Background: Glycyrrhiza glabra(Gg) is a herbal medicine used in the treatment of different syndromes. Recently, the effect of this plant on the reproductive system has been studied. However, its role in in vitro still not clear. Objectives: The present study aims to investigate the possibility of using Glycyrrhiza glabra extract for in vitro sperm direct activation, and its effect on in vivo fertilization rate (FR) and early embryonic development (ED) following intra-peritoneal insemination (IPI). The mice were used as an experimental model for mammals. Materials and Methods: Glycyrrhiza glabra extract (2 mg/ml culture medium) was used for in vitro direct sperm activation technique. The female mice were divided into two groups: spontaneously ovulated (SOM) and superovulated groups (SuOM). Each group was divided into two subgroups, the first: the IPI was accomplished by epididymal sperm activated in vitro by adding 20%Gg to the culture medium. The second subgroup: the IPI is accomplished without adding Gg to the culture medium. Results: In vitro activation of epididymal sperms with 20% Gg has shown positive effects on sperm concentration, sperm motility, and grade activity of progressive forward movement. There was a significant (P<0.05) increase in FR with adding 20%Gg compared with that free of Gg of spontaneously ovulated groups after 24 hours of insemination. The FR in the right oviduct showed highly significant (P<0.001) increment compared to the left. Embryonic development rate significantly ( (P<0.05) increased after 24 and 48 hours of insemination in grouping with adding Gg. The study showed that the quantity and quality of embryos generated from IPI with adding Gg were higher than that of groups inseminated without adding Gg. Conclusion: It is concluded from the results of the present study that adding the 20% Gg to the culture medium of the epididymal sperm and IPI leads to an improvement in certain sperm function parameters and supports the FR in SOM and SuOM with an increase in the embryonic development rate. These data can be utilized for artificial insemination programs.
Male infertility is a global health issue affecting over 20 million males. A spermatogenesis-related cell differentiation abnormality causes teratozoospermia (TZS). Extensive research has linked TZS to poor fertilization outcomes at assisted reproductive technology (ART) centers as well as to spontaneous pregnancies. This study used Glycyrrhiza glabra (Gg) and Platelet Rich Plasma (PRP) media to establish a culture medium that aimed to improve certain sperm functions of teratozoospermic men in vitro. A total of fifty semen samples were examined. The World Health Organization 2021 and 1999 standards were used to evaluate certain sperm parameters and the DNA fragmentation index. Platelet-rich plasma and Gg extract were utilized as a medium for in vitro preparation and activation of semen samples complaining of teratozoospermia factor. Following activation by the utilization of the density gradient approach and the combination of Gg and PRP medium, there was a statistically significant ( p <0.001) improvement in certain sperm function metrics. These improvements included a higher percentage of morphologically normal sperm, a rise in active motility, and a drop in the DNA fragmentation index. The density gradient technique, regardless of the medium used in this investigation, resulted in a significant enhancement of morphologically normal sperms with other certain sperm parameters and a decrease in the DNA fragmentation index.
Background: Many studies found that the Glycyrrhiza glabra (G.glabra) showed a positive effect on the activation of sperm in vitro specifically improve the activity of progressive movement. However, it was not known the impact of this stimulant on the certain sperms parameters in vasal obstructive mammals. Objectives: This experiment was designed to examine the possibility of fertilization ova by epididymal sperm of vasectomized mice activated by G. glabra and study the early embryonic development, as a model for obstructive azoospermia. Methods: Glycyrrhiza glabra aqueous extract (0.3 mg/ml culture medium) was used for in vitro direct sperm activation technique. The superovulated female mice (SUO) were divided into two groups, the first group; the insemination of flushed ova was performed with epididymal sperm of vasectomized male mice activated by adding30% of G. glabra to the culture medium (treated group).While the second group,the ova were inseminated by epididymal sperm of vasectomized male mice activated by G. glabra - free medium (control). Results: This study showed a highly significant (P <0.01) improvement in certain sperm function parameters i.e the sperm forward movement, morphologically normal sperms (MAS). There was a significant (P<0.05) increase in fertilization rate (FR) of SUO mice oocyte using 30% G. glabra medium(75.84%) compared to G. glabraFree medium (Hams -F12 medium) alone (54.43%).By adding 30% G. glabra to the medium, the number of 3-cell and 4-cell embryonic stage of SUO mice was significantly (P <0.05) higher than that of grouping G. glabra- Free medium mice embryo(62.10% and 65.78% vs. 58.88 and 66.29%, respectively). Conclusion: This study showed a highly significant (P <0.01) improvement in certain sperm function parameters i.e the sperm forward movement, morphologically normal sperms (MAS). There was a significant (P<0.05) increase in fertilization rate (FR) of SUO mice oocyte using 30% G. glabra medium(75.84%) compared to G. glabraFree medium (Hams-F12 medium) alone (54.43%).By adding 30% G. glabra to the medium, the number of 3-cell and 4-cell embryonic stage of SUO mice was significantly (P <0.05) higher than that of grouping G. glabra- Free medium mice embryo(62.10% and 65.78% vs. 58.88 and 66.29%, respectively).
Background: Many studies conducted on the pentoxifylline (PX) and Glycyrrhiza glabra (G.glabra) as motility stimulants showed positive effect on sperm preparation in vitro improving the progressive forward movement. However, it was not known the impact of these stimulants on the nature of genetic material, especially after cryopreservation. Objective: The experiment was designed to find out any harmful effect of a medium containing a mixture of PX and aqueous extract of G.glabra on the sperm DNA before and after cryopreservation. Materials and Methods: Fifty mature fertile male mice 8-12 weeks old were used. Epididymal sperms were obtained from caudal region and prepared in vitro by direct activation technique using four media namely; PX alone, G.glabra alone, a mixture of PX with G.glabra and Ham's F-12 (as control medium). Certain sperm function characteristics were examined as well as evaluation of DNA integrity by acridine orange test and comet assay before and after cryopreservation. Results: The results showed a highly significant (P<0.001) improvement in the sperm forward movement, with a highly significant (P<0.001) decrease in DNA abnormality using acridine orange test and comet assay, following in vitro activation by the four media compared to before activation. Following in vitro activation and cryopreservation, the results of epididymal sperm showed a highly significant (P<0.001) decrease in DNA abnormality by using PX medium alone, G.glabra medium alone and the mixture of PX and G.glabra compared to control medium and the results before cryopreservation. Conclusion: According to the results of present study, it has been found no toxic effect of medium containing PX and/or G.glabra on the epididymal sperms DNA after activation in vitro and cryopreservation using acridine orange test and comet assay. The best result obtained when using a medium containing a mixture of PX+ G.glabra together. These data could be applied for preparation and activation of epididymal and testicular sperms obtained from obstructive azoospermic men.