化瘀温胆汤对糖耐量低减大鼠胰岛素抵抗及AMPK信号通路的影响

发布时间：2018-05-09 20:09

本文选题：糖耐量低减 + 胰岛素抵抗　；参考：《黑龙江中医药大学》2017年博士论文

【摘要】：研究目的建立单纯高脂饮食诱导的IGT大鼠模型,在此基础上观察化瘀温胆汤对高脂诱导大鼠外观表现和医学病理特征的影响。阐明化瘀温胆汤改善I R和能量代谢的机制,为探寻其作用靶点提供实验依据。研究方法健康Wistar雄性大鼠,随机分两组,正常组给予普通饲料喂养,其余大鼠给予高脂饲料喂养,共持续20w,诱导IGT模型。观察大鼠外观表现,行IPGTT检测血糖,测定FPG及2hPG。模型复制成功后,随机分为模型组、中药组、西药组,中药组给予5.2g/kg·d化瘀温胆汤,西药组给予99 mg/kg·d二甲双胍溶液,正常组和模型组给予同体积生理盐水。连续灌胃给药4w。实验结束后,腹主动脉取血,分离血清,ELISA法检测Fins、G SP、ADPN、LEP,计算IAI、IRI,生化分析仪检测HDL-C、LDL-C。称取肝脏质量,计算肝脏指数。肝组织匀浆,检测TG、TC、LPO、GSH-Px水平。行肝脏糖原染色观察糖原储备情况,图像分析软件分析阳性目标面密度和平均灰度。Realtime PCR法检测肝脏AMPK信号通路中AMPKα、A CCα mRNA的表达。Western Blot法检测AMPKα、p-AMPKα、ACCα、p-ACCα蛋白的表达。研究结果1.血糖、血脂、胰岛素相关指标:化瘀温胆汤减轻体重,降低大鼠F PG、2hPG,与模型组比较有统计学意义(p0.05或p0.01),降低FPG存在时效性。化瘀温胆汤降低Fins、IRI、GSP水平,提高IAI水平,与模型组比较有统计学意义(p0.05)。化瘀温胆汤提高血清HDL-C水平、降低L DL-C水平,与模型组比较有统计学意义O?0.01)。2.细胞因子:化瘀温胆汤提高血清ADPN水平、降低LEP水平,与模型组比较有显著性差异(p0.01)。3.肝脏糖脂代谢及氧化应激指标:化瘀温胆汤降低肝脏质量、肝脏指数,肝脏TG、TC水平,与模型组比较有统计学意义(p0.05或p0.01)。化瘀温胆汤降低LPO水平,提高GSH-Px水平,与模型组比较有统计学意义(p0.05或p0.01)。化瘀温胆汤提高阳性目标面密度和平均灰度值,与模型组比较有显著性差异(p0.01),增加肝脏糖原储备。4.肝脏AMPK信号通路:化瘀温胆汤上调肝脏组织AMPKα mRNA表达,与模型组比较差异有统计学意义(p0.05),对ACCα mRNA表达无影响(p0.05)。化瘀温胆汤上调肝脏组织AMPKα、p-AMPKα、p-ACCα蛋白表达,与模型组比较有统计学意义(p0.05或p0.01),对ACCα蛋蛋表达无明显影响(p0.05)。结论1.化瘀温胆汤能够降低模型大鼠血糖及血脂水平,纠正糖脂代谢紊乱,改善IR。2.化瘀温胆汤提高模型大鼠ADPN水平,降低LEP水平,改善能量代谢状态。3.化瘀温胆汤能够减轻模型大鼠肝脏脂肪沉积,改善氧化应激状态,增加肝脏糖原储备,起到保护肝脏作用。4.化瘀温胆汤能够提高模型大鼠肝脏AMPK活性,抑制ACC活性,发挥抑制脂肪合成,增加脂肪酸氧化,改善能量代谢状态,增加胰岛素敏感性作用。5.化瘀温胆汤影响模型大鼠肝脏AMPK-ACC信号通路,是其改善IGT IR的作用靶点之一。
[Abstract]:Objective to establish IGT rat model induced by high fat diet and observe the effect of Huayu Wendan decoction on the appearance and medical pathological characteristics of rats induced by high fat diet. To elucidate the mechanism of Huayu Wendan decoction (Huayu Wendan decoction) in improving I R and energy metabolism. Methods healthy male Wistar rats were randomly divided into two groups: normal group were fed with normal diet and other rats were fed with high fat diet for 20 weeks to induce IGT model. The appearance of rats was observed, blood glucose was detected by IPGTT, FPG and 2 h PG were measured. After successful replication, the model group was randomly divided into three groups: model group, traditional Chinese medicine group, western medicine group, 5.2g/kg d Huayu Wendan decoction, western medicine group 99 mg/kg d metformin solution, normal group and model group were given the same volume of normal saline. The drug was continuously administered by gavage for 4 weeks. At the end of the experiment, the abdominal aorta blood was taken and the serum was separated by Elisa for the detection of Finsberg SPN ADPNN LEP, the IAI IRI was calculated, and the HDL-CU LDL-C was detected by biochemical analyzer. The liver mass was measured and the liver index was calculated. Liver tissue homogenate was used to detect the level of GSH-Px in TGV TCU. The glycogen reserve was observed by liver glycogen staining, and the expression of AMPK 伪 -A CC 伪 mRNA in liver AMPK signaling