發(fā)布時(shí)間：2018-04-04 01:41

  本文選題：肝臟內(nèi)質(zhì)網(wǎng)應(yīng)激　切入點(diǎn)：鈣激活中性蛋白酶2　出處：《上海師范大學(xué)》2017年碩士論文

【摘要】：內(nèi)質(zhì)網(wǎng)應(yīng)激(endoplasmic reticulum stress,ERS)是指由于各種原因?qū)е碌募?xì)胞內(nèi)質(zhì)網(wǎng)(endoplasmic reticulum,ER)功能發(fā)生紊亂,蛋白質(zhì)合成受到影響的病理、生理過(guò)程。而肝臟作為人體內(nèi)重要的代謝器官,其出現(xiàn)ERS可誘發(fā)一系列代謝性疾病,肝臟ERS已成為諸多疾病的重要風(fēng)險(xiǎn)因素,危害到人類(lèi)健康。Calpain-2是鈣激活中性蛋白酶,最新的研究表明,Calpain-2在人體內(nèi)的激活可能參與到某些病理活動(dòng)中去,如肝臟ERS。但Calpain-2在肝臟ERS中的作用及相關(guān)機(jī)制目前尚不清楚。研究目的:本研究通過(guò)對(duì)SD大鼠進(jìn)行8周高脂飲食喂養(yǎng)后成功建立大鼠肝臟ERS動(dòng)物模型,并對(duì)高脂飲食組大鼠實(shí)施8周游泳運(yùn)動(dòng)干預(yù),探究Calpain-2在運(yùn)動(dòng)改善肝臟ERS中的作用及相關(guān)機(jī)制,為運(yùn)動(dòng)作為非藥物療法治療相關(guān)疾病提供理論依據(jù)。研究方法:30只雄性SD大鼠隨機(jī)分成對(duì)照組(C組)、高脂飲食ERS模型組(HF組)、高脂飲食ERS模型運(yùn)動(dòng)組(HE組),每組10只。其中,C組大鼠用基礎(chǔ)飼料(碳水化合物65%,蛋白質(zhì)22%,脂肪13%)喂養(yǎng),HF組和HE組大鼠均使用高脂飼料喂養(yǎng)(碳水化合物33%,蛋白質(zhì)22%,脂肪45%),HE組大鼠在喂養(yǎng)高脂飼料的同時(shí)施以8周無(wú)負(fù)重游泳運(yùn)動(dòng)干預(yù)。8周后對(duì)三組大鼠進(jìn)行取材,用比色法(enzymatic colorimetric assay)檢測(cè)大鼠血清游離脂肪酸(free fatty acids,FFAs)含量;用免疫印跡法(westernblotting)檢測(cè)大鼠肝臟Calpain-2、自噬相關(guān)基因7(autophagy related gene,Atg7)的蛋白表達(dá)變化;用實(shí)時(shí)熒光定量PCR(RT-qPCR)檢測(cè)大鼠肝臟Calpain-2、Atg7、CCAAT/增強(qiáng)子結(jié)合蛋白同源蛋白(CCAAT/enhancer binding protein homologus protein,Chop)、葡萄糖調(diào)節(jié)蛋白78(glucose regulated protein78,Grp78)的基因表達(dá)水平。研究結(jié)果:(1)與C組相比,HF組大鼠肝臟組織中Grp78、Chop、EIF2αmRNA顯著升高(p0.01),說(shuō)明大鼠肝臟ERS模型建立成功。(2)與C組相比,HF組大鼠血漿FFAs含量顯著增加,肝臟組織中Calpain-2mRNA及蛋白表達(dá)水平顯著增加,Atg7 mRNA及蛋白表達(dá)水平顯著降低(p0.01),說(shuō)明高脂飲食可通過(guò)上調(diào)肝臟中Calpain-2的表達(dá),抑制肝臟中Atg7的表達(dá)水平進(jìn)而誘導(dǎo)ERS的標(biāo)志物Grp78、Chop及EIF2α的出現(xiàn)。(3)與HF組相比,HE組大鼠血清FFAs含量顯著下降,肝臟組織中Calpain-2mRNA及蛋白表達(dá)水平顯著降低,Atg7 mRNA及蛋白表達(dá)水平顯著升高(p0.01),繼而使得ERS標(biāo)志物Grp78、Chop及EIF2αmRNA表達(dá)水平下降,說(shuō)明游泳運(yùn)動(dòng)可有效緩解肝臟ERS。結(jié)論:(1)8周高脂飲食成功建立大鼠肝臟ERS模型。(2)運(yùn)動(dòng)可有效緩解肝臟ERS。Calpain-2在運(yùn)動(dòng)緩解肝臟ERS中發(fā)揮重要作用,其可能機(jī)制是:運(yùn)動(dòng)降低血清中FFAs含量,進(jìn)而下調(diào)肝臟組織中Calpain-2的表達(dá)水平,恢復(fù)肝臟中Atg7的表達(dá),Atg7表達(dá)上調(diào)可使ERS標(biāo)志物Grp78、Chop及EIF2α的mRNA表達(dá)水平降低,說(shuō)明肝臟ERS得到有效緩解。
[Abstract]:Endoplasmic reticulum stress (ERS) is a pathological and physiological process in which the endoplasmic reticulum (ER) function is disturbed and the protein synthesis is affected.As an important metabolic organ in human body, liver ERS can induce a series of metabolic diseases. Liver ERS has become an important risk factor for many diseases. Calpain-2 is a calcium activated neutral protease, which is harmful to human health.Recent studies suggest that the activation of Calpain-2 in the human body may be involved in certain pathological activities, such as liver ERSs.However, the role of Calpain-2 in liver ERS and its related mechanisms are unclear.Objective: to establish a rat liver ERS model after feeding SD rats with high fat diet for 8 weeks, and to interfere with swimming exercise for 8 weeks in high fat diet group.To explore the role of Calpain-2 in the improvement of liver ERS by exercise and its related mechanism, and to provide theoretical basis for the treatment of related diseases by exercise as a non-drug therapy.Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: control group (n = 10), high-fat diet ERS model group (n = 10), high-fat diet ERS model group (n = 10) and high-fat diet ERS model exercise group (n = 10).The rats in group C were fed with basic diet (carbohydrate 65, protein 22, fat 13), HF group and HE group were fed with high fat diet (carbohydrate 33, protein 22, fat 4530%) while feeding high fat diet.The rats in the three groups were selected after 8 weeks of weightless swimming intervention.The content of free fatty acid free fatty acids#en0# in serum was detected by colorimetric method, and the protein expression of Calpain-2 and 7(autophagy related gene Atg7 in rat liver was detected by Western blotting method.The expression of CCAAT-2Ag7 / enhancer binding protein homologus protein Chop#en0# and 78(glucose regulated protein 78 in rat liver were detected by real-time fluorescence quantitative PCR- RT-qPCR.The results showed that the expression of CCAAT-2CCAAT-7 / enhancer protein homologus protein and 78(glucose regulated protein 78-Grp78 were detected by real-time fluorescence quantitative PCR.Results compared with group C, the level of plasma FFAs in HF group was significantly higher than that in group C (P < 0.01).The expression of Calpain-2mRNA and protein in liver tissue increased significantly, and the expression of Atg7 mRNA and protein decreased significantly, indicating that high fat diet can up-regulate the expression of Calpain-2 in liver.Inhibiting the expression of Atg7 in liver and inducing the appearance of ERS markers Grp78 Chop and EIF2 偽.) compared with HF group, the serum FFAs content in HE group was significantly lower than that in HF group.The expression of Calpain-2mRNA and protein in liver tissue decreased significantly, and the expression of Atg7 mRNA and protein increased significantly, and the expression level of ERS marker Grp78 Chop and EIF2 偽 mRNA decreased, which indicated that swimming exercise could effectively alleviate liver ERSs.Conclusion ERS model of rat liver was successfully established by high-fat diet for 8 weeks. Conclusion exercise can effectively alleviate the role of liver ERS.Calpain-2 in relieving liver ERS. The possible mechanism is that exercise can reduce the content of FFAs in serum.Furthermore, down-regulation of Calpain-2 expression and up-regulation of Atg7 expression in liver could decrease the mRNA expression of ERS marker Grp78 Chop and EIF2 偽, which indicated that liver ERS was effectively alleviated.
【學(xué)位授予單位】：上海師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R455

