双酚A和染料木黄酮对大鼠糖脂代谢的影响

发布时间：2018-02-27 09:36

  本文关键词： 高脂膳食 双酚A 染料木黄酮 葡萄糖代谢 脂代谢　出处：《华中科技大学》2014年博士论文　论文类型：学位论文

【摘要】：目的：(1)观察BPA和染料木黄酮干预对成年雄性Wistar大鼠糖脂代谢的影响。(2)探索BPA暴露导致血葡萄糖平衡紊乱的可能机制；观察父代大鼠长期“安全剂量”BPA暴露对子代大鼠出生结局及子代大鼠成年后糖脂代谢的影响。(3)探索BPA和染料木黄酮对大鼠肝脏脂代谢影响的可能机制。 方法：体重为150-180g的雄性Wistar大鼠,适应性喂养一周后,随机分为：普通饲料组(STD),普通饲料+50μg/kg BPA组(STD-BPA50),普通饲料+50μg/kg BPA+10mg/kg染料木黄酮组(STD-(BPA50+G)),普通饲料+10mg/kg染料木黄酮组(STD-G)；高脂对照组(HFD),高脂饲料+50μg/kg BPA组(HFD-BPA50),高脂饲料+50μg/kg BPA+10mg/kg染料木黄酮组(HFD-(BPA50+G)),高脂饲料+10mg/kg染料木黄酮组(HFD-G)。在干预的第0周和第21周末测定血糖、胰岛素、血清TG和TC水平；在干预的30周末进行腹膜内葡萄糖耐量试验(IPGTT),在干预的35周末,处死大鼠,测定生化指标。干预21周末,选取STD、STD-BPA50、HFD和HFD-BPA50组大鼠作为父代,与健康成年雌性Wistar大鼠合笼。子代大鼠出生后计算性别比、记录出生窝重和每窝子代个数。子代断奶后给予普通饲料直至10周龄后处死。用免疫荧光法测定胰腺β细胞面积；用实时荧光定量PCR检测大鼠胰腺组织LC3、Beclin-1和Bip的(?)mRNA表达水平；采用免疫组化和蛋白免疫印迹法检测胰腺LC3的蛋白表达水平；用激光共聚焦显微镜检测p细胞上的LC3蛋白表达。测定子代大鼠血清生化指标,计算HOMA指数和胰岛素敏感性指数。用HE染色观察父代大鼠肝脏组织的病理变化；用实时定量PCR检测肝脏中脂代谢关键基因的表达水平；用蛋白免疫印记法检测肝脏中PPARα、PPARγ和LC3蛋白表达水平。 结果：(1)干预35周后,HFD-BPA50组大鼠血糖水平在明显高于HFD组(P0.01)；STD-BPA50组胰岛素水平明显高于STD组(P0.05)和STD-G组(P0.01)。在IPGTT实验中,HFD-BPA50组在给予葡萄糖后的第15min和30min血糖水平和血糖曲线下面积明显高于HFD组(P0.05)。此外,在干预35周后STD-BPA50组的HOMA指数明显高于STD组和STD-G组(P0.01),并且胰岛素敏感指数明显低于STD组(P0.05)。在干预21周后HFD-(BPA50+G)和HFD-G组大鼠的HOMA指数明显低于HFD-BPA50组(P0.05和P0.01)。在干预35周末,HFD-G组的血清TG水平明显低于HFD-BPA50组(P0.05)；并且HFD-G组血清TC水平明显低于HFD组和HFD-BPA50组(P0.05)。在干预35周末,与HFD组相比,HFD-(BPA50+G)组和HFD-G组肝脏中TG和TC水平明显降低(P0.05)。(2)高脂饮食和BPA干预明显增加了父代大鼠的胰腺β细胞数量(P0.01)；高脂饮食和BPA干预还明显增加了父代大鼠胰腺LC3、Beclin-1和Bip的mRNA表达水平；并且高脂喂养和BPA干预的大鼠胰腺LC3蛋白表达水平也明显高于各自的对照组(P0.01)；高脂饮食和BPA干预增加了胰腺β细胞上的LC3表达水平(P0.01)。子代大鼠的出生性别比、窝重和窝别大小、血糖和血脂无明显差别(P0.05)。(3)HFD组、HFD-(BPA50+G)组和HFD-G组大鼠肝脏SREBP-1C的mRNA表达水平明显高于STD组(P0.05)；HFD组大鼠肝脏PPARγ的mRNA表达水平明显高于STD组、HFD-(BPA50+G)组和HFD-G组(P0.05),并且HFD组大鼠肝脏的PPARγ蛋白质表达水平明显高于STD组、HFD-(BPA50+G)组和HFD-G组(P0.05)；HFD组大鼠肝脏LC3的mRNA和蛋白质表达水平低于STD组(P0.01,P0.05)。 结论：(1)长期BPA暴露导致了大鼠的胰岛素抵抗和葡萄糖不耐受；染料木黄酮干预能明显改善高脂喂养大鼠的血脂紊乱。(2)父代大鼠BPA暴露导致的胰腺β细胞功能紊乱可能与上调的内质网应激和自噬水平相关；父代大鼠长期BPA暴露并没有导致子代大鼠的出生结局及成年后糖脂代谢发生改变。(3)BPA干预并没有影响肝脏脂代谢相关基因和自噬水平；染料木黄酮干预可能通过下调肝脏PPARγ的表达进而缓解高脂造成的脂代谢紊乱。
[Abstract]:Objective: To observe the BPA (1) and genistein intervention effects on lipid metabolism of adult male Wistar rats. (2) to explore the possible mechanisms leading to BPA exposure to blood glucose balance disorders; observe parent rats long-term safe dose BPA exposure affects lipid metabolism in offspring rats and offspring birth outcome adult rat. (3) the possible mechanism of BPA and explore the effects of genistein on liver lipid metabolism in rats.
