Personalized antihypertensive treatment guided by pharmacogenomics in China.

Background: The implementation of genotyping for anti-hypertensive drugs in clinical practice remains a challenge. We conducted this study to analyze the distribution of polymorphisms of antihypertensive drug-related genes in Changsha County in China and compare the clinical effectiveness of genotype-guided and clinical experience-guided antihypertensive therapy in hypertensive individuals. Methods: A total of 9,933 essential hypertensive participants from Changsha County were consecutively enrolled in our study, and 7 genetic polymorphic loci (CYP2D6*10, ADRB1, CYP2C9*3, AGTR1, ACE, CYP3A5*3 and NPPA) were detected by a polymerase chain reaction (PCR)-fluorescence probe. From an available sample of 660 hypertensive participants, 495 cases were randomly identified by genotype-guided therapy and 165 cases by clinical experience-guided therapy. We performed 24-hour ambulatory blood pressure (BP) monitoring on each of these cases, pre- and post-intervention. Results: In the enrolled 9,933 cases, the mutation frequencies of CYP2C9*3, ADRB1(1165G>C), AGTR1(1166A>C), CYP2D6*10, ACE(I/D), CYP3A5*3 and NPPA(2238T>C) were 4.41%, 74.60%, 5.55%, 57.08%, 30.94%, 69.03% and 1.19%, respectively. In both genotype-guided and clinical experience-guided groups, the comparisons of intra-group pre-and post-treatments showed significant decreases in diastolic blood pressure (DBP) (P<0.01) and significant increases in the control rate of BP (47.1% vs. 38.6% and 37.5% vs. 33.9%, P<0.05) in response to adjusted antihypertensive agents. Correspondingly, the extent of the reduction of systolic blood pressure (SBP; 3.52±11.72 vs. 0.92±9.14 mmHg), the extent of the increase in the rate of BP control (8.5% vs. 3.6%) and the percentage rate of decrease of grades 2 and 3 hypertensive individuals were more significant in the genotype-guided group than that in the clinical experience-guided group (P<0.01). Conclusions: While prescribing anti-hypertensive drugs, appropriate dosage and type adjustments should be made according to the gene mutation frequency and individual circumstances. Pharmacogenomics-guided personalized treatment of hypertensive patients is likely to be a more effective strategy, especially in those with significantly elevated SBP.

Downregulation of lncRNA SNHG16 inhibits vascular smooth muscle cell proliferation and migration in cerebral atherosclerosis by targeting the miR-30c-5p/SDC2 axis.

Atherosclerosis (AS) is the basic lesion underlying the occurrence and development of cerebrovascular diseases. Abnormal proliferation of vascular smooth muscle cells (VSMCs) plays a crucial role in AS. We aimed to explore the role of SNHG16 in AS and the molecular mechanism of VSMC involvement in the regulation of AS. The expression levels of SNHG16, miR-30c-5p and SDC2 were detected by qRT-PCR. CCK-8, wound healing and Transwell assays were used to assess ox-LDL-induced VSMC proliferation, migration, and invasion, respectively. Western blot analysis was used to detect SDC2 and MEK/ERK pathway-related protein levels. A dual-luciferase reporter assay confirmed the binding of SNHG16 with miR-30c-5p and miR-30c-5p with SDC2. SNHG16 and SDC2 expression was upregulated in patients with AS and ox-LDL-induced VSMCs, while miR-30c-5p was downregulated. Ox-LDL-induced VSMC proliferation and migration were increased, and the MEK/ERK signalling pathway was activated. MiR-30c-5p was targeted to SNHG16 and SDC2. Downregulating SNHG16 or upregulating miR-30c-5p inhibited ox-LDL-induced VSMC proliferation and migration and inhibited MEK/ERK signalling pathway activation. In contrast, downregulating miR-30c-5p or upregulating SDC2 reversed the effects of downregulating SNHG16 or upregulating miR-30c-5p. Furthermore, downregulating SDC2 inhibited ox-LDL-induced proliferation and migration of VSMCs and inhibited activation of the MEK/ERK signalling pathway, while upregulating lncRNA SNHG16 reversed the effects of downregulating SDC2. Downregulation of SNHG16 inhibited VSMC proliferation and migration in AS by targeting the miR-30c-5p/SDC2 axis. This study provides a possible therapeutic approach to AS.

