Reduction of AMPK activity and altered MAPKs signalling in peripheral blood mononuclear cells in response to acute glucose ingestion following a short-term high fat diet in young healthy men

Background

Peripheral blood mononuclear cells (PBMCs) are known to respond to systematic changes in nutrient availability. The impact of a short-term high fat diet (HFD), with and without acute glucose ingestion, on the energy-sensing enzyme 5’ AMP-activated protein kinase (AMPK) as well as mitochondrial oxidative phosphorylation (OXPHOS) proteins in PBMCs is currently unknown.

Methods

Nine healthy, lean young males participated in a 7 day HFD intervention, designed to induce transient glucose intolerance. The phosphorylation status and total protein content of AMPK and inflammatory mitogen-activated protein kinases (MAPKs), and total OXPHOS protein in PBMCs, along with circulating cytokines, were assessed in the fasted state and following an oral glucose tolerance test (OGTT) before and after the HFD.

Results

One week of HFD resulted in relative glucose intolerance. The HFD resulted in a reduction of AMPK phosphorylation under fasting basal conditions and following the OGTT (both P < 0.05), while there were no differences in OXPHOS protein expression. Although the short-term HFD had no effect on basal phosphorylation of p38, JNK or ERK1/2, the activation of MAPKs signalling in response to glucose ingestion was attenuated post-HFD as compared to pre-HFD (P < 0.05 for all). Circulating cytokines were not significantly affected by the HFD.

Conclusions

We conclude that impaired glucose tolerance in response to 7 day HFD resulted in decreased AMPK activity and impaired glucose-stimulated MAPK activation following glucose ingestion in vivo in PBMCs from young, lean subjects. Further studies are warranted to explore how dietary manipulations impact interplay between AMPK and inflammatory signalling, along with immune function, in PBMCs.     

西罗莫司对高脂饮食诱导的NAFLD大鼠外周血Treg/Th17细胞的影响

目的 研究西罗莫司对高脂饮食诱导的非酒精性脂肪性肝病(NAFLD)大鼠外周血Treg/Th17细胞的影响。方法 随机将60只SD大鼠分为西罗莫司干预组、模型组和对照组,每组20只。采用高脂饮食喂养12周建立NAFLD模型,在造模成功后,在其中20只动物,给予西罗莫司干预2周。采用ELISA法检测血清白介素-10(IL-10)、IL-17和肿瘤坏死因子-α(TNF-α)水平,使用全自动生化分析仪检测血生化指标,使用FACSVia型流式细胞仪检测外周血Treg和Th17细胞百分比。结果 干预后,西罗莫司干预组血清AST和ALT水平分别为(134.9±15.3)U/L和(109.4±18.2)U/L,显著低于模型组【(151.1±28.9)U/L和(128.9±17.7)U/L,P<0.05】;血清TG、TC和LDL-C水平分别为(0.9±0.3)mmol/L、(2.6±0.8)mmol/L和(3.3±1.2)mmol/L,显著低于模型组【(1.1±0.5)mmol/L、(3.6±1.3)mmol/L和(5.4±1.8)mmol/L,P＜0.05】,而血清HDL-C水平为(1.3±0.5)mmol/L,显著高于模型组【(0.7±0.4) mmol/L,P＜0.05】;血清IL-10水平为(68.9±26.6)pg/ml,显著高于模型组【(37.8±17.2)pg/ml,P＜0.05],而血清IL-17和TNF-α水平分别为(13.2±3.3)pg/ml和(0.7±0.2)ng/ml,显著低于模型组【(22.6±4.0)pg/ml和(1.2±0.4)ng/ml,P＜0.05】;西罗莫司干预组外周血Treg细胞百分比为(4.5±0.7)%,Th17细胞百分比为(1.7±0.6)%,Treg/Th17比值为(2.6±0.7),与模型组比,差异显著【(3.8±0.9)%、(2.8±1.1)%和(1.5±0.7),P＜0.05]。结论 应用西罗莫司干预高脂饮食诱导的NAFLD大鼠能减轻炎症反应,纠正外周血Treg/Th17细胞比值失衡状态,可能有助于阻止NAFLD进展,值得进一步研究。     

