本文選題：2型糖尿病 + 大鼠　； 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:探討GLP-1類似物(利拉魯肽)對2型糖尿病大鼠肝臟脂肪變性及JNK通路的影響。方法:SPF級健康雄性SD大鼠90只,隨機(jī)分為正常對照組(NC組)20只和實驗組70只。實驗組采用高脂高糖飲食聯(lián)合腹腔注射小劑量鏈脲佐菌素誘導(dǎo)建立2型糖尿病大鼠模型。成模大鼠隨機(jī)化分為3組:利拉魯肽低劑量(400μg/kg)干預(yù)組(LR-L)18只,利拉魯肽高劑量(800μg/kg)干預(yù)組(LR-H)18只,糖尿病對照組(DM)19只。干預(yù)組按其劑量每日皮下注射利拉魯肽1次,DM組和NC組以等量生理鹽水皮下注射,共4周。每周監(jiān)測各組大鼠的血糖、體重并記錄,同時觀察其飲食情況以及精神狀態(tài)。干預(yù)4周后,所有大鼠3%戊巴比妥鈉腹腔注射麻醉,收集大鼠血清,檢測大鼠血糖、血脂、肝功能、胰島素抵抗相關(guān)指標(biāo)。留取肝組織,酶聯(lián)免疫法測定肝組織TNF-α;HE染色觀察2型糖尿病大鼠肝臟病理學(xué)改變,免疫組化法檢測JNK1及P-JNK1的表達(dá);熒光定量PCR測定JNK1mRNA在肝臟中的表達(dá)。結(jié)果:利拉魯肽干預(yù)治療4周末:①體重:NC組DM組LR-H、LR-L組(分別為:337.77±28.64、250.29±19.02、220.25±22.49、206.73±26.50,P0.05);②血糖及血脂:FBG:DM 組LR-H、LR-L 組NC 組(分別為:24.14±4.11、18.99±3.65、19.35±3.94、5.51±1.18,P0.05);TG:DM組LR-H、LR-L 組、NC 組(分別為:0.64±0.10、0.51±0.17、0.51±0.13、0.24±0.08,P0.05);TC:DM組LR-H、LR-L組NC 組(分別為:1.74±0.88、1.23±0.13、1.16±0.21、0.85±0.22,P0.05);③HOMA-IR 及 ISI:干預(yù)前 HOMA-IR:DM、LR-H、LR-LNC 組(分別為:33.37±8.74、32.58±5.42、32.09±5.20、5.82±1.13,P0.05);干預(yù)前 ISI:NC 組DM、LR-H、LR-L 組(分別為:-4.85±0.20、-6.59±0.26、-6.58±0.17、-6.57±0.17,P0.05);干預(yù)4周后HOMA-IR均明顯較前下降,ISI有所升高(P0.05),干預(yù)后HOMA-IR:DM 組LR-H、LR-L 組NC 組(分別為:33.79±6.74、26.46±4.01、26.32±5.27、6.16±1.87,P0.05);干預(yù)后 ISI:NC 組LR-H、LR-L 組DM 組(分別為:-4.88±0.33、-6.61±0.21、-6.34±0.17、-6.37±0.19,P0.05)④ALT:DM 組NC 組、LR-H 組LR-L 組(分別為:153.33±51.32、103.80±40.17、109.44±36.24、144.75±45.91,P0.05);⑤肝組織切片HE染色及NAS積分:光鏡下觀察NC組大鼠肝細(xì)胞排列整齊,肝小葉規(guī)則,細(xì)胞中央有大而圓的核,細(xì)胞質(zhì)均勻,無脂肪變性及炎癥細(xì)胞浸潤。DM組肝細(xì)胞排列不規(guī)則,伴有肝細(xì)胞腫脹、脂肪變性,存在炎細(xì)胞浸潤,部分出現(xiàn)點、灶狀壞死。LR-H組及LR-L組肝細(xì)胞病理改變明顯減輕:NAS積分:DM組LR-H、LR-R 組NC 組(分別為:4.29±0.95、2.13±0.99、2.18±1.25、0.92±0.64,P0.05);⑥TNF-α及 JNK1mRNA:DM 組LR-H、LR-L 組NC 組(TNF-α分別為:1490.40±75.11、1282.09±133.31、1261.58±178.20、1108.11±110.41;JNK1mRNA分別為:2.02±0.23、1.41±0.26、1.41±0.27、1.01±0.16,P0.05);⑦JNK1 蛋白及P-JNKI蛋白組內(nèi)及各組間差異均無統(tǒng)計學(xué)意義(P0.05);結(jié)論:①高脂高糖喂養(yǎng)SD大鼠5周后腹腔注射STZ35mg/kg,可誘導(dǎo)具有高血糖、高血脂、高胰島素血癥和胰島素抵抗特征的2型糖尿病非酒精性脂肪肝病(NAFLD)模型。②利拉魯肽可降低2型糖尿病大鼠體重、空腹血糖,改善胰島素抵抗,增加胰島素敏感性,降低肝指數(shù)、血清TC、TG水平。③利拉魯肽干預(yù)可改善2型糖尿病NAFLD肝臟脂肪變性及NAS積分。④利拉魯肽可降低TNF-α水平,下調(diào)JNK1 mRNA表達(dá)。⑤利拉魯肽改善2型糖尿病肝臟脂肪變性的作用可能是通過降低肝細(xì)胞炎癥因子TNF-α水平,從而抑制JNK信號通路的活性來實現(xiàn)的。
[Abstract]:Objective: To investigate the effect of GLP-1 analogues (lyacinin) on hepatic steatosis and JNK pathway in type 2 diabetic rats. Methods: 90 SPF healthy male SD rats were randomly divided into normal control group (NC group) 20 and experimental group 70. The experimental group was induced by high fat and high glucose diet combined with small dose streptozotocin to induce type 2 diabetes mellitus. Rat model rats were divided into 3 groups: 3 groups of Liraru peptide low dose (400 g/kg) intervention group (LR-L), high dose of Liraru peptide (800 mu g/kg) intervention group (LR-H) 18 and 19 diabetic control group (DM). The intervention group was subcutaneous injection of Liraru peptide every day at its dose, and group DM and NC group were subcutaneously injected with normal saline for 4 weeks. The blood sugar, weight and record of rats were measured and the diet and mental state were observed at the same time. After 4 weeks, 3% pentobarbital sodium was injected into the rat serum to detect