Transcriptome profile of rat genes in injured spinal cord at different stages by RNA-sequencing.

BACKGROUND: Spinal cord injury (SCI) results in fatal damage and currently has no effective treatment. The pathological mechanisms of SCI remain unclear. In this study, genome-wide transcriptional profiling of spinal cord samples from injured rats at different time points after SCI was performed by RNA-Sequencing (RNA-Seq). The transcriptomes were systematically characterized to identify the critical genes and pathways that are involved in SCI pathology. RESULTS: RNA-Seq results were obtained from total RNA harvested from the spinal cords of sham control rats and rats in the acute, subacute, and chronic phases of SCI (1 day, 6 days and 28 days after injury, respectively; n = 3 in every group). Compared with the sham-control group, the number of differentially expressed genes was 1797 in the acute phase (1223 upregulated and 574 downregulated), 6590 in the subacute phase (3460 upregulated and 3130 downregulated), and 3499 in the chronic phase (1866 upregulated and 1633 downregulated), with an adjusted P-value <0.05 by DESeq. Gene ontology (GO) enrichment analysis showed that differentially expressed genes were most enriched in immune response, MHC protein complex, antigen processing and presentation, translation-related genes, structural constituent of ribosome, ion gated channel activity, small GTPase mediated signal transduction and cytokine and/or chemokine activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the most enriched pathways included ribosome, antigen processing and presentation, retrograde endocannabinoid signaling, axon guidance, dopaminergic synapses, glutamatergic synapses, GABAergic synapses, TNF, HIF-1, Toll-like receptor, NF-kappa B, NOD-like receptor, cAMP, calcium, oxytocin, Rap1, B cell receptor and chemokine signaling pathway. CONCLUSIONS: This study has not only characterized changes in global gene expression through various stages of SCI progression in rats, but has also systematically identified the critical genes and signaling pathways in SCI pathology. These results will expand our understanding of the complex molecular mechanisms involved in SCI and provide a foundation for future studies of spinal cord tissue damage and repair. The sequence data from this study have been deposited into Sequence Read Archive ( http://www.ncbi.nlm.nih.gov/sra ; accession number PRJNA318311).

Temporal kinetics of CD8+ CD28+ and CD8+ CD28- T lymphocytes in the injured rat spinal cord.

This study aims to explore the temporal changes of cytotoxic CD8+ CD28+ and regulatory CD8+ CD28- T-cell subsets in the lesion microenvironment after spinal cord injury (SCI) in rats, by combination of immunohistochemistry (IHC) and flow cytometry (FCM). In the sham-opened spinal cord, few CD8+ T cells were found. After SCI, the CD8+ T cells were detected at one day post-injury (dpi), then markedly increased and were significantly higher at 3, 7, and 14 dpi compared with one dpi (p < 0.01), the highest being seven dpi. In CD8+ T cells, more than 90% were CD28+ , and there were only small part of CD28- ( < 10%). After 14 days, the infiltrated CD8+ T cells were significantly decreased, and few could be found in good condition at 21 and 28 dpi. Annexin V and propidium iodide (PI) staining showed that the percentages of apoptotic/necrotic CD8+ cells at 14 dpi and 21 dpi were significantly higher than those of the other early time-points (p < 0.01). These results indicate that CD8+ T cells could rapidly infiltrate into the injured spinal cords and survive two weeks, however, cytotoxic CD8+ T cells were dominant. Therefore, two weeks after injury might be the "time window" for treating SCI by prolonging survival times and increasing the fraction of CD8+ regulatory T-cells. © 2016 Wiley Periodicals, Inc.

Co-transplantation of MRF-overexpressing oligodendrocyte precursor cells and Schwann cells promotes recovery in rat after spinal cord injury.

Oligodendrocyte (OL) replacement is a promising treatment strategy for spinal cord injury (SCI). However, the poor survival of transplanted OLs or their precursors and inhibition of axonal regeneration are two major challenges with this approach. Our previous study showed that Schwann cells (SCs) promoted survival, proliferation, and migration of transplanted OL progenitor cells (OPCs) and neurological recovery. Remyelination is an important basis for functional recovery following spinal cord injury. It has been reported that myelin gene regulatory factor (MRF), a transcriptional regulator which specifically is expressed in postmitotic OLs within the CNS, is essential for OL maturation and CNS myelination. In the present study, we investigated whether co-transplantation of MRF-overexpressing OPCs (MRF-OPCs) and SCs could improve functional recovery in a rat model of contusional SCI. MRF overexpression had no effect on OPC survival or migration, but stimulated the differentiation of OPCs both in vitro and in vivo. Co-transplantation of MRF-OPCs and SCs increased myelination and tissue repair after SCI, leading to the recovery of neurological function. These results indicate that co-transplantation of MRF-OPCs and SCs may be an effective treatment strategy for SCI.