Melatonin ameliorates high fat diet-induced diabetes and stimulates glycogen synthesis via a PKCζ-Akt-GSK3β pathway in hepatic cells

Abstract:  Low levels of melatonin in circulation had been reported to be related to the development of diabetes. Melatonin administration in animals increases hepatic glycogen content to lower blood glucose. However, the signaling pathway for these effects is still unclear. The present study shows that intraperitoneal injection of 10 mg/kg melatonin ameliorated glucose utilization and insulin sensitivity in high fat diet-induced diabetic mice with an increase in hepatic glycogen and improvement in liver steatosis. We used HepG2 cells to investigate the signaling pathways for the melatonin-stimulated hepatic glycogen increment. Treatment of HepG2 cells with 1 n m melatonin markedly increased glycogen synthesis which was blocked by the melatonin receptor antagonist luzindole. In addition, melatonin increased the phosphorylation of subcellular signals at the level of protein kinase C ζ (PKCζ), Akt, and glycogen synthase kinase 3β (GSK3β) while the increase in glycogen synthesis induced by melatonin was inhibited by PKCζ pseudo-peptide. However, 3',5'-cyclic adenosine monophosphate-activated protein kinase (AMPK) was not influenced by melatonin treatment. Taken together, melatonin improves glucose intolerance and insulin resistance in high fat diet-induced diabetic mice and stimulates glycogen synthesis via a PKCζ-Akt-GSK3β pathway in HepG2 cells.     

Effect of high protein and fat diet on postprandial blood glucose levels in children and adolescents with type 1 diabetes in Cairo,Egypt

Background and aimsTo determine the effect of high protein and high fat meals on post prandial glycemia in patients with type 1 diabetes.MethodsThis study included 51 children and adolescents with type 1 diabetes who were following up at Diabetes, Endocrine and Metabolism Pediatric Unit (DEMPU), Abo Elrish Children’s hospital, Cairo University.Post prandial blood glucose levels were recorded and compared following three breakfast meals with varying protein and fat content (standard carbohydrate meal, high fat meal, and high protein meal) over a period of 5 hours on 3 consecutive days.ResultsHigh protein meal resulted in hyperglycemia with the peak level at 3.5 hours and continued for 5 hours post prandial while high fat meal caused early hyperglycemia reached the peak at 2 hours then declined towards 5 hours.Comparison of the three different breakfast meals revealed statistically significant difference regarding the postprandial glycemia at 30, 60, 90,120, 180, 210, 240, 270, 300 min.ConclusionMeals high in protein caused sustained increase in postprandial glucose levels over a period of 5 h. However, high fat meals caused early postprandial hyperglycemia. Protein and fat content of meals affect the timing and values of the peak blood glucose as well as the duration of postprandial hyperglycemia. Therefore, fat/protein unit should be taken in consideration while calculating the bolus insulin dose and anticipating the postprandial glucose response.     

Crucial role of insulin receptor substrate-2 in compensatory beta-cell hyperplasia in response to high fat diet-induced insulin resistance

In type 2 diabetes, there is a defect in the regulation of functional beta-cell mass to overcome high-fat (HF) diet-induced insulin resistance. Many signals and pathways have been implicated in beta-cell function, proliferation and apoptosis. The co-ordinated regulation of functional beta-cell mass by insulin signalling and glucose metabolism under HF diet-induced insulin-resistant conditions is discussed in this article. Insulin receptor substrate (IRS)-2 is one of the two major substrates for the insulin signalling. Interestingly, IRS-2 is involved in the regulation of beta-cell proliferation, as has been demonstrated using knockout mice models. On the other hand, in an animal model for human type 2 diabetes with impaired insulin secretion because of insufficiency of glucose metabolism, decreased beta-cell proliferation was observed in mice with beta-cell-specific glucokinase haploinsufficiency (Gck(+/) (-)) fed a HF diet without upregulation of IRS-2 in beta-cells, which was reversed by overexpression of IRS-2 in beta-cells. As to the mechanism underlying the upregulation of IRS-2 in beta-cells, glucose metabolism plays an important role independently of insulin, and phosphorylation of cAMP response element-binding protein triggered by calcium-dependent signalling is the critical pathway. Downstream from insulin signalling via IRS-2 in beta-cells, a reduction in FoxO1 nuclear exclusion contributes to the insufficient proliferative response of beta-cells to insulin resistance. These findings suggest that IRS-2 is critical for beta-cell hyperplasia in response to HF diet-induced insulin resistance.     

