Legumain: a potential biomarker for atherosclerosis.

Atherosclerosis is a lipid-driven chronic inflammatory disease that poses a serious threat to health. Legumain (LGMN), also known as asparagine endonuclease, is a new type of cysteine proteases that can specifically hydrolyze substrate molecules containing asparagine residues. It has anti-apoptotic effects in mammals and plays an antigen-presenting role in inflammatory response. Several studies have found that LGMN can activate multiple signal pathways to promote cell apoptosis and migration, inflammatory response, and the development of atherosclerosis. Importantly, LGMN exerts pro-atherogenic effects by participating in a variety of pathophysiological mechanisms of atherosclerosis, including vascular remodeling, inflammatory response, plaque stability, and the degradation of extracellular matrix. In the present review, we describe the LGMN distribution, structure, generation, and functional partners. Furthermore, we summarize the relationship between LGMN and atherosclerosis. Based on the relationship between LGMN and atherosclerosis, LGMN may be a potential biomarker for atherosclerosis.

Microtubule affinity regulating kinase 4: A promising target in the pathogenesis of atherosclerosis.

Microtubule affinity regulating kinase 4 (MARK4), an important member of the serine/threonine kinase family, regulates the phosphorylation of microtubule-associated proteins and thus modulates microtubule dynamics. In human atherosclerotic lesions, the expression of MARK4 is significantly increased. Recently, accumulating evidence suggests that MARK4 exerts a proatherogenic effect via regulation of lipid metabolism (cholesterol, fatty acid, and triglyceride), inflammation, cell cycle progression and proliferation, insulin signaling, and glucose homeostasis, white adipocyte browning, and oxidative stress. In this review, we summarize the latest findings regarding the role of MARK4 in the pathogenesis of atherosclerosis to provide a rationale for future investigation and therapeutic intervention.

TIGAR mitigates atherosclerosis by promoting cholesterol efflux from macrophages.

BACKGROUND AND AIMS: TP53-induced glycolysis and apoptosis regulator (TIGAR) is now characterized as a fructose-2,6-bisphosphatase to reduce glycolysis and protect against oxidative stress. Recent studies have demonstrated that TIGAR is associated with cardiovascular disease. However, little is known about its role in atherosclerogenesis. In this study, we aimed to investigate the effect of TIGAR on atherosclerosis and explore the underlying molecular mechanism. METHODS: The Gene Expression Omnibus (GEO) datasets were used to analyze the differential expression of relative proteins. THP-1-derived macrophages were used as an in vitro model and apolipoprotein E-deficient (Apoe-/-) mice were used as an in vivo model. [3H] labeled cholesterol was used to assess the capacity of cholesterol efflux and reverse cholesterol transport (RCT). Both qPCR and Western blot were used to evaluate the mRNA and protein expression, respectively. Lentiviral vectors were used to disturb the expression of TIGAR in vitro and in vivo. Oil Red O, hematoxylin-eosin, and Masson staining were performed to evaluate atherosclerotic plaques in Apoe-/- mice fed a Western diet. Conventional assay kits were used to measure the levels of reactive oxygen species (ROS), plasma lipid profiles and 27-hydroxycholesterol (27-HC). RESULTS: Our results showed that TIGAR is increased upon the formation of macrophage foam cells and atherosclerosis. TIGAR knockdown markedly promoted lipid accumulation in macrophages. Silencing of TIGAR impaired cholesterol efflux and down-regulated the expression of ATP-binding cassette transporter A1 (ABCA1) and ABCG1 by interfering with liver X receptor α (LXRα) expression and activity, but did not influence cholesterol uptake by macrophages. Additionally, this inhibitory effect of TIGAR deficiency on cholesterol metabolism was mediated through the ROS/CYP27A1 pathway. In vivo experiments revealed that TIGAR deficiency decreased the levels of ABCA1 and ABCG1 in plaques and aorta and impaired the capacity of RCT, thereby leading to the progression of atherosclerosis in Apoe-/- mice. CONCLUSIONS: TIGAR mitigates the development of atherosclerosis by up-regulating ABCA1 and ABCG1 expression via the ROS/CYP27A1/LXRα pathway.

Long non-coding RNA PCA3 inhibits lipid accumulation and atherosclerosis through the miR-140-5p/RFX7/ABCA1 axis.

