Rosuvastatin acts on the lymphatic system to improve atherosclerosis / 中华心血管病杂志

Objective: To investigate whether rosuvastatin acts on lymphatic system and influences lymphatic system-mediated reverse cholesterol transport to play an anti-atherosclerosis role. Methods: Forty-eight apolipoprotein E-/- mice fed a high fat diet were used to construct the atherosclerosis model. They were randomly divided into 4 groups with 12 rats in each group. They were treated with rosuvastatin, vascular endothelial growth factor-C (VEGF-C) and rosuvastatin+VEGF-C inhibitors as experimental group, and no intervention measures were given in control group. After 8 weeks, aortic plaque area, high density lipoprotein cholesterol (HDL-C) content in lymph fluid, the function of popliteal lymphatic drainage of peripheral Evans blue, and the ability of lymphatic system to transport peripheral cell membrane red fluorescent probes to label high-density lipoprotein (HDL) were detected. Subsequently, the effects of rosuvastatin on proliferation, migration and tubular function of lymphoendothelial cells and the expression of scavenger receptor class B type 1 (SR-B1) on lymphoendothelial cells at different concentrations were detected. Results: Compared with the control group, Rosuvastatin and VEGF-C could reduce the area of aortic atherosclerotic plaque (P<0.05). In addition to rosuvastatin plus VEGF-C inhibitor, the intra-aortic plaque area increased (P<0.05). Compared with the control group, Rosuvastatin could increase the content of HDL-C in lymphatic fluid (P<0.05), enhance the drainage function of lymphatic vessels, and enhance the capacity of HDL in the transport tissue fluid of lymphatic system. Compared with the control group, VEGF-C increased the content of HDL-C in mouse lymph fluid (P<0.01), enhanced the drainage function of popliteal lymphatic canal, and enhanced the ability of lymphatic system to transport HDL. With the addition of VEGF-C inhibitor on the basis of rosuvastatin, the content of HDL-C in lymph fluid was reduced, the drainage of popliteal lymphatic canal was interrupted, and the ability of lymphatic system to transport HDL was reduced. Western blotting showed that rosuvastatin increased the protein expression of SR-B1. Conclusion: Rosuvastatin can promote the proliferation, migration and tube formation of lymphatic endothelial cells. At the same time, SR-B1 expression on lymphatic endothelial cells is promoted, thus enhancing the lymphatic system mediated cholesterol reversal transport and playing the role of anti-atherosclerosis.

Short-term outcome of complex coronary lesions treated by excimer laser coronary atherectomy / 中国介入心脏病学杂志

Objective To investigate the feasibility, safety and efficacy of excimer laser coronary atherectomy used in complex lesions, including in-stent restenosis, non-crossable or nonexpandable lesions, heavily calcified lesions without successful wire-exchange and saphenous vein grafts lesions. Methods From Jul 24, 2017 to Aug 24, 2018, 22 cases with 24 lesions were treated with excimer laser coronary atherectomy in Peking University People's Hospital, combined with or without IVUS/OCT, rotational atherectomy or other percutaneous coronary intervention instrument, and with or without stent implantation. Results The procedural success rate was 23/24. There was no complications in all cases. Drug-eluting stents were implanted in 19/24 of lesions. There were no major advent cardiovascular events, including death, acute ST-segment-elevation myocardia and pericardial tamponade recorded. Conclusions Excimer laser coronary atherectomy used in complex lesions is feasible, safe and efficient with satisfactory in-hospital short-term outcome.

