沙苑子、泽泻降脂作用的比较研究

【作者】 张文涛；

【导师】 张建军；

【作者基本信息】 北京中医药大学 ， 临床中药学， 2012， 硕士

【摘要】 沙苑子味甘性温，归肝、肾经，具有温补肝肾、固精、缩尿、明目等功效，用于肾虛腰痛、遗精早泄、白浊带下、小便余沥、眩暈目眩等病症。本实验室前期研究发现沙苑子醇提物、沙苑子总黄酮具有较好的降脂作用。泽泻为性寒，味甘、淡，归肾、膀胱经，有利水渗湿、泄热之功效，临床常用于治疗小便不利、水肿胀满、痰饮眩暈及淋浊等病症。本实验室前期研究发现泽泻提取物泽泻总$^类物质具有较好的降脂作用。传统医学认为高脂血症与脾贤阳虛，痰浊阻滞关系密切，故本研究基于前期研究的基础上，采用沙苑子、泽泻的降脂有效成分，沙苑子总黄酮及泽i写总萜类物质探讨沙苑子及泽》写的降脂作用，比较二者降脂作用的特点，并从甘油三酯合成及分解的角度探讨其机理。1、文献综述检索2001-2011年间发表的治疗高脂血症的文献，发现传统医学根据高脂血症相关病因病机、临床表现、并发症等，认为本病存在于中医"痰浊"、"血瘀"、"湿油"、"肥胖，，、"眩暈"、"中风"、"心悸"、"胸痹"等病证之中，病机多为本虛标实，标实主要表现为痰浊、瘀血阻滞，本虛主要责之脾肾功能失调或虛损，一些临床调查及体质研究也发现，脾肾阳虛及痰湿是高脂血脂的重要病机，尤其是随着年龄增长，脾肾阳虛所占比例升高，故在高脂血症治疗中补阳药及利湿药运用较多。具有降脂效果的补阳药有：菟丝子、鹿茸、淫羊藿、杜仲、肉桂、补骨脂、金樱子、沙苑子、肉;^蓉等，具有降脂作用的利湿药有：泽泻、茯苓、车前子、茵陈、玉米须、虎杖等。综述二探讨了沙苑子、泽泻药理作用的研究概况，发现沙苑子具有抗肝损伤、调节血脂、抗肿瘤、抗氧化、抗疲劳、抗衰老药理作用，且对生殖系统具有一定调控作用；泽泻的主要药理作用有：降血脂及抗脂肪肝作用、利尿作用、抗肾结石形成作用、降血压作用等。在研究现代医学对高脂血症的认识时发现，胆固醇酰基转移酶（DGAT)可促进甘油二酯合成甘油三酯，是甘油三酯合成的关键限速酶。DGAT的表达主要是胰岛素应答，DGAT的活性与胰岛素抵抗关系密切。甘油三酯脂肪酶（ATGL)主要水解底物是甘油三酯，是甘油三酯水解的关键限速酶。ATGL在高脂血症、糖尿病、肥胖，非酒精性脂肪肝等疾病中扮演了重要角色。ATGL的发现为脂代谢研究开辟了一条新的道路，进一步完善了对脂肪分解过程的认识，研究药物对ATGL活性的影响，可能是调节脂代谢的一个新方向。2、实验研究’方法：通过建立高脂饮食诱导建立大鼠高脂血症模型，造模成功后，模型组根据血清TC水平随机分为模型组，阳性药组，沙苑子高剂量、中剂量、低剂量组，泽泻高剂量、中剂量、低剂量组、配伍高剂量组、配伍低剂量组。