Research On Shuganjianpi Gusui Recipe Affect T Lymphocyte Subsets And Inflammatory Factors In Multiple Sclerosis

Multiple sclerosis is an autoimmune disease of the T cell-mediated, inflammatory cytokines secreted by T cells execute inflammatory procession, they occupy an important role in the clinical symptoms of relapse and remission. Shuganjianpi Gusui Recipe as experience recipe is the clinical treatment of multiple sclerosis, suggesting a good effect in the clinical studies before. The study was designed to investigate the effects of Shuganjianpi Gusui Recipe on Multiple Sclerosis T cell subsets and its downstream inflammatory factors, and to investigate the pharmacological target for the treatment of multiple sclerosis combined with the network.1. Study of Shuganjianpi Gusui Recipe affected T cell subsets in multiple sclerosis patientsObjective:To study the the change of T cell subsets before Shuganjianpi Gusui Recipe Treatment of multiple sclerosis and after treatment.Methods:Quantification TCM syndromes with a questionnaire characterizing TCM syndromes in all patients with multiple sclerosis at each visit, statistics the frequency of occurrence of all of the symptoms, studies Syndrome characteristics of multiple sclerosis. T cell subsets of peripheral blood in all patients at each visit carried out by flow cytometry, treating with hormone therapy of the acute period, were tested before and after treatment, with Shuganjianpi Recipe of stable period every 3 months tested once.Results:Male and female patients age compared with no significant difference (P=0.493, P> 0.05), the number of male and female prevalence ratio is about 1:3, no significant difference in the sex ratio of all ages (X2=2.418,P=0.659, P>0.05). Numbness is the most common symptoms of neurological deficit with MS patients, accounting for 66.7%; followed by limb weakness and paresthesia, each with 60.3%.3 Clinical symptoms with MS patients have more than 50% frequency, lassitude (65.1%). soreness-tired of waist and knee(55.6%), depression (50.8%); 5 symptoms in occurrence rate of 40-50%, hyperhidrosis(47.6%), afraid of cold (46%), dreaminess(46%), vexation(41.3%), amnesia(41.3%); 30-40% occurrence rate of symptoms has 6, respectively is, listlessness, dry mouth and throat, stool dry, stool sticky, dazed head, chest tightness; tongue appears most is big fat tongue (12.7%), pale tongue (60.3%), thin fur(76.2%); pulse appears most is thready pulse (52.4%), followed by deep pulse (44.4%).CD3+ CD4-CD8-T cells subsets reduced 41% after hormone therapy compared to before hormone therapy in patients with multiple sclerosis, the difference was significant (P=0.010, P＜0.05); the rest of the T cell subsets was no significant difference (P>0.05); CD4+/CD8+T cell ratio had no significant change before and after hormone treatment (P=0.345,,P>0.05). CD3+CD4-CD8-T cells subsets before Shuganjianpi Gusui Recipe treatment had a 12% reduction compared with after therapy, the difference was significant(P=0.000, P<0.05); CD3+CD4+CD8-T cells subsets absolute value increased 5% after treatment than before treatment, but no significant difference (P=0.097,.P>0.05); the rest of the T cell subsets was no significant difference (P>0.05); CD4+/CD8+T cell ratio had no significant change before treatment and after (P=0.206, P>0.05).Conclusion:The number of patients the presence of men and women of all ages are significant differences in female patients significantly more than men, male to female ratio is about 1:3.2, in line with the relevant research literature.The highest frequency of clinical syndromes is deficiency of the kidney essence, followed by the liver depression, spleen deficiency, tongue and pulse showing the main performance of deficiency. Suggesting syndrome manifestations of multiple sclerosis is the main essence deficiency, liver depression, spleen deficiency, provided Syndrome basis Shuganjianpi Gusui Recipe Treatment of Multiple Sclerosis.CD3+CD4-CD8-T cell subsets with significantly reduced after treatment with hormones and Shuganjianpi Gusui Recipe than before, suggesting CD3+CD4-CD8-T cell subsets promote inflammation occurs in the course of MS, and the hormones and Shuganjianpi Gusui Recipe anti-inflammatory effect of significantly infleuence the CD3+CD4-CD8T cell subsets at different stages of the disease.(4) CD3+CD4+T cell subsets as T helper cells, represents the degree of activation of the immune response, CD3+CD4+T absolute value tended increase after Shuganjianpi Gusui Recipe trearment to before the treatment, but not statistically significant. 