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[Objective]To investigate the effects of different doses of anthocyanins Cy-3-g on serum lipids,platelet-derived chemokines,including MIF and CXCL12 together with their receptors CXCR4 and CXCR7.[Methods]Male ApoE-/-mice were distributed to 5 groups(n=15 per group),and fed with standard diet,high fat diet(HFD),HFD with 200 mg/kg Cy-3-g(low),HFD with 400 mg/kg Cy-3-g(middle),HFD with 800 mg/kg Cy-3-g(high)respectively for 16 weeks. The changes of body weight and food intake were recorded weekly. At the end of the experiment term,the serum lipids(triglyceride,cholesterol,HDL-C,LDL-C)were detected by kits. Arteries were separated to determine plaque histology by Oil-Red-O stain.MIF,CXCL12 and CCL2 in serum were detected by ELISA kits.The expression of CXCR4 and CXCR7 on the surface of leukocytes were tested via flow cytometry. Tail bleeding time was measured mean-while.[Results]Compared with the HFD group,the levels of serum lipids in medium(400 mg/kg)and high(800 mg/kg)Cy-3-g groups were significantly decreased(P<0.05). The plaque area of carotid artery was decreased in high Cy-3-g group(P<0.05).Cy-3-g at all doses significantly reduced the serum concentrations of CXCL12 and CCL2(P<0.002).Cy-3-g of medium(400 mg/kg)and high(800 mg/kg)dose significantly inhibited the expression of CXCR4 and CXCR7 on leukocyte surface(P<0.05). Cy-3-g does not prolong the tail bleeding time.[Conclusions]Anthocyanin Cy-3-g inhibits chemokine CXCL12,CCL2 in serum,and the expression of CXCR4 and CXCR7 on leukocytes without bleeding risk in ApoE-/-mice.
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OBJECTIVE: Platelet factor 4 tetramers (CXCL4 chemokine) form complexes with ß2glycoprotein I (ß2GPI), recognized by anti-ß2GPI antibodies leading to platelet activation in antiphospholipid syndrome (APS), either primary (PAPS) or secondary (SAPS). Increased plasma levels of CXCL4 may favor this process; therefore we measured plasma levels of CXCL4, a CXCL4 variant (CXCL4L1) and as controls, platelet-derived chemokines CXCL7 (NAP-2) and CCL5 (RANTES), in APS, and disease controls such as patients with systemic lupus erythematosus (SLE) coronary artery disease (CAD) and healthy donors (HDs). METHODS: Plasma samples and platelets were isolated from patients with APS (n = 87), SLE (n = 29), CAD (n = 14) and 54 HDs. Plasma levels of CXCL4, CXCL4L1, CXCL7 and CCL5 as well as intracellular platelet CXCL4 and CXCL4L1 were measured using ELISA. Platelet CXCL4 and CXCL4L1 RNA levels were determined by RT-PCR. RESULTS: CXCL4, CXCL7 (NAP-2) and CCL5 (RANTES) plasma levels were significantly higher in patients with APS compared to both control groups (SLE, CAD) and HDs. CXCL4L1 plasma levels were also significantly higher in APS than in SLE and HDs, but lower from that of CAD patients. Statistically significant concordance was detected between CXCL4 and CXCL7 (p < 0.0001) or CCL5 (p < 0.0001) plasma levels in patients with APS, either PAPS or SAPS. CXCL4L1 plasma levels were inversely correlated with CXCL4 (P = 0.0027), CXCL7 (p = 0.012) and CCL5 (p = 0.023) in PAPS and positively with CXCL4 (p = 0.0191), CCL5 (p < 0.0001) and CXCL7 (P < 0.0001), in SAPS. Levels of CXCL4, CXCL4L1, CXCL7 and CCL5 were divided in "high" (exceeding a level defined as the mean of HDs and 3 SD) and "low" (below this level); The "CXCL4L1 high" group was characterized by increased IgG aCL, (p = 0.0215), double antibody positivity (either aCL or anti-ß2GPI plus LA), (p = 0.0277), triple antibody positivity (aCL plus anti-ß2GPI plus LA), (p = 0.0073) and thrombocytopenia (p = 0.0061), as well as with at least 1 thrombotic event or the last 5 years (p = 0.0001), or more than 3 thrombotic events ever (p = 0.0151). CONCLUSIONS: Chemokines associated with platelet activation and immune cell chemotaxis were found to be elevated in APS patients' plasma and may contribute to the pathogenesis of the syndrome. High CXCL4L1 plasma levels are associated with the clinical expression of APS and should be prospectively evaluated as a biomarker.
