二甲双胍或饮食干预对NAFLD大鼠肝脏脂联素及其受体表达的影响
发布时间：2018-07-13 12:43
【摘要】：背景目的：非酒精性脂肪性肝病（NAFLD）是一种与肥胖密切相关的肝脏疾病，目前仍缺乏特异性的治疗方法。脂联素是一种具有抗炎、抗糖尿病、抗动脉粥样硬化作用的细胞因子。本实验旨在研究脂联素在NAFLD发病机制中的作用，并探讨二甲双胍或饮食干预对NAFLD的疗效。 方法：将48只雄性SD大鼠随机分为6组，分别为：8周正常饮食组（NC1组）、8周高脂饮食组（HFD1组）、16周正常饮食组（NC2组）、16周高脂饮食组（HFD2组）、8周高脂饮食+8周正常饮食干预组（DIET组）、8周高脂饮食+继续8周高脂饮食的同时给予二甲双胍干预组（HFD+M组）。采用ELISA检测血清脂联素水平及血清生化指标，免疫组织化学法、Western Blotting法、RT-qPCR法分别检测肝脏组织脂联素及其受体蛋白和mRNA表达水平。 结果：HFD2组大鼠肝脏NAS评分与HFD1组大鼠肝脏NAS评分相比（3.7±0.52vs6.57±0.79，P0.001）明显增高，反映了伴随高脂饮食时间延长，NAFLD的组织学进展过程。根据NAS评分标准，HFD2组的大鼠可诊断为NASH。肝脏脂联素及其受体的免疫组化评分与肝脏的炎症等级、脂肪百分比、气球样变程度均呈显著负相关。与HFD2组相比，饮食干预或者二甲双胍治疗均明显改善NAFLD大鼠肝组织的脂肪变性和气球样变，另外饮食干预(0.86±0.38vs2.14±0.38, P0.001)减轻了肝脏组织的炎症反应，但二甲双胍治疗后肝脏炎症反应与HFD2组相比并无明显差异(2±0.63vs2.14±0.38, P0.05)。与NC组相比，HFD大鼠肝组织的脂联素及其受体蛋白、mRNA的表达水平均降低，，同时伴随着肝酶升高、血脂异常和胰岛素抵抗等变化。与HFD2组相比，饮食干预可以上调脂联素及其受体的蛋白、mRNA表达水平，同时改善血清生化指标；然而，二甲双胍治疗对脂联素及其受体的表达无明显影响，并且增高了血清ALT的水平。 结论：高脂饮食可以成功建立大鼠NAFLD模型。从单纯性脂肪变进展到NASH过程中，脂联素及其受体均发挥重要的作用。饮食干预可能通过上调脂联素及其受体的表达发挥对NAFLD的治疗作用。二甲双胍治疗对脂联素及其受体的表达无明显的影响，可能与二甲双胍治疗组大鼠肝组织炎症反应无明显改善有关。饮食干预应作为NAFLD的基础治疗策略。
[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 卤0. 79 卤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 卤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 卤0.63vs2.14 卤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.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.5
本文编号：2119407
[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 卤0. 79 卤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 卤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 卤0.63vs2.14 卤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.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.5
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本文编号：2119407
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