The role of glycyrrhiza glabra root aqueous extract in modulating the genotoxic effects of methotrexate in albino male mice and acute lymphoblastic leukaemia patients. +CD

The present study was carried out to shed light on the chemical analysis of Glycyrrhiza glabra for different active compounds, and on the cytogenetic effects of methotrexate(MTX) and aqueous extract of G. glabra in laboratory mice (in vivo) and human blood lymphocytes (in vitro). The study also aimed to investigate the role of G. glabra aqueous extract in reducing the cytogenetic effects of MTX in mouse through the parameters mitotic index, and chromosomal aberration in bone marrow, the total and differential counts of leukocytes, and sperm head and tail morphology. The cytogenetic analyses were extended to include blast index, mitotic index, replicative index, sister chromatid exchanges and chromosomal aberrations in human blood lymphocytes in vitro as a result of G. glabra aqueous extract and MTX treatment. The cytogenetic effects of the drug and plant extract were investigated after five days of treatment in mice with three doses for each. For MTX, the doses were 1.2, 1.6, and 2 mg/kg, while the doses of G. glabra were 400, 600, and 800 mg/kg. Using human blood lymphocyte (72hour) culture, the tested concentrations of MTX were 0.4, 2, and 4 µg/ml, and for G. glabra aqueous extract were 50, 100, and 200 µg/ml. An interaction between plant extract and MTX was carried out through two types of treatments (before, and after MTX treatment) to determine the activity of G. glabra aqueous extract in reducing the drug side effects in vivo. The following results were obtained: 1. Chemical analyses showed that G. glabra roots contain different active compounds such as saponins and flavonoids. 2.Methotrexate revealed clear effects in reducing mitotic index and increased chromosomal aberrations in mouse bone marrow cells. In human blood lymphocytes, the drug caused a reduction in blast index, mitotic index, replicative index, while the sister chromatid exchanges and chromosomal aberrations were increased. 3.Aqueous extract of G. glabra had a genotoxic effects at high doses in human blood lymphocytes culture (in vitro). 4.Methotrexate in human blood lymphocytes from patients with leukaemia(ALL) caused reduction in blast index, mitotic index, replicative index, but the increasing observed in chromosomal aberration, and sister chromatid exchanges in vitro. 5.In human blood lymphocytes of leukaemia patients (in vitro), extract caused reduction in blast index, mitotic index, sister chromatid exchanges, and chromosomal aberration, but the increasing observed in replicative index, except the concentration (200µg/ml) because of its genotoxic effect. 6.Aqueous extract of G. glabra at high dose, however, showed a protective value against the genotoxic effect of MTX in mouse bone marrow, this was more pronounced in pre-treatment than post-treatment. 7.Treatment with Glycyrrhiza glabra extract led to increase in the total and differential count of leukocytes, through increasing the number of ymphocytes and monocytes. 8.Methotrexate revealed clear reduction in the total and differential count of leukocytes by the reduction in lymphocytes and monocytes number, while neutrophils, basophils, and eosinophils were increased. 9.The induction in the abnormalities of sperm head and tail morphology were observed in MTX treatment especially after 35 days of treatment. 10.Treatment with G. glabra aqueous extract resulted in reduction of sperm abnormal morphology especially after 35 days of treatment.