本文選題：他克莫司　切入點(diǎn)：嘌呤霉素　出處：《山西醫(yī)科大學(xué)》2014年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的：通過(guò)觀(guān)察他克莫司對(duì)體外嘌呤霉素誘導(dǎo)的小鼠足細(xì)胞凋亡率和凋亡相關(guān)基因、蛋白表達(dá)的影響，探討他克莫司保護(hù)足細(xì)胞的可能機(jī)制。 方法：體外培養(yǎng)小鼠腎小球足細(xì)胞，設(shè)立CON（對(duì)照）組，，PAN(嘌呤霉素)組，F(xiàn)K506（他克莫司）組。CON組用無(wú)血清RPMI-1640培養(yǎng)液培養(yǎng)，PAN組加入50μg/mLPAN作用48小時(shí), FK506組預(yù)先加入1μmol/L他克莫司作用30min后加入50μg/mL PAN共同作用48小時(shí)。采用流式細(xì)胞儀檢測(cè)足細(xì)胞凋亡率，RT-PCR檢測(cè)各組Bax、Bcl-2的mRNA表達(dá)量，Western印跡法檢測(cè)各組Bax、Bcl-2、ERK1/2、p-ERK1/2的蛋白表達(dá)量。 結(jié)果：1、流式細(xì)胞術(shù)檢測(cè)PAN組足細(xì)胞凋亡率明顯高于CON組和FK506組（P0.05）。2、RT-PCR結(jié)果顯示PAN組足細(xì)胞Bax的mRNA表達(dá)量高于CON組和FK506組（P0.05），Bcl-2的mRNA表達(dá)量低于CON組和FK506組（P0.05），Bax/Bcl-2比值高于CON組和FK506組（P0.05）。3、根據(jù)WesternBolt結(jié)果，PAN組足細(xì)胞Bax的蛋白表達(dá)量高于CON組和FK506組（P0.05），Bcl-2的蛋白表達(dá)量低于CON組和FK506組（P0.05），Bax/Bcl-2比值高于CON組和FK506組（P0.05）。4、根據(jù)Western Bolt結(jié)果，PAN組p-ERK1/2/ERK1/2蛋白相對(duì)表達(dá)量低于CON組和FK506組（P0.05）。 結(jié)論：他克莫司可以下調(diào)Bax的表達(dá)，上調(diào)Bcl-2的表達(dá)，降低Bax/Bcl-2比值，抑制PAN誘導(dǎo)的足細(xì)胞的凋亡，而其作用機(jī)制可能與ERK1/2通路的激活相關(guān)。
[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.

【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R692

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