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OBJECTIVE To investigate the effects of β-sitosterol on the function of rheumatoid arthritis （RA） fibroblastic synoviocytes MH7A cells and its mechanism. METHODS Network pharmacology was adopted to screen the targets of β-sitosterol and the targets for the treatment of RA. After the intersection of them， topological analysis was performed to find the most critical target in the treatment of RA. MH7A cells were treated with different concentrations （0， 5， 10， 20， 40 μmol/L） of β-sitosterol， and CCK-8 was used to assay cell viability for screening the optimal concentration of β-sitosterol. MH7A cells were induced by 10 ng/mL TNF-α in vitro and treated with β-sitosterol （the optimum concentration）. CCK-8 and EdU were used to detect the ability of cell proliferation. Scratch experiment and Transwell invasion assay were used to analyze cell migration and invasion. The levels of interleukin-1β （IL-1β） and IL-6 in cell supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expressions of peroxisome proliferator-activated receptor α （PPARα） were measured by qRT-PCR and Western blot， respectively. The siRNA targeting PPARα was transfected into MH7A cells， and the effects of β-sitosterol on cell proliferation， migration， invasion， the secretion of inflammatory factors and the expression of PPARα after PPARα knockdown were detected by the above experimental methods. RESULTS PPARα was the most critical target of β-sitosterol in the treatment of RA. The optimal concentration of β-sitosterol was 20 μmol/L. Compared with model group， β-sitosterol decreased the viability of MH7A cells， and the number of proliferating cells also decreased significantly （P＜0.05）； the cell migration rate and the number of cell invasion decreased significantly （P＜0.05）. The levels of IL-1β and IL-6 were also significantly decreased （P＜0.05）， and the mRNA 15 and protein expression levels of PPARα were significantly increased （P＜0.05）. Compared with negative control small interfering RNA group， after PPARα knockdown， the cell viability increased by about 35.6% （P＜0.05）， the number of cell proliferation， the cell migration rate and the number of cell invasion increased significantly （P＜0.05）， and the levels of IL-1β and IL-6 also increased significantly （P＜0.05）. CONCLUSIONS β-sitosterol could effectively inhibit the proliferation， migration， invasion and secretion of inflammatory factors in MH7A cells， the mechanism of which may be associated with activating PPARα pathway.
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Hyperlipidemia is a dyslipidemia caused by dyslipidemia of lipid metabolism， which can be divided into primary and secondary types. The current clinical diagnostic criteria are mainly changes in lipid levels， which are the inducers of high-risk cardiovascular diseases such as atherosclerosis， pancreatitis and coronary heart disease. As a key target in lipid metabolism， peroxisome proliferator-activated receptor α （PPARα） is involved in a variety of metabolic activities， including fatty acid degradation， synthesis， transport， storage， lipoprotein metabolism， etc. Activation of PPARα can maintain the balance of lipid metabolism through a variety of ways， which is an important way to treat hyperlipidemia. At present， chemical drugs such as statins and bettes are mainly used in the clinical treatment of hyperlipidemia. Although they can slow down the disease to a certain extent， there are many adverse reactions and drug resistance. By reviewing the literature in recent years， the author found that the activation of PPARα pathway by traditional Chinese medicine in the treatment of hyperlipidemia has significant effect and small adverse reactions. The lipid-lowering active ingredients include flavonoids， alkaloids， phenols， terpenoids and other compounds. These active components mainly affect the expression of downstream effectors through the activation of PPARα pathway， thereby inhibiting the synthesis of total cholesterol and promoting fatty acid oxidation， and play a role in the treatment of hyperlipidemia. In this paper， we systematically reviewed the structure types and mechanism of active components of traditional Chinese medicine that activate PPARα pathway， so as to provide guidance for the rational development and clinical application of lipid-lowering traditional Chinese medicine new drugs.
