Yuling Tian

Activation of cannabinoid CB2 receptor ameliorates atherosclerosis associated with suppression of adhesion molecules.

Objective: Adhesion molecules have been implicated in the development and progression of atherosclerosis. Cannabinoids have been reported to modulate the migration and adhesion molecules expression of various cell types. Here we examined the effects of WIN55212-2, a cannabinoid receptor 1 (CB1-R)/cannabinoid receptor 2 (CB2-R) agonist on the development of atherosclerotic lesions in apolipoprotein E-deficient (ApoE-/-) mice, which are vulnerable because of their high plasma cholesterol and triacylglycerol levels, focusing on the expression of endothelial adhesion molecules. Methods and Results: In the aorta of ApoE-/- mice, WIN55212-2 significantly reduced aortic root plaque area. The mechanism for this seemed to be reduced infiltration of macrophages into the atherosclerotic plaque which was also associated with reduced expression of vascular cellular adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and P-selectin in the aorta. In vitro studies revealed reduced cell adhesion of a monocytic cell line (U937) to human umbilical vein endothelial cells after incubation with WIN55212-2. The reduction in macrophage adhesion also correlated with significant reductions in the expression of VCAM-1, ICAM-1, and P-selectin, indicating that reduced infiltration of macrophages in atherosclerotic plaques may occur as a result of the direct effect of WIN55212-2 on adhesion molecules in macrophages and endothelial cells. Conclusion: In conclusion, WIN55212-2 seems to have direct anti-atherosclerotic effects in an animal model of atherosclerosis. These effects were at least partly due to effects on the expression of VCAM-1, ICAM-1, and P-selectin, which led to reduced macrophage adhesion and infiltration. Furthermore, the protective effects completely blocked by the highly selective CB2 receptor antagonist AM630 suggest that these beneficial effects of WIN55212-2 may be mediated through the CB2 receptor.

High Glucose Promotes Intracellular Lipid Accumulation in Vascular Smooth Muscle Cells by Impairing Cholesterol Influx and Efflux Balance

AIMS High glucose promotes macrophage-derived foam cell formation involved in increased influx or reduced efflux of lipids. The aim of this study is to investigate the influence of hyperglycaemia on foam cell transformation of vascular smooth muscle cells (VSMCs) and possible mechanisms contributing to these effects. METHODS AND RESULTS The results showed that high glucose increased the expression of CD36, a regulator of lipid influx, and suppressed the expression and activity of the adenosine triphosphate-binding cassette (ABC) transporter ABCG1, a regulator of cholesterol efflux to high-density lipoprotein, in a dose- and time-dependent manner. However, cholesterol efflux to lipid-free apoAI was not impaired. VSMCs exposed to high glucose readily developed into lipid-loaded cells, as demonstrated by Oil Red O staining and cholesterol content analysis. In addition, high glucose-induced down-regulation of ABCG1 was reversed by nuclear factor-kappaB (NF-kappaB) inhibitors BAY 11-7085 and tosyl-phenylalanine chloromethyl ketone and by the antioxidant N-acetyl-L-cysteine (NAC). This reversal was accompanied by reduced cellular lipid content. Also, NAC and NF-kappaB inhibitors can effectively block the high glucose-induced activity of NF-kappaB binding to DNA and/or peroxide production. CONCLUSION These results suggested that hyperglycaemia-induced foam cell formation in VSMCs was related to the imbalanced lipid flux by increasing CD36-mediated modified low-density lipoprotein uptake and reducing ABCG1-regulated cellular cholesterol efflux. Moreover, this effect was associated with increased oxidative stress and activated NF-kappaB pathway signalling.

Metformin ameliorates the proinflammatory state in patients with carotid artery atherosclerosis through sirtuin 1 induction

Metformin is a widely used classic antidiabetic drug. However, its clinical pharmacologic mechanism remains poorly understood. In the present study, we investigated the anti-inflammatory effects of metformin on circulating peripheral blood mononuclear cells (MNCs) of patients with carotid artery atherosclerosis (AS). A total of 42 patients with carotid artery AS were randomly assigned to metformin (500 mg twice a day; Met; n = 21) or placebo control (Con; n = 21) groups. After 12 weeks of treatment, plasma concentrations of high-sensitivity C-reactive protein (hs-CRP), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α) significantly decreased in the Met group compared with the Con group. In addition, treatment with metformin significantly reduced the expression of IL-6 and TNF-α at the messenger RNA level and attenuated nuclear factor kappa B (NF-κB) DNA binding activity in MNCs. Intriguingly, metformin did not alter the expression of NF-κB p65 subunit, but markedly inhibited its acetylation. Furthermore, metformin significantly induced sirtuin 1 (SIRT1) expression in MNCs. Moreover, we found that metformin treatment dramatically induced SIRT1 expression, blocked p65 acetylation, and inhibited NF-κB activity and the expression of inflammatory factors in MNCs in vitro. We conclude that metformin has a novel direct protective role to ameliorate the proinflammatory response through SIRT1 induction, p65 acetylation reduction, NF-κB inactivation, and inflammatory inhibition in peripheral blood MNCs of patients with carotid artery AS.

