【摘要】：背景與目的:近幾十年來,伴隨著人類生活方式的改變和社會經(jīng)濟(jì)發(fā)展,全球范圍內(nèi)的肥胖率呈現(xiàn)持續(xù)上升趨勢。肥胖是一種慢性疾病,是指由于機體脂肪細(xì)胞體積肥大或數(shù)量增加導(dǎo)致的機體體重超過標(biāo)準(zhǔn)體重的20%及以上或者體重指數(shù)25kg/m2的病理狀態(tài)。是糖尿病、心腦血管疾病、骨關(guān)節(jié)病、癌癥等慢性病和社會心理障礙的重要危險因素,被世界衛(wèi)生組織列為威脅人類健康的十大疾病之一。因此,肥胖需要積極預(yù)防和治療。Embelin(恩貝素)是從紫金�？�(Myrsinaceae.Isoln)植物中提取的一種醌類化合物。已有文獻(xiàn)研究表明其對于高脂飲食誘導(dǎo)的肥胖及其相關(guān)并發(fā)癥有預(yù)防作用。本研究旨在于探討Embelin是否通過抑制脂肪生成從而抑制肥胖。方法:1.分別用二甲基亞砜(DMSO)和不同濃度的Embelin處理骨髓基質(zhì)細(xì)胞系ST2和間充質(zhì)干細(xì)胞系C3H10T1/2,而后對處理過的細(xì)胞進(jìn)行成脂誘導(dǎo),通過油紅O染色、q RT-PCR和Western blotting等方法從細(xì)胞分化程度、脂肪生成相關(guān)基因m RNA表達(dá)水平及相關(guān)蛋白質(zhì)表達(dá)水平來評估embelin對ST2和C3H10T1/2細(xì)胞脂肪生成的影響;2采用4周齡的雄性C57BL/6小鼠,通過飼喂高脂飼料構(gòu)建小鼠肥胖模型,喂養(yǎng)同時皮下注射不同劑量的Embelin,造模過程中記錄各組小鼠體重,造模成功后處死實驗組與對照組小鼠,分離腹股溝脂肪組織稱取并記錄其重量,以此確定Embelin對肥胖的影響;3、分離實驗組與對照組小鼠附睪脂肪組織,提取RNA,逆轉(zhuǎn)錄,通過q RT-PCR檢測脂肪組織中脂肪生成關(guān)鍵轉(zhuǎn)錄因子和標(biāo)記基因的表達(dá)水平;4、各組小鼠尾靜脈取血進(jìn)行口服糖耐量試驗,以此來評估embelin能否改善肥胖相關(guān)的糖耐量減低;5、檢測小鼠血清胰島素和葡萄糖濃度,進(jìn)行HOMA-IR分析,以確定embelin能否改善肥胖相關(guān)的胰島素抵抗。結(jié)果:1、油紅O染色表明Embelin對骨髓基質(zhì)細(xì)胞系ST2和間充質(zhì)干細(xì)胞系C3H10T1/2細(xì)胞脂肪生成有抑制作用,q RT-PCR和Western blotting結(jié)果表明Embelin能抑制脂肪生成標(biāo)記基因固醇調(diào)節(jié)元件結(jié)合蛋白1(sterol regulatory element-binding protein-1,Srebp-1)、已酰輔酶A羧化酶1(acetyl-Co A carboxylase 1,Acc1),脂肪酸合成酶1(fatty acid synthase,Fasn)和硬脂酰輔酶A脫氫酶1(stearoyl-Co A desaturase,Scd1)的表達(dá),且有一定劑量依賴性;2、模型組(即高脂飼料喂養(yǎng)組)小鼠較對照組(即普通飼料喂養(yǎng)組)小鼠體重明顯增加,造模成功,而飼喂高脂飼料伴隨embelin皮下注射組小鼠的體重較對照組明顯降低,同時對照組小鼠腹股溝脂肪墊重量明顯重于高脂飼料喂養(yǎng)伴隨皮下注射embelin組,且有一定的劑量依賴關(guān)系,表明embelin能抑制小鼠肥胖;3、注射Embelin組脂肪生成關(guān)鍵轉(zhuǎn)錄因子和標(biāo)記基因srebp-1、Acc1、Fasn、Scd1表達(dá)水平均較對照組低,說明Embelin能夠在體內(nèi)抑制脂肪生成;4、各組小鼠尾靜脈取血口服糖耐量試驗顯示模型組空腹血糖較對照組顯著升高,血糖峰值亦明顯升高,T120min時間點血糖值仍明顯高于對照組,說明肥胖小鼠存在糖耐量損傷。而Embelin處理組血糖濃度變化與對照組相比無顯著差異。表明Embelin可改善肥胖小鼠的糖耐量損傷;5、各組小鼠HOMA-IR(穩(wěn)態(tài)胰島素評估指數(shù))分析顯示,模型組HOMA-IR大于embelin組,說明embelin能改善肥胖小鼠的胰島素抵抗。結(jié)論:1、Embelin對ST2細(xì)胞和C3H10T1/2細(xì)胞脂肪生成有抑制作用;2、Embelin能抑制脂肪生成從而抑制肥胖;同時Embelin能改善肥胖相關(guān)的糖耐量減低和胰島素抵抗。
[Abstract]:BACKGROUND & OBJECTIVE: In recent decades, with the change of human life style and the development of social economy, the obesity rate has been on the rise all over the world. Obesity is a chronic disease, which means that the body weight exceeds 20% or more of the standard body weight or body mass index (BMI) due to the hypertrophy or increase of adipocytes. The pathological state of 25 kg/m2 is an important risk factor for diabetes, cardiovascular and cerebrovascular diseases, osteoarthropathy, cancer and other chronic diseases and psychosocial disorders. It is listed by the World Health Organization as one of the ten diseases threatening human health. Therefore, obesity needs active prevention and treatment. Embelin (Embelin) is a plant from Myrsinaceae. Isoln. A kind of quinone compound was extracted from human bone marrow stromal cells (BMSCs). It has been reported that it can prevent obesity induced by high fat diet and its related