Adiponectin stimulates Rho-mediated actin cytoskeleton remodeling and glucose uptake via APPL1 in primary cardiomyocytes

Objective

Adiponectin is known to confer its cardioprotective effects in obesity and type 2 diabetes, mainly by regulating glucose and fatty acid metabolism in cardiomyocytes. Dynamic actin cytoskeleton remodeling is involved in regulation of multiple biological functions, including glucose uptake. Here we investigated in neonatal cardiomyocytes whether adiponectin induced actin cytoskeleton remodeling and if this played a role in adiponectin-stimulated glucose uptake.

Materials/methods

Primary cardiomyocytes were treated with full-length and globular adiponectin (fAd and gAd, respectively).

Results

Both fAd and gAd increased RhoA activity, phosphorylation of the Rho/ROCK signaling target cofilin and actin polymerization to form filamentous actin as determined by rhodamine–phallodin immunofluorescence and quantitative analysis of filamentous to globular actin ratio. Scanning electron microscopy also demonstrated structural remodeling. Adiponectin stimulated glucose uptake, was significantly abrogated in the presence of inhibitors of actin cytoskeleton remodeling (cytochalasin D) and Rho/ROCK signaling (C3 transferase, Y27632). We showed that adiponectin increased colocalization of actin and APPL1 and that actin remodeling, phosphorylation of AMPK, p38MAPK and cofilin, glucose uptake and oxidation were all attenuated after siRNA-mediated knockdown of APPL1.

Conclusion

We show that adiponectin mediates Rho/ROCK-dependent actin cytoskeleton remodeling to increase glucose uptake and metabolism via APPL1 signaling.     

高迁移率族蛋白1在乙型重型肝炎患者中的表达水平及其临床意义

目的探讨血清高迁移率族蛋白1(high mobility group-box protein 1,HMGB1)在重型肝炎患者中的表达水平及其临床意义。方法对32例乙型重型肝炎患者、22例慢乙肝初治患者、10例急性乙肝患者血清HMGB1水平进行检测分析,与16名健康人进行对照研究,分析其与患者肝功能生物化学指标的相关性。结果重型肝炎患者血清HMGB1水平高于健康人,差异具有统计学意义(P0.05),重型肝炎组与慢乙肝初治组、重型肝炎组与急性乙肝组、慢乙肝初治组与急性乙肝组、慢性乙肝组与正常人组、急性乙肝与正常人组均无统计学意义(P0.05),重型肝炎患者血清HMGB1水平与ALT水平呈正相关,差异具有统计学意义(r=0.942,P0.05)。结论重型肝炎患者血清HMGB1水平较正常人高,可能成为评价重型肝炎的一个指标,这可能具有一定的临床意义。     

