The molecular mechanism of ginkgolide B regulating the expression of long-chain fatty acid metabolism-related proteins and antioxidant therapy for non-alcoholic fatty liver disease / 药学学报

This study investigated the effects of ginkgolide B on the long-chain fatty acid metabolism-related enzyme protein peroxisome proliferators-activated receptors α (PPARα), long-chain specific acyl-CoA dehydrogenase (LCAD), carnitine palmitoyl transterase-1 (CPT-1), and acyl coenzyme A oxidase 1 (ACOX1) expression in the liver of rats with non-alcoholic fatty liver disease (NAFLD). All the animal welfare and experimental procedures are in accordance with the regulations of the Animal Ethics Committee of Yunnan University of Traditional Chinese Medicine. After successfully building the rat model of non-alcoholic abnormal liver disease, the rats were divided into the model group, the simvastatin group, and the low-dose, middle-dose, and high-dose groups of ginkgolide B according to random number method, and were given corresponding drug treatment 4 weeks. We detected liver pathological indicators and determined blood lipids, transaminase and anti-oxidation indexes. Western blot and RT-PCR assays were used to detect the protein and mRNA levels of PPARα, LCAD, CPT-1, and ACOX1 in livers. The results showed that: ① the liver histopathology showed that the liver slices of the model group had obvious structural disorder, the nucleus was squeezed, and there were obvious fat vacuoles. The treatment groups improved significantly compared with the model group; ② compared with the normal group, the liver function and blood lipid indexes of the model group increased significantly, while the anti-oxidation indexes decreased significantly. Compared with the model group, each treatment groups were significantly improved; ③ compared with the normal group, the protein and mRNA expression levels of PPARα, ACOX1, CPT-1, and LCAD in the model group were significantly reduced, compared with the model group, those indexes in the treatment groups were significantly up-regulated. This study found that ginkgolide B could regulate the expression of long-chain fatty acid metabolism-related proteins PPARα, ACOX1, CPT-1, and LCAD, meanwhile improve the body's antioxidant capacity, thereby reduce blood lipids, further improve liver function and protect the liver.

Effect of Ginsenoside Rg1 on Lipid Uptake and Oxidation in HepG2 Cells of Nonalcoholic Fatty Liver Disease / 中国实验方剂学杂志

Objective:To investigate the effect and mechanism of ginsenoside Rg1（G-Rg1）in ameliorating lipid uptake and oxidation in HepG2 cells induced by free fatty acids (FFA). Method:HepG2 cells were divided into normal group, model group，low-dose ginsenoside Rg1 group (25 μmol·L-1) and high-dose G-Rg1 group (50 μmol·L-1). HepG2 cells were treated with 1 mmol·L-1 free fatty acid for 24 h to construct the NAFLD cell model, and then treated with 25，50 μmol·L-1 G-Rg1 for 24 h. The effect of G-Rg1 on HepG2 cell activity was determined by cell counting kit-8（CCK-8） assay. The level of triglyceride (TG) was detected by micro method. The accumulation of lipid droplets was observed by oil red O staining. Quantitative real-time fluorescence polymerase chain reaction (Real-time PCR) and Western blot were used to detect the alterations of key genes and proteins relating to lipid uptake and metabolism. Result:Compared with the normal group, the intracellular TG level and the absorbance of the oil red O staining in the model group were significantly increased (P<0.01). Compared with the model group, G-Rg1 reduced TG and lipid deposition were significantly reduced (P<0.01).Results of Real-time PCR and Western blot showed that compared with normal group, model group peroxisome proliferators-activated receptors gamma（PPARγ），fatty acid binding protein 1（FABP1），fatty acid transport protein 2/5（FATP2/5）and fatty acid translocase（CD36）expressions increased（P<0.05），whereas peroxisome proliferators-activated receptors α（PPARα），carnitine palmitoyltransferase 1（CPT1）and peroxisomal acyl-coenzyme A oxidase 1（ACOX1）expressions decreased（P<0.05）. Compared with the model group, the expressions of PPARγ, FABP1, FATP2, FATP5 and CD36 in the G-Rg1 group were decreased (P<0.05，P<0.01), while the expressions of PPARα, CPT1 and ACOX1 were increased (P<0.05，P<0.01). Conclusion:G-Rg1 can ameliorate lipid deposition in NAFLD cell model by reducing lipid uptake and increasing lipid oxidation.

