【作者】 白春艳；

【导师】 卓廉士；

【作者基本信息】 重庆医科大学 ， 中医临床基础， 2010， 硕士

【摘要】 非酒精性脂肪肝病(nonalcoholic fatty liver disease,NAFLD)是一种肝组织病理学改变的临床综合病变。与酒精性肝病相类似,但无过量饮酒史。因为其具有患病率高、隐匿性、肝纤维化、肝硬化等发展特点.所与对NAFLD的防治工作已经受到国内外的广泛关注。关于脂肪肝的发病机制,众多研究者一致认为与脂联素(Adi)、瘦素(Lep)、Lep受体有一定联系。Lep是由脂肪细胞分泌的一种蛋白质。在正常的生理情况下, Lep分泌的多少,均可反馈到下丘脑中枢系统,调节副交感神经,以适应饥饿或者摄食的反应[1]脂联素是一种以脂肪组织为唯一来源的活性多肽,它在机体肥胖时表达下调大量研究提示[2-3],脂联素和Lep在NAFLD的发生发展中起到了重要的作用.下丘脑区域是机体内分泌和能量代谢活动的重要结构区。尤其下丘脑-神经肽通路系统在摄食反馈环路中起到了关键作用。Lep及其受体是传入饱食信号的信使因子作用于下丘脑黑色素-4受体及其配体黑色素细胞刺激素,而Lep发挥信使作用的关键则依靠下丘脑的中枢调节效应。大量实验证实,电针可以调整患者体内的内分泌系统,这是针刺效应的作用机制之一.电针治疗NAFLDA已经得到大量实验的证实,关于其作用机制也有通过干预肝细胞色素表达和氧化抗氧化作用进行治疗的说法[4]。中医纯天然疗法——针灸疗法,已被纳入世界医学的组成部分。对针灸作用机理的研究也倍受关注。故本研究通过检测电针治疗后的大鼠检测Lep、脂联素等相关指标,探讨电针对NAFLD的治疗机制。目的检测机体脂联素水平及下丘脑Lep、瘦素受体基因(mRNA)表达,探讨电针治疗非酒精性脂肪肝的作用机制。方法将远交群大鼠(Sprague Dawley,SD)32只随机为模型组22只及空白组10只。普通饲料喂养空白组,高脂饲料喂养模型组制备SD大鼠模型。8周后再将模型组大鼠随机分为:对照组(11只)、针刺组(11只)。治疗针刺组,取“丰隆”、“足三里”、“阴陵泉”、“三阴交”等穴进行针刺并加以频率10 Hz,强度1. 5 V的连续波;治疗4周后,检测各组大鼠血清甘油三酯(Trglyceride,TG)、血清胆固醇(serum cholesterol,TC)、Lep、脂联素,观察肝组织病理学变化,并用逆转录转胺霉链聚合反应( Reverse Transcription-Polymerase Chain Reaction,RT-PCR)方法检测下丘脑组织中Lep和瘦素受体mRNA的表达。结果1一般情况:各组大鼠在实验期间正常饮水摄食,饮水量,摄食量无明显异常。对照组大鼠皮毛有光泽、体态活泼;模型组大鼠体重明显增加、形体肥胖、皮毛发黄、蓬乱不顺滑。解剖观察空白组大鼠肝脏肉眼观察呈鲜红色,边缘锐利,表面光滑。模型组大鼠肝脏体积增大,边缘变钝,颜色呈土黄色,切面呈油腻状。2 HE染色:模型组大鼠:肝细胞胞浆内有大量脂肪空泡呈透明状,细胞核被脂肪空泡挤到肝细胞缘,病理证实脂肪肝形成造模成功。电针组经过治疗后,肝脂肪组织变性程度有所减轻,肝细胞结构趋于正常。3 Lep、脂联素指标:与模型组比较电针组肝组织匀浆中Lep水平显著上升、脂联素水平显著降低、血清中TC、TG值显著下降。4 RT-PCR实验结果:RT-PCR检测显示:模型组大鼠Lep和瘦素受体的mRNA表达明显下降,电针组大鼠下丘脑Lep及瘦素受体mRNA表达上调。见图4、5,图像分析系统分析显示:电针对NAFLD大鼠下丘脑Lep、瘦素受体mRNA表达水平升高。结论:经过8周的高脂饲料喂养的NAFLD大鼠造模成功。电针可以影响实验大鼠下丘脑Lep、瘦素受体mRNA表达水平,缓解脂肪代谢紊乱情况,达到对NAFLD的治疗作用,推测电针治疗NAFLD病的作用机制可能是通过“针刺效应”作用于神经-内分泌调控系统,调整Lep、瘦素受体mRNA表达和脂联素水平。更多还原

