CYTOTOXICITY AND TOXICOLOGICAL CHARACTERISTICS OF THE NEW LEUKOCYTAR POLYPEPTIDE

DOI: https://doi.org/10.29296/25877313-2020-05-08
Issue: 
5
Year: 
2020

T.A. Grishina Post-graduate Student, Department of Chemistry and Biotechnology, Perm National Research Polytechnic University (Perm) E-mail: tatyana_grishina_1990@mail.ru A.G. Volkov Ph.D. (Med.), Department of Pharmacology, Perm State Medical University name acad. E.A. Vagner (Perm) L.V. Volkova Dr.Sc. (Med.), Professor, Department of Chemistry and Biotechnology, Perm National Research Polytechnic University (Perm)

The aim of the study is to assess the safety of a low molecular weight leukocyte protein-peptide complex (LPPC) at the stage of preclinical studies. The object of the study was an experimental sample of a new leukocyte protein-peptide complex (LPPK) obtained by ultrasonic treatment of human leukocytes in vitro. LPPK cytotoxicity was assessed on the cell line of pig embryonic kidney epithelial cells (SPEV) and in vitro MTT test on human tu-mor cell lines HCT 116 (colorectal carcinoma) and MS (melanoma). A proliferation index was used to assess the effect on the growth of cells of the leukocyte protein-peptide complex (LBPC). Experiments on the study of acute toxicity of LPPK were performed on white nonlinear mice, which were in-jected intraperitoneally. To assess the results used the method of determining the average lethal dose according to V. Prozorovsky. When studying cy-totoxicity, it was found that the addition of an experimental sample of the leukocyte protein-peptide complex to the growth medium for culturing the SPEV cell line favorably affects cell growth. The proliferation index increased from the first to the fourth passage by 1.09 times, while in the control sample the increase in PI occurred by 1.03 times. A study of the cytotoxicity of LPPK showed that the sample did not exhibit a cytotoxic effect on tu-mor cells HCT 116 and MS, since the IC50 concentrations calculated for them in all cases were significantly lower than the reference drug Doxorubicin. The results of the assessment of acute toxicity in experimental animals showed that the test sample with intraperitoneal administration is low toxic according to the classification of GOST 12.1.007.-76 (LD50 3500.0 mg / kg). The article demonstrates the results of determining the toxicological and cytotoxic characteristics of the protein-peptide complex obtained from human leukocytes. It was shown that the studied protein-peptide complex did not exhibit toxic effects on the studied cell lines, as well as laboratory animals (mice).

Keywords: 
cytotoxicity
SPEV
leukocyte protein-peptide complex
acute toxicity
mice

References: 
  1. Engalucheva G.N., Syubaev R.D., Vasilev R.D., Snegireva A.A., Verstakova O.L. Otsenka farmakologicheskoi bezopasnosti lekarstvennykh sredstv v doklinicheskikh issledovaniyakh. Vedomosti Nauchnogo tsentra ekspertizy sredstv meditsinskogo primeneniya. 2013; 1: 10–13.
  2. Mironov A.N. Rukovodstvo po provedeniyu doklinicheskikh issledovanii lekarstvennykh sredstv. M.: Grif i K, 2012. S. 347–69.
  3. Eropkin M.Yu., Eropkin E.M. Fermentativnye metody otsenki tsitotoksichnosti na modeli kletok cheloveka v kulture: dinamika laktatdegidrratazy i glytationdegidratazy pod deistviem razlichnykh kontsentratsii DDS-Na. Toksilogicheskii vestnik. 1997; 5: 26–32.
  4. Morita M., Naito Y., Yoshikawa T., Niki E. Plasma lipid oxidation induced by peroxynitrite, hypochlorite, lipoxygenase and peroxyl radicals and its inhibition by antioxidants as assessed by diphenyl-1-pyrenylphosphine. Redox Biology. 2016; 8: 127–135.
  5. Romanova M.A., Dodonova A.Sh. Izuchenie tsitotoksichnosti biologicheski aktivnykh soedinenii v kulture kletok. Molodoi uchenyi. 2016; 18: 110–114.
  6. Volkova L.V., Grishina T.A., Volkov A.G. Fraktsionnyi sostav leikotsintarnogo lizata i ego biologicheskie svoistva. Sovremennye problem nauki i obrazovaniya. 2019; 1. URL: http://science-education.ru/ru/article/view?id=28527.
  7. Bernas T., Dobrucki J. Mitochondrial and nonmitochondrial reduction of MTT: interaction of MTT with TMRE, JC-1, and NAO mitochondrial fluorescent probes. Cytometry. 2002; 47(4): 236–242.
  8. Bolshakov O.P., Nrznanov N.G., Babakhyan R.V. Didakticheskie i eticheskie aspekty provedeniya issledovanii na biomodelyakh i na laboratornykh zhivotnykh. Kachestvennaya klinicheskaya praktika. 2002; 1: 58–61.
  9. Prozorovskii V.B., Prozorovskaya M.N., Demchenko V.M. Ekspress-metod opredeleniya srednei effektivnoi dozy i ee oshibki. Farmakologiya i toksikologiya. 1978; 4: 497–502.
  10. Freshni R.Ya. Kultura zhivykh kletok. M.: Laboratoriya znanii, 2018. S. 145–157.