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Objective: Management of low-density lipoprotein cholesterol (LDL-C) and triglyceride (TG) is important for patients with type 2 diabetes merger hyperlipidemia. Pemafibrate (PF) has different characteristics from conventional fibrates. In this study, we retrospectively compared the efficacy and safety of PF and bezafibrate (BF) in patients with type 2 diabetes merger hypertriglyceridemia.Methods: Patients who were administered PF (0.2 mg/day) or BF (400 mg/day) for 24 weeks or longer were included. Twenty patients in each group were extracted using propensity score matching (PS). PS was calculated using the patient background (before the start of administration) of PF or BF. We investigated lipid-related parameters (TG, high density lipoprotein cholesterol [HDL-C], and LDL-C) and other laboratory test parameters pre administration and 24 weeks post administration.Results: TG decreased significantly in both groups (p<0.05). However, there were no significant differences between the two groups in the TG treatment target (<150 mg/dL) achievement rate (p =1.00), TG change rate (p=0.84), and TG change amount (p=0.77). In addition, there were no significant changes in HDL-C and LDL-C in both groups. In the PF group, alanine transaminase (ALT) (p< 0.05), alkaline phosphatase (p<0.05) decreased. In the BF group, ALT (p<0.05) and γ-GTP (p<0.05) decreased. Both groups showed improvement in liver function after 24 weeks. eGFR (p<0.05) significantly decreased only BF group. There were no significant changes in renal function, creatine kinase (CK), or hemoglobin A1c (HbA1c) in either group.Conclusion: Our study suggests that there is no difference in the TG lowering effect and safety of PF and BF in type 2 diabetic patients.
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OBJECTIVE： To study the effects of selenium-enriched Ganoderma lucidum crude extract on lipid metabolism， liver function and inflammatory response in type 2 diabetic model rats. METHODS： Totally 120 rats were randomly divided into normal control group （n＝20， normal saline） and model group （n＝100）. Normal control group was fed with normal diet， and model group was fed with high-fat diet. 4 weeks later， model group was given intraperitoneal injection of Streptozotocin solution （30 mg/kg） to induce T2DM model. After modeling， 90 rats were randomly subdivided into model control group （normal saline）， positive control group （metformin， 200 mg/kg） and selenium-enriched G. lucidum crude extract low-dose， medium-dose and high-dose groups （300， 600， 1 200 mg/kg， calculated by extract）， with 18 rats in each group. They were given medicine intragastrically， once a day， from Monday to Saturday. Half of rats in each group were selected 4， 8 weeks after medication； the serum levels of glucose and insulin were detected， and islet resistance index were calculated. The serum levels of liver function indexes （AST， ALT， AKP）， blood lipid indexes （FFA， TC， TG， LDL-C） and inflammatory factors （TNF-α， IL-6， IL-1β） were detected by ELISA. After HE staining， the histopathological changes of liver tissue were observed by microscopy. mRNA and protein expressions of peroxisome proliferator activated receptor α （PPARα） and peroxidase acyl coenzyme A oxidase 1 （ACOX1） in liver tissue were detected by RT-qPCR and Western blot assay. RESULTS： Compared with normal control group， glucose， insulin serum levels and islet resistance index were significantly increased （P＜0.01）； serum liver function indexes， blood lipid indexes and inflammatory factor levels of model control group were increased significantly in model control group after 4 and 8 weeks medication （P＜0.05 or P＜0.01）. The hepatocyte swelling of model control group was round and the volume was significantly larger than that of blank control group. The liver had different degrees of steatosis and vacuolization， accompanied by a small amount of inflammatory cell infiltration. mRNA and protein expressions of PPARα and ACOX1 in liver tissue were decreased significantly （P＜0.05 or P＜0.01）. Compared with model control group， except that there was no significant decreased in islet resistunce index and AST， ALT， IL-6， IL-1β serum levels after 4 weeks of medication， and glucose， insulin， ALT serum levels after 8 weeks of medication and the levels of 4 blood lipid indexes after 4 and 8 weeks of medication in selenium-enriched G. lucidum crude extract low-dose group （P＞0.05）， above serum indexes of other groups were decreased significantly after 4 and 8 weeks of medication （P＜0.05 or P＜0.01）. After 4 and 8 weeks of medication， the pathological changes of liver tissue in rats were alleviated in varying degrees. protein and mRNA expressions of PPARα and ACOX1 in liver tissue were increased significantly after 4 and 8 weeks of medication （P＜0.05 or P＜0.01）. CONCLUSIONS： Selenium-enriched G. lucidum crude extract can up-regulate protein and mRNA expressions of PPARα and ACOX1 in liver tissue， promote the excretion of accumulated fatty acid and significantly improve fatty acid metabolism， inflammatory response and liver function in T2DM model rats.