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陳娜子;姜潮;李校X;;內(nèi)質(zhì)網(wǎng)應(yīng)激與疾病[J];中國(guó)生物工程雜志;2016年01期

2 梁麗娜;戰(zhàn)麗彬;胡守玉;隋華;陳靜;;滋補(bǔ)脾陰方藥調(diào)節(jié)下丘腦中自噬及內(nèi)質(zhì)網(wǎng)應(yīng)激改善脾陰虛糖尿病認(rèn)知功能障礙機(jī)制研究[J];世界科學(xué)技術(shù)-中醫(yī)藥現(xiàn)代化;2015年06期

3 謝汝佳;韓冰;楊婷;溫靜靜;楊勤;;鈣中性蛋白酶2及Bax在肝纖維化大鼠肝組織中的表達(dá)變化及意義[J];中國(guó)病理生理雜志;2013年09期

4 劉效磊;牛燕媚;傅力;;自噬——運(yùn)動(dòng)改善胰島素抵抗的新機(jī)制[J];生理科學(xué)進(jìn)展;2013年03期

5 代景友;張新晨;楊維良;吳德全;;內(nèi)質(zhì)網(wǎng)應(yīng)激介導(dǎo)的細(xì)胞自噬與凋亡研究進(jìn)展[J];中華實(shí)驗(yàn)外科雜志;2012年07期

6 向波;易梅;李小玲;李桂源;;細(xì)胞自噬在腫瘤發(fā)生發(fā)展中的作用[J];生物化學(xué)與生物物理進(jìn)展;2012年03期

7 王志鵬;吳靜;景友玲;門(mén)秀麗;;內(nèi)質(zhì)網(wǎng)應(yīng)激在2型糖尿病胰島素抵抗中的作用[J];生理科學(xué)進(jìn)展;2012年01期

8 溫悅萌;張?zhí)鞖W;謝嵐;艾華;管又飛;;一次性力竭運(yùn)動(dòng)對(duì)小鼠肝臟和骨骼肌內(nèi)質(zhì)網(wǎng)應(yīng)激的影響[J];中國(guó)運(yùn)動(dòng)醫(yī)學(xué)雜志;2012年02期

9 張勇;李之俊;;運(yùn)動(dòng)與脂肪動(dòng)員研究進(jìn)展[J];中國(guó)運(yùn)動(dòng)醫(yī)學(xué)雜志;2012年01期

10 陳潔;熊吉;陳瀟迪;牟歌;王軍;樊麗琳;陳東風(fēng);;Calpain 2調(diào)節(jié)自噬相關(guān)基因ATG7的表達(dá)在非酒精性脂肪性肝病中的作用[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2011年20期

相關(guān)博士學(xué)位論文 前3條

1 崔巍;游離脂肪酸通過(guò)Ca~(2+)/calpain-2途徑誘導(dǎo)β細(xì)胞內(nèi)質(zhì)網(wǎng)應(yīng)激和凋亡的機(jī)制研究[D];第四軍醫(yī)大學(xué);2013年

2 程夢(mèng)婕;肝臟內(nèi)質(zhì)網(wǎng)應(yīng)激與飲食誘導(dǎo)肥胖大鼠胰島素抵抗和肝臟脂肪變性的關(guān)系研究[D];華中科技大學(xué);2012年

3 寧波;內(nèi)質(zhì)網(wǎng)應(yīng)激在非酒精性脂肪性肝病肝細(xì)胞損傷中的作用及機(jī)制研究[D];重慶醫(yī)科大學(xué);2009年

，

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