OBJECTIVE: The purpose of this study was to explore the role of long noncoding RNA (lncRNA) prostate cancer antigen 3 (PCA3) in atherosclerosis and the underlying mechanism. METHODS: The Gene Expression Omnibus (GEO) datasets were used to divide differentially expressed lncRNAs, microRNAs (miRNAs), and mRNAs. The expression of PCA3, miR-140-5p, RFX7 and ABCA1 were determined by qPCR or Western blot in ox-LDL-treated macrophages. Macrophage lipid accumulation s was evaluated using the Oil Red O staining and high-performance liquid chromatography. Target relationships among PCA3, miR-140-5p, RFX7, and ABCA1 promoter area were validated via dual-luciferase reporter gene assay or chromatin immunoprecipitation assay. The apoE-/- mouse model in vivo was designed to evaluate the effect of PCA3 on the reverse cholesterol transport (RCT) and atherosclerosis. RESULTS: PCA3 was down-regulated in foam cells, whereas miR-140-5p was highly expressed. Overexpression of PCA3 promoted ABCA1-mediated cholesterol efflux and reduced lipid accumulation in macrophages. Besides, RFX7 bound to the ABCA1 promoter and increased ABCA1 expression. Targeted relationships and interactions on the expression between miR-140-5p and PCA3 or RFX7 were elucidated. PCA3 up-regulated ABCA1 expression by binding to miR-140-5p to up-regulate RFX7 and ABCA1 expression in macrophages. PCA3 promoted RCT and impeded the progression of atherosclerosis by sponging miR-140-5p in apoE-/- mice. Meanwhile, miR-140-5p also inhibit ABCA1 expression via downregulation of RFX7 to impede RCT and aggravate atherosclerosis. CONCLUSIONS: lncRNA PCA3 promotes ABCA1-mediated cholesterol efflux to inhibit atherosclerosis through sponging miR-140-5p and up-regulating RFX7.

Artesunate inhibits atherosclerosis by upregulating vascular smooth muscle cells-derived LPL expression via the KLF2/NRF2/TCF7L2 pathway.

Lipoprotein lipase (LPL) plays a central role in hydrolyzing triglyceride and its deficiency leads to atherosclerosis. Artesunate (ART), a derivative of artemisinin, has been demonstrated that ART reduces the formation of atherosclerotic plaques. However, it remains unclear whether ART-alleviated atherosclerotic lesion is involved in regulating lipid metabolism. ApoE-/- mice were fed a high-fat diet to form atherosclerotic plaques and then injected with artesunate or not. Oil Red O, HE and Masson staining were performed to assess atherosclerotic plaques. Both Western blot and qRT-PCR were applied to detect protein expression. The Luciferase reporter gene and Chromatin immunoprecipitation assays were used to assess the interaction between proteins. Immunofluorescence assay was performed to show the localization of target proteins. In vitro, our data shown that ART increased LPL expression and inhibition of NRF2 blocked the binding of TCF7L2 to LPL promoter region in VSMCs. Downregulated Klf2 could decrease the nuclear enrichment of NRF2, TCF7L2 and LPL expression. In vivo, ART decreased atherosclerotic plaque formation and increased VSMC counts and LPL expression within atherosclerotic plaques. We observed the reduced tendency of serum lipids, and increased in serum LPL activity in mice. In support of vitro data, the markedly increased KLF2, TCF7L2 and LPL expression have been detected in aorta. Our study suggests that ART may be a novel therapeutic drug for inhibition of atherosclerotic plaque formation. The molecular mechanism may involve in upregulation of LPL expression via the KLF2/NRF2/TCF7L2 pathway in VSMCs.

[Protective effects of intermedin preconditioning on hypoxic injury in rat's cardiac myocytes].

OBJECTIVE: To observe the effects of intermedin preconditioning on hypoxic injury in rat's cardiac myocytes and to provide the hypothetical mechanism of sudden cardiac death in the field of forensic pathology. METHODS: The H9c2 cultured rat cardiac myocytes were randomly divided into control group, hypoxia group and IMD group. The myocardial cell viability, cellular ultrastructure, intracellular calcium concentration and apoptosis rate were determined by MTT assay, transmission electron microscopy, laser scanning confocal microscope and flow cytometry, respectively. RESULTS: Compared with the control group, cell viability obviously decreased with inner ultrastructure injury in the hypoxia group (P<0.05), while cell viability significantly increased in the IMD group by reducing the hypoxia injury of cardiac myocytes (P<0.05). Compared with the control group, [Ca2+]i (fluorescence intensity) and apoptosis rate significantly increased in the hypoxia group, but decreased in the IMD group (P<0.05). CONCLUSION: IMD increases the cell survival rate and decreases the cell apoptosis inhibited by intracellular calcium overload from hypoxia. This finding may reveal the mechanism of protective effects of myocardial hypoxia, and provide a scientific basis for the identification sudden cardiac death.