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<p><b>OBJECTIVE</b>To investigate the effect of ginsenosides from stems and leaves of ginseng on ethanol-induced lipid deposition in human L02 hepatocytes.</p><p><b>METHODS</b>L02 cells were exposed to ethanol for 36 h and treated with or without ginsenosides. The viability of L02 cells was evaluated by methylthiazolyldiphenyl-tetrazolium bromide assay and the triglyceride (TG) content was detected. Lipid droplets were determined by oil red O staining. Intracellular reactive oxygen species (ROS) production and the mitochondrial membrane potential were tested by flow cytometry. The ATP level was measured by reverse phase high performance liquid chromatography. The expression of cytochrome p450 2E1 (CYP2E1) and peroxisome proliferator-activated receptor α (PPARα) was detected by reverse transcriptase-polymerase chain reaction and Western blotting, respectively.</p><p><b>RESULTS</b>Ethanol exposure resulted in the increase of TG level, lipid accumulation and ROS generation, and the decrease of mitochondrial membrane potential and ATP production in the cells. However, ginsenosides significantly reduced TG content (9.69±0.22 μg/mg protein vs. 4.93±0.49 μg/mg protein, P<0.01), and ROS formation (7254.8±385.7 vs. 5825.2±375.9, P<0.01). Meanwhile, improvements in mitochondrial membrane potential (10655.33±331.34 vs. 11129.52±262.35, P<0.05) and ATP level (1.20±0.18 nmol/mg protein vs. 2.53±0.25 nmol/mg protein, P<0.01) were observed by treatment with ginsenosides. Furthermore, ginsenosides could down-regulate CYP2E1 expression (P<0.01) and upregulate PPARα expression (P<0.01) in ethanol-treated cells.</p><p><b>CONCLUSIONS</b>Ginsenosides could prevent ethanol-induced hepatocyte steatosis in vitro related to the inhibition of oxidative stress and the improvement of mitochondrial function. In addition, the modulation of CYP2E1 and PPARα expression may also play an important role in the protective effect of ginsenosides against lipid accumulation.</p>
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Peroxisome proliferator-activated receptor α(PPARα)is an important subtype in the PPARs family. PPARs are a group of nuclear hormone receptors,which belong to type II nuclear receptor super family.PPARαagonists could be used in the treatment of hyperlipemia in clinic.PPARαagonists mainly include natural type and synthetic type,and according to the structure,the synthetic PPARαagonists can be divided into phenyl-heterocy-clic derivatives,ureide derivatives,amides derivatives,phenyloxazole or phenylthiazole derivatives,etc.So far, many PPARαagonists have been approved or in clinical development,and a series of novel PPARαagonists with higher activity and selectivity are being developed.This review will survey the progress in PPARαagonists.
