| Objective:Guillain-Barrés syndrome(GBS)is an autoimmune disease and is one of the most important causes of acute limb stagnation in the world.The current treatment are mainly plasma exchange(PE)and intravenous immunoglobulin(IVIg).Despite the above treatment,there is still a poor prognosis or serious sequelae,so the exploration of new treatment methods of GBS is still an urgent problem to be solved in neurology.In recent years,ginkgolide has been increasingly studied for the role and mechanism of autoimmune diseases,but there has been no research in the treatment of GBS.This study intends to observe the role of ginkgolides in EAN and its possible mechanism and to lay a theoretical foundation for the future exploration of new methods for GBS treatment.Through the establishment of its animal model,experimental autoimmune neuritis(EAN),the clinical scores of EAN,sciatic nerve pathology and immunological indicators were used to evaluate the potential therapeutic effects of ginkgolide on GBS.Methods:(1)Experimental animals and grouping: Male C57BL/6 mice,6-8 weeks old,weighing about 18-20 g,were purchased from Beijing Weitong Lihua Experimental Animal Technology,and the animals were randomly divided into control(EAN+PBS)group(n=7),EAN+low dose of ginkgolide(5mg/kg/d)group(n=7),EAN+ high dose of ginkgolide(10mg/kg/d)group(n=7).(2)Establishment of EAN model and clinical score: Pertussis toxin(PTX)was dissolved in PTbuffer and injected into the tail vein,we emulsified peptide P0180-189,incomplete Freund's adjuvant and mycobacterium tuberculosis together fully,then we injected them into the back.(3)Intervention of experimental animals: On the sixth day of EAN model,PBS or ginkgolides was intraperitoneally injected according to grouping.Three groups of mice were simultaneously killed at the peak.(4)Experimental materials and observation indicators:(1)Take the mouse spinal sciatic nerve near the spinal cord segment,observe the sciatic nerve inflammation and myelins loss by HE and LFB pathological staining;(2)Separate the spleen of EAE mice,through the flow cytometry,CD4+ T cells were selected and Th1,Th2,Th17,and Treg cells proportion of CD4+T in the mononuclear cells(MNCs)were detected.Furthermore,we evaluated the therapeutic effect of ginkgolide on EAN.Results:(1)Clinical score: In the course of the disease,the clinical scores of the mice in the ginkgolide treatment group were lower than those in the control group.On the 9th day of model establishment,the clinical score of the low-dose group was lower than the control group(p <0.05),and the clinical scores of the low-dose and high-dose ginkgolides treatment groups were both lower than the control group on the 14th-15 th day.(2)Pathological results of sciatic nerve: In the control group,the sciatic neuritis cell infiltration and myelin loss was the most serious,the high dose group was less serious and in the low dose group was the least serious.(3)Determination of CD4+T cells:(1)The low dose group could down-regulate the proportion of Th17 cells in spleen MNCs,which was significantly lower than the control group(P<0.05),it could also down-regulate the proportion of Th1 cells in spleen MNCs and which was lower than the control group(P>0.05)(2)The high dose group could also down-regulate the proportion of Th17 cells in spleen MNCs which was lower than the control group(P>0.05).(3)The proportion of Treg cells in spleen MNCs in both the low dose group and the high dose group were higher than the control group(P>0.05).(4)The proportion of Th2 cells in MNCs of the low dose group was lower than the normal group(P>0.05),and the Th2 cells in the spleen MNCs of the high dose group was higher than the control group(P>0.05).In conclusion:(1)Low-dose ginkgolide can effectively treat mice EAN,reduce clinical score,and reduce peripheral nerve tissue damage such as peripheral nerve inflammatory infiltration and myelin loss.(2)Low-dose ginkgolide can alleviate the clinical symptoms by inhibiting Th17 cell. |
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