Chlorogenic acid (CGA) upregulates the transcriptional expression of PPARγ, LXRα, ABCA1 and ABCG1 in RAW264.7 cells.

<p>Real-time PCR was conducted with gene specific oligonucleotide primers. The amplification of β-actin served as the internal control. Values are means ± SEM of at least three experiments. *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001 vs. control.</p

Chlorogenic Acid Protects against Atherosclerosis in ApoE^−/− Mice and Promotes Cholesterol Efflux from RAW264.7 Macrophages

<div><p>Chlorogenic acid (CGA) is one of the most abundant polyphenols in the human diet and is suggested to be a potential antiatherosclerotic agent due to its proposed hypolipidemic, anti-inflammatory and antioxidative properties. The aim of this study was to evaluate the effect of CGA on atherosclerosis development in ApoE^−/− mice and its potential mechanism. ApoE^−/− mice were fed a cholesterol-rich diet without (control) or with CGA (200 and 400 mg/kg) or atorvastatin (4 mg/kg) for 12 weeks. During the study plasma lipid and inflammatory parameters were determined. Treatment with CGA (400 mg/kg) reduced atherosclerotic lesion area and vascular dilatation in the aortic root, comparable to atorvastatin. CGA (400 mg/kg) also significantly decreased plasma levels of total cholesterol, triglycerides and low-density lipoprotein-cholesterol as well as inflammatory markers. Supplementation with CGA or CGA metabolites-containing serum suppressed oxidized low-density lipoprotein (oxLDL)-induced lipid accumulation and stimulated cholesterol efflux from RAW264.7 cells. CGA significantly increased the mRNA levels of PPARγ, LXRα, ABCA1 and ABCG1 as well as the transcriptional activity of PPARγ. Cholesterol efflux assay showed that three major metabolites, caffeic, ferulic and gallic acids, significantly stimulated cholesterol efflux from RAW264.7 cells. These results suggest that CGA potently reduces atherosclerosis development in ApoE^−/− mice and promotes cholesterol efflux from RAW264.7 macrophages. Caffeic, ferulic and gallic acids may be the potential active compounds accounting for the <i>in vivo</i> effect of CGA.</p></div

The serum lipid profile.

<p>TC: total cholesterol; TG: triglyceride; HDL-c: high-density lipoprotein-cholesterol; LDL-c; low-density lipoprotein-cholesterol; Atorva: atorvastatin; CGA: chlorogenic acid.</p><p>^*<i>p</i><0.05.</p><p>^**<i>p</i><0.01 v.s. ApoE.</p>−/−<p>mice (n = 6).</p><p>The serum lipid profile.</p

Chlorogenic acid (CGA) inhibits oxidized low-density lipoprotein (oxLDL)-elicited foam cell formation in RAW264.7 cells.

<p>RAW264.7 cells were elicited by oxLDL for 24 h with or without supplementation of CGA or atorvastatin. Cells were then stained with Oil red O, and the representative staining pictures (A), the aborptance at 358 nm (B), and intracellular totale cholesterol content (C) were acquired. Bar  =  50 µm. Values are means ± SEM of at least three experiments. ^###<i>p</i><0.001 oxLDL group vs. blank group; *<i>p</i><0.05, **<i>p</i><0.01 test group vs. oxLDL group. Atorv.  =  atorvastatin, CGA  =  Chlorogenic acid,</p

Chlorogenic acid (CGA) metabolites-containing serum from CGA-treated normal mice decreases lipid accumulation and stimulates cholesterol efflux from RAW264.7 cells.

<p>Normal C57BL/6J mice were orally gavaged with 400 mg/kg of CGA or equal volume of distilled water for 3 days and blood was collected at 45 min after the final treatment. 1% (v/v) of serum from CGA-treated normal mice (S_CGA) significantly decreased oxLDL-induced neutral lipid accumulation (A) and total cholesterol (B), and stimulates cholesterol efflux (C) in RAW264.7 cells as compared with that from distilled water-treated animals (S_NC). Values are means ± SEM of at least three experiments. **<i>p</i><0.01, ***<i>p</i><0.001.</p

A new diterpene from <i>Clinopodium chinense</i>

<div><p>A new abietane diterpene, named as 3β-hydroxy-12-<i>O</i>-β-d-glucopyranosyl-8,11,13-abietatrien-7-one (<b>1</b>), together with four known flavonoids, was isolated from the hot water extract of the aerial parts of <i>Clinopodium chinense</i>. Their structures were determined by analysing the spectroscopic data including 1D, 2D NMR and HR-ESI-MS. Compound <b>1</b> tested against HepG-2 and A549 cancer cell lines expressed weak cytotoxicity. Cardioprotective effects of compounds <b>2</b>–<b>5</b> against H₂O₂-induced apoptosis in H9c2 cells were also evaluated; compounds <b>2</b> and <b>3</b> exhibited moderate cardioprotective effect.</p></div

Caffeic acid, ferulic acid and gallic acid are active in promoting cholesterol efflux from RAW264.7 cell mediated by HDL.

<p>Cells were equilibrated with NBD-cholesterol for 12 h then incubated in serum-free DMEM medium containing HDL and 10 µM of respective compound for 6 h. Cholesterol efflux was expressed as a percentage of fluorescence in the medium relative to the total amounts of fluorescence detected in cells and the medium. Rosiglitazone (5 µM) was used as positive control. Values are means ± SEM of at least three experiments. *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001 vs. control. Rosigli  =  rosiglitazone.</p

Treatment with chlorogenic acid (CGA) reduces intracellular levels of IL-1β (A), IL-6 (B) and TNFα (C) elicited by LPS in RAW264.7 cells.

<p>Values are means ± SEM of at least three experiments. ^###<i>p</i><0.001 LPS+vehicle group vs. vehicle group; *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001 test group vs. LPS+vehicle group. Atorv.  =  atorvastatin, CGA  =  Chlorogenic acid, LPS  =  lipopolysaccharides.</p

Treatment with chlorogenic acid (CGA) for 12 weeks reduces the serum levels of interleukin-6 (IL-6) (A), interleukin-8 (IL-8) (B), tumor necrosis factor α (TNFα) (C) and monocyte chemotactic protein-1 (MCP-1) (D) in ApoE^−/− mice.

<p>Values are means±SEM. *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001 vs. ApoE^−/− group. Atorv.  =  atorvastatin, CGA  =  Chlorogenic acid.</p

Chlorogenic acid (CGA) increases the transcriptional activity of PPARγ.

<p>The transcriptional activity of PPARγ was assessed by transactivation reporter assay in 293T cells. Rosiglitazone (5 µM) was used as positive control. Values are means ± SEM of at least three experiments. *<i>p</i><0.05, **<i>p</i><0.01 vs. control. Rosigli  =  rosiglitazone, CGA  =  Chlorogenic acid.</p