Differential effects of myelin basic protein-activated Th1 and Th2 cells on the local immune microenvironment of injured spinal cord.

Myelin basic protein (MBP) activated T cells (MBP-T) play an important role in the damage and repair process of the central nervous system (CNS). However, whether these cells play a beneficial or detrimental role is still a matter of debate. Although some studies showed that MBP-T cells are mainly helper T (Th) cells, their subtypes are still not very clear. One possible explanation for MBP-T immunization leading to conflicting results may be the different subtypes of T cells are responsible for distinct effects. In this study, the Th1 and Th2 type MBP-T cells (MBP-Th1 and -Th2) were polarized in vitro, and their effects on the local immune microenvironment and tissue repair of spinal cord injury (SCI) after adoptive immunization were investigated. In MBP-Th1 cell transferred rats, the high levels of pro-inflammatory cells (Th1 cells and M1 macrophages) and cytokines (IFN-γ, TNF-α, -ß, IL-1ß) were detected in the injured spinal cord; however, the anti-inflammatory cells (Th2 cells, regulatory T cells, and M2 macrophages) and cytokines (IL-4, -10, and -13) were found in MBP-Th2 cell transferred animals. MBP-Th2 cell transfer resulted in decreased lesion volume, increased myelination of axons, and preservation of neurons. This was accompanied by significant locomotor improvement. These results indicate that MBP-Th2 adoptive transfer has beneficial effects on the injured spinal cord, in which the increased number of Th2 cells may alter the local microenvironment from one primarily populated by Th1 and M1 cells to another dominated by Th2, Treg, and M2 cells and is conducive for SCI repair.

The effect of endothelial progenitor cells on angiotensin II-induced proliferation of cultured rat vascular smooth muscle cells.

Previous studies have demonstrated that endothelial progenitor cells (EPCs) could delay the progress of vascular remodeling in blood vessel-proliferating diseases. The proliferation of vascular smooth muscle cells (VSMCs) is a pivotal factor in cardiovascular diseases. In this study, we investigated whether EPCs could inhibit the Angiotensin II (Ang II)-induced proliferation of VSMCs. The effect of early EPC-conditioned medium (E-EPC-CM), late EPCs-CM (L-EPC-CM), and HUVEC-CM on Ang II-induced proliferation of VSMCs was assessed by BrdU incorporation, total protein content, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, and flow cytometry. Reverse transcriptase-polymerase chain reaction and Western blot were performed to analyze the effect of different CMs on Ang II-induced phosphorylations of ERK, JNK, p38, and NF-κB subunit p65 and the expressions of c-myc and c-fos. E-EPC-CM, L-EPC-CM, and HUVEC-CM significantly inhibited the Ang II-induced DNA synthesis, total protein expression, cell survival, and cell cycle progress of VSMCs. Furthermore, E-EPC-CM significantly inhibited the Ang II-induced phosphorylation of ERK, JNK, p38, and p65 (nuclear translocation of p65) and the expressions of c-myc and c-fos. Taken together, these data suggested that EPCs may delay the progress of vascular remodeling in blood vessel-proliferating diseases by inhibiting Ang II-induced proliferation of VSMCs through inactivating MAPKs and NF-κB signaling pathways and by reducing the expressions of c-myc and c-fos.

Calcitonin gene-related peptide released from endothelial progenitor cells inhibits the proliferation of rat vascular smooth muscle cells induced by angiotensin II.

