本文選題：肥胖 + 2型糖尿病�。� 參考：《天津醫(yī)科大學》2017年碩士論文

【摘要】：目的:肥胖是一種表現(xiàn)為體內(nèi)脂肪堆積過多和(或)分布異常、體重增加的慢性代謝性疾病。糖尿病是以血糖升高為特征的代謝性疾病,90%以上為2型糖尿病(T2DM)。肥胖和T2DM的共同特征為胰島素抵抗,而胰島素抵抗與系統(tǒng)性慢性低度炎癥密切相關(guān)。近年來研究顯示,肥胖和T2DM個體的脂肪組織中M1型(炎性)巨噬細胞增多。半胱氨酸天冬氨酸特異性蛋白酶(caspase)是細胞程序性死亡過程中的關(guān)鍵酶,炎性caspase通過激活炎癥小體,促進炎性細胞因子IL-1β和IL-18的成熟和釋放。本研究通過檢測血漿中單核細胞caspase活性,分析其與體重指數(shù)BMI、腰圍、血糖、血脂、血中炎性細胞因子濃度之間的相關(guān)性,同時體外高糖刺激人THP-1單核細胞,檢測單核細胞活性,探討單核細胞caspase與肥胖和T2DM的關(guān)系,為肥胖和T2DM的發(fā)病和治療機制新視角。內(nèi)容:第一部分:招募健康人和T2DM病人,測量身高、體重、腰圍,計算BMI,根據(jù)BMI和腰圍分為3組:正常組、T2DM組、T2DM合并肥胖組。每名志愿者各抽取2ml EDTA抗凝血和2ml分離膠促凝血,檢測空腹血糖(FPG)、糖化血紅蛋白(HbA1c)、甘油三酯(TG)和總膽固醇(TC)水平。提取抗凝血中的單個核細胞,檢測單核細胞caspase活性,分析單核細胞caspase活性與以上各指標的相關(guān)性第二部分:收集正常組、T2DM組、T2DM合并肥胖組分離膠促凝血,檢測血清中IL-1β、TNF-α、脂多糖(LPS)、游離脂肪酸(NEFA)水平,并分析單核細胞caspase活性與以上各指標的相關(guān)性。第三部分:體外培養(yǎng)人THP-1單核細胞株,首先用含33mM葡萄糖的1640培養(yǎng)THP-1,分別培養(yǎng)12小時、24小時、36小時、48小時,檢測單核細胞caspase活性;然后分別用含25mM、33mM葡萄糖的1640培養(yǎng)12小時和24小時,檢測細胞上清液中乳酸脫氫酶(LDH)水平,以含11mM葡萄糖1640培養(yǎng)的THP-1單核細胞作為對照,進一步驗證高血糖對于單核細胞活性的影響。方法:招募健康人和T2DM病人共37人,測量身高、體重、腰圍,計算BMI,根據(jù)BMI和腰圍分為正常組、T2DM組、T2DM合并肥胖組,生化全自動分析儀檢測血液中FPG、TG、TC,糖化血紅蛋白分析儀檢測HbA1c,流式細胞術(shù)檢測單核細胞caspase活性,ELISA檢測血清IL-1β、TNF-α、LPS水平,游離脂肪酸試劑盒檢測血清中NEFA水平。在體外用含33mM葡萄糖的1640培養(yǎng)THP-1,分別培養(yǎng)12小時、24小時、36小時、48小時,流式細胞術(shù)檢測單核細胞caspase活性,然后分別用含25mM、33mM葡萄糖的1640培養(yǎng)THP-1單核細胞12小時和24小時,細胞毒性試劑盒檢測細胞上清液中LDH。結(jié)果:第一部分結(jié)果顯示:T2DM合并肥胖組總單核細胞、炎性單核細胞以及經(jīng)典單核細胞caspase活性均顯著高于正常組,總單核細胞caspase活性與BMI、腰圍顯著正相關(guān),與FPG、HbA1c、TG、TC不相關(guān)。第二部分結(jié)果顯示:T2DM合并肥胖組IL-1β水平顯著高于正常組和T2DM組,T2DM組NEFA水平顯著高于正常組,但T2DM合并肥胖組NEFA水平與正常組無顯著差異�？倖魏思毎鹀aspase活性、炎性單核細胞caspase活性均與IL-1β顯著正相關(guān),與TNF-α、LPS、NEFA不相關(guān)。第三部分結(jié)果顯示:與含11mM葡萄糖的1640培養(yǎng)的THP-1單核細胞相比,高糖的培養(yǎng)條件并未影響單核細胞的caspase活性以及細胞上清液中LDH水平。結(jié)論:肥胖和T2DM可能在外周血單核細胞caspase活性升高中發(fā)揮協(xié)同促進作用。
[Abstract]:Objective: obesity is a chronic metabolic disease characterized by excessive fat accumulation and (or) distribution and increasing body weight. Diabetes is a metabolic disease characterized by elevated blood sugar, more than 90% is type 2 diabetes (T2DM). Obesity and T2DM are characterized by insulin resistance, and insulin resistance and systemic chronic low inflammation are closely related to insulin resistance. In recent years, studies have shown that M1 (inflammatory) macrophages in fat tissues and T2DM individuals have increased. Cysteine aspartate specific protease (caspase) is the key enzyme in the process of programmed cell death. Inflammatory caspase promotes the maturation and release of inflammatory cytokines IL-1 beta and IL-18 by activating inflammatory cells. The relationship between the caspase activity of monocyte in plasma and the concentration of body mass index (BMI), waist circumference, blood sugar, blood lipid and inflammatory cytokines in blood was analyzed, and the activity of THP-1 mononuclear cells was stimulated by high glucose in vitro, and the relationship between the mononuclear cell caspase and obesity and T2DM was explored, and the pathogenesis of obesity and T2DM and the pathogenesis of T2DM were analyzed. New perspective of treatment mechanism. Content: first part: recruit healthy people and T2DM patients, measure height, weight, waist circumference, calculate BMI, divide into 3 groups according to BMI and waist circumference: normal group, T2DM group, T2DM combined obesity group. Each volunteer takes 2ml EDTA anticoagulant and 2ml separation gel prothrombin, fasting blood glucose (FPG), glycated hemoglobin (HbA1c), glycerin Three ester (TG) and total cholesterol (TC) level. Extract the