Role of miR-106b-5p in the regulation of gene profiles in endothelial cells / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the role of miR-106b-5p in the regulation of gene expression in endothelial cells.@*METHODS@#The Taqman low-density microRNAs (miRNAs) array (TLDA) was used to identify miRNA expression profiles in the plasma of patients with atherosclerotic coronary artery disease (CAD) (atherosclerosis group, n=9) and individuals without atherosclerotic CAD disease (control group, n=9). A weighed and undirected miRNA coexpression network analysis was performed to investigate the interactions among miRNAs in the two groups. MiR-106b-5p, whose coexpression pattern in atherosclerosis group was most different from that of control group, was further studied. Human umbilical vein endothelial cells (HUVEC) were transfected with miR-106b-5p mimic or negative control mimic, and Affymetrix GeneChip Human Transcriptome Array 2.0 was used to screen the differential gene expression profiles after transfection. And the signal transduction pathway of differential gene profiles was further analyzed in Kyoto Encyclopedia of Genes and Genomes (KEGG) signal pathway database. After parsing the whole KEGG database, all differentially expressed genes involved pathways were extracted, and the hypergeometric distribution was used to calculate the pathway enrichment.@*RESULTS@#The coexpression pattern of the patients with atherosclerosis (140 nodes, 1 154 edges) differed from that of the non-atherosclerosis control group (140 nodes, 612 edges). The analysis of array data with significant analysis of microarray (SAM) identified 746 significantly deregulated genes (fold change ≥ 1.5 and false discovery rate < 0.01) altered by overexpression of miR-106b-5p with miR-106b-5p mimic in HUVEC. By calculating the pathway enrichment, we found that multiple signaling pathways enriched in differential gene profiles were closely related to the process of formation and rupture of atherosclerotic plaque, including phosphatidylinositol-3 kinase (PI3K)/ protein kinase B (PKB, also called Akt), mammalian target of rapamycin (mTOR), transforming growth factor-β (TGF-β), janus kinase / signal transducer and activator of transcription (Jak-STAT), tumor necrosis factor (TNF), toll like receptor (TLR) and hypoxia-inducible factor 1α (HIF-1α) and other signal pathways.@*CONCLUSION@#The coexpression pattern of miRNAs in plasma of patients with atherosclerosis is more significantly changed than that of individuals without atherosclerotic disease. MiR-106b-5p, which shows the most significant difference between groups, targets multiple signal pathways in vascular endothelial cells, and might play an important role in the regulatory network of atherosclerotic gene expression.

Effect of Endothelial Microparticles Induced by Hypoxia on Migration and Angiogenesis of Human Umbilical Vein Endothelial Cells by Delivering MicroRNA-19b / 中华医学杂志(英文版)

Background@#Microparticles (MPs) are small extracellular plasma membrane particles shed by activated and apoptotic cells, which are involved in the development of atherosclerosis. Our previous study found that microRNA (miR)-19b encapsulated within endothelial MPs (EMPs) may contribute to the upregulation of circulating miR-19b in unstable angina patients. Hypoxia is involved in atherosclerosis as a critical pathological stimulus. However, it still remains unclear whether the increase of miR-19b levels in EMPs is related to hypoxia and if the effect of miR-19b - wrapped within EMPs - stimulates hypoxia on vascular endothelial cells. This study aimed to explore the changes of miR-19b in EMPs induced by hypoxia as well as their effects on endothelial cells.@*Methods@#Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and arranged to harvest EMPs in two parts: the first part consisted of EMP and EMP and the second part included EMP, EMP, and EMP. Cell migration was detected by scratch migration and transwell chamber migration. Angiogenesis was assessed by tube formation assays. Furthermore, we predicted the target gene of miR-19b by bioinformatics analysis, and luciferase assay was used to verify the targeted gene of miR-19b. Data were analyzed by one-way analysis of variance. Student's t-test was used when two groups were compared.@*Results@#Compared with EMP- and EMP-inhibited migration of cells by scratch migration assay (80.77 ± 1.10 vs. 28.37 ± 1.40, P < 0. 001) and transwell chamber migration assay (83.00 ± 3.46 vs. 235.00 ± 16.52, P < 0.01), the number of tube formations was markedly reduced by 70% in the EMP group (P < 0.001) in vitro analysis of HUVECs. Meanwhile, a strong inhibition of migration and tube formation of HUVECs in the presence of miR-19b-enriched EMP was observed. This effect might be due to the delivery of miR-19b in EMPs. Transforming growth factor-β2 (TGFβ2) was predicted to be one of the target genes of miR-19b, and we further confirmed that TGFβ2 was a direct target gene of miR-19b using the luciferase assay. The expression of TGFβ2 in HUVECs was inhibited by treatment with EMP and EMP.@*Conclusions@#MiR-19b in EMPs induced by hypoxia could reduce endothelial cell migration and angiogenesis by downregulating TGFβ2 expression, which may have inhibited the progression of atherosclerosis.

