Effect of berberine on apoptosis of human coronary artery endothelial cells induced by intermittent high glucose / 中草药

Objective: To investigate the effect of berberine on the apoptosis of human coronary artery endothelial cells (HCAECs) induced by intermittent high glucose in vitro. Methods HCAECs were isolated, identified, cultured, and divided into five groups: Group A was the normal glucose group (5.5 mmol/L glucose, NG); Group B was the persistent high glucose group (25 mmol/L glucose, PHG); Group C was the intermittent high glucose group (5.5 mmol/L and 25 mmol/L glucose fluctuated every 24 h, IHG); Group D was in a volatile hyperosmotic environment (5.5 mmol/L and 25 mmol/L mannitol alternating every 24 h to maintain the same osmotic pressure as IHG, OC); Group E was the fluctuation of hyperglycemia + berberine intervention group (5.5 mmol/L and 25 mmol/L glucose + 50 μmol/L berberine fluctuated every 24 h, IHG + BBR). The cells of each group were changed every 24 h, and the culture was stopped after 7 d, the cell viability and apoptosis were detected in each group. The expression of the apoptosis related protein Caspase-3 was determined by qRT-PCR and Western blotting. Results Compared with the persistent high glucose group, the apoptosis of HCAECs in the intermittent high glucose group was more significant (P < 0.05), but the apoptosis of HCAECs in the fluctuation of hyperglycemia + berberine intervention group was more significantly reduced than that in the intermittent high glucose group (P < 0.05). Berberine significantly reduced the expression of Caspase-3 induced by intermittent high glucose (P < 0.05) Conclusion Intermittent high glucose decreased the activity of HCAECs more easily than the persistent high glucose,and promoted the apoptosis of HCAECs. But berberine significantly reduced the apoptosis of HCAECs under intermittent high glucose, which had a significant protective effect on HCAECs.

Proteomics study of rapamycin-injured human coronary artery endothelial cells based on two dimension fluorescence differential gel electrophoresis / 重庆医学

Objective To explore the molecular mechanisms of the effect of eluting stent drug rapamycin for injuring human coronary artery endothelial cells(HCAECs)by using the proteomics method.Methods HCAECs were treated with rapamycin,and the differentially expressed proteins were analyzed by two dimension fluorescence differential gel electrophoresis(2D-DIGE).The changed proteins were identified by MALDI-ToF-ToF.Results At least 85 differential protein spots were found,including 49 up-regulated and 36 down-regulated protein spots.Twenty-six proteins were identified by MALDI-ToF-ToF,including the endoplasmic reticulum protein,mitochondrial protein,molecular chaperones,ubiquitin system related protein,structural protein and oxidative stress related proteins,etc.Conclusion The changes of specific proteins of HCAECs injury induced by rapamycin are investigated by the proteomic method.

Interleukin-27 Enhances TNF-α-mediated Upregulation of Inflammatory Cytokine and Chemokine / 现代生物医学进展

Objective:Immune cells and inflammatory mediators play important roles in the development of atherosclerotic vascular inflammation.This study focuses on the direct effect ofIL-27 on human coronary artery endothelial cells.Methods:In this study,the effects of IL-27 and TNF-α on inflammatory cytokines and chemokines were investigated.Results:IL-27 could significantly enhance the TNF-α-mediated upregulation of inflammatory cytokine IL-6 and chemokines CCL5 from human coronary artery endothelial cells.The release of IL-6 and CCL5 was significantly suppressed by specific signaling molecule inhibitors.Conclusion:The induction of these mediators from the human coronary artery endothelial cells could be differentially regulated by the c-Jun N-terminal kinase,p38 mitogen-activated protein kinase,and nuclear factor-κB pathways.These results provided new insights into the effect of IL-27 on the TNF-α mediated activation of human coronary artery endothelial cells in atherosclerotic vascular inflammation.

Gene expression of human coronary artery endothelial cells in response to Porphyromonas endodontalis invasion / 대한치과보존학회지

During the last two decades, there has been an increasing interest in the impact of oral health on atherosclerosis and subsequent cardiovascular disease (CVD). To date, some periodontal pathogens including Porphyromonas gingivalis (P. gingivalis) have been reported to be relevant to CVD. Porphyromonas endodontalis (P. endodontalis), which shares approximately 87% sequence homology with P. gingivalis, is mostly found within infected root canals. However, recent studies reveal that this pathogen also resides in the dental plaque or periodontal pocket in patients with periodontitis. It has been shown that P. endodontalis invades human coronary artery endothelial cells (HCAEC) and coronary artery smooth muscle cells (CASMC). To evaluate whether P. endodontalis can participate in the progression of atherosclerosis and CVD, we examined the changes in transcriptional gene expression profiles of HCAEC responding to invasion by P. endodontalis in this study. The following results were obtained. 1. Porphyromonas endodontalis was invasive of HCAEC. 2. According to the microarray analysis, there were 625 genes upregulated more than two-folds, while there were 154 genes downregulated by half. 3. Upregulated genes were relevant to inflammatory cytokines, apoptosis, coagulation and immune response. Enhanced expression of MMP-1 was also noticeable. 4. The transcription profiles of the 10 selected genes examined by real-time PCR agreed well with those observed in the microarray analysis. Thus, these results show that P. endodontalis presents the potential to trigger and augment atherosclerosis leading to CVD.

Endogenous nitric oxide mediates lipoteichoic acid induced preconditioning on reoxygenation injury of cultured human coronary artery endothelial cells / 药学学报

Aim To explore the effects of lipoteichoic acid (LTA) induced delayed preconditioning (PC) on hypoxia-reoxygenation (H/R) injury of cultured human coronary artery endothelial cells (HCAECs), and to investigate the potential role of endogenous nitric oxide (NO) participated in the protective mechanism. Methods HCAECs were incubated for 2 h in a hypoxic atmosphere and reoxygenated for 4 h in a normoxic atmosphere. The delayed PC was induced by pretreatment with LTA assessed by the percentage of cellular injury with Trypan blue exclusion and by the amount of lactate dehydrogenase (LDH) in culture media. The NO level of the culture media was measured detect the expression of eNOS mRNA by RT-PCR method after cells were recovered from different points.Results LTA pretreatment significantly decreased the percentage of the killed cell and the concentration of LDH in media. Also, LTA pretreatment obviously raised the concentrations of NO in culture media. The protective effects of LTA were abrogated by pretreatment with N-monomethyl-L-arginine (L-NMMA).Moreover, the expression of eNOS mRNA was significantly upregulated after HCAECs exposure to LTA for 4 h following 2 h or 4 h recovery. Conclusion LTA could induce the delayed protection against H/R induced endothelial injury and dysfunction of cultured HCAECs. NO produced by eNOS acts initially as a trigger and subsequently as a mediator of delayed PC.