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Objective To examine the effect and mechanism of different targets of glucose control on liver damage in rats with sepsis .Methods The rat sepsis model was established by cecal ligation and puncture (CLP) .Forty Sprague‐Dawley rats were randomly divided into five groups (eight rats to each group):sham operation (sham group) ,sepsis (CLP group) ,glycemic control A group (glucose target 4 .6‐6 .1 mmol/L ) ,glycemic control B group (glucose target 6 .2‐8 .3 mmol/L ) and glycemic control C group (glucose target 8 .4‐10 .0 mmol/L) .The animals were sacrificed 12 hours after CLP .Venous blood was sampled for testing alanine transaminase (ALT ) , aspartate transaminase (AST ) and free fatty acid (FFA ) . Peroxisome proliferator activated receptor‐α (PPAR‐α) and liver carnitine palmitoyltransferase 1 (CPT‐1 ) protein were determined by immunohistochemistry .The pathological changes of liver tissue was observed under an optical microscope .Results The levels of ALT ,AST and FFA in venous blood and the pathological tissue injury score in sepsis groups were higher than those in sham group and all glycemic control groups (P0 .05) .The levels of PPARαand liver CPT‐1 were significantly higher in group A than in group B or group C (P<0 .05) ,and lower in group C than in group B(P<0 .05) .Conclusions The lowest target of glucose control(4 .6‐6 .1 mmol/L)shows better protective effects on liver damage in rats with sepsis ,the mechanism of which may be related to upregulation of PPARα and liver CPT‐1 expression .
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Objective To investigate the effects of peroxisome proliferator-activated receptor α( PPARα) on macrophage-mediated inflammatory responses with the interference of lipopolysaccharide and the possible mechanism.Methods The bone marrow stem cells were isolated from the femora of mice.The granulocyte-macrophage colony stimulating factor ( GM-CSF) was used to stimulate the in vitro differentiation from bone marrow stem cells into primary macrophages.An in vitro model with cultured cells expressing in-flammatory cytokines was established by treating the primary macrophages with lipopolysaccharide ( LPS) .A specific chemical agonist, Wy-14643, was used to activate PPARα. Autophagy inhibitors including 3-methyladenine (3-MA) and small interfering RNA against Atg7 ( Atg7 siRNA) were used to inhibit the autophagy.Western blot assay was performed to detect the expression of autophagy-related proteins ( Atg5, Atg7, Beclin-1 and LC3).The transcriptional levels of TNF-α, IL-1β, IL-6, Atg5, Atg7 and Beclin-1 were analyzed by qRT-PCR.Results Compared with the macrophages treated with LPS alone, those pretreated with various concentrations of Wy-14643 (10 μmol/L, 25 μmol/L and 50 μmol/L) showed inhibited ex-pression of proinflammatory cytokines ( TNF-α,IL-1βand IL-6) and enhanced expression of autophagy-relat-ed proteins (Atg5, Atg7 and Beclin-1) at mRNA level in a dose-dependent manner.The expression of auto-phagy-related proteins (Atg5, Atg7, Beclin-1 and LC3) by macrophages was promoted with the pretreatment of Wy-14643 as indicated by Western blot assay.The transcriptional levels of TNF-α, IL-1βand IL-6 were increased in Wy-14643 pretreated-macrophages after stimulation with 3-MA or Atg7 siRNA .Conclusion PPARαsuppressed the macrophage-mediated inflammatory responses by promoting autophagy, suggesting that the PPARα-autophagy pathway might be one of the signaling pathways regulating LPS induced-inflamma-tory responses.
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AIM@#The potential of Trifolium pratense (red clover) extract in the prevention of lipid disorder has attracted increasing attention in recent years. In this study, the aim was to determine whether and how red clover extract affected the development of murine diet-induced non-alcoholic steatohepatitis.@*METHODS@#Non-alcoholic steatohepatitis was induced in C57BL/6 mice by feeding mice with a methionine-choline-deficient (MCD) diet. Hematoxylin and eosin staining was used for histological analyses. Real-time PCR was used to analyze the mRNA expression levels.@*RESULTS@#Hepatic steatosis and necroinflammation was observed in MCD diet-fed mice, and this diet-induced steatosis was significantly attenuated, whereas liver inflammation was not significantly attenuated, by red clover extract treatment. Consistent with the results of H&E staining, the MCD diet-induced increase of liver triglycerides and cholesterol levels were significantly reduced by red clover extract treatment. However, with the improvement in hepatic steatosis, mRNA levels of acetyl CoA oxidase, carnitine palmitoyl transferase-1, and liver fatty acid-binding protein, three genes regulated by peroxisome proliferator-activated receptor (PPAR) α, were unaffected.@*CONCLUSION@#Red clover extract alleviated MCD diet-induced hepatic steatosis, but did not ameliorate liver inflammation in C57BL/6 mice, and the improvement in hepatic steatosis was not through activating PPARα.