MiR-106b-5p Inhibits Tumor Necrosis Factor-α-induced Apoptosis by Targeting Phosphatase and Tensin Homolog Deleted on Chromosome 10 in Vascular Endothelial Cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Apoptosis of endothelial cells (ECs) plays a key role in the development of atherosclerosis and there are also evidence indicated that phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a viable target in therapeutic approaches to prevent vascular ECs apoptosis. Aberrant miR-106b-5p expression has been reported in the plasma of patients with unstable atherosclerotic plaques. However, the role and underlying mechanism of miR-106-5p in the genesis of atherosclerosis have not been addressed. In this study, we explored the anti-apoptotic role of miR-106-5p by regulating PTEN expression in vascular ECs.</p><p><b>METHODS</b>Real-time reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the expression levels of miR-106b-5p in human atherosclerotic plaques and normal vascular tissues. Human umbilical vein endothelial cells (HUVEC) were transfected with miR-106b-5p mimic or negative control mimic, and apoptosis was induced by serum starvation and tumor necrosis factor-α (TNF-α) treat. Western blotting and real-time RT-PCR experiments were used to detect PTEN expression levels and TNF-α-induced apoptosis was evaluated by the activation of caspase-3 and cell DNA fragmentation levels in HUVEC.</p><p><b>RESULTS</b>The expression of miR-106b-5p was significantly downregulated in plaques than in normal vascular tissues. TNF-α significantly downregulated miR-106b-5p expression levels and upregulated activation of caspase-3 and cell DNA fragmentation levels in HUVEC. Overexpression of miR-106b-5p with miR-106b-5p mimic inhibited PTEN expression and TNF-α-induced apoptosis in HUVEC. Luciferase reporter assays confirmed that miR-106b-5p binds to PTEN mRNA 3' untranslated region site.</p><p><b>CONCLUSION</b>MiR-106b-5p could inhibit the expression of PTEN in vascular ECs, which could block TNF-α-induced activation of caspase-3, thus prevent ECs apoptosis in atherosclerosis diseases.</p>

High density lipoprotein suppresses lipoprotein associated phospholipase A2 in human monocytes-derived macrophages through peroxisome proliferator-activated receptor-γ pathway / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Lipoprotein-associated phospholipase A2 (Lp-PLA2) is mainly secreted by macrophages, serving as a specific marker of atherosclerotic plaque and exerting pro-atherogenic effects. It is known that high-density lipoprotein (HDL) plays an important role against atherosclerosis by inhibiting pro-inflammatory factors, however, the relationship between HDL and Lp-PLA2 remains elusive.</p><p><b>METHODS</b>In this study, reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and a platelet-activating factor (PAF) acetylhydrolase assay were performed to determine the Lp-PLA2 mRNA level, protein expression and activity in human monocyte-derived macrophages upon HDL treatment of different concentrations and durations. To investigate the underlying mechanism of HDL-induced Lp-PLA2 action, pioglitazone, a peroxisome proliferator-activated receptor-γ (PPARγ) ligand, was introduced to human monocyte-derived macrophages and mRNA and protein levels of Lp-PLA2, as well as its activity, were determined.</p><p><b>RESULTS</b>Lp-PLA2 mRNA levels, protein expression and activity were significantly inhibited in response to HDL treatment in a dose and time dependent manner in human monocyte-derived macrophages. Pioglitazone treatment (1 - 10 ng/ml) upregulated the Lp-PLA2 mRNA level, protein expression and activity in human monocyte-derived macrophages, while the effects were markedly reversed by HDL. In addition, pioglitazone resulted in a significant increase in PPARγ phosphorylation in human monocyte-derived macrophages, which could be inhibited by HDL.</p><p><b>CONCLUSION</b>These findings indicate that HDL suppresses the expression and activity of Lp-PLA2 in human monocyte-derived macrophages, and the underlying mechanisms may be mediated through the PPARγ pathway.</p>