【Abstract】 Non-alcoholic fatty liver disease is a type of liver tissue pathology clinical syndrome. And alcoholic liver disease similar to, but no history of excessive drinking. Because of their prevalence is high, the occult, hepatic fibrosis and cirrhosis and other developmental characteristics. and on the prevention and treatment of NAFLD have been affected by the widespread interest both at home and abroad. About the pathogenesis of fatty liver, numerous researchers unanimously and adiponectin (Adi) and leptin (Lep), leptin receptors have a certain contact. .Lep is produced by fat cells and a protein. In a normal physiological situations, Lep quantities, can be fed back to the central system, regulating hypothalamus parasympathetic nervous system, to accommodate the hunger or feeding of reaction[1] adiponectin is a unique source of adipose tissue to polypeptides, its body obesity Downregulation of research tips, adiponectin and leptin in the development of NAFLD played an important role [2] [3].The body of the hypothalamic area is the body of the endocrine and metabolic activity of important structural areas. In particular, the hypothalamus-neuropeptides pathway in the feeding of the feedback loop into play. Leptin and leptin receptors is the Messenger of the incoming signal to the full in the hypothalamus melanin-4 receptors and their ligands melanin-melanocyte-stimulating hormone leptin, and play a key role in the courier and rely on the hypothalamus regulation effect. A large number of laboratory confirmed, electro-acupuncture can be adjusted with the endocrine system, this is the mechanism of the effect of acupuncture. Electroacupuncture NAFLDA has been a large number of experiments confirmed, about its mechanism of action through hepatic cytochrome expression and antioxidant effects for oxidation treatment of [4].Acupuncture as a natural therapy, has been incorporated into the world of medicine. At the same time, the mechanism of acupuncture is also widely appreciated. Therefore this study by detecting electroacupuncture of rats detection leptin, adiponectin, and other related indicators, on treatment of electroacupuncture NAFLD.Objective:Inspection body adiponectin levels and hypothalamus leptin (Lepton, Lep), its receptor gene expression (mRNA), on the treatment of non-alcoholic fatty liver disease (nonalcoholic fatty liver disease, NAFLD) of the mechanism of action.MethodThe distant Sprague Dawley rats (Group, SD) 32 random to model group 22 only and empty group 10 only. The normal feed blank group, high fat diet fed model preparation SD rat model group. 8 week will randomly divided into rat model group: a control group (11 only), acupuncture Group (11 only). Treatment of acupuncture group, the "comments", "in", "vaginal mausoleum Springs", "Taichong", "over", and other points to acupuncture and frequency 10 Hz, strength of 1. 5 v CW; treatment of 4 weeks, each group of serum triglyceride (TG), Trglyceride, serum cholesterol (serum cholesterol, TC), serum Lep, adiponectin, observation histopathological changes in the liver, and RT-transfer amine by chain polymerization (Reverse Transcription-Polymerase Chain Reaction, RT-PCR) method for detecting hypothalamus Lep and its receptor mRNA expression.Results1 outward appearance:Each group of rats in the experiment does drinking water during feeding, drinking water, food consumption without obvious exception. The control group of rats fur glossy, posture and lively; model group rats obviously increase, body fat, skin discoloration, apparently not. An empty set of rat liver Visual inspection is bright red, edge sharpness, smooth surface. Model group see rat liver enlargement with the naked eye, edge blunt, washed Khaki, slice is greasy. Join the mouse after making photos2 HE dyeing show: Model group rats: hepatocytes cytoplasmic high levels of fat vacuoles are clear, the nucleus is fat vacuoles crush into liver cell margin, pathology confirmed fatty liver has formed. Electroacupuncture group treatment, liver fatty tissue denaturation far, liver cell structure also tend to be normal.3 Leptin、adiponectin indexThe treatment group and model group compared liver tissue homogenate leptin levels markedly. Liver tissue homogenate adiponectin levels have significantly lower serum TG TC, a marked decline.4 RT-PCRRT-PCR Detection shows: model group rats Lep and its receptor mRNA are noticeably, electro-acupuncture group hypothalamic Lep and leptin receptors increases. Lep optical density Analyzer on NAFLD hypothalamic, its receptor mRNA expression comparison table 2).ConclusionAfter eight weeks of fat rat feeding NAFLD have been making success. Experimental electro-acupuncture can influence hypothalamic Lep, its receptor mRNA expression, mitigation fat metabolism disorders, reached therapeutic effect on NAFLD, speculated that the treatment of non-alcoholic fatty liver disease mechanisms through "effect" effect to neuro-endocrine regulation system, adjustment of the Lep, its receptor mRNA expression and adiponectin levels.更多还原