Comparison Of The Effect Of Ginsenoside Rb1 And Ginsenoside Rg1 On The Cultured Immature Sertoli Cells

Ginseng is one of the most important Chinese traditional medicines, whichhas been widely used for thousands of years. It contains a lot of components, amongwhich ginsenoside Rb1 and ginsenoside Rg1 are the main active ones. GRb1 andGRg1 play an important regulatory role in many human organs, tissues and cells. Inthe present study mouse primary sertoli cells were used as the model, to examineeffects of GRb1 and GRg1 on the proliferation of Cultured Immature Sertoli cells.These results lay foundations of GRb1 and GRg1 pharmacological action andapplications.Sertoli cells from the testes of 3 to 7 dpp of mouse testis were prepared bytwo-step enzymatic digestion. Sertoli cells were highly purified many times bycentrifugation. Vimentin and smooth muscle α-actin antibodies were used to furtherassess the purity of the Sertoli cell preparations.The results indicated that the cultureswere 95% ± 1.2% pure for Sertoli cells.In order to study the effect of GRb1 and GRg1 on cultured Sertoli cells, we firstused cell enumeration by supravital MTT staining assay for cell activity analysis andperformed enumeration of Sertoli cells in S-phase by BrdU labeling for numberanalysis. The results indicate that GRb1 takes part in supporting the living of sertolicells and certain concentration of GRb1 (20μg/ml) can promote the proliferation ofsertoli cells. With the same methods, The results indicate that GRg1 is not evidenceinfluence on the sertoli cells.Glial cell line-derived neurotrophic factor (GDNF) is a distantly related memberof the transforming growth factor-β (TGF-β) superfamily. Recently, there are somereports about the presence of its receptors GFR-α1, c-Ret and NCAM in testis. Inprevious studies GDNF was proved to be important for the development of mouseembryo and proliferation of sertoli cells in newborn rice. In our research, by RT-RCRand immunocytochemistry, it is confirmed that there is no c-Ret but GFRα1 andNCAM expression in sertoli cells. The results suggest that GDNF stimulates theproliferation of sertoli cells via GDNF-GFRα1/NCAM pathway.Our data confirmed that GRg1 did not significantly affect the proliferation ofsertoli cells and GDNF expression. However, GRb1 could stimulate the proliferationof sertoli cells via its pharmacological action to enhance the secret of GDNF insertoli cells. It suggests that the mechanism of the GRb1 stimulation is possiblelyperformed via GDNF/NCAM pathway.In summary, mouse primary sertoli cells were used as the model to examineeffects of GRb1 and GRg1 on the proliferation of cultured immature Sertoli cells andpossible signal transduction pathways. To our knowledge, it is the first report thatGRb1 could stimulate the proliferation of sertoli cells via its pharmacological actionto enhance the secret of GDNF in sertoli cells, but GRg1 did not. These resultsprovided some valuable information to reseach the difference of the GRb1 and GRg1pharmacological action and spermatogenesis.