脾虚状态对高脂血症大鼠肝脏脂质沉积的影响及机制研究

发布时间：2018-05-31 17:54

本文选题：脾虚 + 脂质沉积　；参考：《辽宁中医药大学》2016年硕士论文

【摘要】：目的:中医认为,脾主运化,脾失健运导致膏脂转输障碍。本研究从中医病机切入,在中医理论指导下,复制脾虚高脂血症大鼠模型,从分子水平初步探讨脾虚状态对血脂水平及肝脏脂质沉积的影响及机制。材料与方法:60只SPF级SD雄性大鼠,体重(300±10)g,适应环境7 d,进入实验过程,实验用大鼠按照随机数字表法分为空白对照组20只、高脂血症组20只、脾虚高脂血症组20只,空白对照组继续常规饲养,高脂血症组予高脂饲料喂饲,脾虚高脂血症组采用游泳劳倦过度加饮食不节结合高脂饲料喂饲造模,造模60 d后分别称重取材。采用间苯三酚法测定尿D-木糖排泄率,全自动生化分析仪检测血清TC、TG、LDL-C、HDL-C、AMY水平,采用HE染色观察肝脏形态变化,油红O染色观察肝脏脂质沉积,分别采用COD-CE-PAP法和GPO-PAP法测定肝脏TC和TG含量,采用实时荧光定量反转录-聚合酶链反应(real-time RT-PCR)及蛋白免疫印迹(Western blot)法检测肝脏HMGCR、HL、CYP7A1、LDL-R基因m RNA和蛋白表达。统计学处理:采用SPSS 17.0软件包进行统计分析,采用单因素方差(One-Way ANOVA)进行分析,统计结果以平均值±标准差(x±S)表示,以P0.05为差异有统计学意义。结果:1.脾虚状态对高脂血症大鼠血脂、血清AMY水平及尿D-木糖排泄率的影响实验结果显示,与空白对照组相比,高脂血症组及脾虚高脂血症组大鼠血清TC、LDL-C水平显著升高(均P0.01),血清HDL-C、AMY水平显著降低(P0.05、P0.01)。与高脂血症组相比,脾虚高脂血症组大鼠血清TC、LDL-C水平显著升高(P0.05、P0.01),血清HDL-C、AMY水平及尿D-木糖排泄率显著降低(P0.05、P0.01)。2.脾虚状态对高脂血症大鼠肝脏形态以及肝脏脂质沉积的影响实验结果显示,与空白对照组相比,高脂血症组及脾虚高脂血症组肝细胞出现大面积脂肪变性和脂质沉积。3.脾虚状态对高脂血症大鼠肝脏脂质含量的影响实验结果显示,与空白对照组相比,高脂血症组及脾虚高脂血症组大鼠肝脏TG、TC含量显著升高(P0.05、P0.01)。与高脂血症组相比,脾虚高脂血症组大鼠肝脏TG、TC含量显著升高(均P0.01)。4.脾虚状态对高脂血症大鼠肝脏HMGCR、HL、CYP7A1、LDL-R基因m RNA表达的影响实验结果显示,与空白对照组相比,高脂血症组和脾虚高脂血症组大鼠肝脏HMGCR、HL、CYP7A1及LDL-R基因m RNA表达水平显著降低(P0.05、P0.01)。与高脂血症组相比,脾虚高脂血症组大鼠肝脏HL、CYP7A1及LDL-R基因m RNA表达水平显著降低(P0.05、P0.01),而HMGCR基因m RNA表达水平无显著变化。5.脾虚状态对高脂血症大鼠肝脏HMGCR、HL、CYP7A1、LDL-R基因蛋白表达的影响实验结果显示,与空白对照组相比,高脂血症组和脾虚高脂血症组大鼠肝脏HMGCR、HL、CYP7A1及LDL-R基因蛋白表达水平显著降低(P0.05,P0.01)。与高脂血症组相比,脾虚高脂血症组大鼠肝脏HL、CYP7A1及LDL-R基因蛋白表达水平显著降低(P0.05,P0.01),而HMGCR基因蛋白表达水平无显著变化。结论:1.通过游泳劳倦过度加饮食不节结合高脂饲料的方法成功构建脾虚高脂血症大鼠模型。2.脾虚状态加重高脂血症大鼠血脂紊乱以及肝脏脂质沉积。3.脾虚状态影响肝脏胆固醇摄取及转化相关基因m RNA及蛋白表达。
[Abstract]:Objective: Traditional Chinese medicine (TCM) thinks that the spleen is main transport and the spleen is lost and transported to the barrier of ointment and fat transfer. In this study, the rat model of spleen asthenia hyperlipidemia was replicated under the theory of traditional Chinese medicine, and the effect and mechanism of spleen deficiency on blood lipid level and liver lipid deposition were preliminarily discussed from the molecular level. Materials and methods: 60 SD male rats of SPF grade, The body weight (300 + 10) g, adapted to the environment 7 d, entered the experimental process. The experimental rats were divided into 20 rats in the blank control group according to the random number table method, 20 in the hyperlipidemia group and 20 in the spleen asthenia hyperlipidemia group. The blank control group continued to be kept regularly, the hyperlipidemia group was fed with high fat diet, and the hyperlipidemia group of the spleen deficiency hyperlipidemia group was overfed with excessive diet and diet. A model was fed with high fat feed and the model was weighed after 60 d. The urine D- xylose excretion rate was measured by the method of phenolol and three phenol. The serum TC, TG, LDL-C, HDL-C, AMY level were detected by the automatic biochemical analyzer. The liver morphologic changes were observed by HE staining, the liver lipid deposition was observed by oil red O staining, and the liver was detected by COD-CE-PAP method and GPO-PAP method respectively. The contents of TC and TG were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction (real-time RT-PCR) and protein immunoblotting (Western blot) method for detecting