We have recently demonstrated that endothelial progenitor cells (EPCs) inhibit AngII-induced proliferation of vascular smooth muscle cells (VSMCs) by inactivating MAPKs and NF-κB signaling pathway and reducing expression of oncogene c-myc and c-fos. The inhibitory effect of EPCs on VSMCs is associated with paracrine mechanism. However, the potential mechanism of EPCs on the regulation of AngII-induced proliferation of VSMCs was unknown. Calcitonin gene-related peptide (CGRP) could inhibit AngII-induced proliferation and transformation of VSMCs. However, it has not been known whether CGRP released from EPCs is a potential regulator in regulation of AngII-induced proliferation of VSMCs. Early endothelial progenitor cell-conditioned medium(E-EPC-CM) was pre-incubated with functional blocking antibodies against CGRP for 1 h or VSMCs was preteated with CGRP(837)(CGRP receptor antagonist) for 1 h before VSMCs were pretreated with CM for 30 min. DNA synthesis ability, total protein levels, cell survival, signal transduction, and expressions of c-myc and c-fos of VSMCs induced by AngII (10(-6)mol/l) were detected to assess the role of CGRP in AngII-induced proliferation of VSMCs. E-EPC-CM could significantly inhibit AngII-induced DNA synthesis ability, total protein levels, cell survival, phosphorylation of ERK, JNK, p38, p65, and expressions of c-myc and c-fos compared with the control group(P < 0.05). However, Pretreatment with anti-CGRP antibody and CGRP(837) could significantly weaken the inhibitory effect of E-EPC-CM on proliferation of VSMCs induced by AngII (P < 0.05). EPCs exert anti-proliferative effects on VSMCs mediated by the release of CGRP.

Losartan inhibits monocytic adhesion induced by ADMA via downregulation of chemokine receptors in monocytes.

OBJECTIVE: Asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase (NOS) inhibitor, can induce the adhesiveness of monocytes to vascular endothelium, and chemokines play an important role in this process. The present study was carried out to test whether the inhibitory effect of losartan on ADMA-induced monocytic adhesion is mediated by chemokine receptors. METHODS: Human monocytoid cells (THP-1) were incubated with exogenous ADMA (30 microM) for 4 or 24 h in the absence or presence of losartan. The monocytic adhesion, the levels of chemokines, and the expression of chemokine receptors were determined. The possible signal pathway was also explored. RESULTS: In cultured monocytes, ADMA (30 microM) markedly increased monocytic adhesion to endothelial cells, elevated the levels of monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8), and upregulated the mRNA expression of chemokine receptors CCR2 and CXCR2. Exposure to ADMA (30 microM) significantly induced the generation of intracellular reactive oxygen species (ROS) and activation of nuclear factor (NF)-kappaB. Pretreatment with AT1 receptor blocker (ARB) losartan (1, 3, 10 microM) attenuated monocytic adhesiveness elicited by ADMA and downregulated the expression of CCR2 and CXCR2 mRNA, accompanied by a significant decrease in ROS generation and NF-kappaB activity and expression. CONCLUSION: The present study suggests that the inhibitory effect of losartan on ADMA-induced monocytic adhesion may be related to downregulation of chemokine receptors by inhibiting the ROS/NF-kappaB pathway.

[Change of endothelial progenitor cells from peripheral blood in patients with coronary heart diseases before and after percutaneous coronary intervention].

OBJECTIVE: To investigate the culture of endothelial progenitor cells (EPCs) from peripheral blood in patients with coronary heart diseases (CHD) before and after percutaneous coronary intervention (PCI), and to observe the cells shape and determine the cell number and proliferation activity. METHODS: Ninety-five patients were divided into a CHD group(n=65) and a control group (n=30). The mononuclear cells were isolated from peripheral blood of patients with CHD before, right after and 4 days after PCI by Ficoll-density centrifugation. The isolated cells were cultured in RPMI1640 medium supplemented with VEGF165 and bFGF.EPCs were characterized as adherent cells of double positive for DiL-acLDL uptake and FITC-UEA-I binding by direct fluorescent staining under a fluorescence microscope. The EPCs specific surface mark CD34 and KDR were assessed by fluorescence activated cell sorter analysis. The cell shapes were analysed and the number of colony-forming units(CFU) was counted by phase-contrast microscope. RESULTS: The number of EPCs reduced in patients with CHD before the PCI, but the cell number was significantly increased in patients with CHD after the PCI, and the number reduced in patients with CHD 4 days after the PCI. How-ever, the number of CFUs did not change in patients before and after the PCI. CONCLUSION: PCI can increase endothelial progenitor cells in patients after the PCI; but 4 days after the PCI, this increase will not exist.

