Abstract: Objective: To investigate the improvement effect of ginsenoside Rg1 combined with ginsenoside Rg3 on the reproductive function in the dinbutyl phthalate (DBP)-induced reproductive function injury model mice, to elucidate the mechanism of ginsenoside Rg1 combined with ginsenoside Rg3 in the protection of reproductive function, and to provide the experimental basis for their further development. Methods: Twenty-eight clean grade male C57BL/6 mice were randomly divided into model group (400 mg·kg^-1 DBP),ginsenoside Rg1+DBP group (20 mg·kg^-1 ginsenoside Rg1+400 mg·kg^-1 DBP),ginsenoside Rg3+DBP group (20 mg·kg^-1 ginsenoside Rg3+ 400 mg·kg^-1 DBP) and combination group (20 mg·kg^-1 ginsenoside Rg1+20 mg·kg^-1ginsenoside Rg3+400 mg·kg^-1 DBP); there were 7 mice in each group. DBP dissolved in corn oil was given to the mice by gavage every morning,ginsenoside Rg1 and ginsenoside Rg3 were dissolved in normal saline and injected subcutaneously in the mice every afternoon, and DBP,ginsenoside Rg1 and ginsenoside Rg3 were administered once a day for 35 consecutive days. The sperm density, sperm viability and total malformation rate of the mice in each group were detected by Wljy-9000 sperm quality detection system. HE staining was used to observe the patholmorphology of testis tissue of the mice. The levels of serum testosterone (T), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were detected by enzyme-linked immunosorbent assay (ELISA). The expression level of Cx43 protein in testis tissue was detected by immunofluorescence assay. Results: Compared with model group, the testicular organ coefficient of the mice in combination group was significantly increased (P<0.05), and the sperm densities and the sperm viabilities of the mice in ginsenoside Rg1+DBP group, ginsenoside Rg3+DBP group and combination group were significantly increased (P<0.01). Compared with model group, the levels of serum T and LH of the mice ginsenoside Rg1+DBP group, ginsenoside Rg3+DBP group and combination group were significantly increased(P<0.01) and the FSH levels were significantly decreased in(P<0.01); the expression levels of Cx43 protein were significantly increased (P<0.05). The HE staining results showed that the spermatogenic epithelium of the testis tissue of the mice in model group was relatively thin, and the spermatogenic cells in the duct cavity were seriously exfoliated; compared with model group, the spermatogenic epithelium of the testis tissue of the mice in ginsenoside Rg1+DBP group and ginsenoside Rg3+DBP group became thicker, and fewer exfoliated cells were found in the lumen; the spermatogenic epithelial structure of the testis tissue in combination group was complete, with regular spermatogenic cells at all levels and abundant intracellular sperm. The result of factorial analysis showed that there was an interaction between ginsenoside Rg1 and ginsenoside Rg3, which resulted in the increase of sperm viability(F=6.704, P=0.016), the serum T level(F=4.912, P=0.036)and the expression level of Cx43 protein in testis tissue(F=6.937, P=0.018). Conclusion: Ginsenoside Rg1 and ginsenoside Rg3 used alone or combined can improve the sperm density and the sperm viability, increase the serum T and LH levels, decrease the FSH level in the mice, and reduce the reproductive function injury in the mice, and its mechanism might be related to the protective role in spermatogenesis by up-regulating the expression level of Cx43.Ginsenoside Rg1 combined with ginsenoside Rg3 has a synergistic effect.

Key words: dinbutyl phthalate, ginsenoside 1, ginsenoside 3, combined effect, reproductive function