The Effect Of Acupuncture On The Expression Of IBA-1 And COX-2 In The Spinal Cord Of EAE Mice And The Level Of Serum TNF-?

OBJECTIVEMultiple sclerosis(MS)is an inflammatory demyelinating disease that occurs in the central nervous system.It has the characteristics of disability,recurrence,progress and death,which greatly endangers life and health.There is no cure for MS in the world.In recent years,TCM treatment of MS has shown good clinical efficacy,but the mechanism of TCM treatment of MS is still unclear,affecting its promotion and application in the world.In this study,we observed the effects of acupuncture on the levels of intraspinal calcium junction protein-1(IBA-1),cyclooxygenase-2(COX-2)and serum tumor necrosis factor-?(TNF-a)in experimental autoimmune encephalomyelitis(EAE)mice,and explored the possible mechanism of acupuncture in treating EAE mice.METHODSThirty-six C57BL/6 mice aged 8-10 weeks were randomly divided into four groups:blank control group,model group,hormone group and acupuncture group,with 9 mice in each group.After one week of adaptive feeding,mice in the blank control group were not treated.The rest of the mice used MOG35-55 polypeptide to establish EAE model.From the 14th day after modeling,blank control group and model group:mice were grabbed once a day;hormone group:prednisone acetate intragastric administration;acupuncture group:acupuncture at "Dazhui"acupoint,"Shenshu"acupoint and "Zusanli" acupoint on both sides.The serum TNF-a content,the cumulative integral optical density of COX-2 and IBA-1 positive expression in spinal cord of mice in each group were measured 28 days after the establishment of the model.RESULTS1.Compared with the blank control group,EAE model mice started to show mild clumsy movements,unstable gait and nerve injury symptoms at 12 days.At 14 days,all the model mice in each group presented symptoms of nerve injury,and the neurological function score was statistically different from that of the blank control group(P<0.05).The Kono neural function score of the model mice was between 1 and 4,and the model was successful.2.Compared with the blank control group,the serum TNF-a content,spinal cord Cox-2 and Iba-1 mean optical density in the model group were significantly increased(P<0.01).3.Compared with the model group,EAE mice in the acupuncture group presented neurological impairment symptoms on the 14th day after immunization,later than the model group(symptoms on the 12th day after immunization),and the neurological function score of EAE mice in the acupuncture group was also lower than that in the model group(P<0.01).The serum TNF-a content of EAE mice in the acupuncture group was 22.16±3.12,significantly lower than that of the model group(26.31±2.65,P<0.01).The mean optical density of Cox-2positive expression in the spinal cord of EAE mice in the acupuncture group(2.86±1.18)×10-3 was lower than that in the model group(12.57±3.76)×10-3(p<0.01).In addition,the average optical density of Iba-1 positive expression in the spinal cord of mice in the acupuncture group(4.16±2.01)×10-3was lower than that in the model group(5.47±1.67)×10-3(P<0.05).4.Compared with the model group,EAE mice in the hormone group had a lower neurological function score,and the neurological impairment symptoms appeared in mice on the 14th day after immunization,which was later than that in the model group.The serum TNF-a content in the hormone group was 22.37=1.62,significantly lower than that in the model group(26.31±2.65,P<0.01).The mean optical density of Cox-2 positive expression in the spinal cord of EAE mice in the hormone group(0.74±0.61)×10-3 was lower than that in the model group(12.57±3.76)×10-3(P<0.01).In addition,the average optical density(0.56±0.23)×10-3 of Iba-1 positive expression in the spinal cord of the hormone group was lower than that of the model group(5.47±1.67)×10-3(P<0.01).The levels of TNF-?,Cox-2and Iba-1 in serum,spinal cord and spinal cord of EAE mice in the hormone group were lower than those in the model group,with statistically significant differences(P<0.01).5.Compared with the acupuncture group,the neurologic function score of EAE mice was lower in the hormone group.The serum TNF-? content of EAE mice in the hormone group was slightly higher than that in the acupuncture group,and there was no statistical difference between the two groups(P>0.05).The mean optical density(0.74±0.61)×10-3 of positive Cox-2 expression in the spinal cord of EAE mice in the hormone group was lower than that in the acupuncture group(P<0.05).The average optical density(0.56±10.23)×10-3 of Iba-1 positive expression in the spinal cord of the hormone group was also lower than that of the acupuncture group(P<0.01).CONCLUSIONS1.In this study,the EAE model was successfully established by using MOG35-55 polypeptides and complete freund's adj uvant.2.Acupuncture can alleviate nerve injury in EAE mice and improve neurological function score,and its effect is better than that of hormone group.3.The effect of acupuncture on reducing serum TNF-a content in EAE mice was similar to that of hormone therapy,but the level of positive expression of Cox-2 and Iba-1 in the spinal cord of EAE mice was worse than that of hormone therapy.4.Acupuncture can improve and alleviate nerve injury in EAE mice,which may be related to the fact that acupuncture can reduce the serum tnf-a content in EAE mice and reduce the positive expression of Cox-2 and Iba-1 in the spinal cord.