[Differentiation of 2 endothelial progenitor cells in vitro and inhibitory effect of asymmetric dimethylarginine on its proliferation].

OBJECTIVE: To investigate the Methods for culturing two types of endothelial progenitor cells (EPC) from human umbilical cord blood and study their differentiation traits and the depressant effect of asymmetric dimethylarginine (ADMA) on its proliferation. METHODS: Mononuclear cells were isolated from fresh cord blood by 6% hydroxyethyl starch(HES) and density gradient centrifugation.Isolated cells were cultured in the medium supplemented with vascular endothelial growth factors (VEGF) and basic fibroblast growth factors (bFGF). The growth characteristics and biological features of the cells were observed at different time points and identified by morphology,immunofluorescence staining,reverse transcription polymerase chain reaction (RT-PCR), and flow cytometry.Attached cells were incubated with different concentrations of ADMA (1,5, and 10 micromol/L) for 24,48, and 72 hours. Methylthiazoletetrazolium (MTT) assay and quantified colony forming units (CFUs) were used to assess the proliferation of endothelial progenitor cells. RESULTS: The attached cells were divided into 2 types:early EPC and late EPC. Early EPC changed from small sized round cells to spindle shaped cells and late EPC formed a typical cobblestone-like cells. Fluorescence microscopy showed that EPC were positive for both Dil-acLDL uptake and FITC-UEA-I binding.RT-PCR and FACS showed the difference of endothelial cell-specific,gene expression and changed AC133,CD34, and KDR among different times.Incubation of EPC with ADMA dose and time-dependently decreased the number and the proliferation of EPC. CONCLUSION: There are 2 types of EPC from a source of human umbilical cord blood and ADMA may depress the EPC proliferation, providing a basis for further research.

[Effect of chronic iron overload on atherosclerosis lesion in apolipoprotein E knockout mice].

OBJECTIVE: To explore the effect of chronic iron overload on the lesion of atherosclerosis (AS) in apolipoprotein (apo) E knockout mice. METHODS: Twenty-four ApoE knockout mice were randomly divided into ApoE knockout group (0.1 mL saline for 4 weeks) and iron overload group (10 mg iron dextran for 4 weeks). The levels of serum iron (SI), total iron binding capacity, contents of malondialdehyde (MDA), and activity of superoxide dismutase (SOD) in the liver were measured. Iron deposition in the liver and heart was determined, and atherosclerotic plaque areas of the sinus aortae were analyzed. RESULTS: In the iron overload group, the levels of SI increased by 377.86%, the saturation of transferrin increased by 121.98% and the levels of iron in the liver increased by 2,548.15% (P<0.01). The contents of MDA in the liver increased by 32.51% (P<0.01), and the activity of SOD in the liver decreased by 17.2% in the ApoE knockout group (P<0.05). The level of MDA in the liver increased by 411.15%, and the activity of SOD in the liver decreased by 46.84% in the iron overload group (P<0.01). There was a significant deposition of iron in the liver and heart of mice, and the areas of atherosclerotic plaque of sinus aortae increased markedly in the iron overload group. CONCLUSION: Chronic iron overload may promote the development of AS lesion in the ApoE knockout mice, in which the increased oxidative stress and lipid oxidation may involve.

[Effect of melatonin on the proliferation, apoptosis, and expression of bcl-2 in oxidized low-density lipoprotein-induced endothelial progenitor cells].