Elevated plasma lipoprotein-associated phospholipase A₂ activity is associated with plaque rupture in patients with coronary artery disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) has recently been shown to be positively related to coronary events in patients with coronary artery disease (CAD). However, direct evidence about the relationship between circulation Lp-PLA(2) activity and vulnerable plaque in patients with CAD remains lacking.</p><p><b>METHODS</b>Plasma Lp-PLA(2) activity was determined in 146 consecutive patients with CAD who underwent clinically-indicated coronary angiography and preinterventional intravascular ultrasound (IVUS).</p><p><b>RESULTS</b>Eighty-three patients were included in the final analysis after the initial screening. Sixty (72.3%) were acute coronary syndrome (ACS) patients and 23 (27.7%) were stable angina pectoris (SAP) patients. Plaque rupture occurred in 39 (47.0%) patients, and 34 (87.2%) were from ACS patients and 5 (12.8%) from SAP patients. There were no significant differences in clinical and angiographic characteristics between patients with plaque rupture and those without plaque rupture, except for smoking, high-sensitive C-reactive protein (hs-CRP) level and Lp-PLA(2) activity (all P < 0.05). IVUS measurement uncovered that patients with plaque rupture had more frequent positive remodeling (74.4% vs. 43.2%, P = 0.004), soft plaques (64.1% vs. 36.4%, P = 0.012) and higher remodeling index (1.13 ± 0.16 vs. 0.99 ± 0.11, P = 0.041) as compared with those without plaque rupture. Multivariate Logistic regression analysis showed that plasma Lp-PLA(2) activity was independently associated with plaque rupture after adjusting for smoking, positive remodeling and soft plaque (Model 1: odds ratio (OR) 1.13, 95% confidence interval (CI): 1.06 - 1.20) or adjusting for smoking, hs-CRP level, positive remodeling and soft plaque (Model 2: OR 1.11, 95%CI: 1.04 - 1.19).</p><p><b>CONCLUSIONS</b>Plasma Lp-PLA(2) activity is associated with plaque rupture in patients with CAD, independently of traditional CAD risk factors, hs-CRP level and IVUS parameters. Lp-PLA(2) may be a risk marker for vulnerable plaques.</p>

Construction of recombinant adenovirus vector of calcitonin gene-related peptide gene and transfection to neonatal rat cardiomyocytes / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus vector of calcitonin gene-related peptide (CGRP) by AdEasy system and to validate its expression in myocardial cells.</p><p><b>METHODS</b>The full-length of CGRP gene cDNA was acquired by RT-PCR and cloned into pShuttle-CMV. After linearization with Pme I, the recombinant plasmid (pShuttle-CMV-CGRP) was transformed into E.coli BJ5183 by electroporation to construct the recombinant adenovirus plasmid AdEasy-pShuttle-CGRP. The recombinant adenovirus plasmids were transformed into E.coli XL10-Gold cells to be amplified. Then the recombinant plasmid was digested with Pac I and transfected to 293 cells to package recombinant adenovirus particles. PCR technique was used to detect target gene. The recombinant adenovirus particles were purified by CsC1 density gradient. The purified recombinant adenovirus was transfected to neonatal rat cardiomyocytes,and the recombinant adenovirus production was observed by fluorescent microscope. Expression of CGRP in hearts 7 days after intravenous delivery of adenoviral vectors AV-CGRP was determined by radioimmunoassay.</p><p><b>RESULT</b>The RT-PCR products confirmed a full-length cDNA of CGRP gene in PUC(57) by sequencing. The corresponding double endonuclease and PCR analysis certified the successful cloning of the gene into the pShuttle-CMV. The recombinant adenovirus plasmid AdEasy-pShuttle-CGRP was digested by Pac I endonuclease to form the typical DNA segments, whose length was about 3 kb and 30 kb. PCR analysis and fluorescent microscope observation confirmed that the CGRP gene was inserted into the adenovirus vector with very strong power of transfection. The recombinant adenovirus particles infected neonatal rat cardiomyocytes successfully. Radioimmunoassay showed that delivery of AV-CGRP significantly increased the expression of CGRP in mice hearts.</p><p><b>CONCLUSION</b>The recombinant adenovirus vector of CGRP gene has been constructed,and it can infect neonatal rat cardiomyocytes successfully. Somatic delivery of CGRP gene can significantly increase the expression of CGRP in mice hearts. The results may provide a sound foundation for further study on the value of CGRP as the target for gene therapy in both laboratory and clinical trials.</p>