分组后，模型组及空白组按lOml/kg给予、泽无菌蒸馏水，阳性药组按loomg/kg给予非诺贝特，沙苑子高剂量、中剂量、低剂量组分别按给药量136mg/kg、68mg/kg、34mg/kg,相当于人给药量30g、15g、7.5g,给予沙苑子醇提物，泽泻高剂量、中剂量、低剂量组分别按给药量98nig/kg、49mg/kg、24.5mg/kg,相当于人给药量20g、10g、5g，给予泽泻萜类提取物，配伍高剂量组给予沙苑子34nig/kg，泽泻24.5mg/kg,分别相当于人给药量沙苑子7.5g,泽泻5g,配伍低剂量组给予沙苑子17mg/kg,泽泻12.25ing/kg,分别相当于人给药量沙苑子3.5g，泽湾2.5g。结果：（1)高脂饮食诱导8周后模型组血清ldl-c、tc含量上升（p<0.05)，hdl-c水平显著下降（p<0.05)。(2)给药治疗8周后，和模型组相比，非诺贝特组、沙苑子各剂量组、泽泻各剂量组及配伍低剂量组均能降低血清ldl-c含量（p<0.05),沙苑子高、中剂量组，泽湾中、低剂量组能降低血清tc含量（p<0.05)，其他各中药组也有一定降低血清――-jf€-会量的趋势，沙苑子高剂量能提高血清hdl-c水平（p<0.05)。在肝脏脂质含量检测实验中发现，高脂饮食诱导大鼠高脂血症模型中，大鼠肝脏中甘油三酯及胆固醇含量较空白组明显升高（p<0.05)，存在明显的脂质蓄积观象，而非诺贝特组、沙苑子各剂量组、泽泻各剂量组，配伍组均能有效降低高脂血症模型大鼠肝脏甘油三酯及胆固醇含量（p<0.05)，具有明显的减轻肝脏脂质蓄积的作用。病理结果显示，各中药组有不同程度的减少并减小脂肪空泡，减轻肝细胞炎症浸润的作用，说明沙苑子、泽泻具有减轻肝脏脂肪变性的作用。在肝脏dgat2及atgl含量的检测中发现和空白组相比，模型组dgat2表达升高(p<0.05),atgl表达降低（p<0.05);和模型组相比，非诺贝特可下调肝脏dgat2表达(p<0.05),上调肝脏atgl表达（p<0.05);jo模型组相比，沙苑子高、中、低剂量组及泽泻高、中剂量组均可下调肝脏dgat2表达（p<0.05)，沙苑子对大鼠肝脏atgl表达没有明显影响（p>0.05),泽泻有一定升高大鼠肝脏atgl表达的作用，但未见差异（p>0.05)。结论：1、高脂饮食可以导致大鼠血清tc、ldl-c水平升高，hdl-c水平降低，肝脏tc、tg含量显著上升，肝脏出现脂肪变性，此外高脂饮食还可以提高大鼠肝lf;tdgat2的袭^,膝低atgl表达。2、沙苑子、泽^具有较好的降低血清tc、ldl-c的作用；沙苑子还有升高hdl-c的作用，可能与其补肾固精作用有关；沙苑子、泽泻配伍也具有一定的降脂作用。。3、沙苑子、泽泻及其配伍均可以减轻肝脏脂质沉积，沙苑子、泽^均可以减轻高脂饮食导致的肝脏脂肪变性。4、沙苑子、泽泻均可降低dgat2的表达，减少甘油三酯的合成，沙苑子对肝脏atgl表达没有明显影响，泽萄有一定升高大鼠肝脏atgl表达的作用，说明沙苑子主要降低tg的合成，而泽泻既有降低TG合成的作用，也有一定促进TG分解的作用。5、非诺贝特具有较好的降脂作用，并可以减轻肝脏脂质沉积，并明显减轻肝脏脂肪变性，非诺贝特减轻肝脏脂质沉积的机理可能是遗过降低DGAT2表达，同时也提高ATGL表达。更多还原

