Fenofibrate modifies human vascular smooth muscle proteoglycans and reduces lipoprotein binding

Aims/hypothesis Vascular disease in type 2 diabetes is associated with an up-regulation of atherogenic growth factors, which stimulate matrix synthesis including proteoglycans. We have examined the direct actions of fenofibrate on human vascular smooth muscle cells (VSMCs) and have specifically investigated proteoglycan synthesis and binding to LDL.Methods Proteoglycans synthesised by human VSMCs treated with fenofibrate (30 µmol/l) were assessed for binding to human LDL using a gel mobility shift assay, metabolically labelled with [³⁵S]-sulphate and quantitated by cetylpyridinium chloride. They were then assessed for electrophoretic mobility by SDS-PAGE, for size by gel filtration, for sulphation pattern by fluorophore-assisted carbohydrate electrophoresis, and for glycosaminoglycan (GAG) composition by enzyme digestion.Results Proteoglycans synthesised in the presence of fenofibrate showed an increase in the half-maximum saturation concentration of LDL from 36.8±12.4 µg/ml to 77.7±17 µg/ml under basal conditions, from 24.9±4.6 µg/ml to 39.1±6.1 µg/ml in the presence of TGF-1, and from 9.5±4.4 µg/ml to 31.1±3.4 µg/ml in the presence of platelet-derived growth factor/insulin. Fenofibrate treatment in the presence of TGF-1 inhibited the incorporation of [³⁵S]-sulphate into secreted and cell-associated proteoglycans synthesised by human VSMCs by 59.2% (p<0.01) and 39.8% (p<0.01) respectively. The changes in sulphate incorporation following treatment with fenofibrate were associated with a concentration-related increase in the electrophoretic mobility due to a reduction in GAG length. There was no change in the sulphation pattern; however, there was an alteration in the disaccharide composition of the GAGs.Conclusions/interpretation Fenofibrate modifies the structure of vascular proteoglycans by reducing the length of the GAG chains and GAG composition, resulting in reduced binding to human LDL, a mechanism which may lead to a reduction of atherosclerosis and cardiovascular disease in people with diabetes treated with fenofibrate.     

过氧化物酶体增殖物激活型受体α配体对压力超负荷大鼠心肌细胞凋亡变化的调控

目的探讨过氧化物酶体增殖物激活型受体α(PPARα)配体非诺贝特对压力超负荷致大鼠左心室肥厚过程中动态心肌细胞凋亡的影响。方法雄性Wistar大鼠腹主动脉缩窄致压力超负荷模型,术后48h存活的40只随机分成(1)手术组(CAA组),假手术组(SH组),(2)非诺贝特组(F组)30mg/kg.d,每组又按术后4周、8周两个时相,随机分为4周和8周组2个亚组,每组10只;(3)另取10只Wistar雄性大鼠,只穿线不节扎以作对照。给药干预4周、8周后检测血流动力学参数、心室重塑指标;采用脱氧核苷酸末端转移酶介导的缺口末端原位标记(TUNEL)法,检测心肌细胞凋亡指数(CAI);用West-ern-blot法观察PPARα蛋白的表达变化。结果与假手术组相比,非诺贝特处理4周时对血流动力学、心肌重塑指标及心肌细胞凋亡无明显影响,但8周时显著上调了PPARα蛋白表达,减轻了压力超负荷诱导的心肌肥厚,改善了血流动力学指标,抑制了心肌肥厚过程中的心肌细胞凋亡。结论PPARα配体长期(8周)能减轻压力超负荷大鼠的心肌肥厚,抑制心肌细胞凋亡,对心力衰竭进程中的心肌重塑有改善作用。     