Multiple roles of SOCS proteins: differential expression of SOCS1 and SOCS3 in atherosclerosis.

Pro-inflammatory cytokines play a key pathogenic role in atherosclerosis, which are induced by the Janus kinase/signal transducer and activator of transduction (JAK/STAT) pathway. Furthermore, the JAK/STAT pathway is negatively regulated by the suppressor of cytokine signaling (SOCS) proteins. However, the change in SOCS expression levels and the correlation between SOCS expression and cholesterol levels in atherosclerosis is not yet well understood. To this end, a mouse model of atherosclerosis was established using apolipoprotein-deficient (ApoE(-/-)) mice. The mice were fed either a chow or high-fat diet. The mRNA and protein expression of SOCS1 and SOCS3 in plaque and vessels were determined at different time points. Furthermore, SOCS1 and SOCS3 mRNA expression was detected in the peripheral blood mononuclear cells (PBMCs) obtained from 18 male subjects with no coronary heart disease (non-CHD) population. The expression of SOCS1 in the ApoE(-/-) mice first increased and then decreased and the high-fat diet accelerated the appearance of the peak; the expression of SOCS3 increased with the increased feeding duration, and this trend was more pronounced in the mice fed the high-fat diet. SOCS1/CD68 and SOCS3/CD68 showed opposite trends in expression with the increased duration of the high-fat diet. Interleukin-6 (IL-6) expression in the main aorta of the ApoE(-/-) mice fed the high-fat diet also increased with the increased feeding duration. In the non-CHD population, the total serum cholesterol levels positively correlated with SOCS3 mRNA expression in the PBMCs (r=0.433, P=0.012). These results demonstrate the differential expression of SOCS1 and SOCS3 in atherosclerosis and suggest that SOCS3, together with IL-6 may promote the formation and development of atherosclerosis.

Early use of granulocyte colony stimulating factor improves survival in a rabbit model of chronic myocardial ischemia

BACKGROUND Granulocyte colony stimulating factor (G-CSF) improves the survival of animals with myocardial infarction by inducing bone marrow stem cell mobilization and homing to infarcted areas. However, its precise mechanisms and direct effects on the ischemic myocardium remain unclear. In this study we investigated the direct effects and mechanisms of G-CSF in a rabbit model of chronic myocardial ischemia. METHODS Myocardial ischemia models were created by partial ligation of the left anterior descending coronary artery in Japanese white male rabbits. Rabbits were subcutaneously injected with 10 μg/kg of G-CSF (G-CSF group) or saline (control group) for 6 days after myocardial ischemia. Direct effects of G-CSF were analyzed by immunohistochemistry and terminal dUTP nick end-labeling (TUNEL). RESULTS Rabbits in the G-CSF group exhibited 75% survival compared to 40% in the control group (p<0.05). Immunohistochemistry of the ischemic myocardium showed increased homing of CD34+ cells on day 7 post-surgery and more vessels on day 28 post-surgery by anti-von Willebrand factor staining in the G-CSF group compared with the control group. Furthermore, an increased percentage of CD34+ cells were observed in peripheral blood and upregulation of vascular endothelial growth factor expression in ischemic tissue in the G-CSF group compared with the control group. TUNEL showed that the apoptotic index in the ischemic myocardium decreased in the G-CSF group compared with the control group on day 28 post-surgery. CONCLUSIONS In addition to increasing stem cell mobilization and homing to ischemic myocardium, G-CSF treatment after myocardial ischemia improves survival by accelerating neovascularization and reducing apoptosis.

