Objective To explore how dihydrotestosterone (DHT) affects the expression of anti-Müllerian hormone (AMH) in primary rat follicular granulosa cells, and then provides a new experimental basis for the role of DHT and AMH in follicular development.  Methods  Granulosa cells was extracted from the ovaries of 95 21 days SD female rats for primary culture. Firstly, cell morphology were detected by HE staining and follicke stimulating hormone receptor(FSHR) immunofluorescence detected cell purity after 48 hours of cell culture. Then, cells were randomly divided into control group (without drug intervention), 10-8mol/L DHT group, 10-5mol/L DHT group, 2×10^-5 protein kinase B(Akt) inhibitor (MK-2206 2Hcl) group and 10-8mol/L DHT+ 2×10^-5 mol/L MK-2206 2Hcl group according to different intervention method  and experiments，corresponding concentrations were used in each group to intervene cells for 3 hours. After intervention, sites and expression changes of AMH were detected by immunofluorescence. Changes of AMH, Akt and p-Akt were detected by Western blotting.   Results  Immunofluorescence showed that the purity of primary follicular granule cells in rats was 93.33%±3.09%,and AMH was expressed in the membrane and cytoplasm of primary follicular granule cells in rats; Compared with the control group, both 10-8 mol/L DHT and 10-5 mol/L DHT increased the expression of AMH in granulosa cells (P<0.05),but there was no difference in the effect of these two concentrations of DHT on the expression of AMH in granulosa cells (P>0.05); Compared with the control group, 10^-8 mol/L DHT increased p-Akt and AMH expression, and 2×10^-5MK-2206 2Hcl decreased AMH expression (P<0.05); Compared with the 2×10^-5 MK-2206 2Hcl, AMH expression was increased in the 10^-8 mol/L DHT+2×10^-5 MK-2206 2Hcl treatment group (P<0.05).   Conclusion  DHT can up-regulate the expression of AMH in primary follicular granulosa cells in rats, and this effect may be related to the Akt signaling pathway.