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. 2019 Apr 25;19(1):88.
doi: 10.1186/s12906-019-2500-1.

Superoxide-producing lipoprotein fraction from Stevia leaves: definition of specific activity

A S Isoyan  1 , K V Simonyan  2 , R M Simonyan  3 , M A Babayan  3 , G M Simonyan  3 , V A Chavushyan  1 , M A Simonyan  3
Affiliations

Superoxide-producing lipoprotein fraction from Stevia leaves: definition of specific activity

A S Isoyan et al. BMC Complement Altern Med. .

Abstract

Background: Stevia rebaudiana Bertoni has various pharmacological actions, which includes antidiabetic, antioxidant, anti-inflammatory activities. The superoxide and consequently NADPH oxidase (Nox) are relevant targets involved in biological effects of Stevia. The presence of NADPH-containing superoxide-producing lipoprotein (suprol) in Stevia leaves has not yet been tested. The mechanism of producing superoxide radicals (O2-) by suprol was determined in vitro, which is associated with the electron transfer from NADPH in the composition of suprol by traces of transition metal ions (Fe3+ or Cu2+) to molecular oxygen, turning it into O2-. It is expected that the therapeutic efficacy of Stevia leaves is caused by specific activity of superoxide-producing lipoprotein fraction.

Methods: For the first time, from the dry leaves of Stevia the NADPH-containing superoxide-producing lipoprotein was isolated and purified. The specific content of suprol (milligrams in 1 g of Stevia leaves- mg/g) was determined after desalination of suprol and lyophilization.

Results: According to the method provided, the specific content of the isolated suprol from Stevia's leaves was up to 4.5 ± 0.2 mg / g (yields up to 68.5 ± 4.7%, p < 0.05, n = 6). Nox forms a stable complex with suprol. The optical absorption spectrum of the Nox-suprol complex represents the overlapping suprol and Nox spectra, with a certain background increase and characteristic features of optical absorption for Nox. Due to O2- producing capacity suprol-Nox complex discolors KMnO4 solutions, Coomassie brilliant blue, restores nitrotetrazolium blue to formazan and oxidizes epinephrine to adrenochrome. The oxidation activity of adrenaline is 50.3 ± 5.1 U / mg / ml (p < 0.05, n = 6).

Conclusion: Superoxide-producing lipoprotein fraction-Nox complex from Stevia leaves (membranes) can modulate redox regulated signaling pathways and may play a positive role in type-2 diabetes by means of adrenaline oxidation mechanism.

Keywords: NADPH-containing superoxide-producing lipoprotein (suprol); Stevia leaves.

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Conflict of interest statement

Authors’ information

A. Isoyan, PhD.

K. Simonyan, PhD.

R. Simonyan, PhD.

M. Babayan, PhD.

G. Simonyan, PhD.

V. Chavushyan, PhD, DSc.

M. Simonyan, PhD, DSc, Professor.

Ethics approval and consent to participate

All of the experimental protocols were approved by the Committee of Ethics of the Yerevan State Medical University (YSMU) (Yerevan, Armenia).

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Chromatographic elution graph of suprol after loading on a column with Biogel P-150 at pH 9.5. For further research, the central fraction was used (indicated by arrows) (p < 0.05, n = 6)
Fig. 2
Fig. 2
Optical absorption spectra of suprol dissolved in 0.005 M PPB at pH 9.5 within 2 h (1) and 30 h (2) at 4 °C after obtaining (p < 0.05 n = 6)
Fig. 3
Fig. 3
Optical absorption spectrum of Nox isoforms isolated from erythrocyte (1) and Stevia (2) membranes. Nox isolated from membranes of 10 ml of erythrocytes with a volume of 12 ml and from membranes of 10 g of dry Stevia leaves with a volume of 2 ml. Nox isoforms are dissolved in 0.1 M PPB, pH 7.4 (p < 0.05, n = 6)
Fig. 4
Fig. 4
Kinetic curves for KMnO4 (a-1) and PPB (b-1) discoloration, reduction of nitrotetrazolium blue, with the formation of formazan (c-1) under the influence of 0.2 mg/ml suprol associated fraction of Nox in the absence of (1 -●-) and the presence of 5 × 10(− 8) M Cu, Zn-SOD (2 -■-), at a temperature of + 25 °C
Fig. 5
Fig. 5
Formation of adrenochrome as a result of oxidation of adrenaline by superoxide radicals produced by suprol associated fraction of Nox of different concentrations at pH 7.4 in the absence of (1 -●-) and presence of 5.10− 8 M Cu, Zn-SOD (2 -■-)
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