發(fā)布時(shí)間：2018-06-14 06:06

  本文選題：骨骼肌 + 胰島素抵抗��； 參考：《安徽醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:觀察高脂飲食(high-fat diet,HFD)對雌性SD大鼠骨骼肌的影響,探討非諾貝特(fenofibrate,FF)對高脂及棕櫚酸導(dǎo)致的骨骼肌胰島素抵抗的保護(hù)作用是否與內(nèi)質(zhì)網(wǎng)應(yīng)激(endoplasmic reticulum stress,ERS)有關(guān)。方法:雌性SD大鼠隨機(jī)分為三組:標(biāo)準(zhǔn)飲食組(standard control diet,SCD)、高脂飲食組(HFD)及治療組(HFD+FF)。高脂組和治療組大鼠先予以高脂飲食20wk后,高脂組大鼠繼續(xù)予以高脂飲食8wk,治療組大鼠予以非諾貝特(30mg·kg-1·d-1)灌胃治療8wk。每周監(jiān)測大鼠體重,第28周檢測血清生化指標(biāo)TC、TG、LDL-C、HDL-C,葡萄糖耐量試驗(yàn)(GTT)和胰島素耐量試驗(yàn)(ITT)測定胰島素抵抗(IR),Real-time RT-PCR測定骨骼肌組織葡萄糖調(diào)節(jié)蛋白78(GRP78)、轉(zhuǎn)錄因子GADD153(CHOP)、肌醇酶1α(IRE1α)、真核起始因2的α亞基(e IF2α)和過氧化物酶體增殖物激活受體α(PPARα)基因表達(dá),Western-blot法檢測骨骼肌組織GRP78蛋白的表達(dá)水平。將骨骼肌細(xì)胞C2C12分組:正常對照組(normal control group,NC)、模型組(棕櫚酸組,palmitic acid,PA)、陽性對照藥組(衣霉素組,tunicamycin,TM)、治療組(棕櫚酸+非諾貝特酸,PA+FA),RT-PCR測定GRP78、CHOP、IRE1α和e IF2αm RNA的表達(dá)水平,Western-blot檢測GRP78、Akt和p-Akt蛋白表達(dá)水平,免疫熒光檢測CHOP蛋白表達(dá)水平。結(jié)果:(1)GTT實(shí)驗(yàn)提示HFD組大鼠給予葡萄糖30 min后血糖值達(dá)高峰,且較SCD組、HFD+FF組血糖峰值升高,此后緩慢下降;與HFD組大鼠相比,HFD+FF組大鼠空腹血糖降低,給予葡萄糖后30 min時(shí)峰值降低,血糖波動(dòng)較小。ITT實(shí)驗(yàn)顯示大鼠注射胰島素后,SCD組血糖迅速下降,30 min時(shí)達(dá)最低值后血糖緩慢上升;HFD組血糖緩慢下降,且始終維持在較高水平,90 min時(shí)達(dá)最低值后血糖緩慢上升;HFD+FF組大鼠各點(diǎn)血糖值介于HCD和SCD組之間,注射胰島素后血糖下降至60 min時(shí)達(dá)最低值,隨后緩慢上升。(2)與SCD組大鼠相比,HFD組大鼠體重顯著增加(P0.05),血清TG、TC水平明顯升高,HDL-C水平顯著下降(P0.05);與HFD組大鼠相比,非諾貝特干預(yù)后能夠顯著減少大鼠體重、改善肥胖,降低血清TC、TG水平,增加血清HDL-C水平(P0.05)。(3)與SCD組大鼠相比,HFD組大鼠GRP78、CHOP、IRE1α和e IF2α基因表達(dá)水平均明顯上調(diào),與HFD組大鼠相比,非諾貝特治療后PPARα表達(dá)水平明顯增加,GRP78、CHOP、IRE1α和e IF2α基因表達(dá)水平下調(diào);(4)與NC組相比,PA組骨骼肌細(xì)胞GRP78、CHOP、IRE1α和e IF2α基因表達(dá)水平顯著增加,p-Akt蛋白表達(dá)水平明顯下調(diào);與PA組相比,PA+FA組骨骼肌細(xì)胞GRP78、CHOP、IRE1α和e IF2α基因表達(dá)顯著降低,p-Akt蛋白表達(dá)水平明顯上調(diào)。結(jié)論:高脂飲食成功構(gòu)建雌性高脂血癥伴IR大鼠模型,非諾貝特干預(yù)可顯著控制大鼠體重,調(diào)節(jié)血脂紊亂,改善胰島素抵抗,其作用機(jī)制可能與顯著抑制ERS標(biāo)志物CHOP、GRP78、IRE1α和e IF2α的表達(dá)水平有關(guān)。
[Abstract]:Aim: to observe the effect of high-fat HFDs on skeletal muscle of female SD rats and to investigate whether the protective effect of fenofibrate FFF on insulin resistance induced by hyperlipidemia and palmitic acid is related to endoplasmic reticulum stress (endoplasmic reticulum stress). Methods: female SD rats were randomly divided into three groups: standard diet group, high fat diet group and treatment group. Rats in hyperlipidemia group and treatment group were treated with high-fat diet (20wk). Rats in high-fat group were given high-fat diet for 8 wk, and rats in treatment group were treated with fenofibrate 30mg kg-1 d-1 (gavage) for 8 wk. The body weight of rats was monitored weekly. Determination of Serum biochemical Indexes TCX TGN LDL-Con HDL-C (glucose tolerance Test GTT) and Insulin tolerance Test (ITT) in skeletal muscle tissue by Real-time RT-PCR for determination of Glucose-regulated protein 78GRP78, transcription Factor GADD153, Inositase 1 偽 -IRE1 偽, Eukaryotic initiation Factor 2 偽 Subunit e IF2 偽) and peroxisome proliferator activated receptor 偽 PPAR 偽) were used to detect