A series of tetrahydroisoquinoline derivatives were prepared and their peroxisome proliferator-activated receptor (PPAR) alpha/gamma agonistic activities were evaluated to obtain more potent PPAR agonist. All of them were new compounds, and their structures were confirmed by 1H NMR and HR-MS. Three compounds exhibited higher agonistic activities of PPARgamma than that of the comparison, six compounds exhibited higher agonistic activities of PPARalpha than that of the comparison, and compound 8a was discovered as a highly potent PPARalpha/gamma agonist that is much more active than that of WY14643 and rosiglitazone. The development of potent PPAR agonists may offer a new choice for the treatment of diabetes.
Drug Design ; HEK293 Cells ; Humans ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; pharmacology ; PPAR alpha ; agonists ; metabolism ; PPAR gamma ; agonists ; metabolism ; Structure-Activity Relationship ; Tetrahydroisoquinolines ; chemical synthesis ; chemistry ; pharmacology ; Transfection

A series of novel tetrahydrocarboline derivatives was designed and synthesized in order to discover more potent peroxisome proliferator-activated receptor (PPAR) alpha/gamma dual regulators. The structures of these compounds were confirmed by 1H NMR and HR-MS; their PPAR-regulating activities were evaluated in vitro. Compounds 6h, 6n, 6p and 6q exhibited more potent PPARalpha agonistic activities than the control drug WY14643, while compounds 60, 6g, 6i and 6q exhibited more potent PPARgamma agonistic activities than the control drug rosiglitazone. Compound 6q was discovered as a potent PPARalpha/gamma dual agonist and deserves further investigation.
Animals ; Carbolines ; chemical synthesis ; chemistry ; pharmacology ; Cells, Cultured ; Drug Design ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; pharmacology ; Molecular Structure ; PPAR alpha ; agonists ; PPAR gamma ; agonists ; Peroxisome Proliferator-Activated Receptors ; agonists ; Pyrimidines ; metabolism ; Structure-Activity Relationship ; Thiazolidinediones ; metabolism ; Transfection

OBJECTIVETo investigate the effect of peroxisome proliferator-activated receptor (PPAR)α agonist bezafibrate and oxidized low density lipoprotein (ox-LDL) on fibroblast growth factor 21 (FGF21) expression and apoptosis in cardiac endothelial cells.

METHODSThe mRNA level of FGF21 was determined by real time-PCR and the protein concentration of FGF21 in culture media was detected by enzyme-linked immunosorbent assay in cultured cardiac microvascular endothelial cells (CMECs) incubated with 10, 50, 100 µg/ml ox-LDL, 50, 100 or 200 µmol/L bezafibrate alone or in combination with 100 µg/ml ox-LDL. CMECs apoptosis in various treatment groups was also determined.

RESULTSFGF21 mRNA and protein expressions were significantly upregulated in proportion to increased ox-LDL, and 200 µmol/L bezafibrate alone also significantly upregulated FGF21 expression and CMECs apoptosis was significantly reduced in 200 µmol/L bezafibrate + 100 µg/ml ox-LDL group compared to 100 µg/ml ox-LDL group (P < 0.05).

CONCLUSIONSOur data suggest that bezafibrate and ox-LDL induced upregulation of FGF21 might mediate the protective effect against apoptosis. Endogenous FGF21 could thus play important roles in improving the endothelial function at the early stage of atherosclerosis and slowing the development of coronary heart disease.

Animals ; Apoptosis ; Atherosclerosis ; metabolism ; pathology ; Bezafibrate ; pharmacology ; Cells, Cultured ; Endothelium, Vascular ; cytology ; metabolism ; Fibroblast Growth Factors ; metabolism ; Lipoproteins, LDL ; pharmacology ; PPAR alpha ; agonists ; Rats ; Rats, Wistar

We sought to determine the effects of activation of peroxisome proliferator-activated receptor-gamma (PPAR-gamma) on multilocularization of adipocytes in adult white adipose tissue (WAT). Male C57BL/6 normal, db/db, and ob/ob mice were treated with agonists of PPAR-gamma, PPAR-alpha, or beta3-adrenoceptor for 3 weeks. To distinguish multilocular adipocytes from unilocular adipocytes, whole-mounted adipose tissues were co-immunostained for perilipin and collagen IV. PPAR-gamma activation with rosiglitazone or pioglitazone induced a profound change of unilocular adipocytes into smaller, multilocular adipocytes in adult WAT in a time-dependent, dose-dependent, and reversible manner. PPAR-alpha activation with fenofibrate did not affect the number of locules or remodeling. db/db and ob/ob obese mice exhibited less multilocularization in response to PPAR-gamma activation compared to normal mice. Nevertheless, all adipocytes activated by PPAR-gamma contained a single nucleus regardless of locule number. Multilocular adipocytes induced by PPAR-gamma activation contained substantially increased mitochondrial content and enhanced expression of uncoupling protein-1, PPAR-gamma coactivator-1-alpha , and perilipin. Taken together, PPAR-gamma activation induces profound multilocularization and enhanced mitochondrial biogenesis in the adipocytes of adult WAT. These changes may affect the overall function of WAT.
Adipocytes/*cytology/metabolism ; Adipose Tissue, White/*cytology ; Animals ; Cell Nucleus Division ; Hypoglycemic Agents/pharmacology ; Ion Channels/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Obese ; Mitochondria/metabolism ; Mitochondrial Proteins/metabolism ; PPAR alpha/agonists/metabolism ; PPAR gamma/*agonists/*metabolism ; Phosphoproteins/metabolism ; Receptors, Adrenergic, beta-3/agonists ; Thiazolidinediones/pharmacology ; Trans-Activators/metabolism