complications. The purpose of this study was to investigate whether Embelin can inhibit obesity by inhibiting fat production. Methods: 1. BMSCs were treated with dimethyl sulfoxide (DMSO) and different concentrations of Embelin, respectively. Stem cell line ST 2 and mesenchymal stem cell line C3H10T1/2 were induced to adipogenesis. The effects of Embelin on adipogenesis of ST 2 and C3H10T1/2 cells were evaluated by oil red O staining, Q RT-PCR and Western blotting. Four-week-old male C57BL/6 mice were fed with high-fat diet to establish obesity model in mice. Different doses of Embelin were injected subcutaneously at the same time. The weight of mice in each group was recorded during the modeling process. After successful modeling, the mice in the experimental group and the control group were sacrificed. 3. Isolate the epididymal adipose tissue of experimental group and control group, extract RNA, reverse transcription, detect the expression of key transcription factors and marker genes in adipose tissue by Q RT-PCR; 4. Blood samples from tail vein of each group were taken for oral glucose tolerance test to evaluate whether Embelin can improve obesity-related glucose tolerance. Results: 1. Oil red O staining showed that Embelin could inhibit adipogenesis of bone marrow stromal cell line ST 2 and mesenchymal stem cell line C3H10T1/2 cells, Q RT-PCR and Western blotting results table. Embelin inhibited the expression of sterol regulatory element-binding protein-1 (Srebp-1), acetyl-Co A carboxylase-1 (Acc-1), fatty acid synthase-1 (Fasn) and stearoyl-Co A dehydrogenase-1 (Scd-1) in adipogenic markers. 2. The weight of mice in the model group (i.e. the high-fat diet group) was significantly higher than that in the control group (i.e. the common diet group), and the weight of mice fed with high-fat diet and subcutaneous injection of Embelin was significantly lower than that in the control group. Meanwhile, the weight of groin fat pad in the control group was significantly heavier than that in the high-fat diet group. The expression levels of key transcription factors and marker genes srebp-1, Acc1, Fasn and Scd1 were lower in the Embelin group than in the control group, indicating that Embelin could inhibit fat production in vivo. The blood glucose tolerance test showed that the fasting blood glucose and the peak blood glucose in the model group were significantly higher than those in the control group, and the blood glucose level at T120 min was still significantly higher than that in the control group, indicating that there was impaired glucose tolerance in obese mice. 5. The HOMA-IR analysis showed that HOMA-IR in model group was higher than that in Embelin group, indicating that Embelin could improve insulin resistance in obese mice. Belin can improve obesity related impaired glucose tolerance and insulin resistance.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R589.2

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