Author + information
- Received April 5, 2017
- Revision received August 15, 2017
- Accepted August 21, 2017
- Published online October 9, 2017.
- Yang Dai, PhDa,b,
- Ying Shen, MD, PhDa,
- Qing Run Li, PhDc,
- Feng Hua Ding, MD, PhDa,
- Xiao Qun Wang, MD, PhDa,b,
- Hong Juan Liu, PhDb,
- Xiao Xiang Yan, MD, PhDa,b,
- Ling Jie Wang, MDa,b,
- Ke Yang, PhDb,
- Hai Bo Wang, PhDb,
- Qiu Jing Chen, MSca,
- Wei Feng Shen, MD, PhDa,b,∗ (, )
- Rui Yan Zhang, MD, PhDb,∗ ( and )
- Lin Lu, MD, PhDa,b,∗ ()
- aDepartment of Cardiology, Rui Jin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China
- bInstitute of Cardiovascular Diseases, Shanghai Jiaotong University School of Medicine, Shanghai, China
- cCAS Key Laboratory of Systems Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
- ↵∗Address for correspondence:
Dr. Lin Lu, Dr. Rui Yan Zhang, OR Dr. Wei Feng Shen, Department of Cardiology, Rui Jin Hospital, 197 Rui Jin Road II, Shanghai 200025, P. R. China.
Background Nonenzymatic glycation of apolipoproteins plays a role in the pathogenesis of the vascular complications of diabetes.
Objectives This study investigated whether apolipoprotein (apo) A-IV was glycated in patients with type 2 diabetes mellitus (T2DM) and whether apoA-IV glycation was related to coronary artery disease (CAD). The study also determined the biological effects of glycated apoA-IV.
Methods The authors consecutively enrolled 204 patients with T2DM without CAD (Group I), 515 patients with T2DM with CAD (Group II), and 176 healthy subjects (control group) in this study. ApoA-IV was precipitated from ultracentrifugally isolated high-density lipoprotein, and its glycation level was determined based on Western blotting densitometry (relative intensity of apoA-IV glycation). ApoA-IV NƐ-(carboxylmethyl) lysine (CML) modification sites were identified by mass spectrometry in 37 control subjects, 63 patients in Group I, and 138 patients in Group II. Saline or glycated apoA-IV (g-apoA-IV) generated by glyoxal culture was injected into apoE–/– mice to evaluate atherogenesis, and was also used for the cell experiments.
Results The relative intensity and the abundance of apoA-IV glycation were associated with the presence and severity of CAD in patients with T2DM (all p < 0.05). The experiments showed that g-apoA-IV induced proinflammatory reactions in vitro and promoted atherogenesis in apoE–/– mice through the nuclear receptor NR4A3. G-apoA-IV with mutations (K-A) at high-frequency glycation sites exhibited more weakened proinflammatory and atherogenic effects than did g-apoA-IV both in vitro and in vivo.
Conclusions ApoA-IV glycation is associated with CAD severity in patients with T2DM, and g-apoA-IV induces atherogenesis through NR4A3 in apoE–/– mice.
High-density lipoprotein (HDL) has atheroprotective and antiatherosclerotic effects (1). Apolipoprotein (apo) A-I is the most abundant protein in HDL, followed by apoA-II. The remaining proteins, including apoA-IV, are minor apolipoproteins (2). ApoA-IV is synthesized mainly by the small intestine. After apoA-IV enters the plasma, a portion associates with HDL, whereas a large portion circulates in a lipid-free form and at a considerable concentration in plasma (3–5). Previous studies have demonstrated that apoA-IV modulates lipid and glucose metabolism (3,6). Its regulation of hepatic gluconeogenesis is mediated through the nuclear receptor NR4A1 (7). ApoA-IV has antioxidant, anti-inflammatory, and antiatherogenic properties (8,9).