Effect of AMPK pathway on apoptosis of human thyroid papillary cancer B-CPAP cells under low glucose and hypoxia conditions through CPT1c regulated by PPARα / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the mechanisms of carnitine palmitoyltransferase 1c (CPT1c) expression to affect the proliferation and apoptosis of human thyroid papillary cancer B-CPAP cells through the AMP-dependent/activated protein kinase (AMPK) pathway in the low glucose and hypoxic conditions. Methods: Firstly,humanthyroidpapillarycarcinomaB-CPAP cells were cultured under normal condition or low glucose and hypoxic condition respectively, followed with the treatment of AMPK inhibitor compound C. Western blotting was used to detect the expressions of AMPK, p-AMPK, peroxisome proliferator-activated receptor α (PPARα) and CPT1c; the proliferation and apoptosis were detected by CCK-8 and Flow cytometry, respectively. Then PPARα-siRNA was synthesized and transfected into B-CPAP cells to knock down PPARα, and then the cells were cultured under normal or low glucose and hypoxic condition respectively.Above indicators were also detected to verify the regulation of PPARα on CPT1c. Finally, the human luciferase reporter plasmid containing CPT1c gene promoter was constructed, and the effect of PPARα on the activity of CPT1c promoter luciferase activity was observed by immunofluorescence. Results: The expressions ofAMPK, p-AMPK, PPARα and CPT1c were significantly increased in B-CPAP cells under low glucose and hypoxia condition (P<0.05 or P<0.01), while cell proliferation and apoptosis rate did not change significantly (P>0.05). After the treatment of AMPK inhibitor compound C, the expressions of p-AMPK, PPARα and CPT1c in low glucose and hypoxia group were significantly decreased (P<0.05 or P<0.01), the inhibitory rate on cell proliferation and apoptosis rate were significantly increased (P<0.05). However, the change range was smaller than that in the normal culture + compound C group (P<0.05).After PPARα knockdown, the expressions ofAMPK, p-AMPK, PPARα and CPT1c in cancer cells cultured under normal conditions were significantly decreased (P<0.05 or P<0.01), and the inhibitory rate on cell proliferation and apoptosis rate were significantly increased (P<0.05). While under low glucose and hypoxia condition, the expression of CPT1c in cells after transfection was significantly decreased (P<0.05), and the inhibition rate on cell proliferation and the apoptosis rate were significantly increased (P<0.05); However, the change range was still lower than that of normal condition group after transfection (P<0.05).After PPARα overexpression, the ratio of fluorescence in the empty vector group was not significantly different from that of the blank group (P>0.05), and the ratio of fluorescence was significantly increased in PPARα over-expression group (P<0.05). Conclusions: AMPK can increase the expression of PPARα to promote the expression of CPT1c in thyroid cancer B-CPAP cells under low glucose and hypoxia conditions, thereby inhibiting cell apoptosis and maintaining cell proliferation ability.

