| Objective:To observe the clinical curative effect and safety of Bupiqiangli Compound on treatment of patients with myasthenia gravis(MG).EAMG animal model were induced the following immunization of Lewis rats with the rat sequence 97-116 peptide of the AChR alpha-subunit(R97-119).The changes of weight and the levels of AChR-Ab in EAMG rats were observed,and observed the changes of the levels of IFN-γ、TGF-β、TNF-α、IL-2、IL-4 and IL-17.The mRNA and protein expression levels of these Cytokines receptors on hypothalamus were determined by real-time quantitive PCR and Immunohistochemistry.The effect of these cytokines and their receptors on the expression levels of CRH and the level of serum cortisol were observed.And then making a explanation of the therapeutic mechanism of Bupiqiangli Compound on treatment of patients with MG.Method:①The myasthenia gravis(type,type Ⅱ a,type Ⅱ b)patients were randomly divided into treatment group and control group according to clinical random-comparing study.The control group received prednisone and pyrido-stigmine bromide,based on the control group,the treatment group received Bupiqiangli Compound for 6 months.The curative effect was evaluated by the Absolute and Relative Score of MG(ARS-MG)and the side-effect by Treatment Emergent Symptom Scale(TESS).②EAMG animal model were induced the following immunization of Lewis rats with R97-116.The female Lewis rats were randomly divided into control group、model group、Prednisone group(5.4mg/kg)and different dose of Bupiqiangli Compound groups(9.49g/kg、7.12g/kg、4.75g/kg).These successful EAMG models were treated once a day for 4 weeks.The changes of weight and AChR-Ab with EAMG rats after the treatment were observed.The levels of IFN-γ、TGF-β、TNF-α、IL-2、IL-4 and IL-17 were tested by ELISA,the mRNA and protein expression levels of Cytokine Receptors and CRH were determined by RT-Q-PCR and Immunohistochemistry.And the levels of serum cortisol were detected.Results:1①.By statistical analysis,the treatment group recovered in 2 cases,basically recovered in 4 cases,14 cases had markedly effect,improved in 4 cases and 2 cases failed,the markedly effective rate of 80%,the total efficiency of 92%;the control group recovered in 1 cases,basically recovered in 3 cases,7 cases had markedly effect,improved in 9 cases and 5 cases failed,the markedly effective rate of 44%,the total efficiency of 80%.The overall curative effect evaluation of treatment group is superior to control group.Compared with control group,the treatment group didn’t show obvious side-effect,and the safety level is higher than control group.②.Compared with the model group,the weight of EAMG rats increased and the levels of AChR-Ab decreased significantly after treated with Bupiqiangli Compound(P<0.01).The levels of AChR-Ab in low-dose group were significantly higher than the prednisone group(P<0.05),but no significant differences showed between prednisone group and Bupiqiangli Compound of high or middle-dose group.③.In terms of intervention cytokines,the levels of TNF-α、IFN-γ、IL-2、IL-4 and IL-17 decreased significantly in Bupiqiangli Compound group,compared with model group,which showed extremely significant differences and significant differences respectively(P<0.01,P<0.05),On contrary,the levels of TGF-(3 increased significantly(P<0.01).There were no significant differences between prednisone group and Bupiqiangli Compound group in the levels of IFN-γ、TGF-β、TNF-α and IL-17,but the levels of IL-2 and IL-4 are significantly higher than prednisone group(P<0.05).④.In terms of intervention cytokines and its receptors signal transduction pathways.Acoording to the test results,the IL-2R mRNA didn’ t expressed in the hypothalamus tissue.Compared with control group,the mRNA and protein expression levels of IFN-γR、TβR and IL-17R were increased in model group(P<0.05),the mRNA and protein expression levels of TNF-aR was decreased(P<0.05),but there was no statistical difference with the mRNA expression levels of IL-4R.Compared with model group,the high、moderate and low-dose group can decrease the mRNA and protein expression levels of IFN-yR and increase the TNF-αR(P<0.05).Bupiqiangli Compound can decrease the protein expression levels of T β R and IL-17R,but it showed significant difference in the mRNA expression levels.⑤.In terms of intervention HPA axis,the levels of serum cortisol in model group、the high、moderate and low-dose groups were slightly higher than that of normal group,but they showed no statistical difference.Compared with the control group,the mRNA and protein expression levels of CRH were decreased in model group(P<0.01).Compared with the model group,the mRNA expression levels of CRH showed no significant difference in prednisone group,but it increased in Bupiqiangli compound high、large and middle-dose group(P<0.05).Conclusions:Bupiqiangli Compound can improve clinical symptoms in patients with MG,and no obvious adverse reactions is reported.It was proved to be safe and effective in clinical application.Experimental studies have found that Bupiqiangli Compound can increase the weight of EAMG rats,and reduce the levels of AchR-Ab,which turned out to be effective for curing EAMG rats.Bupiqiangli Compound can reduce the levels of IFN-γ、TNF-α、IL-2、IL-4 and IL-17,and increase the levels of TGF-β,thus the cytokines network disorders were adjusted.It can intervened cytokines and their receptors signal transduction pathways effectively,and impact themRNA and protein expression of CRH in hypothalamus,thus regulating cytokines-neuroendocrine network in an effective way.Which may be one of the important therapeutic mechanisms of the Bupiqiangli Compound in treatment of myasthenia gravis. |
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