The Study Of Yiqichushi Prescription Effecting Serum AChRab And SPD- L1 To Patients With MG And EAMG

Myasthenia gravis is mainly involving the neuromuscular junction of acetylcholine receptors on postsynaptic membrane of autoimmune diseases.The disease is mainly mediated by acetylcholine receptor antibodies, cell-mediated immunity(cell-mediated immunity, CMI) dependent, complement participation.The disease mainly manifested clinically is that after exercise increased, after the break to reduce, the morning light and nighttime severe skeletal muscle fatigue.Myasthenia gravis is a difficult disease all over the world.Currently Western medicine treatment of hormone and acetylcholine inhibitors have different levels of side effects,therefore, exploring curative effect and reducing the side effects of western medicine of traditional Chinese medicine become the urgent matter.YCR is of TCM in the treatment of myasthenia gravis,tutor professor Tan Zihu proposed, from "treat impotent alone take Yang Ming" theory, combined with many physicians treat impotent theory and clinical experience for many years, and the formulas in clinical and experimental study were made definite curative effect. Section One The Mechanism Study of the efficacy of Yiqi Chushi Recipe in Treating Myasthenia Gravis and its effects on Serum antibody of ACh R and Soluble PD- L1 moleculesObjective:To observe the efficacy of Yiqi Chushi Recipe(YCR)in treating myasthenia gravis(MG)patients,including the total clinical curative effect,absolute score,relative score,the curative efficacy of traditional Chinese medicine symptoms and its integral;to observe its effects on Serum ACh R antibody titre and soluble PD-L1 molecular level of myasthenia gravis patients,From the immunological indexes to judge the mechanism research of The improvement of YCR to the symptoms of MG patients.Methods: To observe 52 cases of patients with MG included in the standard.Acco-rding to the random sequence table prepared to divide them into the treatment group(26 cases) and the control group(26 cases) by 1:1 proportion.Patients in the control group were treated by Prednisone and Pyridostigmine bromide,while those in the treatment group were based on the use YCR treatment.The duration of these two groups were both eight weeks.All patients were tested before treatment and after treatment,including interrogation, physical examination, and the relevant checks.Fill out the form of clinical observation to evaluate the efficacy and observe the safety. Observe the total clinical curative effect,absolute score,relative score,the curative efficacy of traditional Chinese medicine Symptoms and its integral before treatment and after treatment,then we did statistic analysis.Serum ACh R antibody titre of myasthenia gravis patients were measured by Human acetylcholine receptor(ACh Rab) ELISA Kit while the PDCD1LG1 ELISA Kit was used to measure the serum soluble PD-L1 molecular level and both of the data Analyzedstatistically.Results:YCR could significantly improve clinical symptoms of patients with MG,satistics from the total score in the clinical efficacy :The significant effective rate of treatment group was 96.15%, whereas 88.46% in the control group.Obviously, the treatment group curative effect was better than the control group,there was significant difference.From the point of absolute score,after the teratment,the rate of treatment group was 5.54±4.12, whereas 7.71±1.78 in the control group.Thereore,the effect in treatment group was better than that in control group,which had significant difference(P<0.05).From the point of relative score,the rate of treatment group was 53.59±25.33, whereas the control group was37.89±19.93.Obviously,the effect in treatment group was better than that in control group,which had significant difference(P < 0.05).Score of TCM symptoms:the effective rate of the treatment group was 88.46%, while the control group was 57.69%,and the treatment group was higher than the control group, the difference was statistically significant(P<0.05).It showed that on the improvement of TCM syndrome, the treatment group was superior to the control group.The research also found that 2 cases in the treatment group had slight nausea,but these did not affect the progression of the treatment. No extraordinary changes were found in the treatment group such as in cardiovascular,liver,kidney, fasting blood glucose and femoral head.However,2 cases in the control group had been found mild hepatic dysfunction and the fasting glucose of 1 case risen,and 1 case become central boesity.They had notable differences statistically.So the treatment group was much more safety than the control group.The levels of serum ACh R antibody had increased significantly than that in pre-treatment in both groups,and the difference was statistically significant(P<0.05);The level of soluble PD-L1 molecular had decreased than that in pre-treatment in both groups,and the difference was statistically significant(P<0.05).Conclusion:Combined YCR and western medicine could alleviate the clinical symptoms,TCM symptoms,absolute and relative scores of myasthenia gravis patients,which was more superior to the basic