| OBJECTIVEMultiple sclerosis(MS)is an immune-mediated central nervous system disease with myelin loss,myelin regeneration,axonal injury,and neurodegeneration,Mainly pathological changes of white matter in the brain and spinal cord,lead to severe disability,its pathophysiological mechanism is complex,and there is still no specific effective treatment strategy.The MS classical animal model is an experimental autoimmune encephalomyelitis(EAE)model.In this study,female C57BL/6 mice experimental autoimmune encephalomyelitis(EAE)model was used to explore the therapeutic effect of acupuncture"Da zhui"(GV14),bilateral "Shen shu"(BL23),bilateral "Zu san li"(ST36),and its therapeutic mechanism:① The expression of JAK2 and p-STAT3 in brain tissue.② Changes in the expression of the inflammatory cytokine interleukin-17(IL-17)in serum.In order to deeply understand the pathogenesis of multiple sclerosis,and provide new ideas and theoretical support for clinical acupuncture treatment of multiple sclerosis.METHODForty female C57BL/6 mice were weighed and numbered,and divided into blank control group,EAE model group,hormone treatment group and acupuncture treatment group according to the random number table,with 10 rats in each group.Female C57BL/6mice was treated with myelin oligodendrocyte glycoprotein(myelin oligodendrocyte glycoprotein 35-55,MOG35-55)mixed complete Freund’s adjuvant(CFA)combined with pertussis toxin(PTX)to induce into the EAE model.The blank control group and the EAE model group were grasped and fixed once a day on the 14th day after immunization,and were intragastrically administered with 0.1 ml/10g/d normal saline until the 28th day after immunization.The acupuncture treatment group began on the 14th day after immunization,and the acupuncture treatment was performed at the same time every day(9:00 am)for"Da zhui",bilateral "Shen shu" and bilateral"Zu san li"points.Each time the needle was left for 15minutes,and it was ended on the 28th day after immunization,a total of 15 times.The hormone treatment group started on the 14th day after immunization,and at the same time every day(9:00 am),the mice were subjected to hormone shock therapy,each time acetic acid Prednisone suspension(0.6mg/ml,prednisone acetate tablets prepared with physiological saline),gavage,0.1ml/10g/d,continuous treatment for 15 days.From the 0th day after immunization until On the 28th day after immunization the end of the experiment,the behavioral changes(such as coat color,behavior and diet)of the mice were observed,neurological deficit score was recorded,at the same time every day(14:00-16:00 pm),and the body of the mice was weighed every other day.On the 28th day after immunization,Taking blood from the eyeball and taking the brain,the degree of demyelination was observed by myelin staining of Luxol fast blue(LFB),and the expression of JAK2 and p-STAT3 in brain tissue was detected,and the expression of IL-17 in serum.RESULT1.Body mass and neurological deficit scoreOn the 10th day after immunization,EAE model group,the hormone treatment group,and the acupuncture treatment group,the mice in the three groups were successively onset,gradually appearing to be wilting,loss of appetite,and weight loss,tail weakness,limb paralysis,incontinence and occasional back skin ulceration,which gradually worsened.The blank control group mice had no disease,the body weight continued to increase,and the activity was normal.On the 0th day after immunization,there was no significant difference in the body mass and neurological deficit scores between the groups(P>0.05).On the 14th day after immunization,compared with the blank control group,the EAE model group,the hormone treatment group,and the acupuncture treatment group,the weight of the mice continued to decrease(P<0.01),and the scores of neurological deficits were significantly increased(P<0.01),On the 20th day after immunization,compared with the EAE model group,the hormone treatment group and the acupuncture treatment group,the body weight of the mice was increased(P<0.01),and the neurological deficit scores were significantly lower(P<0.01).Compared with the hormone treated group,there was no significant difference in the body weight of the acupuncture treated group(P>0.05),neurological deficit score increased(P<0.01);on the 28th day after immunization,compared with the EAE model group,the body weight of the mice in the hormone treatment group and the acupuncture treatment group increased(P<0.01,P<0.01),neurological deficit scores were decreased(P<0.01,P<0.05).The two treatment groups was compared,the body weight of the acupuncture treatment group was lower(P<0.05),and the neurological deficit score was not statistically different(P>0.05).2.Brain tissue LFB myelin stainingThere was no abnormality in brain tissue of the blank control mice.In the EAE model group,the hormone treatment group and the acupuncture treatment group,the brain tissue of the mice with different sizes showed myelin loss,which showed vacuolization,and was most obvious around the lateral ventricle.Among the EAE model group,the myelin loss was the most obvious,and there were a lot of vacuolization changes.The hormone treatment group and the acupuncture treatment group had less myelin loss than the EAE model group,and there were a few vacuolization changes.3.Brain tissue immunohistochemistry JAK2,p-STAT3The positive expression of JAK2 and p-STAT3 in brain tissue was yellow.Compared with the blank control group,the expression levels of JAK2 and p-STAT3 in the brain of EAE model group,hormone treatment group and acupuncture treatment group were significantly different(P<0.01);compared with EAE model group,The expression levels of JAK2 and p-STAT3 in the brain tissue of the hormone treatment group and the acupuncture treatment group were significantly lower(P<0.01).The acupuncture treatment group and the hormone treatment group was compared,of mice,there was no significant difference in the expression level of JAK2、p-STAT3 in the brain tissue(P>0.05).4.Serum IL-17Compared with the blank control group,the serum ELISA IL-17 expression levels of the EAE model group,the hormone treatment group and the acupuncture treatment group were significantly different(P<0.01);compared with the EAE model group,the hormone treatment group and the acupuncture treatment group,The serum ELISA IL-17 expression level was significantly lower(P<0.01,P<0.05).The acupuncture treatment group and the hormone treatment group was compared,the serum ELISA IL-17 expression level was higher in the acupuncture treatment group(P<0.05).CONCLUSION1.Acupuncture and hormone can significantly increase the body weight of female C57BL/6 mouse EAE model and reduce the neurological deficit score,suggesting that acupuncture and hormone can improve the symptoms of female C57BL/6 mouse EAE model,and effect is the same.2.The expression of JAK2 and p-STAT3 in the brain tissue of EAE model group,acupuncture treatment group and hormone treatment group increased,suggesting that JAK2-STAT3 signal transduction pathway affects the pathogenesis of EAE.Acupuncture and hormone can significantly down regulate female C57BL/6 mice EAE model brain tissue immune histochemical JAK2,p-STAT3 expression,suggesting that acupuncture,hormone regulated JAK2-STAT3 signal transduction pathway to improve the pathogenesis of EAE,and effect is the same.3.Acupuncture can significantly down-regulate the serum IL-17 level in female C57BL/6 mice EAE model,suggesting that acupuncture can reduce the degree of inflammatory response,the mechanism of action may be by regulating the JAK2-STAT3 signal transduction pathway pathway,affect the secretion of IL-17. |
