动物实验,单纯疱疹病毒 z-bo 2020-08-27 543

　　OBJECTIVE: To construct a herpes simplex virus type 1 vector containing a specific RNA interference sequence targeting vesicular glutamine transporter 3 (Vglut3), and observe its effect on reducing the pain caused by touch in mice after sciatic nerve inoculation.

　　Method: First, construct a recombinant herpes simplex virus type 1 vector carrying Vglut3-specific short hairpin RNA (shRNA). The recombinant vector was inoculated through the sciatic nerve to test its analgesic effect in a mouse model of tactile pain. The von Freyksilk and Hargreaves experiments were used to test mechanical tactile and thermal hyperalgesia in mice, and to evaluate the expression of VGLUT3 in the dorsal root ganglia by immunohistochemistry and Western blotting.

　　Result: We have successfully constructed the recombinant herpes simplex virus type 1 vector HSV-1-shvglut3. After sciatic nerve inoculation, the vector can retrogradely transport Vglut3-specific shRNA to the dorsal root ganglia. Two weeks after the HSV-1-shvglut3 vector was inoculated into tactile pain model mice, the expression of VGLUT3 in the dorsal root ganglia was inhibited, and the withdrawal threshold of the mechanical paw of the mice was raised to the basal level. This analgesic effect lasted for more than 2 weeks without obvious systemic side effects, and the thermal pain threshold of the mice did not change significantly.

　　Conclusion: Vglut3 expressed in the dorsal root ganglia is a promising target for blocking tactile pain. The HSV-1-shvglut3 vector constructed by us has specific tactile pain, safe, effective and lasting analgesic effect through peripheral inoculation.