR2012L35), China.