This study was aimed to investigate the influence of PPARalpha agonist on the expression of TF (tissue factor) in THP-1 cells. THP-1 cells were pretreated with different concentrations of PPARalpha agonist (fenofibrate) for definite time. Lipopolysaccharide (LPS)-induced TF mRNA and protein levels were detected by RT-PCR and Western blot respectively. The results showed that fenofibrate decreased tissue factor protein and mRNA expression in supernatants of LPS-stimulated human monocytes in a concentration-dependent manner (P < 0.05 - 0.01, n = 5). It is concluded that fenofibrate inhibit TF expression induced by LPS in THP-1 cells, which may be involved in the anti-atherosclerotic effects of PPARalpha agonist.
Depression, Chemical ; Fenofibrate ; pharmacology ; Humans ; Leukemia, Monocytic, Acute ; metabolism ; pathology ; Lipopolysaccharides ; antagonists & inhibitors ; pharmacology ; PPAR alpha ; agonists ; RNA, Messenger ; biosynthesis ; genetics ; Thromboplastin ; biosynthesis ; genetics ; Tumor Cells, Cultured

OBJECTIVETo investigate the effects of peroxisome proliferator-activated receptor (PPAR) &alpha;/γ agonist on atherosclerotic plaque stabilization in diabetic LDL receptor knockout (LDLr-/-) mice.

METHODSFemale 4-week-old LDLr-/- mice fed with high-glucose and high-fat diet for 4 weeks were randomly divided into three groups (n = 15 each): control group (only fed with high-glucose and high-fat diet), diabetic group [induced by high-glucose and high-fat diet combined with a low-dose of streptozotocin (STZ)] without tesaglitazar and with tesaglitazar (20 µg/kg oral treatment). After 6 weeks, the mice were sacrificed, body weight, fasting blood glucose (Glu), total cholesterol (TC), triglyceride (TG) levels were measured. The expression of ICAM-1, VCAM-1, MCP-1 in the brachiocephalic atherosclerotic lesions were determined by Western blot and immunohistochemistry, respectively. Brachiocephalic artery was prepared for morphologic study (HE, oil red O, Sirius red staining) and immunohistochemical analysis (macrophage surface molecule-3, &alpha;-smooth muscle actin), respectively.

RESULTSSerum TC [(32.34 ± 3.26) mmol/L vs. (16.17 ± 1.91) mmol/L], TG [(3.57 ± 0.99) mmol/L vs. (2.21 ± 0.11) mmol/L] and Glu [(15.21 ± 4.67) mmol/L vs. (6.89 ± 0.83) mmol/L] levels were significantly higher in diabetic group than in the control group (all P < 0.01). The expression of ICAM-1 (2.31 ± 0.35 vs.1.34 ± 0.21), VCAM-1 (1.65 ± 0.14 vs.0.82 ± 0.26), MCP-1 (2.27 ± 0.16 vs.1.56 ± 0.23) were significantly upregulated in diabetic group compared with control group (all P < 0.01). Brachiocephalic atherosclerotic plaque area [(4.597 ± 1.260)×10(3) µm(2) vs. (0.075 ± 0.030)×10(3) µm(2)], lipid deposition [(47.23 ± 2.64)% vs. (9.67 ± 1.75)%], Mac-3 positive area [(19.15 ± 3.51)% vs. (1.72 ± 0.16)%], &alpha;-smooth muscle actin [(5.54 ± 1.17)% vs. (2.13 ± 0.41)%] and collagen content [(4.27 ± 0.74)% vs. (0.43 ± 0.09)%] were all significantly larger/higher in diabetic LDLr-/- mice than in the control group (all P < 0.01). While tesaglitazar treatment significantly reduced serum TC [(30.47 ± 3.18) mmol/L], TG [(3.14 ± 0.71) mmol/L] and Glu [(7.92 ± 1.28) mmol/L] levels (all P < 0.01). Similarly, the expression of ICAM-1 [(1.84 ± 0.22)], VCAM-1 [(1.27 ± 0.11)], MCP-1 [(1.83 ± 0.24)], brachiocephalic atherosclerotic lesion area[(1.283 ± 0.410)×10(3) µm(2)], lipid deposition[(23.52 ± 1.39)%] were also significantly reduced by tesaglitazar (all P < 0.05). Moreover, tesaglitazar increased &alpha;-smooth muscle actin [(9.46 ± 1.47)%] and collagen content [(6.32 ± 1.15)%] in diabetic LDLr-/- mice (all P < 0.05). In addition, lipid deposition and Mac-3 positive areas [(10.67 ± 0.88)% vs. (15.83 ± 1.01)%] in the aortic root were also reduced in tesaglitazar treated diabetic LDLr-/- mice (P < 0.01).