非诺贝特联合双环醇治疗非酒精性脂肪性肝病患者肝组织PPARγ2 mRNA水平变化

目的 探讨应用非诺贝特联合双环醇治疗非酒精性脂肪性肝病(NAFLD)患者的疗效及肝组织过氧化物酶体增殖物激活受体γ₂(PPARγ₂)水平变化。方法 2018年2月～2019年12月我院收治的NAFLD患者66例,采用随机数字表法分为对照组33例和观察组33例,分别给予双环醇片口服或在此基础上给予非诺贝特片口服治疗,两组均治疗3个月。肝穿刺获得肝组织,采用PCR法检测肝组织PPARγ₂ mRNA水平,采用化学发光免疫分析法检测血清肿瘤坏死因子-α(TNF-α)和白介素-6(IL-6)水平,采用免疫比浊法检测血清超敏C反应蛋白(hs-CRP)水平,使用FC500型流式细胞仪检测外周血CD3⁺、CD4⁺和CD8⁺细胞百分比。结果 在治疗结束时,观察组总有效率为97.0%,显著高于对照组的66.7%(P<0.05);观察组肝组织PPARγ₂ mRNA相对水平为(1.5±0.1),显著高于对照组【(1.1±0.2),P<0.05】;观察组血清TNF-α水平为(26.4±3.1)pg/mL,显著低于对照组【(32.8±6.9)pg/mL,P<0.05】,血清IL-6水平为(35.9±4.9)ng/L,显著低于对照组【(46.4±5.8)ng/L,P<0.05】,血清hs-CRP水平为(5.1±1.4)mg/L,显著低于对照组【(8.4±1.9)mg/L,P<0.05】;观察组外周血CD3⁺细胞百分比为(80.2±11.5)%,显著高于对照组【(60.5±7.7)%,P<0.05】,外周血CD4⁺细胞百分比为(44.7±5.8)%,显著高于对照组【(31.4±4.8)%,P<0.05】,CD4⁺/ CD8⁺细胞比值为(1.5±0.4),显著高于对照组【(1.1±0.3),P<0.05】。结论 联合应用非诺贝特和双环醇治疗NAFLD患者有短期疗效,可能与提高了肝组织PPARγ₂水平,降低了血清炎症细胞因子水平有关,值得进一步研究。     

Effect of multifactorial treatment on non-alcoholic fatty liver disease in metabolic syndrome: a randomised study

ABSTRACT

Background: Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of the metabolic syndrome (MetS). There is no established treatment for NAFLD.

Aim: To evaluate a multifactorial intervention in the treatment of NAFLD.

Methods: A prospective, open-label, randomised study in non-diabetic patients (n = 186) with MetS (follow-up: 54 weeks). All patients had both biochemical and ultrasonographic evidence of NAFLD at baseline. Other causes of liver disease were excluded. Patients received lifestyle advice and treatment for hypertension (mainly inhibitors of the renin–angiotensin system), impaired fasting glucose (metformin), obesity (orlistat) and dyslipidaemia [randomly allocated to atorvastatin 20?mg/day (n = 63) or micronised fenofibrate 200?mg/day (n = 62) or both drugs (n = 61)]. Liver ultrasonography was assessed at baseline and at the end of the study.

Results: At the end of treatment, 67% of patients on atorvastatin, 42% on fenofibrate and 70% on combination treatment no longer had biochemical plus ultrasonographic evidence of NAFLD (?p < 0.05 vs. baseline for all comparisons). The percentage of patients who no longer had evidence of NAFLD was significantly higher (?p < 0.009) in the atorvastatin and combination groups compared with the fenofibrate group. This effect was independently related to drug treatment, as well as to reductions in high-sensitivity C-reactive protein, waist circumference, body weight, triglycerides, low-density lipoprotein-cholesterol, total cholesterol, systolic blood pressure and glucose. Four patients discontinued treatment because of adverse effects.

Conclusions: Multifactorial intervention in MetS patients with both biochemical and ultrasonographic evidence of NAFLD offsets surrogate markers of NAFLD (i.e. elevated aminotransferase plus echogenic liver).     

氟伐他汀联合非诺贝特治疗混合性高脂血症的可行性探讨

目的观察氟伐他汀联合非诺贝特治疗混合性高脂血症的临床效果。方法选择2007年1月至2008年12月南通市老年康复医院心内科门诊混合性高脂血症患者180例,随机分为氟伐他汀组、非诺贝特组、联合用药组各60例;氟伐他汀组每晚口服40mg氟伐他汀、非诺贝特组每晚口服非诺贝特200mg、联合用药每日清晨口服非诺贝特200mg,晚口服氟伐他汀20mg,均用至12周。观察血脂参数变化、疗效评价、药物主要不良反应。结果三组TC、LDL-C、TG治疗前无差异(P>0.05),氟伐他汀组治疗后TC、LDL-C均有显著性下降(P<0.05),TG变化不明显(P>0.05);非诺贝特组治疗后TG有显著性下降(P<0.05),TC、LDL-C变化不明显(P>0.05);联合用药组TC、LDL-C、TG均有显著性下降(P<0.05)。联合用药组临床控制(显效、好转)83.3%(50/60)高于氟伐他汀组的68.3%(41/60)和非诺贝特组的65.0%。三组患者血清CK、肝肾功能等参数均无明显变化,未出现肌病症状,无1例退出或终止。结论氟伐他汀联合非诺贝特对混合性高脂血症具有良好的安全性与耐受性,具有临床应用可行性。     

