Influence Of Electroacupunture Combined With Magnetic Stimulation Therapy On The Expression Of AQP-4and MCP-1in Rat Spinal Cord Injury Model

Objective1. To verify whether acupuncture and magnetic stimulation the spinal cord can reduce the swelling and inflammation by observing the inflammatory changes and the changes of water content of spinal cord tissue after acute spinal cord injury and after giving acupuncture and magnetic stimulation;2.Observing the changes of the distribution and expression of AQP-4and MCP-1in rat spinal cord injury model;3Observing the changes of AQP-4and MCP-1expression after EA and magnetic stimulation to explore the mechanism of treatment of spinal cord injury of EA and magnetic stimulation by regulating the expression of AQP-4and MCP-1.MethodsForty SD rats were randomly divided into5groups:normal group, model group, electroacupuncture group, magnetic stimulation group, and combination therapy group. The spinal cord injury model was prepared by heavy objects fall to combat method. The rats are anesthetized with10%chloral hydrate by4ml/kg intraperitoneal injection, then they are fixed on the operation table with back hair removal and skin disinfection. Pursuant to the floating rib connecting to the as positioning signs, the location of spinous process of T8is determined by signing up three thoracic vertebra from the13th vertebrae. Then put the T8spinous process as the center, made a vertical midline incision about3cm. Next we exposed the T6-T10vertebral body, bited off the vertebral lamina of T8on both sides. Then we fully exposed the dura mater, let a lOg weight free falling down from a height of3cm and impacted on a homemade probe on dura mater whose length and diameter are both2mm. That the double lower limbs and body of the rat retracting and fluttering, rat tail appearing spastic swing proved that the model is successfully made. The normal group and the model group were not treated. The acupuncture point of "Guanyuan" was stimulated by electroacupuncture in acupuncture group by a density wave whose frequency is2/15Hz, strength is1mA,20min each time, once a day, lasting for10days. We placed a circular coil whose diameter is the40mm on the sacrum skin of rat of the magnetic stimulation group (to stimulate the S1-S4nerve root). The frequency of the stimulation was0.5Hz, the maximum output intensity is70%, each time gave30stimulation impulses. Each two stimulus had a2minutes interval, once a day treatment lasted for10days. Combination therapy group used electroacupuncture combined with magnetic stimulation therapy. The treatment methods, frequency, time are the same as acupuncture group and magnetic stimulation group. At the end of the treatment,4%paraformaldehyde solution are perfused for cardiac perfusion fixation. The tissue of spinal cord is removed and put into a4%paraformaldehyde solution. After a48hours fixation, Immunohistochemical methods were adopted to quantify the AQP-4and MCP-1positive cells of spinal cord.ResultsCompared with the normal group, the model group rat’s AQP-4and MCP-1expression density of the spinal cord was significantly enhanced which means the AQP-4, MCP-1protein expression was significantly increased and the difference was statistically significant (P<0.01); Acupuncture, magnetic stimulation group and combined treatment group rat spinal cord AQP-4, MCP-1expression density was significantly enhanced which means the AQP-4, MCP-1protein expression was significantly increased, the difference was statistically significant (P<0.01). Compared with the model group, acupuncture group, magnetic stimulation group and combined treatment group rat’ s spinal cord AQP-4, MCP-1expression density was significantly reduced which means the AQP-4, MCP-1protein expression was significantly reduced, and the difference was statistically significant (P <0.01). Compared with the combined treatment group, acupuncture group and magnetic stimulation group rat’s spinal cord AQP-4, MCP-1expression density was significantly enhanced which means AQP-4, MCP-1protein expression was significantly increased, and the difference was statistically significant (P<0.01).Magnetic stimulation group and the acupuncture group have no significant group differences (P>0.05).ConclusionEA and magnetic stimulation treatment can significantly reduce AQP-4, MCP-1levels in spinal cord injury, reduce the inflammation of spinal cord edema, and to some extent prevent secondary spinal cord injury. Electroacupuncture combined with magnetic stimulation therapy have a better effect.