2. Experimental study of Shuganjianpi Gusui Recipe affected anti-inflammatory in EAEObjective:To investigate the change of serum inflammatory factors in EAE mouse model of multiple sclerosis with Shuganjianpi Gusui Recipe treatment.Methods:Establishment the hormone group, EAE model group and Shuganjianpi Gusui Recipe groups. Observation change of inflammatory factors with Shuganjianpi Gusui Recipe intervention EAE, the use of high-throughput protein liquid microarray inflammatory factor at different times, these factors are:IL-23, IL-17E/IL-25, GM-CSF, IL-12p70, IL-17F, IL-17A, TNF-a, IFN-y, IL-4, IL-10, to explore the impact of Shuganjianpi Gusui Recipe to EAE inflammatory factors.Result:(1)In this study, the chronic persistent type of animal model was successfully prepared by induced C57BL/6 mice with MOG35-55 polypeptide fragments, and we observed its full disease course, and observed relapsing phenomenon.(2)There is no vascular proliferation and inflammatory cell infiltration in normal brain and spinal cord tissue. EAE spinal cord tissue biopsy shows infiltration of inflammatory cells, vascular proliferation and showed changes in cuff-like inflammation,and the spinal cord inflammation significantly greater serious to brain. Hormone group and Shuganjianpi Gusui Recipe group brain and spinal cord tissue sections showed the proliferation of capillaries, but Shuganjianpi Gusui Recipe microvessel proliferation is more obvious, and the brain nerve cell swelling and vacuolizing.(3)Change of inflammatory factors in each group at different times:There were no significant differences at different times in EAE model group; no significant differences at different times Shuganjianpi Gusui Recipe groups, the expression of TNF-a significantly decreased on 4 weeks compared with on 0 week in the hormone group (P=0.030,.P＜0.05), no significant difference of the inflammatory factor in the rest at different times.(4)Change of inflammatory factors at the same time in each group:Inflammatory factors had no significant difference before modeling (0 weeks) between EAE group, hormone group and Shuganjianpi Gusui Recipe groups. TNF-a was significantly higher (P=0.002, P<0.05) in Shuganjianpi Gusui Recipe group on 4th week compared to hormone group. IFN-y was significantly reduced hormone group compared to the EAE group on 4th week (P=0.029,P＜0.05). IFN-y was significantly reduced in Shuganjianpi Gusui Recipe group compared to EAE group on 4th week (P=0.021, P<0.05), the rest flammatory cytokines had no significant change.Conclusion:(1)We selected the 14th day after the EAE models as the start of drug treatment, through long-term observation of EAE, and determined the TNF-a, IFN-y on 4th week as sensitive indicators and detector time.(2)It suggesting Shuganjianpi Gusui Recipe and hormones produced anti-inflammatory effect through the inhibition of the expression of IFN-y.(3)lt suggesting hormones may produce anti-inflammatory effects by decreasing the expression of TNF-a. The anti-inflammatory mechanism Shuganjianpi Gusui Recipe may was different with hormones, hormones of TNF-a decreased, while TNF-a was increased with Shuganjianpi Gusui Recipe, but their effects both were weak (they showed no differences with EAE group), suggesting that the role of TNF-a in the EAE may be more complex. 3. Study of Shuganjianpi Gusui Recipe gene network pharmacologyObjective:To study the gene target of Shuganjianpi Gusui Recipe Treatment of multiple sclerosis.Methods:Using the "Multiple sclerosis" as the name of the disease, to find genes associated with the OMIM database; composition based drugs of Shuganjianpi Gusui Recipe, find the appropriate targets in the database in TCMSP. Retained verification target in DrugBank database, and then the disease-related genes and various prescription targets are mapped to the STRING database with network construction, capabilities enrichment analysis each network by use of software DAVID of, from KEGG Pathway point of view, further functional analysis for the results of retention P＜0.05.Result:Tnf, Stat3, Vegfa,1117a and 112ra have a high number of network referrals and networking degree, have an important role in multiple sclerosis-associated gene network. The target gene Tnf, Vegfa had a high number of referrals and networking degree both in Shuganjianpi Gusui Recipe-associated genes and in multiple sclerosis-associated genes.Conclusion:Shuganjianpi Gusui Recipe-affect target genes overlap and multiple sclerosis-associated genes, the overlapping genes are likely to be onset point on Shuganjianpi Gusui Recipe treatment of multiple sclerosis. Tnf, Vegfa in multiple sclerosis-associated genes also had a high degree of networking and referral number, suggesting that these two target genes may be the key sites of Shuganjianpi Gusui Recipe treatment of multiple sclerosis.