In diabetes, glycation of apoA-I impairs its anti-inflammatory function and is associated with coronary artery disease (CAD) (10,11). Given that a considerable percentage of apoA-IV circulates as a lipid-free protein and at a considerable concentration in plasma (4,5), we hypothesized that apoA-IV may be glycated in diabetic conditions and glycated apoA-IV (g-apoA-IV) may exert atherogenic effects.
The present study tested this hypothesis. Because NƐ-(carboxylmethyl) lysine (CML) modifications predominate in advanced glycation end products in vivo in diabetes (12), we evaluated glycation of apoA-IV (CML protein adducts) in HDL. We used immunoprecipitation-Western blotting and mass spectrometry (liquid chromatography–mass spectrometry [LC-MS]) to verify apoA-IV glycation and to identify CML glycation sites in healthy control subjects and patients with type 2 diabetes mellitus (T2DM) with or without CAD. To investigate the biological effects of g-apoA-IV and the mechanisms responsible for these effects, we treated endothelial cells and macrophages with saline, human apoA-IV, or glycated human apoA-IV. We also intraperitoneally injected these preparations into apoE–/– mice and NR4A3–/– and apoE–/–- double-knockout mice every other day for 6 months, after which we performed histological examinations.
This study complied with the Declaration of Helsinki. The local hospital ethics committee approved the study protocol, and we obtained written informed consent from all the participants.
We enrolled a total of 1,885 consecutive patients with T2DM and chest pain on exertion who were referred for diagnostic coronary angiography between January 2014 and November 2016 in this study. The diagnoses of T2DM, hypertension, and dyslipidemia were made according to published guidelines from the American Diabetes Association and American Heart Association (13–15). After excluding noneligible subjects, we grouped the patients with T2DM into Group I (n = 204, without CAD; no coronary artery stenosis or stenosis <30%) and Group II (n = 515, with CAD; coronary artery stenosis ≥70%) based on the angiographic examination results (Online Appendix).
A total of 176 subjects without evidence of cardiovascular diseases or diabetes served as control subjects (control group). The control subjects were citizens from several districts near our hospital who received annual physical checkups. We evaluated each of them by taking a detailed medical and family history and by collecting fasting blood samples during an outpatient visit. History taking and data collection revealed that the subjects had no history of any cardiovascular diseases (including angina or myocardial infarction) and exhibited normal chest x-ray, carotid artery ultrasound, exercise stress test, and coronary computed tomography angiography findings.
Coronary angiography and analysis
Human tissue sample management
We obtained coronary endarterectomy tissue specimens from 19 consenting patients with severe CAD who were undergoing coronary artery bypass grafting. We also obtained surplus segments of nonatherosclerotic internal mammary arteries from 16 patients undergoing coronary artery bypass grafting (Online Appendix).
Materials and antibodies
For details regarding the materials and antibodies used in this experiment, see the Online Appendix.
Isolation of HDL, immunoprecipitation of apoA-IV, and quantification of apoA-IV glycation
To circumvent the interference and masking effects of albumin and immunoglobulins in plasma, we obtained apoA-IV by immunoprecipitating it from isolated HDL. We analyzed apoA-IV glycation through Western blotting densitometry or LC-MS. The detailed steps through which these procedures were performed are provided in the Online Appendix. Briefly, HDL was isolated from 50 ml of fresh plasma by ultracentrifugation, as previously described (17). The HDL solution was then incubated with an anti–apoA-IV antibody and Protein A/G PLUS-Agarose (SC-2003, Santa Cruz, California) to precipitate apoA-IV. The apoA-IV–containing immunoprecipitate was then subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Immunoprecipitation-Western blotting was repeated 3 times for each HDL sample. Glyoxal-modified recombinant apoA-IV was used as an interassay standard to normalize patient samples and was run with each gel (information about this recombinant protein is provided in the following section). The absolute intensity of apoA-IV glycation and the content of apoA-IV were calculated as density values corresponding to the number of pixels for each band of apoA-IV and glyoxal-modified apoA-IV. To reduce intergel differences, we normalized the absolute intensity of apoA-IV glycation in the patient samples by dividing this parameter by the absolute glycation intensity of glyoxal-modified apoA-IV, and we normalized the content of apoA-IV in the patient samples by dividing the parameter by the content of glyoxal-modified apoA-IV. The relative intensity of apoA-IV glycation in the patient samples was subsequently calculated as the ratio of the normalized absolute intensity of apoA-IV glycation to the normalized content of apoA-IV ([apoA-IVsample glycation/apoA-IVstandard glycation]/[apoA-IVsample/apoA-IVstandard]). The relative intensity of apoA-IV glycation was determined in all participants. CML modifications of apoA-IV were analyzed by LC-MS in 37 control subjects and 63 and 138 patients who were randomly selected from the control group and Groups I and II, respectively.
