Objective: To study the effect of the serum containing Zhibai Dihuang Decoction (ZDD) on the energy metabolism of spermatogenic cells infected with Ureaplasma urealyticum (UU) in rats and its action mechanism.
Methods: Healthy male SD rats were randomly divided into six groups, normal control, UU-infection (UUI) model control, doxycycline, and low-, medium- and high-dose ZDD-containing serum. After successful establishment of the UUI model in vivo in the latter five groups, the rats in the normal control group were treated with simple serum and those in the latter five with respective agents. Then primary spermatogenic cells were harvested from the rats for examination of the apoptosis of spermatogenic cells, contents of lactate dehydrogenase (LDH) and adenosine triphosphate (ATP), glucose disposal rate (GDR) and expressions of AMPK and PARα proteins in the spermatogenic cells, and other related parameters.
Results: The apoptosis rate of the spermatogenic cells was dramatically increased in the UUI model controls compared with that in the normal controls (［49.24 ± 0.86］% vs ［10.09 ± 0.52］%, P < 0.01), but significantly decreased in the doxycycline and low-, medium- and high-dose ZDD groups (［11.21 ± 1.02］%, ［30.64 ± 0.99］%, ［35.54 ± 1.17］% and ［42.95 ± 1.31］%) in comparison with that in the UUI model control group (P < 0.01).The content of LDH in the spermatogenic cells was also remarkably increased in the UUI model controls compared with that in the normal controls (［201.12 ± 2.88］ vs ［60.72 ± 1.83］) mU/ml, P < 0.01), but significantly decreased in the doxycycline and low-, medium- and high-dose ZDD groups (［90.66 ± 1.61］, ［94.74 ± 1.20］, ［101.24 ± 2.03］ and ［111.04 ± 3.35］ mU/ml) in comparison with that in the UUI model control group (P < 0.01). The GDR in the spermatogenic cells was markedly reduced in the UUI model controls compared with that in the normal controls (［49.42 ± 1.70］% vs ［99.86 ± 1.26］%, P < 0.01), but significantly elevated in the doxycycline and low-, medium- and high-dose ZDD groups (［86.90 ± 2.03］%, ［84.14 ± 1.21］%, ［80.30 ± 1.37］% and ［75.18 ± 1.76］% in comparison with that in the UUI model control group (P < 0.01). The content of ATP was also dramatically decreased in the UUI model controls compared with that in the normal controls (［19.76 ± 1.46］ vs ［58.94 ± 1.95］ μmol/L, P < 0.01), but significantly increased in the doxycycline and low-, medium- and high-dose ZDD groups (［48.34 ± 1.34］, ［42.82 ± 1.30］, ［38.70 ± 2.03］ and ［34.78 ± 0.82］ μmol/L) in comparison with that in the UUI model control group (P < 0.01). The mitochondrial membrane potential was remarkably elevated in the UUI model controls compared with that in the normal controls (［8.53 ± 0.71］% vs ［2.43 ± 0.25］%, P < 0.01), but markedly reduced in the doxycycline and low-, medium- and high-dose ZDD groups (［3.92 ± 0.36］%, ［4.43 ± 0.27］%, ［4.65 ± 0.22］% and ［4.88 ± 0.10］% in comparison with that in the UUI model control group (P < 0.01). The phosphorylation levels of AMPK and PPARα proteins were significantly up-regulated in the UUI model controls compared with that in the normal controls (P < 0.01), but down-regulated in a dose-dependent manner in the ZDD groups.
Conclusions: Zhibai Dihuang Decoction can significantly improve the damage to the mitochondrial structure and inhibit UU infection-induced apoptosis of spermatogenic cells and secretion of LDH by increasing the ATP content and GDR and regulating the phosphorylation of AMPK and PARα signaling pathway.
Keywords: Zhibai Dihuang Decoction; asthenospermia； rat; spermatogenic cells; Ureaplasma urealyticum.