the expression of GRP78 protein in skeletal muscle by Western-blot. C2C12 skeletal muscle cells were divided into normal control group (normal control group), model group (palmitic acidopyridine), positive control group (tunicamycin TMN), treatment group (fenofibrate palmitate PA FAFA), RT-PCR for the detection of GRP78 CHOPIRE1 偽 and e IF2 偽 mRNA expression levels by Western-blot. The expression level of GRP78, Akt and p-Akt protein, The expression of chop protein was detected by immunofluorescence. Results the blood glucose level of HFD group reached the peak after 30 min glucose administration, which was higher than that of SCD group, and then decreased slowly, and the fasting blood glucose in HFD FF group was lower than that in HFD group. The peak value of glucose decreased at 30 min after glucose administration, and the blood glucose fluctuated slightly. ITT test showed that the blood glucose of the SCD group decreased rapidly after insulin injection and reached the lowest level at 30 min. The blood glucose of the HFD group slowly increased after 30 min, and the blood glucose of the HFD group decreased slowly. The blood glucose level of HFDFF group was between HCD group and SCD group, and the blood glucose level decreased to 60 min after insulin injection. Compared with the rats of SCD group, the weight of HFD group increased significantly (P 0.05), serum TGN TC level increased significantly and HDL-C level decreased significantly compared with HFD group, fenofibrate could significantly reduce body weight and improve obesity. Compared with SCD group, the expression of GRP78 CHOPIRE1 偽 and e IF2 偽 gene in HFD group was significantly up-regulated, compared with that in HFD group, and that in HFD group was significantly higher than that in HFD group, and that in HFD group was significantly higher than that in HFD group, and the expression of IRE1 偽 and e IF2 偽 in HFD group was significantly higher than that in HFD group. After fenofibrate treatment, the expression of PPAR 偽 increased significantly (P < 0.05). The expression levels of GRP78-CHOPOP-IRE1 偽 and eIF2 偽 decreased significantly (P < 0.01). Compared with NC group, the expression levels of GRP78-CHOPON-IRE1 偽 and eIF2 偽 in skeletal muscle cells of PA group increased significantly, and the expression levels of p-Akt protein decreased significantly. Compared with PA group, the expression of GRP78 CHOPIRE1 偽 and eIF2 偽 gene in skeletal muscle cells in PA FA group was significantly lower than that in PA group. Conclusion: female hyperlipidemia with IR rat model was successfully constructed by high-fat diet. Fenofibrate intervention can significantly control the weight of rats, regulate the disorder of blood lipids, and improve insulin resistance. The mechanism may be related to the significant inhibition of the expression levels of ERS markers CHOP-GRP78, IRE1 偽 and eIF2 偽.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R589

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本文編號：2016391