LC-MS analysis, data acquisition, and database searching
The detailed procedures by which the LC-MS analysis was performed, the data were acquired, and the databases were searched are provided in the Online Appendix.
Preparation of the glycated apoA-IV recombinant protein
The Pet30a-Human-apoA-IV plasmid was kindly provided by Prof. Patrick Tso from the University of Cincinnati (Cincinnati, Ohio). The detailed process through which the glycated apoA-IV recombinant protein was prepared is provided in the Online Appendix. We performed apoA-IV glycation by incubating apoA-IV in a solution containing 0.01% ethylenediamenetetraacetic acid, 0.01% sodium azide and 20 mmol/l glyoxal for 7 days, after which we dialyzed the solution. Using LC-MS, we found that the abundance of CML modification and the number of glycation sites in apoA-IV were approximately linearly correlated with the glyoxal concentration (Online Figure 1, Online Table 1). The g-apoA-IV prepared with the glyoxal culture was comparable with the endogenous apoA-IV observed in the diabetic patients with CAD (Group II) regarding the abundance of glycation and the number of glycation sites. The glycation of g-apoA-IV was examined by Western blotting after the immunoprecipitation of the protein from the plasma of apoE–/– mice that had been intraperitoneally injected with g-apoA-IV (10 μg/each) every other day for 1 month (Online Figure 2). The concentrations of this human protein in mouse serum were comparable with those of endogenous mouse apoA-IV after the mice had been injected with g-apoA-IV at the indicated dose for 6 months.
Animal experiments and quantification of atherosclerotic lesions
The detailed procedures through which the animal experiments were performed and information pertaining to the NR4A3–/– and receptor for advanced glycation end products (RAGE)–/– mice are provided in the Online Appendix. Male mice 8 to 10 weeks of age received intraperitoneal injections of saline, apoA-IV (10 μg each), or g-apoA-IV (10 μg each) every other day for 24 weeks.
Histology and immunostaining
Cell culture and in vitro experiments
The cell culture and in vitro experiments performed in the study are detailed in the Online Appendix.
Continuous variables are presented as mean ± SD or mean ± SEM, as appropriate, and categorical data are summarized as frequencies or percentages. We evaluated the differences in categorical variables between groups with a chi-square test. We evaluated the normality of distribution with the Kolmogorov-Smirnov test and applied logarithmic or square-root transformations to continuous variables showing a non-normal distribution. We analyzed the differences between patients with and without CAD by Student t test. We determined the correlations between variables by the Pearson or Spearman correlation tests, as appropriate. We constructed models for multivariable logistic regression to assess the independent determinants of CAD. We included traditional CAD risk factors but not the parameters for apoA-IV glycation in 719 diabetic patients (Model 1A) and 201 diabetic patients who were examined by LC-MS (Model 1B) in Model 1. We adjusted the analysis for the relative intensity of apoA-IV glycation (Model 2A) and the abundance of apoA-IV CML glycation (Model 2B) in Model 2. Receiver-operator characteristic analysis of the CAD risk factors and the biohumoral measurements was performed with and without the addition of these 2 apoA-IV glycation parameters. We used the C statistic to analyze the discriminatory capacity of Model 1 and Model 2 and performed risk reclassification with the method devised by Pencina et al. (18). We determined the net reclassification improvement and integrated discrimination improvement with the same method (18,19). The calibration of the model was verified using the Hosmer-Lemeshow chi-square test. For the cell experiments, the data represent the means from 6 experiments. For the animal experiments, we performed analysis of variance and post hoc analysis to assess the significance of the differences between the experimental groups and the control subjects. We performed the Mann-Whitney U test for multiple comparisons between 2 groups. All analyses used 2-sided tests with an overall significance level (alpha) of 0.05, and all tests were performed with SPSS 19.0 for Windows (IBM Corporation, Armonk, New York) and SAS Version 9.1 (SAS Institute, Cary, North Carolina) software.
