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Effect Of 6-gingerol On T Cell Proliferation And Differentiation And Experimental Autoimmune Encephalomyelitis

Posted on:2021-05-07Degree:MasterType:Thesis
Country:ChinaCandidate:S S YangFull Text:PDF
GTID:2404330602988519Subject:Biology
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Objective:1.To study the effects of 6-gingerol on the proliferation and differentiation of T cells.2.Further apply 6-gingerol to the prevention and treatment of T cellsmediated multiple sclerosis ideal model,an experimental autoimmune encephalomyelitis mouse model,to verify its efficacy and explore its mechanism of actionMethods:1.C57 BL / 6 mice were sacrificed and spleen and lymph nodes were isolated,and use ground glass to make tissue homogenate.After separating and removing connective tissue with a centrifuge,lysing with red blood cell lysate and separating and removing red blood cells again to obtain a spleen cell suspension.A T cell suspension can be obtained after sorting using a T cell sorting kit for culturing and experiments in a cell culture plate,and a CD4 + T cell suspension can be obtained after using CD4 + T cell sorting kit.T cell suspension was used to initially test the effect of 6-gingerol on T cell proliferation,and the peak state of cell proliferation was detected using flow cytometry.CD4 + T cells were treated with different cytokines and the optimal addition protocol was optimized.The iTreg cells were obtained from cell culture plates.6-gingerol was applied to the above cytokine-treated culture process,and the peak of cell proliferation and the number of cell differentiation were detected using flow cytometry,and its effect on the proliferation and differentiation of iTreg cells was understood and analyzed.2.Myelin oligodendrocyte glycoprotein MOG35-55 polypeptide,pertussis toxin and Mycobacterium tuberculosis were used in combination to construct a mouse experimental allergic encephalomyelitis model.C57 BL / 6 mice were anesthetized and injected with the combination.Pertussis toxin was injected intraperitoneally at the time of construction and 48 hours after construction.Myelin oligodendrocyte glycoprotein was mixed with Mycobacterium tuberculosis using incomplete Freund’s adjuvant.And at the time of construction,the back of the forelimbs and hips of the hindlimbs were injected on the subcutaneous at joints.After successfully constructing a mouse experimental allergic encephalomyelitis model,the effect of 6-gingerol on the progression of experimental allergic encephalomyelitis was tested,and the optimal dosage control scheme was optimized.Results:1.CFSE stain can reflect cell proliferation,but at the same time it has certain cytotoxicity.After several adjustments and optimization of the added concentration,it was found that CFSE has a better staining effect at about 0.5 μM,and has less toxicity without causing excessive cell death.6-gingerol was used in CD4 + T cell proliferation culture experiments to determine its effect on cell proliferation.The experimental results showed that 6-gingerol had a dose-dependent inhibitory effect on CD4 + T cells.2.After optimizing cytokine addition conditions and cell culture conditions based on previous experiments,optimal iTreg differentiation conditions were established,and the foundation for cell experiments were lay.After applying 6-gingerol to iTreg differentiation effect experiments,it was found that low-dose 6-gingerol had a certain inhibitory effect on iTreg differentiation,but its inhibitory effect was eliminated by increasing the dose.3.After analyzing the experimental mouse allergic encephalomyelitis model across the world,we selected C57 BL / 6 mice as the target and optimized the construction conditions.Finally,we successfully constructed an experimental animal model with a 100% incidence rate for animal experiments.Laying the foundation for animal experiments.After applying 6-gingerol to the experimental course of EAE animal model,it was found that 6-gingerol significantly inhibited the pathogenesis of EAE.Conclusion:1.6 Gingerol has a dose-dependent inhibitory effect on T cell proliferation,so it has potential therapeutic effect on experimental autoimmune encephalomyelitis caused by excessive T cells.iTreg cells play an important role in the body’s immune balance and inhibit the excessive proliferation of other T cells,while low concentrations of 6-gingerol have a certain inhibitory effect on the differentiation and proliferation of iTreg cells.But by adjusting the optimized dose,its inhibitory effect can be eliminated.2.With the progress of the experimental model of experimental allergic encephalomyelitis in mice,the mice will gradually develop symptoms of quadriplegia and death.Applying 6-gingerol to the experimental model of experimental allergic encephalomyelitis in mice,and optimizing the dosage and dosing schedule can significantly inhibit its pathogenesis.
Keywords/Search Tags:Multiple sclerosis, T cells, 6 gingerol, EAE, ROS, autoimmune disease
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