17β-Estradiol Suppresses the Macrophage Foam Cell Formation Associated with SOCS3

Evidence from clinical trials and animal experiments has shown that estrogen has anti-atherosclerotic effects when administered to young women or experimental animals. The mechanisms involve the modulation of vascular inflammation, growth factor expression, and oxidative stress injured arteries. However, whether estrogen modulates the foam cell formation in plaque remains unknown. Here, we investigated the effects of 17β-estradiol (E2) on cholesterol efflux in vivo and in vitro. ApoE null mice underwent an ovariectomy at 5(th) week of age and then were treated with E2 or vehicle for the following 8 weeks. Compared with the vehicle-treated mice, the serum total cholesterol level, atherosclerotic plaque size, and lipid deposits were decreased and meanwhile ATP-binding cassette transporter A1 (ABCA1) expression in the plaque was increased in mice with E2 treatment. E2 also increased suppressor of cytokine signaling 3 (SOCS3) expression in the atherosclerotic plaques and in RAW264.7 cells. In vitro, E2 treatment reversed janus kinase/signal transducers and activators of transcription (JAK/STAT)-inhibited ABCA1 expression in RAW264.7 cells but had no effect on ABCA1 expression in SOCS3 knockdown cells. SOCS3 overexpression elevated ABCA1 expression through the inhibition of JAK2/STAT3 phosphorylation. Finally, we also found that E2 enhanced the cholesterol efflux to apoA I in RAW264.7 cells. In summary, E2 reduces atherosclerosis in ApoE null mice associated with upregulating ABCA1 expression and modulating the cholesterol efflux, which are dependent on SOCS3 upregulation. These results provide new insight into the athero-protective effects of estrogen.

CD34+ cell mobilization and upregulation of myocardial cytokines in a rabbit model of myocardial ischemia

BACKGROUND Studies have suggested that myocardial infarction may induce bone marrow stem cell mobilization and homing to the infarcted area, contributing to myocardial repair and tissue regeneration. Despite some encouraging results using stem cell therapy for myocardial regeneration in humans and animals, the mechanisms behind this activity remain unclear. In this study, we investigate stem cell mobilization and homing in ischemic myocardium, and investigate the involvement of cytokines TNFα and VEGF in this process. METHODS AND RESULTS Myocardial ischemia models were created by partial ligation of the left anterior descending coronary artery in Japanese white male rabbits. Immunohistochemistry analysis of ischemic myocardium showed the presence of VEGF and TNFα along with homing of CD34 positive (CD34+) cells to the region in the 7 days following surgery. During the same period, an increase in percentage of CD34+ cells in peripheral blood and in VEGF and TNFα mRNA expression in ischemic tissue was observed in animals that underwent partial LAD ligation. Terminal dUTP nick end-labeling (TUNEL) showed that cell apoptosis in the ischemic myocardium decreased between days 7 and 28 following surgery. None of these changes were observed in animals that underwent sham operation. CONCLUSIONS In the early stages of myocardial ischemia, bone marrow stem cells are mobilized and home to ischemic myocardium with a concomitant increase in expression of cytokines VEGF and TNFα. Furthermore, cell apoptosis occurs in the ischemic myocardium, possibly due to the activity of TNFα which is thought to induce cardiomyocyte apoptosis.

Pioglitazone modulates the balance of effector and regulatory T cells in apolipoprotein E deficient mice.

BACKGROUND AND AIMS Pioglitazone (PIO) affects T cell-mediated immunity through actions of peroxisome proliferator activated receptor γ (PPARγ). Effector and regulatory T cells control the development of atherosclerosis, a chronic inflammatory disease affecting the arterial blood vessels. The aim of this study was to examine whether PIO ameliorates atherosclerosis by altering the balance of effector and regulatory T cells. METHODS AND RESULTS To explore the effect of PIO on early and advanced atherosclerosis, apolipoprotein E deficient (ApoE-/-) mice were fed western diet and received PIO (20 mg/kg/day) by gastric gavage at 6 or 14 weeks of age, respectively for 8 weeks. Data showed PIO markedly inhibited early fatty streak formation. Further, although the advanced fibrofatty plaque sizes were not significantly reduced, the numbers of smooth muscle cells within lesions were increased and higher collagen concentrations were produced. In general, macrophage expression in lesions was decreased. Additionally, the expression of Foxp3(+) cells was increased in lesions and spleens in mice at all PIO treatment stages, whereas the CD4(+)IFN-γ(+)/CD4(+)IL-4(+) cell ratios were reduced. CONCLUSION PIO inhibited early atherosclerotic lesion formation and increased the stability of advanced atherosclerotic plaques in ApoE-/- mice, which was associated with altering the balance of effector and regulatory T cells.