【作者】 王昌儒；

【导师】 向楠；

【作者基本信息】 湖北中医药大学 ， 中医内科学， 2014， 博士

【摘要】 目的：通过查阅近几十年来有关代谢综合征的中、西医研究文献，并结合中医药理论及临床实践研究，探讨代谢综合征的中西医发病机制，为临床应用中医理论治疗代谢综合征提供理论支撑。利用高脂高糖饲料复制代谢综合征动物模型，研究温肾健脾化痰方对代谢综合征大鼠向心性肥胖、血脂指标、血压、胰岛素抵抗、肿瘤坏死因子、过氧化物酶体增殖物激活受体-γ及AMPK信号通路的干预作用，探讨温肾健脾化痰方药对代谢综合征大鼠肥胖、血压、胰岛素抵抗及能量代谢的影响，从分子水平及药理学方面研究温肾健脾化痰对代谢综合征的作用机制，期望通过此研究，能从分子生物学及药理学层面解释温肾健脾化痰方对于代谢综合征的作用机制，从而为温肾健脾化痰法治疗代谢综合征提供理论依据。方法：将30只雄性SD大鼠适应性喂养1周后，随机分成空白组、代谢综合征模型组及温肾健脾化痰方药物组。代谢综合征模型组及温肾健脾化痰方药物组利用高糖高脂饲料喂养12周，复制出代谢综合征大鼠动物模型。在造模同时，空白组及代谢综合征模型组给予生理盐水灌胃，温肾健脾化痰方药物组给予同等剂量的温肾健脾化痰方药物。在第12周未，测量各组大鼠体重、身长、尾动脉收缩压并进行心脏采血、取肾周脂肪组织，应用生化分析仪测定血脂、空腹血糖，运用酶联免疫吸附法（ELISA）测血清胰岛素，并计算HOMA-IR指数。药物干预后除检测上述指标外，还采用ELISA法测血清中肿瘤坏死因子（TNF-a），利用免疫组化法测定大鼠脂肪组织腺苷酸活化蛋白激酶（AMPK）、肝激酶B1（LKB1）蛋白表达，同时釆用聚合酶链式反应（RT-PCR）检测大鼠脂肪组织过氧化物酶体增殖物激活受体-γ(PPAR-γ)表达情况，蛋白质印迹法（Westernblot）及免疫组化法检测大鼠脂肪组织AMPK、LKB1蛋白表达。结果:⑴代谢综合征模型组大鼠经应用高糖高脂饲料喂养12周后，大鼠的体重、尾动脉收缩压、低密度脂蛋白（LDL）、总胆固醇（TC）、甘油三酯（TG）、空腹血糖、血清胰岛素及胰岛素抵抗指数等指标与空白组大鼠及温肾健脾化痰方药物组相比明显升高，而高密度脂蛋白（HDL）明显降低，以上指标相比较均具有统计学意义(P＜0.05)。⑵药物干预后，温肾健脾化痰方药物组大鼠的体重、收缩压、低密度脂蛋白（LDL）、总胆固醇（TC）、甘油三酯（TG）、空腹血糖、血清胰岛素及胰岛素抵抗指数，与模型组相比均明显降低，而高密度脂蛋白（HDL）显著升高，此两组大鼠以上指标相比较有统计学意义(P＜0.05)。⑶代谢综合征模型组肿瘤坏死因子（TNF-a）水平明显高于空白组及温肾健脾化痰方药物组，有统计学意义(P＜0.01)；药物干预后，温肾健脾化痰方药物组大鼠的肿瘤坏死因子（TNF-a）水平与代谢综合征模型组相比明显降低，二者相比较有统计学意义(P＜0.01)。⑷代谢综合征模型组过氧化物酶体增殖物激活受体-γ（PPAR-γ）表达明显低于空白组及温肾健脾化痰方药物组，有统计学意义(P＜0.01)；应用温肾健脾化痰方药物干预后，温肾健脾化痰药物组大鼠脂肪组织过氧化物酶体增殖物激活受体-γ（PPAR-γ）表达显著高于代谢综合征模型组大鼠，二者相比较有统计学意义(P＜0.01)。⑸代谢综合征模型组LKB1，AMPK表达明显低于空白组及温肾健脾化痰方药物组，有统计学意义(P＜0.01)；药物干预后，温肾健脾化痰方药物组大鼠脂肪组织LKB1，AMPK表达显著高于代谢综合征模型组大鼠，两组以上指标相比较有统计学意义(P＜0.01)。结论:⑴经过温肾健脾化痰方的治疗，能够使代谢综合征大鼠的体重、血脂水平、尾动脉血压、胰岛素抵抗指数出现明显下降。证明温肾健脾法能通过减轻代谢综合征大鼠的向心性肥胖，改善其高脂血症、降低其血压、提高其胰岛素敏感性而对代谢综合征起到良好治疗作用。⑵通过本研究证明，肿瘤坏死因子与代谢综合征的发生有关联，当发生代谢综合征时，体内的肿瘤坏死因子会增加。经过温肾健脾化痰方干预后，代谢综合征大鼠体内的肿瘤坏死因子出现了下降，证明温肾健脾法能减少代谢综合征大鼠体内肿瘤坏死因子的生成，从而对代谢综合征起到治疗作用。⑶本研究发现，过氧化物酶体增殖物激活受体-γ(PPAR-γ)也与代谢综合征的形成密切相关，其在代谢综合征大鼠的体内出现了下降，证明当代谢综合征产生时，过氧化物酶体增殖物激活受体-γ会出现下降。经温肾健脾化痰方干预后，过氧化物酶体增殖物激活受体-γ在代谢综合征大鼠体内出现了明显增加，证明温肾健脾化痰法能阻止过氧化物酶体增殖物激活受体-γ的减少，从而对代谢综合征起到治疗作用。⑷本研究发现，在代谢综合征形成过程中，LKB1/AMPK信号通路在代谢综合征大鼠体内的表达出现了明显减弱，证明代谢综合征的发生与LKB1/AMPK信号通路密切相关。温肾健脾化痰方能使代谢综合征大鼠体内LKB1/AMPK信号通路的表达得到加强。证明温肾健脾化痰法能使代谢综合征患者体内的能量代谢紊乱得到改善。以上的实验结果表明，温肾健脾化痰方能有效的干预代谢综合征的形成，对代谢综合征的治疗是明确有效的。更多还原