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Animals , Male , Mice , Cholesterol , Metabolism , Choline Deficiency , Diet , Disease Models, Animal , Inflammation , Drug Therapy , Metabolism , Liver , Metabolism , Methionine , Mice, Inbred C57BL , Non-alcoholic Fatty Liver Disease , Drug Therapy , Metabolism , PPAR gamma , Metabolism , Phytotherapy , Plant Extracts , Pharmacology , Therapeutic Uses , RNA, Messenger , Metabolism , Trifolium , Triglycerides , MetabolismABSTRACT
[ABSTRACT]AIM:ToinvestigatethedifferenteffectsofERK1/2/PPARα/SCAD(short-chainacyl-CoAdehy-drogenase) signal pathways on the cardiac hypertrophy induced by insulin-like growth factors 1 ( IGF-1) or phenylephrine ( PE) .METHODS:The neonatal rat cardiomyocytes induced by IGF-1 were used as the model of physiological cardiac hypertrophy , and those induced by PE were used as the model of pathological cardiac hypertrophy .The surface area of the cardiomyocytes, the expression of p-ERK1/2, PPARαand SCAD, the activity of SCAD and the content of free fatty acid in the cardiomyocytes were measured .RESULTS:Compared with the control cells , the surface area of the cardiomyocytes in-duced by IGF-1 and PE were both increased .Compared with the controls , the expression of SCAD and PPARα, and the activity of SCAD in the cardiomyocytes induced by IGF-1 were increased , while the expression of p-ERK1/2 was de-creased.However, the cardiomyocytes treated with PE showed decreased expression of SCAD and PPARα, decreased activ-ity of SCAD and increased expression of p-ERK1/2.Meanwhile, the decrease in free fatty acid in IGF-1-induced cardio-myocytes and the increase in PE-induced cardiomyocytes indicated that the fatty acid utilization was increased in the cardio -myocytes induced by IGF-1, but decreased in the cardiomyocytes induced by PE .CONCLUSION: The changes of p-ERK1/2, PPARαand SCAD in the cardiac hypertrophy induced by IGF-1 or PE indicate that the effects of ERK 1/2/PPARα/SCAD signal pathways are different between physiological cardiac hypertrophy and pathological cardiac hypertro -phy , and that SCAD may be a molecular marker of these 2 different cardiac hypertrophies and a potential therapeutic target for pathological cardiac hypertrophy .
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Objective To investigate the association between the peroxisome proliferator-activated receptor-or (PPARα) polymorphism rs1800206 (c.484C ＞ G,L162V) and the risk of metabolic syndrome (MES)Methods There were 1184 subjects aged 40-60 years recruited from our hospital between March 2010 and March 2011.The PPARα polymorphism 484C ＞ G was genotyped using TAQMAN assay by real-time PCR. The relationship between PPARα polymorphism and MES risk were investigated.Results The genotype frequencies were 91.4％,8.4％ and 0.2％ for the PPARα CC,CG and GG,respectively.This SNP was in Hardy-Weinberg equilibrium(P =0.845).Compared with the most common CC genotype,the variant genotypes(CG + GG) had higher fasting glucose((5.82 ± 1.59) mmol/L vs (5.49 ± 1.17) mmol/L,t =2.630,P =0.009) and LDL-C levels((3.53 ± 1.03) mmol/L vs (3.36 ± 0.65) mmol/L,t =2.376,P =0.018) and lower HDL-C levels ((1.56 ± 0.35) mmol/L vs (1.65 ± 0.43) mmol/L,t =2.430,P =0.015).Furthermore,the 484G variant genotypes(CG + GG)was associated with the risk of MES after adjusting for age,gender,education,BMI and lifestyle (smoking and alcohol status) (adjusted OR =1.89 ;95％ CI =1.09-3.23,P =0.012).Condusion PPARα polymorphism rs1800206 is a significant independent predictor of the MES in Southern Chinese.
