我国菰米中膳食纤维、类黄酮的分析及其对非酒精性肝脂肪变性HepG2细胞作用的研究

发布时间：2018-06-14 18:44

本文选题：菰米 + 膳食纤维　；参考：《扬州大学》2012年硕士论文

【摘要】：目的： (1)提取菰米、玉米、苹果、芹菜膳食纤维及菰米、玉米类黄酮,对其含量进行测定及比较,探讨各类膳食纤维及类黄酮产生生物学效应的物质基础。 (2)观察各种提取的膳食纤维、类黄酮对油酸诱导的HepG2(?)(?)酒精性脂肪肝细胞浆内的甘油三酯(TG)、丙二醛(MDA)、超氧化物歧化酶(SOD)的含量的影响,探讨不同种类的膳食纤维、类黄酮对非酒精性肝脂肪变性细胞作用的效果差异。 (3)观察非酒精性肝脂肪变性细胞内过氧化物酶体增殖体激活受体-α(PPAR-α)、肉碱脂酰转移酶-1A(CPT-1A)在mRNA水平上的影响,探讨菰米、玉米、芹菜、苹果膳食纤维及菰米、玉米类黄酮对非酒精性肝脂肪变性细胞的作用机制。方法： (1)用酶-化学法提取菰米、玉米中膳食纤维,用化学法提取苹果、芹菜的膳食纤维,按国标法对其含量进行测定；用浸提回流法提取菰米、玉米的类黄酮,用比色法对其含量进行测定。 (2)用油酸诱导HepG2细胞建立体外非酒精性肝脂肪变性模型,通过MTT、油红O染色及检测细胞浆内TG的含量,来确定油酸的浓度。 (3)油酸诱导HepG2细胞脂肪变性,加入菰米、玉米、芹菜、苹果膳食纤维及菰米、玉米类黄酮,观察对细胞浆内TG、MDA、SOD含量的变化；同时测定细胞内PPAR-a及CPT-1AmRNA的表达。结果： (1)菰米膳食纤维总膳食纤维(TDF)的含量7.242g/100g,玉米TDF的含量9.13g/100g,均高于大米；苹果TDF占预处理苹果粉的含量为13.52%,芹菜TDF占预处理芹菜粉的含量为38.6%；菰米类黄酮的含量3.674g/100g,显著高于玉米类黄酮的含量。 (2)0.5mM的油酸对细胞活性无显著影响,且细胞内TG显著升高,细胞内脂滴蓄积明显,是诱导脂肪变性的最佳浓度。 (3)100μg/ml菰米、玉米膳食纤维能降低HepG2非酒精性肝脂肪变性细胞浆中TG、MDA含量,升高SOD含量,对非酒精性肝脂肪变性具有一定的改善作用,芹菜膳食纤维的作用次之,而苹果膳食纤维的作用不明显；100μg/ml菰米、玉米类黄酮也降低模型细胞浆内的TG含量。 (4)与非酒精性肝脂肪变性细胞模型组相比,菰米、玉米、芹菜膳食纤维及菰米、玉米类黄酮处理的细胞内PPAR-a及CPT-1A mRNA的表达显著增高(P0.05),而苹果膳食纤维PPAR-a及CPT-1A mRNA的表达稍有增高,但没有统计学意义(P0.05)。结论： (1)0.5mM的油酸与HepG2细胞孵育24小时,能诱导脂肪变性,形成非酒精性肝脂肪变性细胞模型。 (2)菰米、玉米膳食纤维及类黄酮对非酒精性肝脂肪变性细胞具有一定的改善作用,作用的机制可能是激活PPAR-a,促进CPT-1A的表达,增加脂肪酸分解,减少体脂含量,延缓脂肪肝形成。 (3)芹菜膳食纤维也能改善非酒精性肝脂肪变性,而苹果膳食纤维的作用则较小。可能的原因是,膳食纤维的不同来源,所含的可溶性膳食纤维和不可溶性的膳食纤维的含量不同,膳食纤维相结合的植物化学物不同,而导致的作用有所差异。
[Abstract]:Objective: to extract Zizania latifolia, corn, apple, celery dietary fiber, Zizania rice and corn flavonoids, and compare them with each other. Study on the material basis of biological effects of dietary fiber and flavonoids. The effects of triglyceride, malondialdehyde (MDA) and superoxide dismutase (SOD) on the contents of triglyceride (TGG), malondialdehyde (MDA) and superoxide dismutase (SOD) in the cytoplasm of alcoholic fatty liver were studied. The effects of flavonoids on non-alcoholic hepatic steatosis cells were studied. The effects of peroxisome proliferator activator receptor-伪 -PPAR- 伪, carnitine acid-transferase -1AnCPT-1A) on the mRNA level in non-alcoholic hepatic steatosis cells were observed. To investigate the mechanism of effects of Zizania latifolia, maize, celery, apple dietary fiber, Zizania rice and zeatin flavonoids on non-alcoholic hepatic fatty degeneration cells. Methods: 1) extract the dietary fiber from Zizania latifolia, corn, apple and celery by enzymatic chemical method, and extract the flavonoids from Zizania rice and corn by extraction and reflux method, and determine the content of dietary fiber of apple and celery by chemical method, and extract the flavonoids of Zizania rice and corn by extraction and reflux method. HepG2 cells were induced by oleic acid to establish a model