the blood glucose, blood lipid, liver function and insulin resistance related indexes. The liver tissue was retained and the liver tissue TNF- alpha was determined by ELISA; HE staining observation was used to observe the liver tissue. Liver pathological changes in type 2 diabetic rats, immunohistochemical method to detect the expression of JNK1 and P-JNK1; fluorescence quantitative PCR to determine the expression of JNK1mRNA in the liver. Results: lial Lu peptide intervention therapy for 4 weekend: (1) weight: LR-H in group DM group NC, LR-L group (337.77 + 28.64250.29 + 19.02220.25 + 22.49206.73 + 26.50, P0.05); 2. Group FBG:DM LR-H, group LR-L, group NC (24.14 + 4.11,18.99 + 3.65,19.35 + 1.18, P0.05), TG:DM group LR-H, LR-L group, NC group (respectively 0.64 + 1.74 + + 0.22, 0.22, respectively); Pre HOMA-IR:DM, LR-H, and LR-LNC group (33.37 + 8.74,32.58 + 5.42,32.09 + 5.20,5.82 + 1.13, P0.05), before intervention, DM, LR-H, LR-L group (respectively: -4.85 + 0.20, + 0.26, 0.17, 0.17, respectively). Group C (33.79 + 6.74,26.46 + 4.01,26.32 + 5.27,6.16 + 1.87, P0.05), LR-H in group ISI:NC and DM group in group LR-L (respectively: -4.88 + 0.33, -6.61 + 0.21, -6.34 0.17, 0.19 and 45.91, respectively); liver tissue cut HE staining and NAS integral: observed under light microscope, the liver cells in group NC were arranged orderly, liver lobule rule, large and round nucleus in the central cell, homogeneous cytoplasm, fat free degeneration and inflammatory cell infiltration in group.DM, group of hepatocytes were irregular, accompanied by hepatocyte swelling, fatty degeneration, inflammatory cell infiltration, partial appearance, focal necrosis,.LR-H and L The pathological changes of hepatocytes in group R-L were obviously reduced: NAS integral: group DM LR-H, group NC of LR-R group (4.29 + 0.95,2.13 + 0.99,2.18 + 1.25,0.92 + 0.64, P0.05); 6. TNF- alpha and JNK1mRNA:DM group (1490.40 + + 2.02 + 0 respectively); 2.02 + 0 0 respectively .26,1.41 + 0.27,1.01 + 0.16, P0.05), and there was no significant difference between the groups of JNK1 protein and P-JNKI protein group and each group (P0.05). Conclusion: (1) high fat and high glucose feeding of SD rats after 5 weeks of intraperitoneal injection of STZ35mg/kg can induce non alcoholic fatty liver disease of type 2 diabetes with the characteristics of hyperglycemia, hyperlipidemia, hyperinsulinemia and insulin resistance. NAFLD) model. (2) lialuru can reduce the weight of type 2 diabetic rats, fasting blood glucose, improving insulin resistance, increasing insulin sensitivity, reducing liver index, serum TC, TG level. (3) lialurin intervention can improve the NAFLD liver fatty degeneration and NAS score in type 2 diabetes mellitus. (4) lialuru can reduce the level of TNF- alpha and reduce the expression of JNK1 mRNA. The effect of lialurin on hepatic steatosis in type 2 diabetes may be achieved by reducing the level of inflammatory factor TNF- alpha in liver cells and inhibiting the activity of JNK signaling pathway.

【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R587.2;R575

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相關(guān)期刊論文 前4條

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本文編號：1847347