Biology of incretins: GLP-1 and GIP

This review focuses on the mechanisms regulating the synthesis, secretion, biological actions, and therapeutic relevance of the incretin peptides glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1). The published literature was reviewed, with emphasis on recent advances in our understanding of the biology of GIP and GLP-1. GIP and GLP-1 are both secreted within minutes of nutrient ingestion and facilitate the rapid disposal of ingested nutrients. Both peptides share common actions on islet beta-cells acting through structurally distinct yet related receptors. Incretin-receptor activation leads to glucose-dependent insulin secretion, induction of beta-cell proliferation, and enhanced resistance to apoptosis. GIP also promotes energy storage via direct actions on adipose tissue, and enhances bone formation via stimulation of osteoblast proliferation and inhibition of apoptosis. In contrast, GLP-1 exerts glucoregulatory actions via slowing of gastric emptying and glucose-dependent inhibition of glucagon secretion. GLP-1 also promotes satiety and sustained GLP-1-receptor activation is associated with weight loss in both preclinical and clinical studies. The rapid degradation of both GIP and GLP-1 by the enzyme dipeptidyl peptidase-4 has led to the development of degradation-resistant GLP-1-receptor agonists and dipeptidyl peptidase-4 inhibitors for the treatment of type 2 diabetes. These agents decrease hemoglobin A1c (HbA1c) safely without weight gain in subjects with type 2 diabetes. GLP-1 and GIP integrate nutrient-derived signals to control food intake, energy absorption, and assimilation. Recently approved therapeutic agents based on potentiation of incretin action provide new physiologically based approaches for the treatment of type 2 diabetes.     

胰腺癌HMGB1表达及其与血行转移的关系研究

目的 探讨人胰腺癌高迁移率族蛋白B1(hiish mobility group protein B1,HMGB1)表达及其与血行转移的关系.方法 应用Western blot法检测68例胰腺癌患者、18例CP和21例健康者血清HMGB1水平,并对其中37例胰腺癌患者手术前后的血清HMGB1水平进行比较;应用免疫组织化学法检测67例胰腺癌组织HMGB1和CD31的表达.结果 胰腺癌、CP及健康者血清HMGB1水平分别为(119.7±54.5)ng/ml、(40.2±25.5)ng/ml和(13.1±4.3)ng/ml,相差非常显著(P<0.001).胰腺癌患者术后血清HMGB1水平为(69.3±5.1)ng/ml,显著低于术前的(120.2±8.2)ng/ml(P<0.001).胰腺癌组织HMGB1表达阳性率为43.6%,HMGB1表达与组织分化、TNM分期及转移有关,P均<0.01;HMGB1表达与血管密度呈显著正相关(r=0.76,P<0.0001),免疫组化显示,HMGB1表达阳性的肿瘤细胞多位于有腔血管周围,位于血管内的肿瘤细胞HMGB1阳性表达率为71%.结论 胰腺癌患者HMGB1呈高表达,表达HMGB1的肿瘤细胞易于进入血管内,与其血行转移有关.     

乙型肝炎患者血清高迁移率族蛋白-1含量的检测及临床意义

目的观察不同病情的乙型肝炎患者血清中晚期炎症介质高迁移率族蛋白-1（HMGB1）的含量，通过与肝炎病情评价指标TBil和PTA进行相关性比对，初步阐明晚期炎症介质HMGB1在慢性重型乙型肝炎患者肝衰竭发生中可能发挥的作用。方法收集慢性重型乙型肝炎患者23例，CHB患者31例，健康对照者10名，抽取外周血10ml，按常规方法检测肝炎病情指标TBil和PTA。超滤管去除血清中的大分子蛋白质，再浓缩血清小分子蛋白，然后用Western blot的方法检测样品中HMGB1。利用纯化出的GST-HMGB1融合蛋白进行相对定量，计算出不同患者血清中HMGB1的水平，最后与肝炎病情评价指标TBil和PTA进行相关性比对。结果23例慢性重型乙型肝炎患者血清中HMGB1检出率为100％（23／23），平均含量为（83．4±21．3）μg／L；10例重度CHB患者血清中HMGB1检出率为90％（9／10），平均含量为（78．1±19．5）μg／L；11例中度CHB患者的血清中HMGB1检出率为55％（6／11），平均含量为（60．6±14．3）μg／L；10例轻度CHB患者及12例恢复患者中均仅1例检出HMGB1，检出率分别为10％和8％，含量分别为28．9μg／L和39．7μg／L；健康对照者血清中未能检出HMGB1。血清中HMGB1的含量与TBil呈正相关，与PTA呈负相关。结论慢性重型乙型肝炎患者血清中HMGB1的含量与疾病的严重程度密切相关，晚期炎症介质HMGB1在慢性重型乙型肝炎患者肝衰竭的发生中可能起重要的作用。     