the m RNA and protein expression of the liver HMGCR, HL, CYP7A1, LDL-R genes. Statistical analysis was carried out by the statistical analysis of 17 software packages and the statistical analysis was carried out by single factor variance. Results the results were statistically significant with the mean value of standard deviation (x + S) and P0.05 as difference. Results: the effects of 1. spleen deficiency on blood lipid, serum AMY level and urine D- xylose excretion rate in hyperlipidemia rats showed that the serum TC and LDL-C levels in the hyperlipidemia group and the spleen asthenia hyperlipidemia group were significantly higher than those in the blank control group (all (all). P0.01), the level of serum HDL-C and AMY decreased significantly (P0.05, P0.01). Compared with the hyperlipidemia group, the serum TC, LDL-C level in the spleen asthenia hyperlipidemia group increased significantly (P0.05, P0.01), the serum HDL-C, AMY level and the excretion rate of urinary D- xylose decreased significantly. The experimental results showed that compared with the blank control group, the effects of large area fatty degeneration and lipid deposition of.3. spleen deficiency on liver lipid content in hyperlipidemia rats showed that the liver TG, T in hyperlipidemia and spleen hyperlipidemia rats were compared with those in the blank control group. C content increased significantly (P0.05, P0.01). Compared with the hyperlipidemia group, the liver TG, TC content in the spleen asthenia hyperlipidemia group increased significantly (P0.01).4. spleen deficiency state on the liver HMGCR, HL, CYP7A1, LDL-R genes in hyperlipidemia rats. The results showed that the hyperlipidemia group and the spleen deficiency hyperlipidemia group were larger than the blank control group. The expression level of M RNA in rat liver HMGCR, HL, CYP7A1 and LDL-R decreased significantly (P0.05, P0.01). Compared with hyperlipidemia group, the liver HL, CYP7A1 and LDL-R genes in the spleen asthenia hyperlipidemia rats were significantly lower than those in the hyperlipidemia group. The effects of HL, CYP7A1 and LDL-R gene protein expression showed that the expression level of HMGCR, HL, CYP7A1 and LDL-R genes in the liver of hyperlipidemia and spleen deficiency hyperlipidemia rats was significantly lower than that in the blank control group (P0.05, P0.01). Compared with the hyperlipidemia group, the liver HL, CYP7A1 and LDL-R gene proteins in the spleen asthenia hyperlipidemia rats' liver were compared with those in the hyperlipidemia group. The expression level of the HMGCR gene protein was significantly reduced (P0.05, P0.01), but the expression level of HMGCR gene protein was not significantly changed. Conclusion: the spleen deficiency hyperlipidemia model of the rat model of spleen asthenia hyperlipidemia was successfully constructed by swimming overwork and diet combined with high fat diet. The effect of.2. on blood lipid disorder in hyperlipidemia rats and the effect of.3. spleen deficiency on liver lipid deposition in rats. Liver cholesterol uptake and transformation related genes m RNA and protein expression.
【学位授予单位】：辽宁中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R259

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