RESUMO
OBJECTIVE: To investigate the mechanism of Xianfang Huoming Decoction (XHD) improving sperm motility in mice with asthenospermia (AS). METHODS: Thirty normal BALB/c mice were randomly divided into six groups, blank control, AS model control, low-dose XHD, medium-dose XHD, high-dose XHD and levocarnitine + vitamin E (LC+VE). The AS model was established in the latter five groups by injection of methotrexate at 0.5 mg/kg once a week, and the mice in the blank control group were injected with the same volume of normal saline, all for 8 weeks. From the ninth week, the animals in the blank control and AS model control groups were treated with PBS at 0.1 ml/d, those in the low-, medium- and high-dose XHD groups with XHD at 7.13, 14,2 and 28.52 g/kg/ d respectively, and those in the LC+VE group with LC+VE (30:1) at 0.55 g/kg/d, all for 4 weeks. Then, the bilateral epididymides were harvested from all the mice for preparation of a sperm suspension and observation of the total numbers of sperm and motile sperm. The testis tissues were obtained for to determination of the expressions of Nrf-2- and HO-1-related mRNA and proteins by fluorescence staining, RT-PCR and Western blot. RESULTS: Compared with the AS model controls, the mice treated with low-, medium- and high-dose XHD showed dramatically increased sperm concentration (ï¼»22.36 ± 16.02ï¼½ vs ï¼»39.04 ± 4.50ï¼½, ï¼»40.76 ± 6.57ï¼½ and ï¼»41.04 ± 8.39ï¼½ ×106/ml, P < 0.01) and motility (ï¼»22.89 ± 14.96ï¼½% vs ï¼»47.98 ± 4.74ï¼½%, ï¼»48.53 ± 6.03ï¼½% and ï¼»49.31 ± 6.24ï¼½%, P< 0.01), decreased level of reactive oxygen species (ROS) (ï¼»16.82 ± 14.96ï¼½% vs ï¼»12.08 ± 3.26ï¼½%, ï¼»10.77 ± 2.21ï¼½% and ï¼»9.56 ± 2.08ï¼½%, P< 0.01), and up-regulated expressions of Nrf-2- and HO-1-related mRNA and proteins in the testis tissue (P < 0.05 or P < 0.01). CONCLUSION: Xianfang Huoming Decoction inhibits the development of oxidative stress by up-regulating the expressions of Nrf-2- and HO-1-related mRNA and proteins in the testis tissue, which has provided theoretical evidence for its clinical application in the treatment of asthenospermia.