JTT-130, a novel intestine-specific inhibitor of microsomal triglyceride transfer protein, suppresses high fat diet-induced obesity and glucose intolerance in Sprague-Dawley rats

Aim: Microsomal triglyceride transfer protein (MTP) takes part in the mobilization and secretion of triglyceride‐rich lipoproteins from enterocytes and hepatocytes. We investigated the effects of JTT‐130, a novel intestine‐specific MTP inhibitor, on high fat diet‐induced obesity and glucose intolerance. Methods: Male Sprague‐Dawley rats were fed a 3.1% fat diet or a 35% fat diet with or without JTT‐130 as a food admixture (0.029%). Food intake, body weight, abdominal fat, hepatic triglyceride, faecal free fatty acids and plasma levels of glucagon‐like peptide‐1 (GLP‐1) and peptide YY (PYY) were assessed. Plasma levels of glucose and insulin were measured during intraperitoneal glucose tolerance tests. In addition, indirect calorimetry was performed on rats fed with a 35% fat diet. Results: JTT‐130 treatment decreased body weights, abdominal fat and hepatic triglyceride with suppression of food intake and elevation of faecal free fatty acids and plasma GLP‐1 and PYY levels in rats fed with the 35% fat diet, whereas no significant effects on these parameters except for increased faecal free fatty acids were observed in rats fed with the 3.1% fat diet. JTT‐130 treatment decreased plasma levels of glucose and insulin during intraperitoneal glucose tolerance tests on rats fed with the 35% fat diet, but not on rats fed with the 3.1% fat diet. JTT‐130‐treated rats showed increased O₂ consumption and CO₂ production on a 35% fat diet. Conclusions: JTT‐130 suppresses high fat diet‐induced obesity and glucose intolerance with suppression of food intake and fat absorption and could be useful for prevention and treatment of obesity and obesity‐related insulin resistance.     

蛋白酪氨酸磷酸酶1B基因多态性与2型糖尿病和肥胖的相关性研究

目的 研究中国人群中蛋白酪氨酸磷酸酶1B（PTP-1B）基因的单核苷酸多态性（SNP）与2型糖尿病及肥胖的相关性。方法 采用直接测序法对PTP—1B基因作SNP筛查，并在夫妻配对样本中对所检出的SNP作基因分型。结果 共检出6个SNPs位点，其中内含子区3个（15／37C→A，16／82A→G，17／301C→T），外显子区3个（E8／45C→T，E9／35G→A，E10／372G→A），其中E9／35G→A为新发现的突变类型；在病例-配偶对照研究中发现，15／37C→A，16／82A→G和17／301C→T等位基因频率在糖尿病患者和正常人配偶中差异有统计学意义（均P〈0．05），其余位点的等位基因频率在两组间的分布则无明显差异。与肥胖的相关性研究中发现15／37C→A和17／301C→T位点与男性的腰臀比（WHR）相关（P〈0．05）。结论 PTP-1B基因的SNP位点15／37C→A，16／82A→G和17／301C→T多态性可能和2型糖尿病的发病相关，其中15／37C→A和17／301C→T与男性的WHR相关。     

Phosphoinositide 3-kinase as a novel functional target for the regulation of the insulin signaling pathway by SIRT1