CONCLUSIONSTesaglitazar has anti-inflammatory effects in the diabetic LDLr-/- mice. Tesaglitazar could reduce lipid deposition, increase collagen and &alpha;-SMA content in the brachiocephalic atherosclerotic lesions, thus, stabilize atherosclerotic plaque in this model.

Actins ; metabolism ; Alkanesulfonates ; pharmacology ; Animals ; Collagen ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Diet, High-Fat ; adverse effects ; Female ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipid Metabolism ; Mice ; Mice, Knockout ; PPAR alpha ; agonists ; PPAR gamma ; agonists ; Phenylpropionates ; pharmacology ; Plaque, Atherosclerotic ; metabolism ; pathology ; Receptors, LDL ; genetics ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo investigate the influence and significance of peroxisome proliferator- activated receptor-gamma (PPAR-gamma) agonist rosiglitazone on the expression of I kappa B kinase-beta(IKK-beta) mRNA and protein induced by LPS in Kupffer cells (KCs) cultured in vitro and to investigate the activity of nuclear factor-kappa B (NF-kappa B) together with the expression of cyclooxygenase-2 (COX-2) in livers of rats with non-alcoholic steatohepatitis (NASH).

METHODS(1) KCs from healthy Wistar rats were isolated and purified with IV collagenase digestion and gradient centrifugalization, and then were incubated in the presence or absence of LPS (1 microg/ml) together with two different concentrations of rosiglitazone (10 nmol/L and 50 nmol/L). (2) Thirty-eight healthy Wistar rats were randomly divided into a normal blank control group (10 rats) fed with a normal diet and a NASH model group (28 rats) fed with a fat-rich diet (10% lard + 2% cholesterol + 5% corn oil). After the NASH model was established successfully and confirmed by pathological examination of the livers of 4 rats, 24 rats that continued with the high fat-rich diet, were divided into three groups (8 rats in each group): a control group fed normal saline (NS), a lower dose rosiglitazone group (1 mg.kg(-1).d(-1)) and a higher dose rosiglitazone group (4 mg.kg(-1).d(-1)) for 12 weeks. The mRNA expression of IKK-beta in KCs and COX-2 in livers were measured using reverse transcription-polymerase chain reaction (RT-PCR). The IKK-beta protein in KCs and the NF-kappa B activity of hepatic tissues were determined respectively by Western blot and electrophoretic mobility shift assay (EMSA). The concentration of tumor necrosis factor alpha (TNF alpha) in the supernatant of KCs cultures and serum of the rats was quantified by enzyme linked immunosorbent assay (ELISA).

RESULTSLPS significantly increased the expression of IKK-beta mRNA and protein in the KCs and the concentration of TNF alpha in the supernatant of the KCs cultures. The expressions of COX-2 mRNA and protein were more obvious in rats with NASH than those in the normal control group, and the binding activity of NF-kB correlated positively with the expression of COX-2 in the livers and the level of serum TNF alpha of model rats as well. Rosiglitazone blocked the expression of IKK-beta mRNA and protein induced by LPS in KCs, and also inhibited NF-kappa B activation and reduced COX-2 expression in the rats.

CONCLUSIONSPPAR-gamma specific agonist rosiglitazone can play an anti-inflammatory role by IKK-beta/I kappa B/NF-kappa B/TNF alpha signal ways, and minimize inflammatory reaction at cellular and molecular levels. This may help to provide a new idea for treating NASH effectively.