ApoA-IV glycation is significantly increased in the HDL of diabetic patients with CAD and is associated with disease severity
Information pertaining to the diabetic patients with or without CAD is detailed in Table 1, and information pertaining to the participants whose samples underwent LC-MS analysis is provided in Online Table 2. The relative intensity of apoA-IV glycation was markedly increased in T2DM patients with CAD (Group II, 13.37 ± 3.78) compared with T2DM patients without CAD (Group I, 9.85 ± 1.33; p < 0.001) (Figures 1A and 1B, Table 1). We organized Group II patients into subgroups according to the number of diseased coronary arteries (Figure 1C) and the extent index, cumulative coronary stenosis score, and SYNTAX score tertiles. After adjustment for sex, age, body mass index, cigarette smoking, history of hypertension, glycosylated hemoglobin, the total-to-HDL cholesterol ratio, the estimated glomerular filtration rate and high-sensitivity C-reactive protein, the relative intensity of apoA-IV glycation was significantly correlated with CAD severity (Spearman’s r = 0.285, r = 0.371, r = 0.329, and r = 0.239 for the number of diseased coronary arteries and the extent index, cumulative coronary stenosis score, and SYNTAX score tertiles, respectively; all p < 0.001) (Online Figures 3A to D).
Likewise, the CML glycation percentage of apoA-IV was significantly higher in the 138 patients from Group II than in the 63 patients from Group I (p < 0.001). After adjustment for the previously mentioned confounders, glycation abundance was also positively correlated with CAD severity. Spearman’s r was 0.311 for the number of diseased coronary arteries, 0.293 for the extent index tertiles, 0.369 for the cumulative coronary stenosis score tertiles, and 0.246 for the SYNTAX score tertiles (all p < 0.01) (Online Figures 3E to 3H).
Immunohistochemistry of the coronary arteries from diabetic patients with severe CAD who died of acute myocardial infarction demonstrated CML and apoA-IV colocalization in atherosclerotic plaques (Figure 1D); however, no glycated apoA-IV was noted in the nonatherosclerotic arteries.
Glycation of apoA-IV is independently associated with CAD in diabetic patients, according to the multivariable regression analysis
To identify the independent determinants of CAD in patients with T2DM, we performed multivariable regression analyses including the traditional cardiovascular risk factors and biomeasurements detailed in Table 1 and Online Table 2. Male sex, age, smoking, history of hypertension, glycated hemoglobin, total-to-HDL cholesterol ratio, estimated glomerular filtration rate, and high-sensitivity C-reactive protein were independent determinants of CAD in Model 1A. In addition to the conventional factors that were included in Model 1A, the relative intensity of apoA-IV glycation remained an independent determinant of CAD when it was included in the multivariable regression analysis in Model 2A (odds ratio: 7.049; 95% confidence interval: 4.615 to 10.768; p < 0.001) (Table 2).
Likewise, in addition to the conventional factors that were included in Model 1B, the abundance of apoA-IV CML glycation remained independently associated with CAD when it was included in the multivariable regression analysis in Model 2B (odds ratio: 2.447; 95% confidence interval: 1.234 to 4.854; p < 0.001) (Table 2).
The calibration of Model 1A with Model 2A and that of Model 1B with Model 2B was good. The addition of the 2 apoA-IV glycation parameters resulted in a significant improvement in goodness-of-fit and predictive performance, with an increase in the Nagelkerke R2, C statistic, net reclassification improvement, and integrated discrimination improvement (Online Appendix).