of non-alcoholic fatty degeneration of liver in vitro. The content of TG in the cytoplasm was detected by MTT, oil red O staining. To determine the concentration of oleic acid. 3) oleic acid induced fatty degeneration of HepG2 cells, adding Zizania latifolia, corn, celery, apple dietary fiber and Zizania rice, corn flavonoids to observe the changes of SOD content in cytoplasm. At the same time, the expression of PPAR-a and CPT 1 A mRNA were measured. Results: (1) the content of total dietary fiber (TDF) of Zizania latifolia was 7.242 g / 100 g, the content of TDF of maize was 9.13 g / 100 g, the content of TDF of apple was 13.52%, the content of TDF of celery was 38.6%, the content of TDF of apple was 13.52%, the content of TDF of celery was 38.6%. The content of flavonoids in Zizania latifolia was 3.674g / 100g, which was significantly higher than that of corn flavonoids. It is the best concentration to induce steatosis. The corn dietary fiber can decrease the content of TG-MDA and increase the content of SOD in the cytoplasm of HepG2 non-alcoholic hepatic steatosis, and it can improve the non-alcoholic liver steatosis. The effect of celery dietary fiber was the second, while the effect of apple dietary fiber was not obvious. Corn flavonoids also decreased the TG content in the cytoplasm of the model cells. The expression of PPAR-a and CPT-1A mRNA in maize, celery dietary fiber and Zizania rice, maize flavonoid treated cells was significantly increased, while the expression of PPAR-a and CPT-1A mRNA in apple dietary fiber was slightly increased, but there was no significant difference in the expression of PPAR-a and CPT-1A mRNA, but there was no statistically significant difference in the expression of PPAR-a and CPT-1A mRNA. Conclusion: 0. 5 mm oleic acid can induce fatty degeneration in HepG2 cells for 24 hours and form a non alcoholic hepatosteatosis cell model. Corn dietary fiber and flavonoids can improve non-alcoholic liver steatosis cells by activating PPAR-a, promoting CPT-1A expression, increasing fatty acid decomposition, and reducing body fat content, the mechanism of which may be the activation of PPAR-a, the increase of CPT-1A expression, the increase of fatty acid decomposition, and the decrease of body fat content. The celery dietary fiber can also improve the non-alcoholic hepatic steatosis, but the apple dietary fiber has little effect. The possible reason is that different sources of dietary fiber, the content of soluble dietary fiber and insoluble dietary fiber, and different phytochemicals combined with dietary fiber lead to different effects.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R151.3

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