N-乙酰半胱氨酸对糖尿病大鼠肝脏成纤维细胞生长因子21表达影响的初步研究

发布时间：2018-04-22 01:40

本文选题：糖尿病 + 1型　；参考：《天津医科大学》2015年硕士论文

【摘要】：研究背景:成纤维细胞生长因子21(FGF21)是成纤维细胞生长因子(FGF)家族的一员,最先由Nishimura等首次于2000年发现,主要在肝脏及脂肪组织合成的内分泌激素,可以以自分泌、旁分泌及内分泌的方式发挥激素样作用,并在糖脂代谢中起重要作用。一项新的研究发现FGF21的产生与氧化应激有关,而N-乙酰半胱氨酸(NAC)可以通过其自身-SH的还原性及脱乙酰基后生成GSH的特性,使组织对抗氧化应激的能力增强。NAC能否通过其抗氧化作用调节糖尿病大鼠肝脏的FGF21的表达尚未见相关报道。研究目的:探讨NAC对于DM大鼠肝脏FGF21表达的影响。研究方法:1.以Wistar大鼠为实验动物,予以一次性注射STZ(45mg/kg)建立DM大鼠模型。随机分为NC组、DM组及DM+NAC干预组,观察周期分别为6W和12W。2.定期检测大鼠一般情况及随机血糖、血脂、肝酶学指标及MDA、SOD等指标变化。3.通过免疫组织化学法观察肝脏中FGF21的表达位置及各组FGF21的相对表达量。4.采用Western Blot法检测6W及12W各组FGF21蛋白在肝脏中的表达变化。5.应用实时定量PCR技术检测6周及12周各组肝脏FGF21mRNA的相对表达量。结果:1.成功建立1型糖尿病大鼠模型。2.与同期NC组大鼠比较,DM组及NAC干预组的大鼠其多饮、多尿、多食及消瘦症状明显,体重下降显著(P0.05),随机血糖升高显著(P0.05)。3.与同期NC组比较,DM组及NAC干预组其肝功血脂均明显升高(P0.01);NAC组较同期DM组相比ALT、TG均有所改善,统计学差异显著(P0.05),AST、TC有下降趋势,但差异不显著。4.与同期NC组比较,DM组及NAC组其MDA水平均明显升高,SOD活性均明显降低(P0.01);NAC同期DM组相比MDA水平有所下降,SOD活性有所上升,差异具有统计学意义(P0.05)。5.免疫组化结果显示FGF21表达于肝细胞胞浆,以中央静脉周围较丰富,与NC组相比DM组表现为棕黄色颗粒的先增加后减少,NAC干预组则表现为棕黄色颗粒的逐渐增加。6.Western-Blot结果显示Western blot结果显示DM 6周组及NAC 6周组FGF21表达量显著高于同期NC组,差异具有统计学意义(P0.05)。DM 12周组FGF21表达量明显低于同期NC组,差异具有统计学意义(P0.05),NAC干预12周组的FGF21表达量则明显高于同期对照组。且随着时间的延长糖尿病组FGF21蛋白表达量呈先升后降趋势,而NAC干预组则呈逐渐上升趋势。7.实时定量PCR结果显示:DM 6W组FGF21mRNA相对表达显著高于同期NC组及NAC干预6W组,统计学差异显著(P0.05);DM 12 W组FGF21表达量明显低于同期NC组及NAC干预组,统计学差异显著(P0.05)。随着观察周期的延长,DM组及NAC干预组FGF21mRNA的表达均呈先升后降趋势。结论:1.糖尿病可造成肝脏损伤,并造成氧化抗氧化系统的不平衡。2.6周及12周正常对照组大鼠其肝脏FGF21mRNA相对表达量及FGF21蛋白表达量均无明显差异。3.与正常对照组相比,糖尿病组及NAC干预组肝脏FGF21的mRNA相对表达量均呈先升后降趋势。4.随糖尿病时间的延长,糖尿病组肝脏FGF21蛋白表达量呈先升后降趋势;干预组肝脏FGF21蛋白表达呈逐渐上升趋势。5.NAC可能通过抗氧化应激作用保护肝脏,增加FGF21的表达,最终使脂代谢紊乱减轻。
[Abstract]:Background: fibroblast growth factor 21 (FGF21), a member of the fibroblast growth factor (FGF) family, was first discovered in 2000 by Nishimura, and was first discovered in 2000. The endocrine hormone, synthesized mainly in the liver and adipose tissue, can play a hormone like role in autocrine, paracrine and endogenous secretions, and play a heavy role in glycolipid metabolism. A new study found that the production of FGF21 is associated with oxidative stress, and N- acetyl cysteine (NAC) can produce GSH by its reducibility and deacetylation of its own -SH, so that the tissue against oxidative stress enhances the ability of.NAC to regulate the expression of FGF21 in the liver of diabetic rats by its antioxidant effect. Objective: To investigate the effect of NAC on the expression of FGF21 in the liver of DM rats. Methods: 1. a rat model of DM rats was established by injecting STZ (45mg/kg) with Wistar rats. The rats were randomly divided into NC group, DM group and DM+NAC intervention group. The observation period was 6W and 12W.2. regularly to detect general and random blood glucose in rats. Changes in blood lipids, liver enzymes, MDA, SOD and other indexes of.3., the expression of FGF21 in the liver and the relative expression of FGF21 in each group.4. were observed by immunohistochemistry. The expression of FGF21 protein in the liver was detected by Western Blot method and the expression of FGF21 protein in the liver was detected by Western Blot method. Results: 1. the results were as follows: 1. the rat model of type 1 diabetic rats was successfully established and compared with the NC group in the same period, the rats in the DM group and the NAC intervention group had more drink, polyuria, polydipsia and emaciation symptoms, the weight loss was significantly decreased (P0.05), the blood glucose increased significantly (P0.05).3. compared with that of the NC group, and the liver function in the DM group and the NAC intervention group increased significantly (P). 