發(fā)布時(shí)間：2018-01-18 03:02

  本文關(guān)鍵詞：二甲雙胍對(duì)妊娠期糖尿病小鼠母代和跨代代謝功能的影響及機(jī)制　出處：《重慶醫(yī)科大學(xué)》2017年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 妊娠期糖尿病 二甲雙胍 母代 跨代 代謝功能

【摘要】：目的:妊娠期糖尿病(Gestational diabetes mellitus,GDM)是指妊娠期間首次發(fā)生或者發(fā)現(xiàn)的糖代謝異常,是一種常見的妊娠期疾病。近年來(lái)我國(guó)GDM發(fā)病率逐漸增高達(dá)17.5%。相對(duì)于快速增長(zhǎng)的發(fā)病率,GDM的臨床治療藥物卻十分有限。二甲雙胍是當(dāng)前全球治療2型糖尿病最廣泛的口服降糖藥之一,但目前二甲雙胍對(duì)母兒的影響尤其是遠(yuǎn)期影響知之甚少,缺乏相關(guān)基礎(chǔ)研究。因此本研究通過(guò)創(chuàng)建GDM小鼠模型,研究二甲雙胍對(duì)GDM小鼠母代及跨代代謝功能影響及機(jī)制。方法:(1)利用Leprdb/+轉(zhuǎn)基因小鼠和C57BL/6J高脂(HFD)喂養(yǎng)小鼠進(jìn)行GDM表型驗(yàn)證。驗(yàn)證模型期間,稱量孕鼠體質(zhì)量,測(cè)量孕鼠口服葡萄糖耐量試驗(yàn)(OGTT)、空腹血糖水平、空腹胰島素水平和瘦素水平,以及解剖孕鼠對(duì)生理表型指標(biāo)進(jìn)行對(duì)比,以確定GDM小鼠模型成功。(2)給予GDM孕鼠二甲雙胍干預(yù),研究二甲雙胍對(duì)GDM孕鼠糖脂代謝的影響及機(jī)制。干預(yù)后稱量GDM小鼠孕期體質(zhì)量,并測(cè)定口服葡萄糖耐量試驗(yàn)以及空腹血糖、胰島素、瘦素、血脂水平;解剖孕鼠對(duì)生理表型指標(biāo)進(jìn)行對(duì)比;肝臟蘇木精-伊紅(HE)染色和油紅-O染色觀察肝臟脂肪變性;Western blotting檢測(cè)肝臟組織和肌肉組織中腺苷酸活化蛋白激酶(AMPK)、乙酰輔酶A羧化酶(ACC)、磷脂酰肌醇-3激酶(PI3K)、蛋白激酶B(Akt)、以及絲裂原活化蛋白激酶(MAPK)家族成員(細(xì)胞外調(diào)節(jié)蛋白激酶(ERK)、c-Jun氨基末端激酶(JNK)、P38)等信號(hào)分子的表達(dá)差異;RT-PCR分析AMPKα1、ACC、Akt1、腫瘤壞死因子α(TNFα)、白介素-1β(IL-1β)和白介素-6(IL-6)m RNA在肝臟組織和肌肉組織中的表達(dá)差異。(3)建立GDM小鼠傳代模型,制造子一代(F1)和子二代(F2)。喂養(yǎng)F1代小鼠至26周,于3周齡和20周齡分別給予高脂飼料(HFD)喂養(yǎng)第二次打擊;喂養(yǎng)F2代小鼠至28周,于22周給予HFD喂養(yǎng)第二次打擊。每周稱量F1和F2代體質(zhì)量,測(cè)量第二次打擊前后OGTT和胰島素耐量試驗(yàn)(ITT),解剖F1代和F2代小鼠對(duì)生理表型指標(biāo)進(jìn)行對(duì)比;測(cè)量F2代小鼠第二次打擊前后動(dòng)脈收縮壓。結(jié)果:(1)Leprdb/+轉(zhuǎn)基因小鼠第一次妊娠和第二次妊娠均無(wú)糖耐量受損、胰島素抵抗等GDM表型;HFD飲食誘導(dǎo)C57BL/6J小鼠妊娠期出現(xiàn)糖耐量異常、胰島素抵抗、超重、胎鼠質(zhì)量/胎盤質(zhì)量比值增高等GDM典型表型。故Leprdb/+轉(zhuǎn)基因小鼠模型失敗,HFD飲食誘導(dǎo)小鼠GDM模型成功。(2)母代研究中:二甲雙胍可明顯改善GDM小鼠葡萄糖耐量水平和胰島素抵抗水平;降低GDM小鼠體質(zhì)量、降低血清甘油三酯、(TG)、游離脂肪酸(f FA)、低密度脂蛋白濃度(LDL)濃度。二甲雙胍可以改善肝臟脂肪變性。二甲雙胍可以增加肝臟和肌肉組織磷酸化AMPK、磷酸化ACC、磷酸化PI3K、磷酸化Akt、磷酸化JNK、磷酸化ERK、磷酸化P38蛋白表達(dá)。二甲雙胍可顯著降低GDM小鼠肝臟和肌肉組織TNFα、IL-β、IL-6等m RNA表達(dá)。(3)子代研究中;二甲雙胍干預(yù)GDM小鼠后可使F1代雄性小鼠HFD打擊后遠(yuǎn)期體質(zhì)量降低、葡萄糖耐量升高和胰島素抵抗降低;不同宮內(nèi)背景的F1代雄性小鼠與相同母系基因背景小鼠交配后發(fā)現(xiàn),HFD打擊后二甲雙胍可跨代降低F2代雄性小鼠體質(zhì)量、增加葡萄糖耐量、減低動(dòng)脈收縮壓等。結(jié)論:(1)Leprdb/+轉(zhuǎn)基因小鼠不再是自發(fā)性GDM小鼠模型;HFD飲食誘導(dǎo)C57BL/6J小鼠可作為GDM小鼠模型。(2)二甲雙胍可以改善GDM小鼠母代糖耐量異常、胰島素抵抗,降低體質(zhì)量和血脂水平,降低炎癥因子,這可能與其肝臟和肌肉AMPK-ACC通路、PI3K-AKT通路以及MAPK通路信號(hào)蛋白磷酸化水平增高有關(guān)。(3)二甲雙胍可改善GDM小鼠F1代和F2代代謝功能、降低體質(zhì)量。