【Abstract】 Objective: To investigate the pathogenesis of metabolic syndrome oftraditional Chinese medicine and Western medicine through theory andliterature research. Effects of Wenshen Jianpi Huatan Decoction onmetabolic syndrome rat body weight， blood lipid， insulin resistance，tumor necrosis factor， peroxisome proliferator-activated receptorgamma and AMPK signaling pathway， to explore the effect of JianpiWenshen Huatan Decoction on MS rat obesity， insulin resistance，energy metabolism，thus study on the mechanism of action of warmingkidney and invigorating spleen and resolving phlegm on metabolicsyndrome.Methods: The30male SD rats were fed for one week later，were randomly divided into the blank control group， MS model groupand Wenshen Jianpi Huatan Decoction group. MS model group andWenshen Jianpi Huatan Decoction group using high-sugar high-fat dietfor12weeks， to replicate the rat model of metabolic syndrome. Inmodeling the same time， the blank control group and model groupwere given normal saline，Wenshen Jianpi Huatan Decoction groupwere given the same dose of the drug Wenshen Jianpi Huatanprescription drugs.At the last day of the rats were feed for12weeks，the rats weight， height， systolic blood pressure were measured and taken the blood from the rats，heart，take the perirenal adipose tissue，application of biochemical analyzer lipids， fasting glucose， seruminsulin using ELISA method to measure and calculate HOMA-IR index.After the detection of drug intervention in addition to the aboveindicators， measured by ELISA in serum TNF-a， measured AMPKand LKB1protein expression in adipose tissue， use ofimmunohistochemistry， and preclude the use of RT-PCR detection ofadipose tissue PPAR-γ expression， Western blot detected adiposetissue AMPK， LKB1protein.Results:⑴model group rats afterapplication of high-sugar high-fat diet for12weeks， body weight ofrats， systolic blood pressure， LDL， TC， TG， fasting bloodglucose， serum insulin and insulin resistance index were significantcompared with the control group of rats increased， while HDLsignificantly reduced， compared to the above indicators werestatistically significant (P＜0.05).⑵After drug intervention， theweight， systolic blood pressure， LDL， TC， TG， fasting bloodglucose， serum insulin and insulin resistance index of Wenshen JianpiHuatan Decoction drugs rats compared with the model group weresignificantly decreased， while HDL significantly rise.The aboveindicators of the two groups of rats compared were statisticallysignificant (P＜0.05).⑶The TNF-a levels of MS model group weresignificantly higher than the control group and Wenshen Jianpi HuatanDecoction drugs group，was statistically significant (P＜0.01). After drugintervention， Compared with the model group， the TNF-a levels ofWenshen Jianpi Huatan Decoction drugs group were significantlylower， the comparison of TNF-a levels between the two wasstatistically significant (P＜0.01).⑷Model group PPAR-γ expressionwas significantly lower than the control group and Wenshen Jianpi Huatan Decoction drugs group， the comparison of PPAR-γ levelsbetween the three groups was statistically significant(P＜0.01); Afterdrug intervention， PPAR-γ expression in adipose tissue of the druggroup was significantly higher than model group， both of which werestatistically significant compared (P＜0.01).⑸Model group P-LBK1，AMPK expression was significantly lower than the blank control groupand Wenshen Jianpi Huatan Decoction drugs group， the comparisonof P-LBK1and AMPK levels between the three groups wasstatistically significant (P＜0.01); After drug intervention， theexpression of P-LBK1， AMPK adipose tissue of Wenshen JianpiHuatan Decoction drugs group was significantly higher than the modelgroup， compared with the two groups was statistically significantindicator (P＜0.01).Conclusions:⑴After the treatment of WenshenJianpi Huatan Decoction， The body weight， lipid levels， bloodpressure and insulin resistance index of the rats which suffering fromillness of metabolic syndrome was significantly decreased.⑵Tumor necrosis factor is a key factor that will lead to metabolicsyndrome， when metabolic syndrome occurring，the body’s tumornecrosis factor will increase， while warming kidney Spleen candecrease the tumor necrosis factor of the rats which take a disease ofmetabolic syndrome.⑶PPAR-γ is another key factor that will lead tometabolic syndrome，when metabolic syndrome occurring，the body’sPPAR-γ will decrease， while warming kidney Spleen can increase thePPAR-γ of the rats which take a disease of metabolic syndrome.⑷Theexpression of LKB1/AMPK signaling pathway in the rats which ake adisease of metabolic syndrome appeared significantly decreased.Sothis proved that the LKB1/AMPK signaling pathways is a factor that hasclosed relationship with metabolic syndrome. Wenshen Jianpi Huatan Decoction can cause the the expression of LKB1/AMPK signalingpathway of MS rats to be raised. The above results suggest that，Wenshen Jianpi Huatan Decoction can prevent the emerge ofmetabolic syndrome， The effectiveness of Wenshen Jianpi HuatanDecoction to treat the metabolic syndrome is clear and effective.更多还原