【摘要】：背景目的：非酒精性脂肪性肝病（NAFLD）是一種與肥胖密切相關(guān)的肝臟疾病，目前仍缺乏特異性的治療方法。脂聯(lián)素是一種具有抗炎、抗糖尿病、抗動脈粥樣硬化作用的細(xì)胞因子。本實(shí)驗(yàn)旨在研究脂聯(lián)素在NAFLD發(fā)病機(jī)制中的作用，并探討二甲雙胍或飲食干預(yù)對NAFLD的療效。 方法：將48只雄性SD大鼠隨機(jī)分為6組，分別為：8周正常飲食組（NC1組）、8周高脂飲食組（HFD1組）、16周正常飲食組（NC2組）、16周高脂飲食組（HFD2組）、8周高脂飲食+8周正常飲食干預(yù)組（DIET組）、8周高脂飲食+繼續(xù)8周高脂飲食的同時(shí)給予二甲雙胍干預(yù)組（HFD+M組）。采用ELISA檢測血清脂聯(lián)素水平及血清生化指標(biāo)，免疫組織化學(xué)法、Western Blotting法、RT-qPCR法分別檢測肝臟組織脂聯(lián)素及其受體蛋白和mRNA表達(dá)水平。 結(jié)果：HFD2組大鼠肝臟NAS評分與HFD1組大鼠肝臟NAS評分相比（3.7±0.52vs6.57±0.79，P0.001）明顯增高，反映了伴隨高脂飲食時(shí)間延長，NAFLD的組織學(xué)進(jìn)展過程。根據(jù)NAS評分標(biāo)準(zhǔn)，HFD2組的大鼠可診斷為NASH。肝臟脂聯(lián)素及其受體的免疫組化評分與肝臟的炎癥等級、脂肪百分比、氣球樣變程度均呈顯著負(fù)相關(guān)。與HFD2組相比，飲食干預(yù)或者二甲雙胍治療均明顯改善NAFLD大鼠肝組織的脂肪變性和氣球樣變，另外飲食干預(yù)(0.86±0.38vs2.14±0.38, P0.001)減輕了肝臟組織的炎癥反應(yīng)，但二甲雙胍治療后肝臟炎癥反應(yīng)與HFD2組相比并無明顯差異(2±0.63vs2.14±0.38, P0.05)。與NC組相比，HFD大鼠肝組織的脂聯(lián)素及其受體蛋白、mRNA的表達(dá)水平均降低，，同時(shí)伴隨著肝酶升高、血脂異常和胰島素抵抗等變化。與HFD2組相比，飲食干預(yù)可以上調(diào)脂聯(lián)素及其受體的蛋白、mRNA表達(dá)水平，同時(shí)改善血清生化指標(biāo)；然而，二甲雙胍治療對脂聯(lián)素及其受體的表達(dá)無明顯影響，并且增高了血清ALT的水平。 結(jié)論：高脂飲食可以成功建立大鼠NAFLD模型。從單純性脂肪變進(jìn)展到NASH過程中，脂聯(lián)素及其受體均發(fā)揮重要的作用。飲食干預(yù)可能通過上調(diào)脂聯(lián)素及其受體的表達(dá)發(fā)揮對NAFLD的治療作用。二甲雙胍治療對脂聯(lián)素及其受體的表達(dá)無明顯的影響，可能與二甲雙胍治療組大鼠肝組織炎癥反應(yīng)無明顯改善有關(guān)。飲食干預(yù)應(yīng)作為NAFLD的基礎(chǔ)治療策略。
[Abstract]:Background: nonalcoholic fatty liver disease (NAFLD) is a liver disease closely related to obesity. Adiponectin is a cytokine with anti-inflammatory, anti-diabetic and anti-atherosclerosis effects. The aim of this study was to investigate the role of adiponectin in the pathogenesis of NAFLD and to explore the therapeutic effects of metformin or dietary intervention on NAFLD. Methods: 48 male SD rats were randomly divided into 6 groups. 8 weeks normal diet group (NC1 group), 8 week high fat diet group (HFD1 group), 16 week normal diet group (NC2 group), 16 weeks high fat diet group (HFD2 group), 8 weeks high fat diet, 8 weeks normal diet intervention group (DIET group), 8 weeks high fat diet and 8 weeks high fat diet. The fat diet was treated with metformin intervention group (HFD M group). The levels of serum adiponectin and serum biochemical indexes were detected by Elisa, and the expression of adiponectin and its receptor protein and mRNA in liver tissue were detected by Western blotting and RT-qPCR, respectively. Results compared with HFD1 group, the liver NAS score of the two groups was significantly higher than that of the HFD1 group (3. 7 鹵0. 79 鹵0. 001), which reflected the histological progression of NAFLD with the prolongation of high fat diet time. According to the standard of NAS, the rats in HFD2 group could be diagnosed as NASH. The immunohistochemical scores of adiponectin and its receptors were negatively correlated with the grade of inflammation, percentage of fat and degree of balloon degeneration. Compared with HFD2 group, diet intervention or metformin treatment significantly improved the steatosis and balloon degeneration of liver tissue in NAFLD rats, and diet intervention (0.86 鹵0.38, P0.001) alleviated the inflammatory response of liver tissue. However, there was no significant difference in hepatic inflammatory response between metformin group and HFD2 group (2 鹵0.63vs2.14 鹵0.38, P0.05). Compared with NC group, the expression of adiponectin and its receptor protein mRNA in the liver of HFD rats was decreased, accompanied by the increase of liver enzyme, abnormal blood lipid and insulin resistance. Compared with HFD2 group, dietary intervention could up-regulate the expression of adiponectin and its receptor protein mRNA and improve serum biochemical indexes, however, metformin treatment had no significant effect on adiponectin and its receptor expression. And elevated serum alt levels. Conclusion: high fat diet can successfully establish NAFLD model in rats. Adiponectin and its receptors play an important role in the progression from simple adipogenesis to Nash. Dietary intervention may play a therapeutic role in NAFLD by up-regulating adiponectin and its receptor expression. The expression of adiponectin and its receptor was not significantly affected by metformin treatment, which may be related to the improvement of hepatic inflammation in metformin treatment group. Dietary intervention should be used as the basic therapeutic strategy of NAFLD.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R575.5

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