G-apoA-IV induces inflammatory reactions in endothelial cells and promotes leukocyte-endothelium interactions in vitro and in vivo
To assess the effect of g-apoA-IV on endothelial cells, we stimulated human aortic endothelial cells (HAECs) with saline, apoA-IV, and g-apoA-IV at concentrations similar to the plasma concentrations of apoA-IV previously reported in healthy control subjects or obese patients (15 to 170 μg/ml) (4,5). G-apoA-IV significantly up-regulated the messenger ribonucleic acid (mRNA) and protein levels of the indicated adhesion molecules (ICAM-1, VCAM-1 and E-selectin) and inflammatory cytokines (tumor necrosis factor alpha and interleukin 1β) in a concentration-dependent manner in HAECs compared with the control treatment (Online Figures 4A to 4K). In contrast, apoA-IV suppressed tumor necrosis factor alpha–induced increments in adhesion molecule expression (Online Figures 5A to 5C). Consistent with these cell experiment results, additional results showed that g-apoA-IV significantly induced the adhesion of THP-1 cells to HAECs (Online Figures 6A and 6B) and enhanced leukocyte-endothelium interactions in vivo (intravital microscopy assay) compared with PBS (Online Figures 6C to 6F).
G-apoA-IV induces atherogenesis in apoE–/– mice
To assess the influence of g-apoA-IV on atherogenesis, we intraperitoneally injected g-apoA-IV, apoA-IV, or saline into apoE–/– mice every other day for 24 weeks (Online Figures 7A to 7H). After completion of the experiment, apoE–/– mice injected with g-apoA-IV displayed a significantly increased atherosclerotic lesion area throughout the entire aorta and in the aortic root compared with mice injected with saline. However, apoA-IV injection mildly decreased the atherosclerotic lesion area in apoE–/– mice compared with saline injection (Figures 2A to 2D).
We examined the aortic tissues from these mice further. Adhesion molecule and inflammatory cytokine levels were elevated in the aortic tissues of apoE–/– mice injected with g-apoA-IV compared with those injected with saline, findings consistent with the pathology of atherosclerosis (Figure 2E, Online Figures 8A to 8E).
Glycated apoA-IV up-regulates NR4A3 expression in HAECs, as shown by mRNA microarray analysis, and NR4A3 expression is increased in atherosclerotic plaques
To decipher the pathogenic mechanism through which g-apoA-IV exerts its effects, we treated HAECs with g-apoA-IV or saline overnight and then performed mRNA microarray analysis. We found that g-apoA-IV significantly up-regulated NR4A3 (35-fold increase vs. saline), a member of the NR4A nuclear receptor subfamily (Figure 3A and Online Appendix). This subfamily has broad functions, as it regulates the genes involved in metabolism, atherosclerosis, and vascular remodeling (20). In particular, NR4A3 up-regulates the expression of adhesion molecules in endothelial cells (20). The results of our in vitro experiments verified the results of the mRNA microarray analysis, as they showed that g-apoA-IV significantly increases NR4A3 expression (Online Appendix, Figures 3B and 3C, Online Figures 9A to 9D). Moreover, severely atherosclerotic coronary arteries (n = 19) from diabetic patients exhibited higher NR4A3 expression than nonatherosclerotic internal mammary arteries (n = 16; p < 0.01) (Figure 3D). Immunohistochemistry showed that NR4A3 expression was significantly elevated in atherosclerotic plaques from diabetic patients compared with nonatherosclerotic arteries and that NR4A3 colocalized with the endothelial cells of microvessels as represented by CD31+ (Figure 3E).
NR4A3 knockdown or deletion suppresses glycated apoA-IV–induced inflammation in endothelial cells and monocytes or macrophages
Using small interfering RNA to knockdown NR4A3 in endothelial cells or macrophages and using NR4A3–/–– or RAGE–/––derived cells, we showed that NR4A3, as well as RAGE, mediates g-apoA-IV–induced proinflammatory reactions (Online Appendix, Online Figures 10 to 16).