The protein deacetylase SIRT1, and its activator resveratrol, exert beneficial effects on glucose metabolism. Different SIRT1 targets have been identified, including PTP1B, AMPK, FOXO, PGC-1α and IRS2. The latter may underscore a tight link between SIRT1 and insulin signaling components. However, whether SIRT1 has a direct effect on insulin resistance and whether resveratrol acts directly or indirectly in this context is still a matter of controversy and this question has not been addressed in muscle cells. Here, we show that SIRT1 protein expression is decreased in muscle biopsies and primary myotubes derived from type 2 diabetic patients, suggesting a contribution of diminished SIRT1 in the determination of muscle insulin resistance. To investigate the functional impact of SIRT1 on the insulin pathway, the activation of insulin downstream effector PKB was evaluated after SIRT1 inactivation by RNAi, SIRT1 overexpression, or resveratrol treatments. In muscle cells and HEK293 cells, downregulation of SIRT1 reduced, while overexpression increased, insulin-induced PKB activatory phosphorylation. Further molecular characterisation revealed that SIRT1 interacts in an insulin-independent manner with the PI3K adapter subunit p85. We then investigated whether resveratrol may improve insulin signaling in muscle cells via SIRT1, or alternative targets. Incubation of muscle cells with resveratrol reverted the insulin-resistant state induced by prolonged TNFα or insulin treatment. Resveratrol-dependent improvement of insulin-resistance occurred through inhibition of serine phosphorylation of IRS1/2, implicating resveratrol as a serine kinase inhibitor. Finally, a functional interaction between PI3K and SIRT1 was demonstrated in C. elegans, where constitutively active PI3K - mimicking increased IIS signaling - lead to shortened lifespan, while removal of sir-2.1 abolished PI3K-induced lifespan shortening. Our data identify SIRT1 as a positive modulator of insulin signaling in muscle cells through PI3K, and this mechanism appears to be conserved from C. elegans through humans.     

Molecular determinants of angiotensin II type 1 receptor functional selectivity

The angiotensin AT₁ receptor is an important pharmacological target in the treatment of cardiovascular disorders, such as hypertension, diabetic nephropathy, cardiac hypertrophy, arrhythmia and failure. Simultaneously, the AT₁ receptor has emerged to be a prominent model for the emerging concept that receptors may attain multiple active states with differentiated functional outcomes. Two major signalling pathways are employed by the AT₁ receptor, namely 1) the canonical G_q protein-dependent activation of inositol phosphate turnover and intracellular calcium release, and 2) G protein-independent recruitment of β-arrestin-scaffolded signalling complexes that activate protein kinase pathways. Different states of receptor activation with preference for individual downstream pathways (functional selectivity) have been demonstrated in mutational studies of the AT₁ receptor and by pharmacological probing with analogues of angiotensin II. These studies also provide clues about the conformational changes that underlie different functional outcomes. In this review, we evaluate current knowledge of the molecular determinants of AT₁ receptor activation, which may distinguish G protein-dependent and -independent behaviour. While G protein activation is known to be detrimental, G protein-independent signalling by the AT₁ receptor has been associated with phenotypes such as cell survival and renewal, regulation of cardiac contraction and cell migration. It is therefore currently hypothesized that selective blockade of G protein actions and simultaneous activation of G protein-independent signalling will prove to be a feasible strategy for improved cardiovascular therapy. The pharmacological perspectives of functional selectivity by receptors, such as the AT₁ receptor, urge the elucidation of molecular mechanisms that govern disparate signalling events.     

