Objective Although the clinical implementation of antiretroviral therapy ( cART) has significantly prolonged the lifetime of AIDS patients, HIV-associated neurocognitive disorders (HANDs) remain a critical concern in a considerable number of HIV-positive individuals. HIV-1Tat protein is a viral protein released from HIV-infected cells, which may be one of the underlying mechanisms behind HANDs. Methamphetamine (METH) can penetrate the blood-brain barrier, damaging nerve cells and increasing the risk of HANDs. The combination of HIV-1Tat and METH can induce autophagy and apoptosis in nerve cells, but the regulatory mechanism behind this has remained unclear. Methods In this study, the primary midbrain neurons of tree shrews were used to determine the expression of ATG5 / 7 proteins and genes induced by HIV-1Tat and METH via Western blot and real-time PCR. Results HIV-1Tat protein and METH induced autophagy in tree shrews’ primary midbrain neurons and had a synergistic effect. ATG5/7genes were involved in this autophagy. Conclusions ATG5/7 genes are involved in HIV-1Tat protein and METH-induced autophagy of nerve cells, which provides a new theoretical basis for studying the relationship between HANDs and autophagy.