NR4A3 deletion attenuates glycated apoA-IV-induced atherogenesis in C57BL/6 and apoE–/– mice
Determination of the glycation sites of apoA-IV from HDL by LC-MS
To determine the CML modification sites of apoA-IV in diabetic patients, we analyzed apoA-IV by LC-MS in 37 control subjects, 63 patients from Group I, and 138 patients from Group II (see the Online Appendix for details). We identified a total of 19 CML modification sites in apoA-IV in diabetic patients (Figures 5A and 5B, Online Tables 3 and 4, Online Figure 18). Of these sites, K189, K123, K65, and K361 had the highest frequencies (particularly, the frequency of K189 was 23% in Group II patients). However, we found no glycation sites in healthy control subjects. Group II patients had significantly more CML modification sites (19 sites) than did Group I patients (9 sites).
The pathogenic effects of G-apoA-IV are attenuated by mutations at major glycation sites (K65A, K123A, K189A, AND K361A)
To explore the potential contributions of key apoA-IV glycation sites (K65, K123, K189, and K361) to atherogenesis, we constructed a mutated apoA-IV plasmid (apoA-IV-M: apoA-IV with mutations at K65A, K123A, K189A, and K361A) and prepared a g-apoA-IV-M recombinant protein, as described in the Online Appendix. Using an anti-CML antibody, we detected much milder CML modification of g-apoA-IV-M than g-apoA-IV after both proteins were exposed to the same glyoxal-conditioned medium (Online Figures 19A and 19B). The cell experiments showed that g-apoA-IV-M had significantly weakened proinflammatory effects in HAECs compared with g-apoA-IV (Online Appendix, Figure 6A, Online Figures 20A to 20M). Consistent with these findings, we found that mice receiving g-apoA-IV-M injections displayed markedly attenuated atherogenesis after 6 months compared with those receiving g-apoA-IV injections (Figures 6B to 6F, Online Figures 21A to 21J).
In the present study, we found that: 1) apoA-IV glycation is significantly increased in patients with T2DM with CAD compared with T2DM patients without CAD and is associated with CAD severity; 2) g-apoA-IV induces inflammatory reactions in vitro and in vivo, and intraperitoneal injections of g-apoA-IV induce atherogenesis in apoE–/– mice; 3) NR4A3 mediates g-apoA-IV–induced inflammatory reactions and atherogenesis; and 4) 19 glycation sites in apoA-IV are present in diabetic patients, and g-apoA-IV with major glycation site mutations has weakened proinflammatory and atherogenic effects in in vitro experiments and in apoE–/– mice. Collectively, these findings suggest that apoA-IV glycation is associated with CAD in diabetic patients and induces diabetic atherosclerosis mainly through NR4A3 in mice (Central Illustration).
The NR4A subfamily includes 3 members, NR4A1, NR4A2, and NR4A3. NR4A3 induces the expression of adhesion molecules in endothelial cells, and atherogenesis is attenuated in apoE–/–/NR4A3–/– double-knockout mice (20). In our study, g-apoA-IV up-regulated NR4A3 expression in vascular endothelial cells and macrophages. NR4A3 levels were increased in atherosclerotic plaques from diabetic patients. Using small interfering RNA, NR4A3–/– mouse–derived cells and apoE–/–/NR4A3–/– mice, we showed that NR4A3 mediates the proinflammatory reaction and atherogenesis induced by g-apoA-IV. Moreover, we showed that g-apoA-IV–induced NR4A3 up-regulation and inflammation were significantly suppressed in aortic endothelial cells and peritoneal macrophages from RAGE–/– mice. Based on these data, we propose that g-apoA-IV promotes inflammation and atherogenesis mainly through the RAGE-NR4A3 axis.
