In vitro effects of L-arginine on lysosomal cysteine proteases activity in isolated experiment and in the state of oxidative stress
- Authors: Fomina MA1, Kudlaeva AM1
-
Affiliations:
- Ryazan State Medical University after I.P. Pavlov, Ryazan, Russia
- Issue: Vol 96, No 5 (2015)
- Pages: 876-882
- Section: Experimental medicine
- URL: https://journals.rcsi.science/kazanmedj/article/view/2047
- DOI: https://doi.org/10.17750/KMJ2015-876
- ID: 2047
Cite item
Full Text
Abstract
Aim. Assessment of direct influence of arginine on lysosomal cysteine proteases activity in vitro, in isolation as well as the stimulation of oxidative stress.
Methods. The study was conducted on the 72 female conventional mature Wistar rats 280-320 g divided into 6 series of 12 rats each. Lysosome slurries were isolated from the liver of intact animals with a subsequent in vitro incubation in a sucrose solution, in the presence of L-arginine, as well as in the presence of L-arginine accompanied by the stimulation of oxidative stress. Samples of control groups were exposed in vitro with the addition of isolate and oxidant, respectively. Each batch was reproduced three times, incubation was performed at 37 °C in a water bath for 1, 2 and 4 hours. The activity of cathepsins B, L and H was studied using spectrofluorimetric method in two fractions - intra- and extralysosomal. Acid phosphatase activity was used as the main marker of membrane labialization.
Results. One hour Incubation with 5 mM arginine in vitro led to inhibition of the cathepsin H activity and lysosomal membrane damage, however, further increase in incubation time led to its stabilization. In vitro exposure to 5 mM H2O2 caused an increase in activity of cathepsines B and L and the drop in the cathepsin H activity without obvious changes in the distribution of enzymes between extra and intralysosomal fractions. In a state of oxidative stress 2-hour in vitro incubation with 5 mM arginine reduced the permeability of lysosomal membranes for cathepsines B, H and L; while 4-hour incubation led to the destabilization of lysosomal membranes.
Conclusion. The direct effect of arginine at a concentration of 5 mM within the 1,2 and 4-hour time intervals leads to a distinct change as a lysosomal cysteine protease activity and stability of lysosomal membranes.
Keywords
About the authors
M A Fomina
Ryazan State Medical University after I.P. Pavlov, Ryazan, Russia
Author for correspondence.
Email: anyakudlaeva@mail.ru
A M Kudlaeva
Ryazan State Medical University after I.P. Pavlov, Ryazan, Russia
Email: anyakudlaeva@mail.ru
References
- Арапова А.И., Фомина М.А. Эффекты l-аргинина на состояние лизосомального цистеинового протеолиза сердечной и скелетной мышц // Фундаментал. исслед. - 2014. - №10. - С. 1269-1273.
- Васильева О.С. Комплексное участие цистеиновых катепсинов в раковой прогрессии [Электронный ресурс] // Электрон. научн. ж. «ИССЛЕДОВАНО В РОССИИ» 677. - http://zhurnal.ape.relarn.ru/articles/2009/055.pdf (дата обращения: 07.12.2014).
- Дилакян Э.А., Цветкова И.В. Лизосомные цистеиновые протеиназы при неопластической трансформации // Биомед. хим. - 2005. - Т. 51, вып. 5. - С. 485-500.
- Пупышев А.Б. Пермеабилизация лизосомных мембран как апоптогенный фактор // Цитология. - 2011. - Т. 53, №4. - С. 313-324.
- Степанов Ю.М., Кононов И.Н., Журбина А.И., Филиппова А.Ю. Аргинин в медицинской практике (обзор литературы) // Ж. АМН України. - 2004. - Т. 10, №1. - С. 340-352.
- Barrett A.J., Kirschke H. Cathepsin B, cathepsin H, cathepsin L // Methods in Enzymol. - 1981. - Vol. 80. - P. 535-561. http://dx.doi.org/10.1016/S0076-6879(81)80043-2
- Felbor U., Dreier L., Bryant R.A. et al. Secreted cathepsin L generates endostatin from collagen XVIII // EMBO J. - 2000. - Vol. 19, N 6. - P. 1187-1194. http://dx.doi.org/10.1093/emboj/19.6.1187
- Hamel F.G., Upward J.L., Siford G.L. et al. Inhibition of proteasome activity by selected amino acids // Metabolism. - 2003. - Vol. 52, N 7. - Р. 810-814. http://dx.doi.org/10.1016/S0026-0495(03)00094-5
- Horn M., Pavlik M., Doleckova L. et al. Arginine-based structures are specific inhibitors of cathepsin C. Application of peptide combinatorial libraries // Eur. J. Biochem. - 2000. - Vol. 267, N 11. - Р. 3330-3336. http://dx.doi.org/10.1046/j.1432-1327.2000.01364.x
- Shaw E., Kettner C. The specificity of cathepsin B // Acta Biol. Med. Ger. - 1981. - Vol. 40, N 10-11. - Р. 1503-1511.
- Terman А., Kurz T., Gustafsson B. et al. Lysosomal labilization // IUBMB Life. - 2006. - Vol. 58, N 9. - P. 531-539. http://dx.doi.org/10.1080/15216540600904885
- Zhang L., Lockwood T.D. Phenylalaninylargininylarginine: a novel tripeptide exerting Zn(2+)-dependent, insulin-mimetic inhibitory action on myocardial proteolysis // Biochem J. - 1993. - Vol. 293, pt. 3. - Р. 801-805. http://dx.doi.org/10.1042/bj2930801