【Abstract】 Hyperlipidemia (HLP) is a high level of serum total cholesterol (TC) or tirglyceride (TG) and(or) low levels of high density lipoprotein cholesterol (HDL-C). Evidence-Based Medicine studyfound that hyperlipidemia is caused by an independent risk factor for cardiovascular andcerebrovascular diseases. Due to differences in diet, environment and other aspects of foreignhypercholesterolemia more common, more study of cholesterol metabolism, tirglyceridemetabolism, the relative lack of About90million people in China suffering rfom hyperlipidemia,elevated triglyceirdes are relatively common, so lowering tirglyceirdes is increasingly becomingone of the keys of the lipid-loweirng drugs. Therefore, emphasis on research on triglyceirdes isnecessary and valuable. Western medicine lipid-loweirng drugs such as statins can causerhabdomyolysis, ifbrates can lead to liver damage, elevated aminotransferases, and niacin causehigh uric acid, although Western medicine with better lipid-loweirng effects, but also show someside effects the advantages of Chinese medicine, it should carry forward the multi-target,multi-component, multi-channel, the overall regulation of blood lipid metabolism.1， Literature ReviewRetrieval treatment of hyperlipidemia literature published between2001-2011, found thattraditional medicine hyperlipidemia pathogenesis, clinical manifestations, complications, and thatthe disease exists in the Chinese medicine "phlegm","stasis" wet muddy "," obesity "", Vertigo ""，stroke "", palpitations ""， Chest "Syndrome, pathogenesis, and more oriented vacuity and excessmainly of phlegm, blood stasis block the blood, the virtual primary responsibility of the spleen andkidney dysfunction or deficiency, also found a number of clinical investigations and physical,spleen and kidney yang and phlegm fat lipids in pathogenesis, especially with age, spleen kidneyyang deifciency increased the proportion, and therefore more to BHD drugs and the dampnessdrug use in the treatment of hyperlipidemia. Yang drugs with lipid-loweirng effect of: dodder seed,antler, Epimedium, Eucommia, cinnamon, psoralen, Rosa laevigata, complanatus, Cistanche andother liapid-loweirng effects of dampness medicine: Alism, Fori a capillaris, Plantago, Com Silk,Polygonum, etc..Found in the study of modem medical understanding of hyperlipidemia, tirglyceride lipase (ofATGL) main hydrolysis substrate is a triglyceride, the key rate-limiting enzyme for triglyceirdehydrolysis. ATGL plays an important role in the hyperlipidemia, diabetes, obesity, non-alcoholicfatty liver and other diseases. ATGL discovery of lipid metabolism research has opened up a newpath, to further improve the understanding of the process of lipolysis, the impact of drugs on theactivity of ATGL may regulate lipid metabolism in a new direction.Ideal hyperlipidemia animal models is to promote research and development of hyperlipidemia important factors, rfom the perspective of exogenous and endogenous lipid metabolism pathwaysof feeding method to establish a rat model of hyperlipidemia, found that rats strain, sex, feedformulation, such as cholesterol, lard, sugar, whether it contains anti-thyroid hormone drugs,modeling time, feeding practices and other factors can affect blood lipid levels of rats.2, the expeirmental studyMethods: through the establishment of high-fat diet induced rat model of hyperlipidemia atfersuccessful modeling, model group, serum TC levels were randomly divided into model group, thepositive control group, complanatus high-dose, medium dose, low dose group, Alisma high-dose,medium dose, low dose group. Atfer grouping, the model group and control groups according to1Oml/kg given sterile distilled water, positive drug group lOOmg/kg fenoifbrate complanatus highdose, dose, low-dose group were administered amount of136mg/kg68mg/kg,34mg/kg, theamount of crude drug4g/kg,2g/kg, Ig/kg, giving complanatus ethanol extract of Alisma high dose,medium dose, low dose group were administered dose of98mg/kg,49mg/kg,24.5mg/kg, theamount of crude drug2.64g/kg,1.32g/kg,0.66g/kg, giving Alisma terpens extract.Results:(1)8weeks atfer the high-fat diet-induced model group, serum LDL and TC contentincreased (p <0.05)， HDL levels decreased signiifcantly (p <0.05).(2) atfer8weeks of drugtreatment, and compared to model group, the fenoifbrate group, complanatus each dose group,Alisma each dose group are able to reduce serum LDL levels (p less than0.05), complanatus, themiddle dose group, Alisma, low-dose group can reduce serum TC levels (p less than0.05), andother traditional Chinese medicine group must reduce serum TC levels, complanatus high dosescan increase serum HDL levels (p <0.05).(3) The biopsy revealed that fenofibrate with a certainliver injury, but can be significantly reduced hepatic steatosis, improved liver morphology, reduceinflammation and infiltration, and other traditional Chinese medicine group also different degreesof reduction in hepatic steatosis, and reduce and reduce fat vacuoles, improve liver morphology,reducing the role of inflammatory infiltration of the liver cells.Found in the liver lipid content in the detection expeirment, high-fat diet-induced rat model ofhyperlipidemia, and rat liver triglyceride and cholesterol levels compared with blank control groupwas signiifcantly increased (p <0.05), there is a signiifcant lipid accumulation phenomenon, ratherthan the fenofibrate group, complanatus each dose group, Alisma each dose group can effectivelyreduce hyperlipidemia rat model of liver tirglyceride and cholesterol content (p less than0.05),with obvious reduce the role of the liver lipid accumulation.ATGL and blank group, model group to reduce the level found in the liver ATGL and DGAT2content,, detection (p <0.05), DGAT2levels (p <0.05); compared with model group, fenoifbratemay increase the liver ATGL expression (p <0.05) lowered liver DGAT2expression (p <0.05); 6沙苑子泻降脂作用的比较研究、泽compared with model group, the rats of vairous traditional Chinese medicine liver ATGL levelthere was no difference (p>0.05); complanatus high, medium and low-dose group, Alisma highschool dose group, can be lowered liver DGAT2expression (p <0.05).Conclusion: The high-fat diet can increase expression of the rat liver DGAT2and reduced ATGLexpression.2complanatus, Alisma certain lipid-lowering effect, and can reduce liver lipiddeposition, and improve hepatic steatosis.3complanatus each dose group, Alisma, the middle dosegroup can be reduced DGAT2expression, reducing tirglyceirde synthesis, but not in liver ATGLexpression. Fenofibrate has a better lipid-lowering effect, and can reduce the deposition of liverlipids significantly reduce hepatic steatosis. Fenofibrate to reduce liver lipid deposition mechanismmay increase the ATGL expression also reduced DGAT2expression.更多还原