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AIM: To investigate the role and signal mechanism of PPAR-α in the pathogenesis of cardiac hypertrophy. METHODS: Small interfering RNA (siRNA) was applied to efficiently silence the gene expression of PPAR-α in cardiac myocytes. [~3H] leucine incorporation assay was performed to measure protein synthesis. Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the mRNA level of atrial natriuretic factor (ANF) and PPAR-α. Western blotting analysis was performed to investigate the levels of phosphorylation of protein kinase B (PKB/Akt) and glycogen synthase kinase 3β (GSK3β). Immunofluorescence analysis was used to examine the cellular localization of NFATc4. RESULTS: (1)RSS304168 was the most efficient stealth RNAi duplex to specifically inhibit PPAR-α expression. (2)RSS304168 significantly potentiated the ET-1-induced cardiomyocyte hypertrophy and enhanced ET-1-induced protein synthesis and ANF mRNA expression in cardiomyocytes. Moreover, RSS304168 completely reversed the inhibitory effects of fenofibrate on ET-1-induced protein synthesis and ANF mRNA expression. (3)RSS304168 enhanced ET-1-induced phosphorylation of Akt at Ser473 and GSK3β at Ser9. The effects of ET-1 or ET-1 combined with RSS304168 on phosphorylation of Akt/GSK3β were completely blocked by LY294002, a PI3K specific inhibitor. Fenofibrate markedly inhibited ET-1-induced phosphorylation of Akt/GSK3β while RSS304168 abolished these effects of fenofibrate. (4)Fenofibrate prevented the nuclear translocation of NFATc4 induced by ET-1 while RSS304168 abolished this effect of fenofibrate. CONCLUSION: Activation of PPAR-α inhibits ET-1-induced cardiomyocyte hypertrophy through blocking Akt/GSK3β-NFATc4 signaling pathways.
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AIM: To study the role of peroxisome proliferator-activated receptor-α (PPAR-α) signal transduction pathway in cardiac hypertrophy induced by high glucose and insulin (HGI). METHODS: The cultured neonatal rat cardiomyocytes were used to observe the effect of fenofibrate (FF), a selective PPAR-α agonist, on cardiomyocyte hypertrophy induced by HGI (glucose at concentration of 25.5 mmol/L and insulin at 0.1 μmol/L). The cardiomyocyte hypertrophic responses were assayed by measuring the cell surface area, protein content, and mRNA expression of atrial natriuretic factor (ANF). The expressions of mRNA and protein were assayed by real -time PCR and Western blotting. RESULTS: In cultured cardiomyocytes, HGI induced profound change of hypertrophic morphology, the significant increase in cell surface area, protein content and ANF mRNA expression compared to those in vehicle control (P<0.01), but the expressions of PPAR-α mRNA and protein decreased significantly (P<0.05). At the same time, the expression of cyclooxygenase 2 (COX-2), one of the PPAR-α downstream effectors was obviously elevated (P<0.05). However, FF (0.1, 0.3 and 1 μmol/L) inhibited the cardiomyocyte hypertrophy induced by HGI in a concentration-dependent manner (P<0.01). FF at concentration of 0.3 μmol/L increased the expressions of PPAR-α in both mRNA and protein levels (P<0.05) and inhibited the expressions of COX-2 (P<0.05), which were abolished by MK 886 (0.3 μmol/L), a selective PPAR-α antagonist (P<0.05). CONCLUSION: PPAR-α signal transduction pathway and its downstream effector COX-2 might involve in the cardiomyocyte hypertrophy induced by HGI.