Effect of Buyang Huanwu Tang on Myocardial Mitochondrial Energy Metabolism and AMPK/PPARα Signaling Pathway in Rats with Diastolic Heart Failure / 中国实验方剂学杂志

Objective: To study the effect of Buyang Huanwu Tang on myocardial energy metabolism in rats with diastolic heart failure (DHF) based on adenosine monophosphate (AMP)-activated protein kinase (AMPK)/peroxisome proliferators-activated receptors α (PPARα) signaling pathway,and investigate its mechanism of action.Method: The 48 SD rats were randomly divided into sham operation group and model group.DHF rat model was established by abdominal aorta constriction method.The successfully modeled rats were randomly divided into model group,Buyang Huanwu Tang group (12.72 g·kg-1·d-1),metoprolol tartrate group (0.004 5 g·kg-1·d-1),with corresponding drugs in each group by intragastric administration.The sham operation group and model group were given with equal amount of deionized water,once a day.After 8 weeks of continuous drug intervention,the contents of adenosine monophosphate (AMP),adenosine diphosphate (ADP) and adenosine triphophate (ATP) in peripheral blood of rats were determined by enzyme linked immunosorbent assay (ELISA).The changes of myocardial mitochondrial ultrastructure were detected by electron microscope.The protein expression levels of AMPK,PPARα and peroxisome proliferator-activated receptor-γ coactivator-1α(PGC-1α) in rat myocardium were detected by Western blot.Result: As compared with sham operation group,the contents of AMP and ADP in model group were increased significantly,and ATP content was decreased significantly (PPPPα and PGC-1α protein in the model group were decreased significantly (Pα and PGC-1α protein in Buyang Huanwu Tang group and metoprolol tartrate group were increased significantly (PConclusion: Buyang Huanwu Tang may improve the energy metabolism of the failed heart and delay the progression of heart failure by improving the structure and function of mitochondria,activating AMPK and up-regulating the expression of AMPK/PPARα signaling pathway.

The Activation of PPAR-α and Wnt/β-catenin by Luffa cylindrica Supercritical Carbon Dioxide Extract

Luffa cylindrica (LC) is a very fast-growing climber and its fruit have been considered as agricultural wastes. We conducted to check the comparative qualities of ethanol solvent extraction (LCE) and supercritical carbon dioxide extraction (LCS) of L. cylindrica fruit and seed. LCS had higher antioxidant and polyphenol contents than LCE. LCS were significantly increased peroxisome proliferator-activated receptor (PPAR)-a and involucrin expression as epidermal differentiation marker in 3D skin equivalent model. LCS also showed antimicrobial activity against Staphylococcus aureus, a causative bacteria in atopic dermatitis. In addition, LCS inhibited the adipocyte differentiation of 3T3-L1 cells. When treated with the extract at a concentration of 100 µg/mL, the Wnt/β-catenin pathway reporter luciferase activity of HEK 293-TOP cells was increased approximately by 2-folds compared to that of the untreated control group. These results indicate that L. cylindrica supercritical carbon dioxide extract may serve as a cosmeceutical for improving skin barrier function and the treatment of obesity.

Receptor activation of curcumin on human peroxisome proliferators-activated receptors γ1 / 中草药

Objective: To determine whether curcumin is a natural ligand for human peroxisome proliferators-activated receptors γ1 (hPPARγ1) by measuring the combination ability and internal activity. Methods: The combination ability was determined by radioactively labeled ligand binding experiment (RBCA), and the internal activity was estimated by trans-activation reporter gene test. Results: The combination ability of curcumin on hPPARγ1 showed that IC50 was (8.82 ± 0.74) μmol/L, and Ki was 0.72 μmol/L. The internal activity showed that EC50 was 7.3 μmol/L and Emax was 43.3. Conclusion: Curcumin has affinity and intrinsic activity with hPPARγ1, which suggests that curcumin may be a natural ligand of hPPARγ1.

Mechanism progress of bronchial asthma cured by PPARγagonists / 中国基层医药

Peroxisome proliferators -activated receptors γ(PPARγ)agonist is a kind of drugs that is widely used in the treatment of type 2 diabetes,also shows effects on hypertensive disease,regulating blood lipid metabolism, anti -atherosclerosis and inhibiting inflammatory response.In recent years,good effects of PPARγagonist have been found on different animal models of asthma,and production of proinflammatory cytokines can significantly be reduced. Airway remodeling can be reduced,and airway hyper responsiveness can be inhibited by PPARγagonist that provide theoretical basis on new drugs for asthma.The mechanism of bronchial asthma cured by PPARγagonists are reviewed briefly.

