With the aging of the population, more and more patients are suffering from aging-related erectile dysfunction (AED). The activation of JAK2 and its downstream pathway plays an important role in age-related diseases, but has not received attention in aging-related ED. CRISPR interference (CRISPRi) technology is the latest endogenous gene regulation technology developed based on the CRISPR/Cas9 system in recent years. It can inhibit the expression of target genes without changing the genome sequence. In this study, we aimed to investigate the role of JAK2 inhibition in ameliorating aging-related ED with CRISPR interference (CRISPRi) technology.

The lentivirus carrying the CRISPRi system and negative control (NC) lentivirus was constructed. 32 male rats were randomly divided into young group, AED group, AED + NC group and AED + JAK2-CRISPRi group. Phosphate-buffered saline was injected in young and AED group. Four weeks after intracavernous injection of JAK2-CRISPRi or NC lentivirus, the erectile function was evaluated by electrical stimulation. At the same time, corpus cavernosum smooth muscle cells were extracted from normal rats for primary culture and stimulated with H2O2 to simulate the process of cell aging in vitro. JAK2-CRISPRi lentivirus was also used to downregulate JAK2 gene expression in cells. Western blot, Elisa, immunohistochemistry, and immunofluorescence were performed to measure the changes in related molecules.

1. The results showed that the erectile function of aged rats was impaired, and was improved after JAK2 lentivirus injection. It reminded that the inhibition of the JAK2 gene expression can improve the erectile function. 2. In vivo experiments showed that the JAK2/STAT3 pathway was activated in AED group, thereby increasing the level of oxidation stress, apoptosis and autophagy in the penis. However, the above changes were reversed to a certain extent in the AED + JAK2-CRISPRi group. 3. The in vitro experiments obtained similar results to the in vivo experiments. Downregulation of JAK2 gene expression in cells improved the activation of the JAK2/STAT3 pathway, the increase in oxidative stress, apoptosis and autophagy levels caused by H2O2.

JAK2 inhibition with CRISPRi technology could reduce the JAK2/STAT3 pathway, thereby regulating the disturbance of oxidation stress, apoptosis levels and autophagy to play a therapeutic effect on aging-related ED. Our research provides new targets and ideas for the treatment of age -related ED.