[Abstract]:Objective: gestational diabetes mellitus (Gestational diabetes, mellitus, GDM) refers to the first occurrence of pregnancy or abnormal glucose metabolism, is a common disease in pregnancy. The incidence of GDM in our country gradually increased up to 17.5%. with respect to the onset of rapid growth rate in recent years, the clinical treatment of drug GDM metformin is very limited. One of the current global oral hypoglycemic drugs in treatment of type 2 diabetes most widely, but the effect of metformin on mothers and infants especially little is known about the effects of long-term, the lack of relevant basic research. The GDM mice model through the study of metformin on GDM mice generation and cross generation effects and mechanism of metabolic function. Methods: (1) the use of Leprdb/+ transgenic mice and C57BL/6J mice fed high-fat (HFD) GDM phenotype and verified, model, body weight of pregnant rats weighing, weight test oral glucose tolerance (OG pregnant rats of grape TT), fasting blood glucose, fasting insulin and leptin levels, and anatomy of pregnant rats on physiological phenotypes were compared, in order to determine the GDM mouse model successfully. (2) pregnant rats given GDM metformin intervention, study the effects of metformin on glucose and lipid metabolism of GDM rats and the mechanism. The intervention of GDM mice weighing body mass during pregnancy determination, and oral glucose tolerance test and fasting blood glucose, insulin, leptin, lipid level; anatomy of pregnant rats to compare the physiological index of liver phenotype; hematoxylin eosin (HE) staining to observe hepatic fatty degeneration and oil red -O staining; amp activated protein kinase Western blotting detection of liver tissue and muscle tissue (AMPK), acetyl coenzyme A carboxylase (ACC), phosphatidylinositol -3 kinase (PI3K), protein kinase B (Akt), and mitogen activated protein kinase (MAPK) family members (extracellular regulated protein kinase (ERK), c-Jun amino terminal 绔縺閰，

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