HDL和阿托伐他汀对oxLDL刺激下脂肪细胞炎症反应的影响

目的 观察高密度脂蛋白（HDL）及阿托伐他汀对氧化型低密度脂蛋白（oxLDL）刺激下3T3-L1脂肪细胞肿瘤坏死因子-α（TNFα）分泌及mRNA表达的影响,并探讨其可能的作用机制.方法 3T3-L1脂肪细胞促分化成熟后,oxLDL刺激脂肪细胞,给予不同浓度的HDL（10～100 μg/mL）和阿托伐他汀（0.1～10 μM）,及H-89（10 μM）＋HDL（100 μg/mL）干预,收集细胞,测定脂肪细胞TNFα水平、TNFα mRNA表达水平、核因子-κB（NF-κB）活性及NF-κB抑制单位（IκB）蛋白浓度.结果 oxLDL刺激使3T3-L1脂肪细胞TNFα分泌、mRNA表达水平及NF-κB活性明显增强.阿托伐他汀浓度依赖性降低TNFα 分泌及mRNA表达,抑制NF-κB活化.10 μM阿托伐他汀使oxLDL诱导的脂肪细胞TNFα mRNA表达降低56.5%,NF-κB活性减少41.2%.HDL也呈浓度依赖性抑制TNFα分泌及mRNA表达,降低NF-κB活性,减少IκB降解.与oxLDL刺激组比较,100 μg/ml HDL使TNFα mRNA表达降低64.5%,NF-κB活性减少49%,并明显增加IκB蛋白水平.HDL的这些抗炎效应能被蛋白激酶A（PKA）抑制剂（应放在H89第一次出现之处）H-89部分抑制.结论 HDL能抑制oxLDL诱导的3T3-L1脂肪细胞TNFα分泌和mRNA表达,PKA-IκB-NF-κB信号通路可能是其中作用途径之一,该效应不需要HDL与oxLDL的直接接触作用.阿托伐他汀亦通过NF-κB途径抑制oxLDL诱导的3T3-L1脂肪细胞TNFα分泌和mRNA表达.HDL的抗炎作用强度与阿托伐他汀相似.     

丹栀逍遥散对非酒精性脂肪性肝病大鼠LKB1/AMPK/ACC通路及肝脂肪变性的影响

目的:探讨丹栀逍遥散对非酒精性脂肪性肝病(NAFLD)大鼠肝脏激酶B1(LKB1)/腺苷酸活化蛋白激酶(AMPK)/乙酰辅酶A羧化酶(ACC)通路及肝脂肪变性的影响。方法:SD雄性大鼠随机分为空白组、模型组、水飞蓟宾组(26.25 mg·kg^-1)、丹栀逍遥散高、中、低剂量组(38.0、19.0、9.5 g·kg^-1),每组13只。采用高脂饲料连续喂养8周复制非酒精性脂肪肝(NAFLD)大鼠模型。模型复制成功后水飞蓟宾组和丹栀逍遥散各剂量组大鼠开始灌胃给予相应的药物,连续给药4周,给药结束后进行取材和相关指标的检测。给药期间,观察各组大鼠的精神状况、毛色、饮食、活动等一般状态,记录给药前后大鼠体重变化;取大鼠肝、肾、脾脏并称重,计算脏器指数;生化法检测肝组织中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、甘油三脂(TG)、总胆固醇(TC)、游离脂肪酸(NEFA)含量;苏木精-伊红染色(HE)观察各组大鼠肝组织病理变化;Western blot检测肝组织中LKB1、AMPKα1、AMPKα2、ACC及其磷酸化水平。结果:与空白组相比,模型组大鼠毛色发黄暗淡,精神萎靡;体重增量、肝脏指数、肝组织中ALT、AST、TG、TC、NEFA水平均显著升高(P<0.05),肝细胞排列紊乱,胞界不清晰,较多的脂肪空泡;肝细胞中LKB1、p-LKB1、AMPKα1、p-AMPKα1、AMPKα2、p-AMPKα2、p-ACC蛋白表达水平明显降低,ACC蛋白表达升高(P<0.05);与模型组相比,水飞蓟宾组和丹栀逍遥散高、中剂量组大鼠一般状态有不同程度的改善,体重增量、肝脏指数、肝组织中ALT、AST、TG、TC、NEFA水平均显著降低(P<0.05),病理损伤程度减轻;肝细胞中LKB1、p-LKB1、AMPKα1、p-AMPKα1、AMPKα2、p-AMPKα2、p-ACC蛋白表达水平明显升高,ACC蛋白表达降低(P<0.05)。结论:丹栀逍遥散可能通过LKB1/AMPK/ACC信号通路,降低脂肪的合成,改善肝功能,减轻NAFLD模型大鼠肝脂肪变性。