Artemether inhibits proliferation and invasion via the mediation of peroxisome proliferator-activated ;receptor-gamma activation pathway in Lewis lung cancer cells / 国际肿瘤学杂志

mRNA in ARE and GW9662 group were 2.276 ±0.534 and 0.362 ±0.026,respectively.Compared with control group,PPARγmRNA level in both of ARE and GW9662 group reached statistical significance (t =4.785,P =0.001 ;t =2.395,P =0.044).PPARγprotein expression in ARE group,GW9662 +ARE group and control group were 27 688.33 ±3 593.06,21 816.00 ±1 644.07,17 716.33 ±2 273.95,respectively,which was higher in ARE group than that in control and GW+ARE group (t =5.159,P =0.001 ;t =3.038,P =0.016). NF-κB p65 mRNA expression in GW9662 +ARE group was 0.346 ±0.149,which in ARE group and GW9662 group were 0.392 ±0.1 87 and 1 .720 ±0.338,respec-tively.The differences of NF-κB p65 mRNA expression level between ARE,and control or GW9662 group were statistically significant (t =3.592,P =0.007;t =7.851 ,P =0.000).While,the differences of Caspase-3 mRNA and protein expression levels among the four groups were not statistically significant (F =1 .1 81 ,P =0.376;F =0.647,P >0.05).Conclusion ARE may restrain NF-κB through up-regulating PPARγto inhibit the proliferation and invasive potential of LLC in vitro, which suggests that PPAR-γmay be a novel therapeutic target for lung cancer.

Association of peroxisome proliferators-activated receptor-γ coactivator 1 Gly482Ser with apolipoprotein E and the longevity and metabolic traits of Hans in Guangxi Yongfu / 中华老年医学杂志

Objective To explore the association of peroxisome proliferators-activated receptor-γ coactivator-1 (PPARGC1) Gly482Ser with apolipoprotein E (ApoE) variations in longevity (aged above 90 yrs) Hans in Guangxi Yongfu and to explore the potential association between the variations and metabolic traits.Methods Based on our survey in Guangxi Yongfu in 2008-2011,212 elderly cases (aged 90～105 years) were included as longevity group and 207 cases without longevity history were included as control group.By household survey,we collected the longevity related parameters,blood glucose,blood lipid,blood pressure and other related metabolic traits.Peripheral blood was collected to extract DNA,the gene variations of Gly482Ser and ApoE were genotyped,and the database with genome and traits information were set up.By univariate analysis and multivariate genetic statistical analysis,the association between the variations and longevity and metabolic traits was assessed.Results Compared with the control group,the levels of fasting blood glucose,total cholesterol and low density lipoprotein were lower in the longevity group.Gly482Ser was genotyped in all samples and fully fulfilled the Hardy Weinberg equilibrium.After the Bonferroni correction,recessive model failed to find association between GG genotype and longevity.Stratified analyses by ApoEε4 allele revealed that,in the subgroup with no ApoEε4,PPARGC-1 GG genotype was positively associated with longevity in the recessive model,even after Bonferroni correction (OR =1.72,P＜0.05).In addition,longevity group with Gly482Ser GG genotype seemed to have relativelower fasting blood glucose (P ＜ 0.05) and higher high density lipoprotein levels (P ＜ 0.05).Conclusions Longevity Hans in Guangxi Yongfu preserve better metabolic state compared with the control group.GG genotype of Gly482Ser in PPARGC-1 is positively associated with longevity,which depends on not carrying the risk allele of ApoE ε4.

