本文選題：葡萄糖　切入點(diǎn)：同型半胱氨酸　出處：《山西醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：觀察體外高糖和/或高同型半胱氨酸(high homocysteine, hHcy)環(huán)境下，小鼠腎小球足細(xì)胞內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)蛋白：葡萄糖調(diào)節(jié)蛋白78(glucose regulatedprotein78, GRP78/BiP)、生長(zhǎng)阻滯和DNA損傷誘導(dǎo)基因153(growth arrest andDNAdamage inducible gene153, CHOP/GADD153)的表達(dá)，初步探討高糖和/或hHcy誘導(dǎo)內(nèi)質(zhì)網(wǎng)應(yīng)激(endoplasmic reticulum stress, ERS)導(dǎo)致足細(xì)胞凋亡的相關(guān)機(jī)制，觀察抗氧化劑白藜蘆醇(resveratrol, RES)對(duì)足細(xì)胞的保護(hù)作用。 方法：分化后的條件永生化小鼠足細(xì)胞分6組：正常對(duì)照組、高滲對(duì)照組(葡萄糖5mmol/L+甘露醇15mmol/L)、高糖組(葡萄糖20mmol/L)、hHcy組(hHcy200umol/L)、hHcy+高糖組(hHcy200umol/L+葡萄糖20mmol/L)、混合+RES組(RES20umol/L預(yù)先干預(yù)半小時(shí)+hHcy200umol/L+葡萄糖20mmol/L)，作用24小時(shí)后結(jié)束干預(yù)。采用流式細(xì)胞術(shù)AnnexinV-FITC/PI法測(cè)定各組細(xì)胞凋亡率；Real-time PCR技術(shù)分析各組細(xì)胞GRP78、CHOP mRNA的表達(dá)情況；Western blot技術(shù)評(píng)價(jià)各組細(xì)胞GRP78、CHOP蛋白的表達(dá)情況。 結(jié)果：高糖組的凋亡率、GRP78、CHOP mRNA及蛋白表達(dá)明顯高于正常對(duì)照組及高滲對(duì)照組(P 0.05)；hHcy組的凋亡率、GRP78、CHOP mRNA及蛋白表達(dá)顯著高于正常對(duì)照組(P 0.05)；hHcy+高糖組的凋亡率、GRP78、CHOPmRNA及蛋白表達(dá)分別與高糖組、hHcy組比較均顯著升高(P 0.05)；混合+RES組較hHcy+高糖組，凋亡率、GRP78、CHOP mRNA及蛋白表達(dá)顯著下降(P 0.05)，上述各組差異均有統(tǒng)計(jì)學(xué)意義；高滲對(duì)照組較正常對(duì)照組,凋亡率、GRP78、CHOP mRNA及蛋白表達(dá)水平無(wú)顯著變化(P0.05)，差異無(wú)統(tǒng)計(jì)學(xué)意義。 結(jié)論：體外高糖、hHcy環(huán)境能誘導(dǎo)小鼠足細(xì)胞凋亡，hHcy和高糖共同環(huán)境下對(duì)足細(xì)胞損傷及致凋亡作用更明顯，提示hHcy、高糖兩者有協(xié)同作用；內(nèi)質(zhì)網(wǎng)應(yīng)激參與其凋亡過(guò)程，，抗氧化劑RES能抵抗高糖和hHcy共同環(huán)境下誘導(dǎo)的足細(xì)胞凋亡，阻斷內(nèi)質(zhì)網(wǎng)應(yīng)激CHOP通路為其機(jī)制之一。
[Abstract]:Aim: to investigate the expression of high homocysteine (hHcyine) stress associated proteins in glomerular podocyte endoplasmic reticulum (ER): glucose regulated protein 78 (GRP 78 / BiPN), growth arrest and DNA damage inducible gene 153 arrest andDNAdamage inducible genetics 153, CHOP- GADD153) in high glucose and / or hyperhomocysteine (hHcyine) environment in vitro. To investigate the mechanism of endoplasmic reticulum stress induced by high glucose and / or hHcy in podocyte apoptosis, and to observe the protective effect of antioxidant resveratrol on podocyte. Methods: the conditioned immortalized mouse podocytes after differentiation were divided into 6 groups: normal control group. The hypertonic control group (glucose 5 mmol / L mannitol 15 mmol / L), the high glucose 20 mmol / L Hcy group (20 mmol / L hHcy group), the hHcy200 mol / L glucose 20 mmol / L group, the mixed RES group Res 20 mol / L pretreated with hHcy200umol/L 20 mmol / L L / L, and the flow cytometry AnnexinV-FITC/PI / L were used. Methods the expression of GRP78 chop mRNA was analyzed by real-time PCR and Western blot was used to evaluate the expression of GRP78 chop protein. Results: the expression of GRP78 / CHOPmRNA and protein in high glucose group was significantly higher than that in normal control group and hyperosmotic control group. The expression of GRP78 / CHOPmRNA and protein in hyperosmotic control group was significantly higher than that in normal control group (P 0.05 Hcy group) and the expression of GRP78 CHOPmRNA and protein in hyperosmotic control group was significantly higher than that in control group (P 0.05 Hcy high glucose group). Compared with high glucose group, Hcy group significantly increased P 0.05, and mixed RES group was higher than hHcy high glucose group. The expression of GRP78 chop mRNA and protein decreased significantly (P 0.05), and there was no significant difference between the hypertonic control group and the normal control group in the expression of GRP78 chop mRNA and protein (P 0.05), but there was no significant difference in the expression of GRP78 chop mRNA and protein between the hypertonic control group and the normal control group (P 0.05). Conclusion: Hcy in vitro can induce apoptosis of podocyte in mice. Hcy can induce apoptosis and damage of podocyte in vitro, suggesting that hHcyand high glucose have synergistic effect, endoplasmic reticulum stress is involved in the apoptosis process of podocyte. The antioxidant RES can resist the apoptosis of podocyte induced by high glucose and hHcy. Blocking endoplasmic reticulum stress CHOP pathway is one of its mechanisms.
【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
【分類號(hào)】：R587.2;R692

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 朱雪婧;劉伏友;彭佑銘;劉虹;袁曙光;許向青;劉映紅;孫林;凌光輝;;糖尿病腎病病理分型最新國(guó)際標(biāo)準(zhǔn)的臨床應(yīng)用(附37例回顧性分析)[J];中南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2012年02期

2 關(guān)麗英;許彩民;潘華珍;;內(nèi)質(zhì)網(wǎng)應(yīng)激介導(dǎo)的細(xì)胞凋亡[J];生物化學(xué)與生物物理進(jìn)展;2007年11期

3 李金紅;陶建瓴;李航;;足細(xì)胞損傷與糖尿病腎病的研究現(xiàn)狀[J];中國(guó)醫(yī)學(xué)科學(xué)院學(xué)報(bào);2010年05期

本文編號(hào)：1625724