Human umbilical cord-derived mesenchymal stem cells and nitric oxide synthase modulators inhibit apoptosis and activate autophagy in the lungs of mice with an antiphospholipid syndrome model

Abstract

Antiphospholipid syndrome is one of the most common causes of hypercoagulability. Antiphospholipid syndrome is characterized by thrombotic and obstetric complications, the formation of antiphospholipid antibodies. Molecular phospholipid-protein complexes formed during apoptosis are the target of antiphospholipid antibodies. Considering that endothelial dysfunction plays an important role in the pathogenesis of antiphospholipid syndrome, it is relevant to study the effect of NO synthesis modulators, as well as human mesenchymal stem cells, which have regenerative, anti-inflammatory and immunomodulatory effects on the mechanisms of lung damage in this pathology. Purpose – to investigate the effect of human umbilical cord–derived mesenchymal stem cells and nitric oxide synthase modulators (L-arginine, aminoguanidine) on the level of antiapoptotic markers Bcl-xL and Bcl-xL-BAX and autophagy marker BECN1 in the lungs of mice with experimental antiphospholipid syndrome. Material and methods. Antiphospholipid syndrome was modeled in female BALB/c mice by administering cardiolipin 1.2 mg/kg 4 times with an interval of 14 days. For correction, human umbilical cord-derived mesenchymal stem cells (5×106 cells/kg), L-arginine (25 mg/kg) and aminoguanidine (10 mg/kg) were used, which were administered intraperitoneally once a day for 10 days after the development of APS. To separate proteins from lung tissue, electrophoresis was performed in a polyacrylamide gel in the presence of sodium dodecyl sulfate. For the purpose of immunochemical detection of Bcl-xL, Bcl-xL-BAX, BECN1, proteins were transferred from the gel to a nitrocellulose membrane by electroblotting. Densitometric analysis of immunoreactive zones was performed using the Total Lab TL120 software. Results. In the group of animals with APS, the level of Bcl-xL was lower by 29 %, and Bcl-xL-BAX by 30 %, BECN1 was lower by 42 % compared to the indicators in the animals of the control group. In the group of animals where L-arginine and aminoguanidine were used, an increased level of Bcl-xL protein and Bcl-xL-BAX complex by 36 % and 90 %, respectively, was found, the level of BECN1 was 3 times higher compared to the group of animals with APS. In the group of animals with APS, which were corrected with mesenchymal stem cells, the level of Bcl-xL was 149 % higher, the level of BECN1 increased by 133 % compared to animals with pathology. No significant differences were found in the level of Bcl-xL-BAX. In the group of animals with combined administration of corrective substances – nitric oxide synthase modulators (L-arginine, aminoguanidine) and mesenchymal stem cells – the level of Bcl-xL was 112% higher, and the level of BECN1 was 143% higher compared to animals with APS, while the Bcl-xL-BAX did not differ significantly. Conclusion. In the lungs of mice with antiphospholipid syndrome, inhibition of autophagy and reduced levels of antiapoptotic protein markers were found. With the separate and combined use of human umbilical cord-derived mesenchymal stem cells and nitric oxide synthase modulators (L-arginine, aminoguanidine) for the correction of APS, higher values of antiapoptotic markers Bcl-xL and autophagy protein BECN1 were determined.