本文選題：草酸鈣 + 人腎小管上皮細胞��； 參考：《沈陽醫(yī)學院》2017年碩士論文

【摘要】：目的內(nèi)質(zhì)網(wǎng)是蛋自翻譯后修飾的場所,內(nèi)質(zhì)網(wǎng)應激被視為誘導細胞發(fā)生凋亡的第二條信號通路,是誘發(fā)多種慢性、代謝性疾病發(fā)病的重要途徑。本項目研究腎結(jié)石形成與內(nèi)質(zhì)網(wǎng)應激的關系。方法本研究采用100μg/mL的草酸鈣結(jié)晶刺激HK-2人腎小管上皮細胞3h、6h、12h和24h,Oh為對照組,用Real-time PCR法檢測內(nèi)質(zhì)網(wǎng)應激標志基因BIP/GRP78,同時檢測內(nèi)質(zhì)網(wǎng)應激相關信號通路中CHOP、PERK、JNK、P38和CasPase-3等基因mRNA的表達情況,用Western blot法檢測內(nèi)質(zhì)網(wǎng)應激相關蛋白BIP/GRP78、CHOP、P-PERK、P-JNK、P-P38、XBP1、Cleaved-CasPase-3 和CasPase-12的表達情況,用TUNEL法檢測草酸鈣結(jié)晶刺激24h時細胞凋亡的情況。結(jié)果用100μg/mL的草酸鈣結(jié)晶刺激HK-2細胞后,內(nèi)質(zhì)網(wǎng)應激標志基因BIP/GRP78和CHOP基因的mRNA表達升高,內(nèi)質(zhì)網(wǎng)應激信號通路中的PERK、JNK、P38基因的mRNA表達均升高。當用草酸鈣結(jié)晶刺激HK-2細胞6h,BIP/GRP78蛋白表達升高,暴露12h,CHOP蛋白表達升高與對照組Oh相比具有統(tǒng)計學差異,P-PERK、P-JNK、P-P38和XBP1蛋白分別在草酸鈣結(jié)晶刺激3h、3h、6h和3h時蛋白表達與對照組相比升高,Western blot檢測結(jié)果與Real-time RCR結(jié)果一致。TUNEL檢測發(fā)現(xiàn)24小時結(jié)晶暴露后細胞發(fā)生凋亡,凋亡率約達25%。Western blot顯示內(nèi)質(zhì)網(wǎng)應激特異性蛋白CasPase-12和Cleaved-CasPase-3的表達升高。結(jié)論我們研究發(fā)現(xiàn)草酸鈣結(jié)晶誘導腎小管上皮細胞發(fā)生了內(nèi)質(zhì)網(wǎng)應激,結(jié)晶誘導的腎小管細胞凋亡是通過線粒體凋亡通路和內(nèi)質(zhì)網(wǎng)凋亡通路共同引起的。這些發(fā)現(xiàn)提示內(nèi)質(zhì)網(wǎng)應激可能是腎結(jié)石形成過程的重要發(fā)病機制。
[Abstract]:Objective the endoplasmic reticulum (ER) is a place where eggs are modified after translation. Endoplasmic reticulum stress is regarded as the second signal pathway to induce apoptosis, and it is an important way to induce many chronic and metabolic diseases. The purpose of this study was to study the relationship between renal stone formation and endoplasmic reticulum stress. Methods in this study, 100 渭 g/mL calcium oxalate crystals were used to stimulate HK-2 human renal tubular epithelial cells for 12 h and 24 h respectively as control group. The expression of the endoplasmic reticulum stress marker gene BIP-GRP78, the expression of CHOPPERKP38 and CasPase-3 genes in the endoplasmic reticulum stress-related signaling pathway and the expression of BIPP / GRP78 CHOPP-PERKP-JNKP-P38 and XBP1Cleaved-CasPase-3 and CasPase-12 were detected by Real-time PCR method, and the expression of BIP-PERKP-JNKP-38-XBP1Cleaved-Caspase-3 and CasPase-12 were detected by Western blot method. The apoptosis of calcium oxalate after 24 h was detected by TUNEL method. Results after 100 渭 g/mL calcium oxalate was used to stimulate HK-2 cells, the mRNA expression of endoplasmic reticulum stress marker gene BIP/GRP78 and CHOP gene was increased, and the mRNA expression of PERKN JNKN P38 gene in endoplasmic reticulum stress signaling pathway was increased. The expression of BIPP / GRP78 protein was increased in HK-2 cells stimulated by calcium oxalate crystallization for 6 h. There were significant differences in the expression of chop protein between the control group and the control group at 12 h after exposure. The protein expression of P-PERKU P-JNKP-P38 and XBP1 was increased at 3h and 3h after the crystallization of calcium oxalate, respectively. The results of Western blot and Real-time RCR were consistent with those of the control. It was found that cell apoptosis occurred after 24 hours of crystallization exposure. Apoptosis rate up to 25%.Western blot showed increased expression of ER stress-specific proteins CasPase-12 and Cleaved-CasPase-3. Conclusion We found that calcium oxalate crystallization induced endoplasmic reticulum stress in renal tubular epithelial cells. The apoptosis of renal tubular cells induced by calcium oxalate was induced by mitochondrial apoptosis pathway and endoplasmic reticulum apoptosis pathway. These findings suggest that endoplasmic reticulum stress may play an important role in the formation of renal calculi.
【學位授予單位】：沈陽醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R692.4

【參考文獻】

相關期刊論文 前5條

1 姜澤群;姚志華;鄧正亭;蔣鑫超;顏曉靜;陳衛(wèi)平;;涼血化瘀方拮抗內(nèi)質(zhì)網(wǎng)應激引起的L02肝細胞凋亡作用及機制研究[J];中國中藥雜志;2013年20期

2 劉友平;嚴冬梅;陳川寧;段春燕;陳紹坤;李洪;代榮陽;;PI3K/Akt調(diào)控內(nèi)質(zhì)網(wǎng)應激對GRP78的誘導[J];中國病理生理雜志;2011年04期

3 楊欣榮,陳鐘;肝細胞移植中肝細胞凋亡的研究進展[J];國外醫(yī)學.外科學分冊;2005年06期

4 呂游,張示杰;肝臟疾病中肝細胞凋亡的研究進展[J];中國基層醫(yī)藥;2005年10期

5 程法娟;許冬梅;;內(nèi)質(zhì)網(wǎng)應激在腎小球疾病中的研究進展[J];臨床腎臟病雜志;2010年04期

，

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