Effects of short-term rapid atrial pacing on electrophysiological characteristics of atrium in hyperthyroidism / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of short-term rapid atrial pacing on the electrophysiological characteristics of atrium in hyperthyroidism.</p><p><b>METHODS</b>Forty-six adult rabbits were randomly divided into 4 groups: normal control group (n=10), pacing group (n=10), hyperthyroidism group (n=14), hyperthyroidism/pacing group (n=12). Baseline AERP and AERPs after pacing 2, 4, 6 h were determined in all groups at driver cycle length (DCL) of 200 ms, 150 ms and 130 ms.</p><p><b>RESULT</b>In pacing group, AERPs at different DCL (200 ms, 150 ms and 130 ms) were shortened after rapid pacing 2, 4, 6 h when compared with before pacing and control group (P<0.01). AERPs (at DCL of 200 ms, 150 ms and 130 ms) in hyperthyroidism group were shorter than those in control group at all time points (P<0.01). AERPs (at DCL of 200 ms, 150 ms and 130 ms) in hyperthyroidism/pacing group after rapid pacing 2, 4, 6 h were shorter than those in pacing 0 h (P<0.01) and hyperthyroidism group (P<0.05). AERP200-150 and AERP200-130 in pacing group after rapid pacing 2, 4, 6 h were significantly different from at pacing 0 h and control group (P<0.01). AERP200-150 and AERP200-130 in hyperthyroidism and hyperthyroidism/pacing group were significantly different from control group at all time points (P<0.01). No differences were observed in AERP200-150 and AERP200-130 between hyperthyroidism group and hyperthyroidism/pacing group.</p><p><b>CONCLUSION</b>Hyperthyroidism and short-term atrial pacing in the presence of hyperthyroidism can lead to remodeling of atrial electrophysiology.</p>

Effects of hyperthyroidism on the electrophysiological characteristics between atrium and gap junction between atrium and pulmonary vein / 中华急诊医学杂志

Objective To investigate the effects of hyperthyroidism on the electrophysiological characteristics of atrium and gap junction between atrium and pulmonary vein.Methods Twenty-four rabbits were randomly divided into control group and hyperthyroid group.Atrium and pulmonary vein were dissected after the atrial effective refractory period (AERP)was measured.Connexin 43(Cx43)and Counexin 40(Cx40)protein and mRNA were determined by Western blot and semi-quantitative reverse transcription-polymerase chain reaction,respectively.Results In comparison with control group,AERP and rate adaption of AERP in hyperthyroid group were significantly shorter than that of control group( P＜0.01).The concentration of Cx43 protein in hyperthyroid group was significantly higher than that of control group,but Cx40 protein was significantly lower than that of control group(P＜0.05).The expression of Cx43 mRNA in atrium and pulmonary vein was found to be up-regulated in hyperthyroid group as compared with that of control group(P＜0.01). With the level of Cx40 mRNA,there was no significant difference between two groups.Conclusion Thyroid hormone could lead to remodeling of both atrial electrophysiology and gap junction between atrium and pulmonary vein.