0.01): compared with the DM group in the same period, ALT and TG were all improved, and the statistical difference was significant (P0.05), AST, TC had a downward trend, but the difference was not significant between.4. and NC group in the same period, and the MDA level of DM group and NAC group increased obviously. The results of statistical significance (P0.05).5. immunohistochemical staining showed that FGF21 was expressed in the cytoplasm of the liver cell, which was rich around the central vein. Compared with the group NC, the DM group showed a first increase in brown yellow granules, and the NAC intervention group showed a gradual increase in the.6.Western-Blot result of brown yellow granules. The blot result of Western showed that DM 6 weeks group and NAC 6 were shown. The expression of FGF21 in the week group was significantly higher than that in the same period NC group. The difference was statistically significant (P0.05) in the.DM 12 week group, the expression of FGF21 was significantly lower than that in the same period NC group. The difference was statistically significant (P0.05). The FGF21 expression of NAC intervention in the 12 week group was significantly higher than that in the control group, and the expression of FGF21 protein in the diabetic group increased first and then decreased. Trend, while the NAC intervention group showed a gradual increase in.7. real-time quantitative PCR results. The relative expression of FGF21mRNA in DM 6W group was significantly higher than that of NC group and NAC intervention group 6W group, and the statistical difference was significant (P0.05), and DM 12 W group was significantly lower than that of the same period and the intervention group. The expression of FGF21mRNA in the group and the NAC intervention group all increased first and then descended. Conclusion: 1. diabetes can cause liver damage, and cause the imbalance.2.6 weeks and 12 weeks of the oxidative antioxidant system, the relative expression of FGF21mRNA and the expression of FGF21 protein in the liver of the normal control rats were not significantly different from that of the normal control group, the diabetic group and the NAC The mRNA relative expression of FGF21 in the liver of the intervention group increased first and then decreased with the prolongation of the time of diabetes. The expression of FGF21 protein in the liver of the diabetic group increased first and then descended, and the expression of FGF21 protein in the liver of the intervention group was gradually increasing and.5.NAC could protect the liver by antioxidant stress, and the expression of FGF21 was increased, and finally the lipid generation was made. Thanks to the derangement.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1

【参考文献】