Methods: male Wistar rats weighing 150-180g, feeding a week, were randomly divided into normal diet group (STD), +50 g/kg BPA group with normal diet (STD-BPA50), +50 g/kg BPA+10mg/kg normal diet genistein group (STD- (BPA50+G)), normal diet + 10mg/kg genistein group (STD-G); high fat control group (HFD), high fat diet +50 g/kg BPA group (HFD-BPA50), high fat diet +50 g/kg BPA+10mg/kg genistein group (HFD- (BPA50+G)), high fat diet +10mg/kg genistein group (HFD-G). Insulin in zeroth weeks and twenty-first weeks of intervention measure blood glucose, serum TG and TC levels; intraperitoneal glucose tolerance test in the 30 week intervention (IPGTT), in the 35 week intervention, the rats were sacrificed and the determination of biochemical indicators. At the end of the 21 week intervention, including STD, STD-BPA50, HFD and HFD-BPA50 rats as the parent, and healthy adult female Wistar rats rats were mated. The calculation of rats after birth sex ratio, litter birth weight and position of each record generation number. The offspring after weaning was given normal feed until 10 weeks of age were determined. The area of pancreatic beta cells by immunofluorescence; using real-time fluorescence quantitative PCR detection of pancreatic tissue of rats with LC3, Beclin-1 and Bip (?) mRNA expression; expression of pancreatic LC3 detected by immunohistochemistry and Western blotting of protein; expression of P cells was detected by laser confocal microscope. The LC3 protein determination of offspring rats serum biochemical index, HOMA index and insulin sensitivity index. HE staining was used to observe the pathological changes of parent rat liver the expression levels of key genes; using real-time quantitative PCR detection of liver lipid metabolism; PPAR alpha protein by Western blot in the liver, the level of PPAR gamma and LC3 protein expression.
Results: (1) after 35 weeks of treatment, blood glucose levels in rats of HFD-BPA50 group was significantly higher than HFD group (P0.01); insulin levels were significantly higher in STD-BPA50 group than in STD group (P0.05) and STD-G group (P0.01). In IPGTT experiment, HFD-BPA50 group was given glucose after 15min and 30min blood glucose levels and blood glucose curve the next area was significantly higher than HFD group (P0.05). In addition, in the 35 weeks after the intervention group STD-BPA50 HOMA index was significantly higher than that of STD group and STD-G group (P0.01), and the insulin sensitivity index was significantly lower than that of STD group (P0.05). In 21 weeks after the intervention of HFD- (BPA50+G) and HFD-G group rats HOMA index was significantly lower than that of group HFD-BPA50 (P0.05 and P0.01). After 35 weeks, the serum TG level of HFD-G group was significantly lower than that of HFD-BPA50 group (P0.05); and the level of serum TC in HFD-G group was significantly lower than that of HFD group and HFD-BPA50 group (P0.05). After 35 weeks, compared with group HFD, HFD- (BPA50+G) group and HFD-G group of liver TG 鍜孴C姘村钩鏄庢樉闄嶄綆(P0.05).(2)楂樿剛楗�椋熷拰BPA骞查�勬槑鏄惧�炲姞浜嗙埗浠ｅぇ榧犵殑鑳拌吅尾缁嗚優鏁伴噺(P0.01)锛涢珮鑴傞ギ椋熷拰BPA骞查�勮繕鏄庢樉澧炲姞浜嗙埗浠ｅぇ榧犺儼鑵篖C3,Beclin-1鍜孊ip鐨刴RNA琛ㄨ揪姘村钩锛涘苟涓旈珮鑴傚杺鍏诲拰BPA骞查�勭殑澶ч紶鑳拌吅LC3铔嬬櫧琛ㄨ揪姘村钩涔熸槑鏄鹃珮浜庡悇鑷�鐨勫�圭収缁，

本文编号：1542125