Inhibition of monkey sperm hyaluronidase activity and heterologous cumulus penetration by flavonoids

A microplate assay for hyaluronidase and a heterologous cumulus penetration assay were used to determine the effects of four flavonoids (tannic acid, kaempferol, quercetin, and apigenin) on the function of cynomolgus monkey sperm. All four flavonoids inhibited the activity of hyaluronidase extracted from monkey sperm in a concentration-dependent manner over the range of 50-200 μM. Tannic acid and apigenin had lower inhibitory effects than kaempferol and quercetin. Kaempferol, quercetin, and apigenin at 100 μM were shown to significantly inhibit monkey sperm penetration into hamster cumulus. There was a significant linear relationship between the capacity of the flavonoids to inhibit monkey sperm hyaluronidase activity and their inhibitory effects on hamster cumulus penetration (r = 0.97). Tannic acid was observed to reduce sperm motility, and it was not used in the cumulus penetration assay. The other three flavonoids tested in the cumulus penetration assay did not affect sperm motility, nor did they induce acrosome reactions. The results demonstrate that the flavonoids are useful tools for assessing the involvement of hyaluronidase in the functions of monkey sperm that are involved in fertilization.