pathway was detected by image analysis software. The expression of AMPK 伪 -ACC 伪 -p-ACC 伪 protein in liver AMPK signaling pathway was detected by image analysis software and the expression of AMPK 伪 -p-AMPK 伪 -ACC 伪 -p-ACC 伪 protein was detected by Western Blot method. Results 1. Blood glucose, blood lipids, insulin related indexes: Huayu Wendan decoction to reduce body weight, reduce the rat F-PG2 hPG2, compared with the model group, compared with the model group, p0.05 or p0.01a, reduce the time effect of FPG. Huayu Wendan decoction decreased the level of IAI and increased the level of IAI, which had statistical significance compared with the model group. Huayu Wendan decoction increased serum HDL-C level and decreased L DL-C level, which had statistical significance compared with model group. Cytokines: Huayu Wendan decoction increased the level of serum ADPN and decreased the level of LEP, there was significant difference between the model group and the model group. Liver glucose and lipid metabolism and oxidative stress index: Huayu Wendan decoction decreased liver quality, liver index, liver TGG TC level, and compared with model group, there was significant difference between model group and model group (P 0.05 or p 0.01). Huayu Wendan decoction decreased the level of LPO and increased the level of GSH-Px, which had statistical significance compared with the model group (p0.05 or p0.01). Huayu Wendan decoction increased the positive target surface density and average gray value, compared with the model group, the difference was significant (p0.01), and increased liver glycogen reserve. 4. Liver AMPK signal pathway: Huayu Wendan decoction upregulated the expression of AMPK 伪 mRNA in liver tissue, which was significantly different from the model group, but had no effect on the expression of ACC 伪 mRNA. Huayu Wendan decoction upregulated the expression of AMPK 伪 -pACC 伪 protein in liver tissue, which had statistical significance compared with the model group (p0.05 or p0.01), but had no significant effect on the expression of ACC 伪 egg. Conclusion 1. Huayu Wendan decoction can reduce blood sugar and blood lipid level, correct glucose and lipid metabolism disorder and improve IR.2. Huayu Wendan decoction increased ADPN level, decreased LEP level and improved energy metabolism state of model rats. Huayu Wendan decoction can reduce hepatic fat deposition, improve oxidative stress, increase liver glycogen reserve and protect liver. Huayu Wendan decoction could increase AMPK activity, inhibit ACC activity, inhibit fat synthesis, increase fatty acid oxidation, improve energy metabolism and increase insulin sensitivity. Huayu Wendan decoction (Huayu Wendan decoction) affects AMPK-ACC signal pathway in liver of model rats and is one of the targets for improving IGT IR.
【学位授予单位】：黑龙江中医药大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R285.5

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