mononuclear cells in anticoagulant, detect the caspase activity of mononuclear cells, analyze the correlation between the caspase activity of mononuclear cells and the above indexes, to collect the normal group, T2DM group, T2DM combined with the obesity group and promote the coagulation of blood, and detect the serum IL-1 beta, TNF- a, lipopolysaccharide (LPS), free fat. The level of acid (NEFA), and the correlation between the caspase activity of mononuclear cells and the above indexes. Third: in vitro culture, human THP-1 mononuclear cell line was cultured with 33mM glucose 1640 for 12 hours, 24 hours, 36 hours and 48 hours respectively to detect the caspase activity of mononuclear cells, and then with 25mM, 33mM glucose 1640 respectively. The level of lactate dehydrogenase (LDH) in the cell supernatant was detected for 12 hours and 24 hours, and the THP-1 mononuclear cells cultured with 11mM glucose 1640 were used as control. The effect of hyperglycemia on the activity of mononuclear cells was further verified. Methods: 37 people were recruited and T2DM patients were recruited to measure height, weight, waist circumference, BMI, BMI and waistline. For the normal group, the T2DM group and the T2DM group, the biochemical full automatic analyzer was used to detect the FPG, TG, TC, and the glycosylated hemoglobin analyzer to detect the HbA1c, the flow cytometry was used to detect the caspase activity of mononuclear cells. ELISA was used to detect the serum IL-1 beta, TNF- alpha and LPS levels. The free fatty acid kit was used to detect the level of the serum NEFA level. In vitro, 1 of the glucose containing glucose was used. 640 THP-1 was cultured for 12 hours, 24 hours, 36 hours and 48 hours respectively. Flow cytometry was used to detect the caspase activity of mononuclear cells. Then, THP-1 mononuclear cells were incubated with 25mM and 33mM glucose for 12 hours and 24 hours respectively. The cytotoxic kit was used to detect the LDH. results in the cell supernatant. The first part showed that the T2DM combined with the obesity group was total. The activity of monocyte, inflammatory mononuclear cell and classical monocyte caspase were significantly higher than that of normal group. The caspase activity of total monocyte was significantly correlated with BMI, waist circumference, and not related to FPG, HbA1c, TG, TC. The second results showed that the level of IL-1 beta in T2DM combined obesity group was significantly higher than that in normal group and T2DM group, and NEFA level in T2DM group was significantly higher than that in normal group. The NEFA level in the T2DM combined obesity group was not significantly different from that in the normal group. The total monocyte caspase activity and the caspase activity of the inflammatory monocytes were positively correlated with the IL-1 beta, and were not related to TNF- alpha, LPS, NEFA. The third part showed that the high sugar culture conditions were not compared with the THP-1 mononuclear cells containing 11mM glucose of 1640. Caspase activity of mononuclear cells and the level of LDH in cell supernatant. Conclusion: obesity and T2DM may play a synergistic role in promoting high school of caspase activity in peripheral blood mononuclear cells.

【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R587.1;R589.2

【參考文獻】

相關(guān)期刊論文 前1條

1 Chandra Kanti Chakraborti;;Role of adiponectin and some other factors linking type 2 diabetes mellitus and obesity[J];World Journal of Diabetes;2015年15期

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本文編號：1789815