Study of dry clover and chicory extracts with their effect on mouse splenocyte phenotype under methotrexate-induced immunosuppression

Abstract

The study of the quantitative content of phenolic compounds and the expression of antioxidant activity of the studied dry extracts showed that the level of phenolic compounds in the clover extract in terms of rutin was 6.50 ± 0.01%, the expression of antioxidant activity was 47.64 ± 2.15%, in the chicory extract the accumulation of phenolic compounds in terms of chicoric acid was 8.24 ± 0.01%, and the expression of antioxidant activity was 62.67 ± 1.85%. It was determined that dry extracts of clover and chicory do not have pronounced toxicity and are class 5 substances in accordance with GOST 32644-2014, and can be used in further research. A study of the effect of dry extracts on mouse and spleen weight, splenocyte count, splenic index, splenocyte-to-weight/spleen ratio, and splenocyte phenotype demonstrated that administration of the extracts did not cause statistically significant deviations from the intact group and the group of mice receiving the reference drug Immunal. Methotrexate, however, exerted a significant immunosuppressive effect, reducing mouse and spleen weight, splenocyte count, splenocyte-to-weight/spleen ratio, and splenic CD3+, CD4+, CD8+, and CD20+ lymphocyte levels compared to intact animals. Administration of dry extracts and Immunal, combined with methotrexate, resulted in a restoration of the studied parameters to values comparable to those in the intact group, indicating the immunocorrective effect of the dry extracts. The obtained data allow dry extracts of clover and chicory to be considered promising herbal remedies with immunomodulatory potential, as they possess antioxidant and immunotropic properties, as evidenced by the normalization of parameters characterizing the composition of splenic lymphocytes in animals exposed to an immunosuppressant. The effects of the extracts, comparable to those of the well-known drug «Immunal», suggest their potential use in supportive therapy for conditions with impaired immune response..