[Distribution characteristics of soil cadmium in different textured paddy soil profiles and its relevance with cadmium uptake by crops].

Adopting atomic absorption spectrometry (novAA400), the present study investigated the distribution characteristics of soil cadmium (Cd) in different textured paddy soil profiles under rice-wheat rotation and its correlation with Cd uptake by rice and wheat in Chengdu Plain through repeatedly taking soil and plant samples at the fixed sites in the field. The results revealed that Cd in the paddy soil profiles was mainly concentrated at the plough layer (0-15 cm) that obviously featured 'Cd accumulates towards the rooting layer'. Soil total Cd and available Cd (1 mol x L(-1) MgCl2 extraction) in the profile declined with soil depths and its average values at 30-45 cm only accounted for 47.60% and 39.49% of those at 0-15 cm. The potential downward movement of Cd in the different textured soil profiles was observed as sandy loam > heavy loam > loam. There was no significant correlation between soil pH and available Cd(r = - 0.46) at 0-15 cm soil depth, while significantly negative correlations between soil pH and available Cd were observed at 15-30 cm (r = -0.78) and 30-45 cm (r = -0.86). The results further demonstrated that the Cd contents in either grain or straw of rice and wheat were not well correlated with soil total Cd at any soil depth (r = -0.092-0.383 for rice and r = 0.174-0.424 for wheat), but significantly correlated with soil available Cd at 0-15 cm and at 15-30 cm (r = 0.766*-0.953**) despite insignificant correlation at 30-45 cm (r = 0.526-0.584). It is strongly suggested that the soil available Cd can be used as a better criterion than the total soil Cd to rate Cd contaminated soils in relation to safety of agricultural products.

[Methodology improvement in background elimination in plant Cd measurement by graphite furnace atom absorption].

A problem of higher background value and lower measured Cd value exists when Cd is digested by conventional dry digestion method, measured by graphite furnace atomic absorption, and calculated with peak height numeration. This problem results in difficulty for evaluating samples with Cd contents in the neighborhood of critical values. In order to solve the problem, the present paper focused on background measurement with graphite furnace atomic absorption, modified the pre-preparation procedures of the traditional method, and screened the optimal concentration of the modifier. Results showed that addition of Mg (NO3)2 as a modifier into the samples before incineration was preferred. It could not only yield a clear solution but also reduce its background by ten times, considerably eliminating background interference. In addition, using 3% HNO3 in stead of 1 mol x L(-1) HCl as dilution for the incinerated sample could eliminate the background. The improved methods could acquire a zero concentration of measurement for the blank, re-correct the values measured by graphite furnace atomic absorption and obtain more reliable results.

[Effect of different N, P and K fertilizers on soil pH and available Cd under waterlogged conditions].

In order to tackle the problem of Cd pollution in paddy soils and investigate soil available Cd as affected by different fertilizers, incubation experiments were carried out to study the effects of different N, P and K fertilizers and pH by adding acid or base on soil available Cd under waterlogged conditions. Results revealed that soil pH increased sharply after the soil was flooded, especially at the beginning of incubation, and gradually decreased with incubation time and finally tended to approach the neutral values. The patterns of soil pH change were just opposite to those of soil available Cd, a negative correlation observed between the two. Soil flooding made the soil available Cd drop by 58.2%-84.1%. There were significant differences between different fertilizer types/varieties on soil available Cd, being most complex with N fertilizers and followed by K and P fertilizers. Among the fertilizers studied, ammonium chloride showed the unique ability in reducing soil pH and enhancing soil available Cd, and urea, single super phosphate and potassium chloride also promoted to a less extent amounts of Cd extracted from the soil. Ammonium sulfate, potassium sulfate and mono-ammonium phosphate significantly decreased soil available Cd compared to the CK treatment. Whether or not the soil was flooded, soil available Cd was highly negatively correlated with soil pH after adding acid or base (R = - 0.994 without incubation and R = - 0.919 after incubation for 60 d). The results further suggest that in the Cd polluted paddy soil, use of ammonium chloride should be avoided, S bearing fertilizers in combination with alkaline materials can be adopted, and the rice field should be flooded all the time during growing season, all the these practices can effectively lower soil available Cd.