非诺贝特抑制血管紧张素Ⅱ所诱导的心脏成纤维细胞骨桥蛋白表达升高

目的：探讨过氧化物酶体增殖物激活受体α（PPARα）激动剂非诺贝特对血管紧张素Ⅱ（An-gⅡ）诱导的心脏成纤维细胞骨桥蛋白表达升高的影响。方法：分离并传代培养SD大鼠乳鼠心脏成纤维细胞,使用不同浓度（0、25、50、100μmol/L）的非诺贝特预处理1h后,加入An-gⅡ作用24h。分别采用实时定量PCR法和Western blotting技术检测骨桥蛋白mRNA和蛋白表达情况。结果：非诺贝特显著抑制AngII诱导的心脏成纤维细胞骨桥蛋白mRNA及蛋白的表达,并且呈剂量依赖性。结论：非诺贝特对心脏成纤维细胞中骨桥蛋白的表达具有明显的抑制作用,这可能是其在心血管系统发挥抗纤维化和抗炎作用的部分机制。     

高龄老年混合型高脂血症122例的治疗

目的:观察联合应用阿托伐他汀和非诺贝特治疗75岁以上混合型高脂血症的临床疗效及安全性。方法:混合型高血脂症患者122例,随机分为3组,即阿托伐他汀组(10 mg/d,n=40)、每日联合治疗组(阿托伐他汀10 mg/d+非诺贝特200 mg/d,n=40)、隔日联合治疗组(阿托伐他汀10 mg+非诺贝特200 mg隔日交替服用,n=42),治疗12周。观察治疗前、后主要血脂水平的变化率及不良反应。结果:联合用药组血脂参数变化显著,降低甘油三酯和升高高密度脂蛋白胆固醇的能力明显优于单药组(P<0.05);隔日联合治疗组不良反应发生率(7.1%)低于每日联合治疗组(17.5%)(P<0.05)。结论:隔日联合应用阿托伐他汀和非诺贝特治疗75岁以上混合型高脂血症较单药更有效,具有良好的安全性和耐受性。     

Fenofibrate, a peroxisome proliferator–activated receptor-α agonist, exerts anticonvulsive properties

The underlying mechanisms of the ketogenic diet (KD) remain unknown. Involvement of peroxisome proliferator–activated receptor-α (PPARα) has been suggested. The aim of this study was to assess the anticonvulsant properties of fenofibrate, a PPARα agonist. Wistar rats were fed at libitum during 14 days by regular diet, KD, regular diet containing 0.2% fenofibrate (F), or KD containing 0.2% fenofibrate (KD + F). Pentylenetetrazol (PTZ) threshold and latencies to the onset of status epilepticus induced by lithium–pilocarpine were used to assess diet treatments with anticonvulsive effects. Myoclonic and generalized seizure PTZ thresholds were increased in F- and KD-treated animals in comparison to control. No difference was observed between KD + F group and the others groups (control, F, KD). Latencies to the onset of status epilepticus were increased in F and KD groups compared to control. Fenofibrate exerts anticonvulsive properties comparable to KD in adult rats using PTZ and lithium–pilocarpine models. The underlying mechanisms such as PPARα activation and others should be investigated. These findings may provide insights into future directions to simplify KD protocols.     

Effect of fenofibrate on LDL-induced endothelial dysfunction in rats

Previous investigations have demonstrated that asymmetric dimethylarginine (ADMA) is an important factor contributing to endothelial dysfunction, and that fenofibrate has a protective effect on the endothelium in hyperlipidaemic patients. In the present study in rats treated with native low-density lipoprotein (nLDL), we addressed the question of whether the beneficial effect of fenofibrate on endothelial cells is related to reduction of the ADMA concentration. A single injection of nLDL (4 mg/kg, 48 h) markedly reduced endothelium-dependent relaxation in response to acetylcholine and the plasma level of nitrite/nitrate and increased the plasma concentrations of ADMA, malonyldialdehyde (MDA) and tumour necrosis factor- (TNF-). Treatment with fenofibrate (30 or 100 mg/kg) significantly reduced the inhibition of vasodilator responses to acetylcholine, decreased the elevated levels of ADMA, MDA and TNF-, and enhanced the decreased level of nitrite/nitrate in the rats treated with LDL. These results suggest that the protective effect of fenofibrate on endothelial cells in rats treated with LDL may be related to the reduction of ADMA concentration.