| OBJECTIVE:To observe the effects of acupuncture at Dazhui,Shenshu and Zusanli points on neurological injury and phosphorylated mitogen-activated protein kinase p-p38MAPK in spinal cord and serum IL-lbeta in experimental autoimmune encephalomyelitis(EAE)mice,which is a classic animal model for studying multiple sclerosis(MS),to verify the therapeutic effect of acupuncture on multiple sclerosis,explore the mechanism of acupuncture intervention on EAE mice,and provide experimental basis for clinical acupuncture treatment of MS.METHODS:Forty C57BL/6 mice were randomly divided into four groups:blank group,model group,model+acupuncture group(acupuncture group)and model+hormone group(hormone group),with 10 mice in each group.Except for the blank group,mice in other groups were immunized with emulsifier prepared by mixing myelin oligodendrocyte glycoprotein 33-55(MOG33-55)with complete Freund’s adjuvant(the final concentration of Mycobacterium tuberculosis H37Ra was 4mg/mL)to establish EAE model.Starting from the 14th day of modeling,each group was intervened at the same time every morning.The blank group and the model group were grasped and fixed daily,and were intragastrically administered with normal saline[0.1mL/(10g·d)].The hormone group was intragastrically administered with[0.1mL/(10g·d)]of prednisolone acetate solution(0.6mg/mL);In the acupuncture group,"Dazhui" points,bilateral "Zusanli" points and bilateral " Shenshu"points were taken for routine disinfection and acupuncture intervention,each time for 15 min.All groups were treated continuously for 15 days.From the day of modeling,the behavioral changes of mice were closely observed,and the body weight of mice in each group was measured every other day and the neurological function was scored;On the 28th day of immunization,blood was collected from the eyeballs of mice in each group and spinal cord tissue was collected.LFB staining was used to detect the degree of demyelination in spinal cord of mice.The expression of p-p38MAPK protein in the spinal cord tissue of mice was detected by Western blot,and the content of IL-1β in serum of mice in each group was detected by enzyme-linked immunosorbent assay.RESULTS:(1)Changes of body weight in mice:Compared with the blank group,the average body weight of mice in the model group,the acupuncture group and the hormone group decreased significantly after 12 days of immunization;compared with the model group,the average body weight of mice in the acupuncture group and the hormone group increased significantly after 20 days of immunization(P<0.05);and compared with the hormone group,the average body weight of mice in the acupuncture group decreased significantly after 22 days of immunization(P<0.05).(2)Behavioral changes in mice:The mice in the blank group did not show any abnormalities in symptoms and behavior throughout the experiment.After the 10th day of immunization,the mice in the model group,the acupuncture group and the hormone group all developed symptoms,which were characterized by menta l fatigue,dull coat,contracted piles and varying degrees of neurological deficits.Compared with the blank group,the mean neurological function scores of the model group,the acupuncture group and the hormone group were significantly increased from the 14th day after immunization(P<0.05);compared with the model group,the peak period of the disease(On the 20th day after immunization),the average neurological function scores of the acupuncture group and the hormone group were significantly decreased(P<0.05);after the peak period,the symptoms of the model group,the acupuncture group and the hormone group were slightly relieved.But compared with the EAE model group,the mean neurological function scores of the acupuncture group and the hormone group were significantly decreased on the 28th day of immunization(P<0.05),while there was no significant difference between the acupuncture group and the hormone group.Difference(P>0.05).(3)LFB staining results:The myelin sheath of the spinal cord tissue of the normal group was dense and arranged neatly.In the model group,the myelin sheath was lost in multiple parts of the spinal cord tissue,which was mainly caused by white matter invasion.The white matter of the spinal cord is loose,and a number of flaky demyelinating areas of different sizes are seen,which are "vacucal-like"changes.The degree of demyelination was significantly reduced in the acupuncture group and the hormone group,and the loss of myelin was significantly reduced.(4)Detection results of IL-1β:At 28 days of onset,the serum IL-1β expression in the model group was higher than that in the blank group(P<0.05).Compared with the model group,the acupuncture group and the hormone group were compared with the model group.The expression of IL-1β was decreased(P<0.05).There was no significant difference between the acupuncture group and the hormone group(P>0.05).(5)Detection results of p-p38MAPK:At 28 days after onset,the expression of p-p38MAPK in the spinal cord of the model group was increased compared with the blank group(P<0.05).Compared with the model group,the expression of p-p38MAPK protein in the spinal cord of the acupuncture group and the hormone group was decreased.(P<0.05);and there was no significant difference between the acupuncture group and the hormone group(P>0.05).CONCLUSIONS:(1)Acupuncture can significantly improve the performance of neurological damage in EAE mice.(2)Acupuncture can down-regulate the protein content of p-p38MAPK in the spinal cord of EAE mice,suggesting that acupuncture can inhibit the activation of P38MAPK signaling pathway.(3)Acupuncture can significantly down-regulate the expression of serum IL-1β protein in EAE mice,and its regulation is equivalent to that of hormones.(4)This study demonstrated that acupuncture has a therapeutic effect on EAE mice,and its mechanism may be related to the inhibition of p38MAPK protein phosphorylation and the down-regulation of serum IL-1β expression. |
