[Animal experiment] -CaN-NFATc3 pathway in rat abdominal aortic restenosis after balloon dilation
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 type 1 herpes simplex virus 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 Flake Silk and Hargreaves experiments were used to test the mechanical tactile and thermal hyperalgesia of mice, and the expression of VGLUT3 in the dorsal root ganglia was evaluated by immunohistochemistry and Western blotting. \r\nResults: We have successfully constructed the type 1 herpes simplex virus recombinant vector HSV-1-shvglut3. After inoculation with the sciatic nerve, the vector can reverse the Vglut3-specific shRNA to the dorsal root ganglia. Two weeks after inoculating tactile pain model mice with HSV-1-shvglut3 vector, the expression of VGLUT3 in the dorsal root ganglia was inhibited, and the withdrawal threshold of the mechanical paw increased to the basic level. This analgesic effect lasted for more than 2 weeks without obvious systemic side effects. Similarly, the thermal pain threshold of mice did not change significantly. Conclusion: Vglut3 is expressed in the dorsal root ganglia and is a promising target for blocking tactile pain. The HSV-1-shvglut3 vector we constructed has specific tactile pain and has a safe, effective and lasting analgesic effect through peripheral inoculation. \r\nObjective: To study the role of calcineurin (CaN) and its downstream activated T cell nuclear factor (NFATc3) in restenosis after balloon dilation, and to establish a new theoretical basis for the prevention and treatment of vascular restenosis. provide. Methods: Male SD rats were randomly divided into sham operation group (n = 12), balloon group (n = 12) and cyclosporine (CsA) group (n = 12). The rats in the balloon group injured the abdominal aorta due to balloon expansion; the rats in the CsA group were given 12.5 mg/(kg.d)-1 CsA every day from before the operation to the third day before the end of the experiment. Thirty days after the balloon injury, specimens were collected, blood vessel tissues were stained with hematoxylin and eosin (HE), and the level of CaN in the blood vessel wall was detected by immunohistochemistry. Observe the pathological changes under an optical microscope. mRNA expression; Elisa method was used to determine serum MCP-1 level.
Results: After balloon injury, the neointima appeared on the blood vessel wall with uneven thickness; compared with the balloon injury group, the intimal hyperplasia and the intima/media thickness of the CsA group were significantly reduced (P\u003c0.05). Compared with the sham operation group, the expression of CaN protein and mRNA in the vascular wall tissue of the balloon injury group was significantly increased, the mRNA expression of NFATc3 was significantly increased, and the plasma inflammatory factor MCP-1 level was also increased (P\u003c0.05). The above indicators of the CsA group were significantly lower than those of the balloon injury group (P\u003c0.05).
Conclusion: CaN-NFATc3 pathway is involved in the development of restenosis after balloon injury in rats; CsA reduces the formation of restenosis by inhibiting this pathway.
动物实验
z-bo
2020-08-27
495