他克莫司对嘌呤霉素诱导足细胞凋亡的影响及其机制研究

发布时间：2018-03-14 18:01

本文选题：他克莫司　切入点：嘌呤霉素　出处：《山西医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：通过观察他克莫司对体外嘌呤霉素诱导的小鼠足细胞凋亡率和凋亡相关基因、蛋白表达的影响，探讨他克莫司保护足细胞的可能机制。方法：体外培养小鼠肾小球足细胞，设立CON（对照）组，，PAN(嘌呤霉素)组，FK506（他克莫司）组。CON组用无血清RPMI-1640培养液培养，PAN组加入50μg/mLPAN作用48小时, FK506组预先加入1μmol/L他克莫司作用30min后加入50μg/mL PAN共同作用48小时。采用流式细胞仪检测足细胞凋亡率，RT-PCR检测各组Bax、Bcl-2的mRNA表达量，Western印迹法检测各组Bax、Bcl-2、ERK1/2、p-ERK1/2的蛋白表达量。结果：1、流式细胞术检测PAN组足细胞凋亡率明显高于CON组和FK506组（P0.05）。2、RT-PCR结果显示PAN组足细胞Bax的mRNA表达量高于CON组和FK506组（P0.05），Bcl-2的mRNA表达量低于CON组和FK506组（P0.05），Bax/Bcl-2比值高于CON组和FK506组（P0.05）。3、根据WesternBolt结果，PAN组足细胞Bax的蛋白表达量高于CON组和FK506组（P0.05），Bcl-2的蛋白表达量低于CON组和FK506组（P0.05），Bax/Bcl-2比值高于CON组和FK506组（P0.05）。4、根据Western Bolt结果，PAN组p-ERK1/2/ERK1/2蛋白相对表达量低于CON组和FK506组（P0.05）。结论：他克莫司可以下调Bax的表达，上调Bcl-2的表达，降低Bax/Bcl-2比值，抑制PAN诱导的足细胞的凋亡，而其作用机制可能与ERK1/2通路的激活相关。
[Abstract]:Objective: To observe the effect of tacrolimus on the apoptosis rate and apoptosis related genes and protein expression of podocycin induced podocytes in vitro, and to explore the possible mechanism of tacrolimus in the protection of podocytes.
Methods: mouse podocyte cell culture in vitro, the establishment of CON (control) group, PAN (puromycin) group, FK506 (tacrolimus) group.CON group with serum free RPMI-1640 medium, PAN group with 50 g/mLPAN for 48 hours, FK506 group added beforehand 1 mol/L tacrolimus 30min after adding 48 hours together effect of 50 g/mL PAN. The apoptosis rate was detected by flow cytometry, RT-PCR detected the Bax expression of Bcl-2, mRNA, Bax were detected by Western blotting, Bcl-2, ERK1/2, the expression quantity of p-ERK1/2.
Results: 1. The detection rate of podocyte apoptosis in PAN group was significantly higher than that of CON group and FK506 group by flow cytometry (P0.05).2, RT-PCR showed that PAN group Bax of podocyte mRNA expression is higher than that of CON group and FK506 group (P0.05), Bcl-2 mRNA expression was lower than that of CON group and FK506 group (P0.05). The Bax/ ratio of Bcl-2 is higher than that of CON group and FK506 group (P0.05) according to the results of WesternBolt,.3, PAN group the expression of Bax of podocyte protein is higher than that of CON group and FK506 group (P0.05), the expression of Bcl-2 was lower than that of CON group and FK506 group (P0.05), the ratio of Bax/Bcl-2 is higher than that of CON group and FK506 group (P0.05.4) Western Bolt, according to the results, the relative expression of PAN protein in p-ERK1/2/ERK1/2 group was lower than that of CON group and FK506 group (P0.05).
Conclusion: tacrolimus can down regulate the expression of Bax, up regulate Bcl-2 expression, decrease Bax/Bcl-2 ratio and inhibit podocyte apoptosis induced by PAN, and its mechanism may be related to activation of ERK1/2 pathway.

【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692

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