Roles of peroxisome proliferators activated receptors NF-κB and SOD in liver injury of obstructive jaundice in rats / 中华肝胆外科杂志

Objective To determine the expression of peroxisome proliferators-activated receptors (PPARs), nuclear factor-κB (NF-κB) and activity of superoxide dismutase (SOD) in rats with obstructive jaundice induced by bile duct ligation (BDL) and elucidate the molecular regulation mechanisms of hepatic injury due to obstructive jaundi(c)e. Methods All rats were sacrificed on the 7th day and 19th day after BDL and blood and liver tissue samples were obtained. SOD enzyme activity was detected by SOD kit. RT-PCR was performed to determine the mRNA expression of PPARs in all groups. The detection of PPARs protein and activation of NF-κB were performed using an immunohistochemical method. Results The activities of normal SOD and CuZn-SOD were decreased compared to the sham group (P＜0.01), and the decrease on 19th day after BDL were more significant. The level of PPARs expression in the BDL groups liver except the PPARβ in the BDL 7th group was reduced compared to the sham group (P＜0.01), and the level on the 19th day after BDL were more significantly reduced. PPARs protein expression was significantly inhibited (P＜0.01) in the sham group. SOD in BDL groups had significant positive correlation as compared with PPARs protein expression (P＜0.01), but NF-κBp65 protein expression had significant negative correlation as compared with PPARs protein expression (P＜0. 01). Conclusion PPARs are inhibited in expression level, and this inhibition becomes more significant as the pathological process progresses. PPARs might be key regulatory factors for SOD and NF-κB. The low expression of PPARs might be one of the important molecular mechanisms in liver injury due to obstructive jaundice.

Anti-oxidation effect of rosiglitazone in aging rat kidney / 中华肾脏病杂志

Objective To investigate the effect of rosiglitazone (RGTZ) on anti-oxidation in aging rat kidney. Methods Twenty-four-month-old male Sprague-Dawley rats were randomly divided into three groups (n=10): control group (CON), rosiglitazone group (RGTZ) and caloric restriction group (CR). The CON rats were allowed ad libitum access to feed and tap water.The RGTZ rats received intragastric administration of RGTZ (4 mg·kg-1·d-1),and the CR rats were provided with a vitamin and mineral fortified version of the same diet at a level of 40% less food (by weight) than the CON rats. After 12 weeks all the animals were sacrificed by decapitation, and both the body weight and the percentage of kidney and heart in each group were measured.Western blot was performed to analyze the expression of PPARγ protein. The content of MDA and the activity of SOD and GSH-PX in kidney tissue were detected. Besides, frozen sections of kidney tissue were stained for senescence-associated-13 galactosidase (SA-β-Gal). Results The body weight of CR rats decreased obviously, in contrast, which did not change in CON and RGTZ group. Percentage of kidney and heart to body weight was normal in CR or RGTZ group after intervention. Western blot result showed that PPARγ protein expression in rat kidney was significantly higher in RGTZ and CR group as compared to CON group (P<0.05). Compared with RGTZ and CR rats, obviously lower activities of SOD and GSH-Px were noted in CON rats, however, the content of MDA was higher in CON rats. Additionally, the positive staining area of [3-Gal in CR and RGTZ group was significantly smaller than that in CON rats (P<0.05, P<0.01 ). Conclusion RGTZ can defer the kidney aging in senescence SD rat, and the mechanism may be related to amelioration of oxidative damage and enhancement of antioxidation.

Os efeitos do ácido linoléico conjugado no metabolismo animal: avanço das pesquisas e perspectivas para o futuro: [revisão] / Effects of conjugated linoleic acid on animal metabolism: advances in research and perspectives for the future: [review]