western medicine without any poison side effects.Combined YCR and western medicine was believed to increase the content of serum soluble PD-L1,which directly affected the negative costimulatory signals of PD-1/PD-L1 which were mediated by receptor and ligand,and the mutual interaction between PD-1 and PD-L1 were correlated to the inhibition of CD4+ and CD8+T-cell,moreover,it could suppress the secretion of IL-2.All of these could keep cells arrest in G0/G1 phase and suppress the secretion of cytokines such as IFN-γ,IL-10, IL-2,then reduction 0f the synthesis of ACh R antibody worked,at the same time,the combination could enter the blood and participate in regulating the immune function.Increased of soluble PD-L1 molecular levels,as a kind of hindrance molecules,could increase the chances of combining with PD-1 on the mononuclear cells surface,then the probability of these PD-L1 on T lymphocytes surface interacted with PD-1 on some cells surface,such as vascular endothelial cells,tissue cells,and mononuclear cells,increased greatly.Meanwhile these activated T lymphocytes obtained enough strength of negative regulation signal,which could prevented the unremitting immune responses coused by T lymphocytes overactive activation and proliferation,thus reducing the damage of self organize issues.The decrease of ACh R antibody synthesis,could alleviate the suppressive to ACh R,slowed the solubilizing capacity of postsynaptic membrane mediated by complement,meanwhile increased the functional receptor number on poet junctional membranes,thus improving clinical symptoms of patients with MG.Section Two The Mechanism Study of Yiqi Chushi Recipe effect on Serum ACh R antibody of experimental autoimmune myasthenia gravis(EAMG) rats and Soluble PD- L1 moleculesObject:Preparation of rat model of EAMG.By taking YCR, observed the clinical symptoms, serum soluble PD- L1 and ACh R antibody levels of change in rats.To in level,detection and classification Lennon reclaimed symptoms after 8 weeks.Results:Lemon clinical score of 0.5-1 level, and with the time prolonging of myasthenia gravis symptoms were more obvious, by the end of the building clinical scoring at level 1 or above accounted for 75.39% of the total building;Application 5Hz low-frequency electrical stimulation for 10 times in 30 rats were a total of 23 rats that were 1st and 5 percentage attenuation potential of more than 10%, the average of(21.68 ± 4.23), and the remaining seven rats electricity decay rate of less than 10%;Neostigmine test positive had 25;The content of serum ACh R antibody in EAMG rats than in healthy rats was high, and the content of soluble PD- L1 molecular level lower than that in healthy rats,by statistics analysised, there was significant difference(P <0.05).Description serum antibody of ACh R,soluble PD- L1 molecules was of great significance in the pathogenesis of myasthenia gravis. YCR and prednisone all could improve EAMG rats symptoms of muscle weakness, affected the weight of the rats, by statistics analysised, YCR was better than prednisone(P <0.05)。The content of serum ACh R-Ab of YCR group and prednisone group were significantly lower in the model group(P < 0.05). YCR, prednisone could reduce the concentration of serum ACh R antibodies EAMG rats,YCR group to reduce the extent of ACh R antibodies in serum concentrations was less than prednisone group(P> 0.05). YCR, prednisone could increase EAMG rats serum soluble PD- L1 molecular content, the degree of increasing the level of serum soluble PD- L1 molecules of YCR was superior to prednisone(P < 0.05).Conclusion:With subcutaneous injection murine ACh R-α subunit peptides 97-116 EAMG model could be successfully produced.YCR could improve EAMG rats’ weight, muscle strength and exercise capacity, inhibit ACh R antibody in serum concentration of EAMG model rats and increase the serum soluble PD- L1 molecular level.YCR might increase the serum content of soluble PD-L1, a direct impact on PD-1 / PD-L1 negative costimulatory signal receptor and ligand interactions mediated, PD-1/PD-L1 interaction inhibits CD4+ and CD8+T cell proliferation and IL-2 secretion, so that the cell cycle vestigate the pathogenesis of myasthenia gravis and YCR mechanism in the treatment of myasthenia gravis. For replenishing YCR to provide theoretical basis for the clinical application of the treatment of myasthenia gravis.Methods: Forty healthy adult female Lewis rats were randomly selected 10 as the control group(group A), remaining source of rats subcutaneously murine ACh R-α subunit peptides 97-116 which were EAMG building, building after the rats were randomly divided into model control group(group B), the positive drug control group(group C), YCR group(group D),where B, C, D group with 10 rats in each group, a total of 4 groups. Two weeks after successful model, A group and B group given normal saline, group C prednisone orally, D group were fed YCR,conduct ACh R antibody and soluble PD- L1 molecular remained in the G0/G1 phase, and reduced the IFN-γ, IL-10 and IL-2 gamma secreting cells factor, thereby enabling the ACh R antibody synthesis cut back.Ultimately the symptoms of muscle weakness of EAMG rats were improved.