Animals ; Cyclooxygenase 2 ; metabolism ; Fatty Liver ; drug therapy ; metabolism ; pathology ; Hypoglycemic Agents ; therapeutic use ; I-kappa B Kinase ; metabolism ; Male ; NF-kappa B ; metabolism ; PPAR gamma ; agonists ; Rats ; Rats, Wistar ; Signal Transduction ; Thiazolidinediones ; therapeutic use ; Tumor Necrosis Factor-alpha ; metabolism

Novel organic cation transporter-2 (OCTN2), a member of the organic cation transporter family, may transport carnitine and multiple organic cationic drugs. Thus OCTN2 possesses substantial roles in physiology and pharmacology. A number of researches have proven that many factors can regulate the expression and/or function of OCTN2 via different pathways, and then may affect the homeostasis and disposition of drugs. This paper reviews recent progresses in this field.
Animals ; Biological Transport ; Carnitine ; metabolism ; Carrier Proteins ; physiology ; Clofibrate ; pharmacology ; Colitis ; metabolism ; Homeostasis ; drug effects ; Humans ; Mutation ; Organic Cation Transport Proteins ; genetics ; metabolism ; physiology ; PPAR alpha ; agonists ; RNA, Messenger ; metabolism ; Solute Carrier Family 22 Member 5

In the present study, we show that the expression of type 2 glucose transporter isoform (GLUT2) could be regulated by PPAR-gamma in the liver. Rosiglitazone, PPAR-gamma agonist, activated the GLUT2 mRNA level in the primary cultured hepatocytes and Alexander cells, when these cells were transfected with PPAR-gamma/RXR-alpha. We have localized the peroxisome proliferator response element in the mouse GLUT2 promoter by serial deletion studies and site-directed mutagenesis. Chromatin immunoprecipitation assay using ob/ob mice also showed that PPAR-gamma rather than PPAR-alpha binds to the -197/-184 region of GLUT2 promoter. Taken together, liver GLUT2 may be a direct target of PPAR-gamma ligand contributing to glucose transport into liver in a condition when PAPR-gamma expression is increased as in type 2 diabetes or in severe obesity.
Animals ; Cells, Cultured ; Chromatin Immunoprecipitation ; Gene Expression Regulation ; Genes, Reporter ; Hepatocytes/*metabolism ; Liver/metabolism ; Male ; Mice ; Mice, Inbred ICR ; Mice, Transgenic ; Monosaccharide Transport Proteins/*biosynthesis/genetics ; Mutagenesis, Site-Directed ; PPAR alpha/genetics/metabolism ; PPAR gamma/agonists/genetics/*metabolism ; *Promoter Regions (Genetics) ; Protein Isoforms/biosynthesis ; Research Support, Non-U.S. Gov't ; *Response Elements ; Thiazolidinediones/pharmacology

The aim of this study is to establish a simple and stable model like poloxamer 407 (P-407)-induced dyslipidemia of golden hamster model, and investigate the mechanism of lipid metabolism disturbance in this model. PPARalpha agonist and HMG-CoA reductase inhibitor were administrated to validate the efficacy on regulating lipid metabolism in the dyslipidemia golden hamster model. Six weeks male golden hamsters were chosen to inject P-407 intraperitoneally at a bolus dose of 300 mg x kg(-1), an intermittent injection at a dose of 200 mg x kg(-1) every 72 hours after the bolus. The results showed that P-407-induced golden hamster model characterized as increased serum triglyceride (TG), total cholesterol (TC), free cholesterol (free-CHO), cholesteryl ester (CE), free fatty acids (FFA) and apoB levels, and the hyperlipidemia state maintained at a stable level persistently. Meanwhile, augmented malondialdehyde (MDA) and nitric oxide (NO) level was observed. LCAT and SR-B I mRNA levels in liver of model group were down-regulated (expression ratio is 0.426; 0.783), while HMG-CoA reductase mRNA level was up-regulated (expression ratio is 1.493) compared with those of the normal group. The serum cholesterol and triglyceride levels were significantly lower in P-407-induced dyslipidemia hamster model after treated with atorvastatin (Ato) at a dose of 50 mg x kg(1) or fenofibrate (Fen) at 100 mg x kg(-1) for two weeks. These findings suggest that serum lipid distribution in dyslipidemia golden hamster is similar to that of human, and which may be relevant to the disturbance of the enzymes expression involved in lipid metabolism in liver. Results obtained from this study support the concept that dyslipidemia golden hamster may be an adequate animal model to evaluate the efficacy of lipid-lowering agents.
Animals ; Anticholesteremic Agents ; pharmacology ; Atorvastatin Calcium ; CD36 Antigens ; genetics ; metabolism ; Cricetinae ; Disease Models, Animal ; Dyslipidemias ; chemically induced ; metabolism ; Fenofibrate ; pharmacology ; Heptanoic Acids ; pharmacology ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Hypolipidemic Agents ; pharmacology ; Lipid Metabolism ; Liver ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mesocricetus ; Nitric Oxide ; metabolism ; PPAR alpha ; agonists ; Phosphatidylcholine-Sterol O-Acyltransferase ; genetics ; metabolism ; Poloxamer ; Pyrroles ; pharmacology ; RNA, Messenger ; metabolism ; Superoxide Dismutase ; metabolism