Realizou-se uma revisão sistemática, sem restrição de data, sobre os efeitos fisiológicos do ácido linoléico conjugado sobre a regressão da carcinogênese, o estresse oxidativo, o metabolismo de lípides e glicose e a alteração da composição corporal. Objetivando estabelecer o aspecto histórico do avanço da pesquisa em ácido linoléico conjugado, consideraram-se artigos originais resultantes de trabalhos realizados com animais, com cultura de células e com humanos. Quanto às pesquisas sobre o efeito anticarcinogênico do ácido linoléico conjugado foram encontradas inúmeras evidências a esse respeito, especialmente na regressão dos tumores mamários e de cólon, induzida por ambos os isômeros os quais agem de maneiras distintas. Os pesquisadores se empenham em reinvestigar as propriedades antioxidantes do ácido linoléico conjugado. Embora tenham sido investigadas as propriedades antioxidantes, tem-se identificado efeito pró-oxidante, levando ao estresse oxidativo em humanos. Foram poucos os estudos que demonstraram efeito positivo significativo do ácido linoléico conjugado sobre o metabolismo dos lípides e da glicose e sobre a redução da gordura corporal, especialmente em humanos. Estudos sobre efeitos adversos foram também identificados. Há fortes indícios de que a ação deste ácido graxo conjugado sobre uma classe de fatores de transcrição - os receptores ativados por proliferadores de peroxissomo - e sobre a conseqüente modulação da expressão gênica, possa ser a explicação fundamental dos efeitos fisiológicos. Embora incipientes, os mais recentes estudos reforçam o conceito da nutrigenômica, ou seja, a modulação da expressão gênica induzida por compostos presentes na alimentação humana. O cenário atual estimula a comunidade científica a buscar um consenso sobre os efeitos do ácido linoléico conjugado em humanos, já que este está presente naturalmente em alguns alimentos, que, quando consumidos em quantidades adequadas e de forma freqüente...

This systematic review without date restrictions is about the physiological effects of conjugated linoleic acid on regression of carcinogenesis, oxidative stress, glucose and lipid metabolism and change in body composition. The objective was to establish the historical aspect of research advances regarding conjugated linoleic acid, considering original articles reporting work on animals, cell cultures and humans. Regarding the researches on the anticarcinogenic effect of conjugated linoleic acid, innumerous evidences were found in this respect, especially in the regression of mammary and colon tumors induced by both isomers which act distinctively. The researchers devoted considerable effort to reinvestigate the antioxidant properties of conjugated linoleic acid. Although the antioxidant properties have been investigated, pro-oxidant effect has been identified leading to oxidative stress in humans. Few studies demonstrated significant beneficial effects of conjugated linoleic acid on the metabolism of lipids and glucose and on the reduction of body fat, especially in humans. Studies with adverse effects were also identified. There is strong indication that the action of this conjugated fatty acid on a class of transition factors - the peroxisome proliferator-activated receptor - and on the consequent modulation of gene expression can be the fundamental explanation of its physiological effects. The most recent studies reinforce the nutrigenomic concept, that is, the modulation of gene expression induced by compounds present in the foods consumed by humans. This current scenario stimulates the scientific community to seek a consensus on the effects of conjugated linoleic acid in humans, since it is naturally found in some foods; when these foods are consumed regularly and in appropriate amounts, they could help prevent and control innumerous chronic diseases.

STUDY OF GALLIC ACID IN PU-ERH TEA ON THE PEROXISOME PROLIFERATORS ACTIVATED RECEPTORS FUNCTION / 营养学报

Objective To study the effect of gallic acid isolated from Pu-erh Tea on the peroxisome proliferators activated receptors function.Method The appropriate concentration of gallic acid added to three cell models was decided to be 50 ?g/ ml,and the activity of gallic acid on peroxime prolipevators activated receptors PPAR?,PPAR?,PPAR? was studied.Results Gallic acid could activate PPAR?,as high as 2.436 fold and the effect corresponded to that of positive drug which value was 2.438.gallic acid had no effect on PPAR? and PPAR?.Conclusion Gallic acid in Pu-erh Tea had good activity on PPAR? and this could offer scientific basis for study of the anti-diabetes and anti-hyperlipidenmia mechanism of Pu-erh Tea.