| Background: With the enhancement of scientific and technological development and social progress,the pace of people's life is getting faster and faster,which leads to a higher incidence of spinal cord injury(SCI).SCI,a serious wound on the central nervous system accompanied with many complications and high disability rate,has brought great burden to the families of patients and the society due to high cost of treatment.The deficits of sensorimotor function below the injury plane are directly caused by SCI,resulting in people living a difficult life.Since global attention focus on alleviating the dysfunction,it is urgent to find a feasible,economical and effective treatment for SCI.With the development and popularization of traditional Chinese medicine(TCM)treatment,increasing studies have confirmed the good effect of Du meridian electroacupuncture(EA)in the treatment of spinal cord injury,but little is known about the underlying treatment mechanism.As sports medicine has expanded rapidly,rehabilitation therapy appears to be a better way to treat SCI.Though there are numerous ways of rehabilitation therapy applied in common life,no specific training program can be provided,and few studies has been carried out on combination therapy.Objective: To investigate whether EA combined with Wheeled treadmill training could improve the function of rats with spinal cord injury;to study whether EA combined with Wheeled treadmill training exert better effect than treatment with EA or Wheeled treadmill training alone;to elucidate the morphological basis of EA combined with Wheeled treadmill training alleviating neurological dysfunction;to explore the regulatory mechanism so as to provide a new therapeutic target and insight into the clinical treatment of SCI.Methods: Fifty adult female healthy specific pathogen-free(SPF)sprague dawley(SD)rats,weighing 250 ~ 300 g,were selected and randomly divided into 5groups(n=10)as follows: Sham group,Control group,EA group,Wheeled treadmill group,EA+Wheeled treadmill group.A week before the formal trial,20 minutes ofwheeled treadmill adaption training was conducted every day,followed by SCI model establishment.Afterwards,7 consecutive weeks of EA,wheeled treadmill and combined treatment were employed in the indicated groups,with body weight and urine volume measured once every Saturday.BBB score was used to detect the motor function recovery of hind limbs,tail flick latency test for the sensory function recovery.The inclined plane test was conducted every two weeks.Seven weeks later,Magnetic Resonance Imaging was performed to detect the anisotropic fraction and the number of nerve fibers at the injured site of the spinal cord,PET-CT to detect the energy metabolism of injured site,bladder and hindlimb,electrophysiological experiments to test the control ability of cortical motor area to the muscle of the hind limb and the electromyography for the hind limb of the rats after spinal cord injury repair.At last,injured site of spinal cord was harvested for Nissl staining and immunofluorescence staining to observe the recovery of rats after SCI.Results: 1)Rats weight demonstrated varying degrees of decline in each group and rats weight was rebounded as treatment period prolonged.The weight regain degree of rats in the Sham group was significantly higher than that in the control group and the other treatment groups.Rats in control group regained weight obviously compared to EA group,Wheeled treadmill group and EA+Wheeled treadmill group.Compared with EA group and Wheeled treadmill group,rats weight in EA+Wheeled treadmill group regained more notably.2)The urine volume of the rats in each group decreased to different degrees with the prolonging of treatment time.Compared to the control group,the urine volume in EA group,Wheeled treadmill group and EA+Wheeled treadmill group was obviously decreased.Moreover,compared with the single treatment group alone,the combined treatment group showed a more significant decline of urine volume.3)BBB scores in the Sham group remained the highest(21 scores),and all the groups exhibited a gradual upward trend as the treatment period prolonged.Compared with the treatment group alone,BBB scores in EA group,Wheeled treadmill group and EA+Wheeled treadmill group increased significantly compared to Control group,and the combined treatment group showed a higher score than that in the single treatment group alone.4)In the Tail Flick Latency test,the rats tolerance ability of mechanical pain threshold in Shamgroup was obviously higher than that of control group and three treatment groups,and the mechanical pain threshold of rats tolerance ability were significantly elevated as treatment period prolonged.Pain threshold in EA group,Wheeled treadmill group and EA+Wheeled treadmill group decreased significantly compared to Control group,and the combined treatment group demonstrated a marked increase relative to that in the single treatment group alone.5)In the inclined plane test,the maximum gradient at which the rats stayed on the inclined plane in Sham group was obviously higher than that of control group and three treatment groups,and the maximum gradient augmented obviously as treatment period prolonged.Maximum gradient in EA group,Wheeled treadmill group and EA+Wheeled treadmill group rised significantly compared to Control group,and maximum gradient in the combined treatment group demonstrated a marked increase relative to that in the single treatment group alone.6)In the MRI detection,the fractional anisotropy of rats in Sham group was obviously higher than that of control group and three treatment groups,and the fractional anisotropy augmented obviously as treatment period prolonged.Fractional anisotropy in EA group,Wheeled treadmill group and EA+Wheeled treadmill group rised significantly compared to Control group,and fractional anisotropy in the combined treatment group demonstrated a marked increase relative to that in the single treatment group alone.7)In the PET-CT detection,the energy metabolism of injured site,bladder and hindlimb in Sham group was obviously higher than that of control group and three treatment groups,and the energy metabolism of injured site,bladder and hindlimb augmented obviously as treatment period prolonged.Energy metabolism of injured site,bladder and hindlimb in EA group,Wheeled treadmill group and EA+Wheeled treadmill group rised significantly compared to Control group,and energy metabolism of injured site,bladder and hindlimb in the combined treatment group demonstrated a marked increase relative to that in the single treatment group alone.8)As for electromyogram motor-evoked potential amplitude,amplitude difference and area of Sham group was obviously higher than that of control group as well as other three treatment groups.Amplitude difference and area in EA group,Wheeled treadmill group and EA+Wheeled treadmill group rised significantly compared to Control group,and amplitude difference and area in thecombined treatment group demonstrated a marked elevetion relative to that in the single treatment group alone.9)In the morphological detection,the nissl body of neurons in the Sham group presented normal state with large number and plump body.In the control group,nissl body showed abnormal morphology with fewer neurons,visible edema,cavitation and even fragmentation.Compared with the control group,the number of neurons in each treatment group increased,and their morphology improved significantly,especially in combined treatment group.The results of immunofluorescence staining revealed that the number of microglia cells in the Sham group was significantly lower than that in the control group and other three treatment groups,and the number of microglia cells in treatment groups was prominently lower than in the control group.Importantly,the number of microglia cells in the EA+Wheeled treadmill group was markedly lower than that in the EA group and Wheeled treadmill group.Conclusions: 1)EA combined with Wheeled treadmill training significantly could preferably improve motor function,sensory function and autonomic nerve function in rats with spinal cord injury compared with treatment with EA or Wheeled treadmill training alone.2)EA combined with Wheeled treadmill training could improve the nerve fiber bundle of spinal cord after spinal cord injury,repair its nerve conduction function;at the same time,strengthen the exercise of the muscles of the hind limbs,reduce the atrophy of the muscles of the hind limbs,which is conducive to the recovery of motor function.3)EA combined with Wheeled treadmill training could inhibit the activation of microglia cells in injured rats with spinal cord injury by repairing the structure and function of neurons,thereby reducing the scar generation and promoting the recovery of motor function. |
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