In this study, we identified 19 CML modification sites in apoA-IV from diabetic patients. Of these sites, K65, K123, K189, and K361 had the highest frequencies in diabetic patients. ApoA-IV shares several structural characteristics, including amphipathic α-helical repeat sequences, which are crucial for lipid-binding and self-association, with apoA-I and other apolipoproteins (21). Sites K189, K123, and K65 are located in these α-helices. Two variants of apoA-IV (T347S and Q360H, which are close to site K361) exhibits altered apoA-IV lipid binding and cholesterol efflux capacity (22), suggesting that CML modification of these sites results in apoA-IV dysfunction. Our in vitro and in vivo experiments showed that g-apoA-IV with mutations at these sites exhibited attenuated proinflammatory and atherogenic effects compared with g-apoA-IV, a finding indicative of the importance of these glycation sites.
First, our clinical part is a cross-sectional study, thus allowing the detection of associations but not predictions or outcomes. Second, the conclusion of second part that glycated apoA-IV promotes atherogenesis is speculative. In addition, the human g-apoA-IV used herein was exogenous. The pathogenic effects of endogenous apoA-IV glycation should be assessed in future studies to verify our findings.
ApoA-IV glycation is associated with CAD severity in diabetic patients. G-apoA-IV induces atherogenesis in apoE–/– mice through nuclear receptor NR4A3.
COMPETENCY IN MEDICAL KNOWLEDGE: Nonenzymatic glycation of apolipoproteins contributes to the pathogenesis of atherosclerosis in patients with T2DM, and apoA-IV glycation is associated with the severity of coronary disease.
TRANSLATIONAL OUTLOOK: Future studies should evaluate the therapeutic efficacy of inhibiting apolipoprotein glycation on the progression of atherosclerosis in patients with diabetes.
This study was supported by a “973” grant from the National Science and Technology Department of China (2015CBS553600), the National Natural Science Foundation of China (81470392, 81470469, 81400327, and 91539117), and Doctoral Innovation Found Projects from Shanghai Jiao Tong University (BXJ201614). The authors have reported that they have no relationships relevant to the contents of this paper to disclose. Drs. Dai, Shen, and Li contributed equally to this work.
- Abbreviations and Acronyms
- apolipoprotein A-IV
- mutated apolipoprotein A-IV plasmid
- coronary artery disease
- NƐ-(carboxylmethyl) lysine
- glycated apolipoprotein A-IV
- human aortic endothelial cell
- high-density lipoprotein
- liquid chromatography–mass spectrometry
- messenger ribonucleic acid
- receptor for advanced glycation end products
- type 2 diabetes mellitus
- Received April 5, 2017.
- Revision received August 15, 2017.
- Accepted August 21, 2017.
- 2017 American College of Cardiology Foundation
- Rosenson R.S.,
- Brewer H.B. Jr..,
- Ansell B.,
- et al.
- Wang F.,
- Kohan A.B.,
- Lo C.,
- Liu M.,
- Howles P.,
- Tso P.
- Verges B.,
- Guerci B.,
- Durlach V.,
- et al.
- Kohan A.B.,
- Wang F.,
- Lo C.M.,
- Liu M.,
- Tso P.
- Li X.,
- Xu M.,
- Wang F.,
- et al.
- Duverger N.,
- Tremp G.,
- Caillaud J.M.,
- et al.
- Geronimo F.R.,
- Barter P.J.,
- Rye K.A.,
- Heather A.K.,
- Shearston K.D.,
- Rodgers K.J.
- Nobecourt E.,
- Tabet F.,
- Lambert G.,
- et al.
- Pu L.J.,
- Lu L.,
- Zhang R.Y.,
- et al.
- Yan S.F.,
- Ramasamy R.,
- Naka Y.,
- Schmidt A.M.
- American Diabetes Association
- Ren S.,
- Shen G.X.
- Ridker P.M.,
- Paynter N.P.,
- Rifai N.,
- Gaziano J.M.,
- Cook N.R.
- Zhao Y.,
- Howatt D.A.,
- Gizard F.,
- et al.
- Walker R.G.,
- Deng X.,
- Melchior J.T.,
- et al.