OBJECTIVE: To explore the effect of melatonin(Mel) on the proliferation, apoptosis and expression of bcl-2 in oxidized low-density lipoprotein(ox-LDL)-induced endothelial progenitor cells (EPC) from human umbilical cord blood in vitro. METHODS: Total mononuclear cells were isolated from human umbilical cord blood in vitro by Ficoll density gradient centrifugation, and the cells were plated on fibronectin-coated culture dishes. After 7 days, the attached cells were divided into 7 groups: a control group (normal cells), 3 ox-LDL groups[the attached cells were incubated with different concentrations of ox-LDL(5,10,and 20mg/L) for 24 hours], and 3 Mel groups[the attached cells were incubated with different concentrations of Mel (0.5,1.0, and 2.0 mmol/L) respectively for 24 hours before incubation with 10 mg/L ox-LDL]. EPC was identified by examining the expression of CD34, vascular endothelial growth factor receptor-2(VEGFR-2) and CD133 under a laser scanning confocal microscope. We used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to detect the effect of Mel and ox-LDL on the multiplication ability of EPC. Flow cytometry was used to detect the apoptosis. The expressions of Bcl-2 mRNA and protein were detected respectively by RT-PCR and immunohistochemistry technology. RESULTS: After being exposed to the ox-LDL, the proliferation of EPC in the 3 ox-LDL groups was lower, and the apoptosis rate was higher than that in the control group in a dose-dependent manner (P<0.01); Mel was added at different concentrations before the ox-LDL incubation, and the cells in the 3 Mel groups showed higher proliferation and lower apoptosis rate than those of the 3 ox-LDL groups (P<0.01). Expression of Bcl-2 mRNA and protein of EPC in the 3 Mel groups was higher than that in the 3 ox-LDL groups (P<0.01). CONCLUSION: Ox-LDL can inhibit the proliferation of EPC and promote the apoptosis of the cells by down-regulating the bcl-2 expression. Mel can inhibit these effects of ox-LDL.

[Short- and long-term therapeutic effects of combination therapy with perindopril and irbesartan in a rat model of dilated cardiomyopathy].

OBJECTIVE: To evaluate the short-term, and long-term therapeutic effects of combination therapy with perindopril and irbesartan in a rat model of dilated cardiomyopathy (DCM). METHODS: Sprague-Dawley rats were administered adriamycin intraperitoneally to develop DCM. Grouping of rats: Group A contained normal rats, and Group B contained DCM rats. Both Group A and B were not given drug treatment. Group C and D contained DCM rats, however, Group C was administered perindopril 2mg/(kg x d) while Group D was administered perindopril 1mg/(kg x d) and irbesartan 25mg/(kg x d). Brain natriuretic peptide (BNP) was determined by enzyme linked immunosorbent assay; plasma potassium and creatinine were measured; the pathological lesions of cardiac muscle tissues were evaluated after HE staining; and the survival time of each rat during the intervention was recorded. RESULTS: After the three-week intervention, the plasma concentrations of BNP in Group D were lower than those in Group C (P<0.05). In each group, plasma concentrations of potassium and creatinine showed no significant differences between pre-intervention and post-intervention (P>0.05); pathological lesions of cardiac muscle tissues in both Group C and D were attenuated compared with those in Group B (P<0.01), but pathological lesions of cardiac muscle tissues showed no significant differences between Group C and Group D (P>0.05). Log-rank test showed that the life span of Group C was shorter than that of Group D (P<0.05); Cox regression analysis showed that both combination therapy and monotherapy with perindopril could prolong the survival time, but the effect of combination therapy was more obvious. CONCLUSION: Combination therapy with perindopril and irbesartan in a rat model of DCM can more effectively improve the cardiac function and long-term prognosis than those monotherapy with perindopril. Both these two treatment plans can attenuate the pathological lesions of cardiac muscle tissues, without elevating the concentrations of plasma potassium and creatinine.

Role of asymmetric dimethylarginine in inflammatory reactions by angiotensin II.

Previous investigations have demonstrated that angiotensin (Ang) II induces inflammatory reactions and asymmetric dimethylarginine (ADMA), an endogenous NOS inhibitor, might be a novel inflammatory factor. Endothelial cell activation was induced by incubation with Ang II or ADMA. Incubation with Ang II (10(-6) M) for 24 h elevated the levels of ADMA and decreased the levels of nitrite/nitrate concomitantly with a significant increase in the expression of protein arginine methyltransferase and a decrease in the activity of dimethylarginine dimethylaminohydrolase (DDAH). Exposure to Ang II (10(-6) M for 24 h) also enhanced intracellular ROS elaboration and the levels of tumor necrosis factor (TNF)-alpha and interleukin (IL)-8, upregulated chemokine receptor CXCR2 mRNA expression, increased adhesion of endothelial cells to monocytes and induced a significant increase in the activity of nuclear factor (NF)-kappaB, which was attenuated by pretreatment with the Ang II receptor blocker losartan (1, 3 and 10 muM). Exogenous ADMA (30 microM) also increased ROS generation and the levels of TNF-alpha and IL-8, decreased the levels of nitrite/nitrate, upregulated CXCR2 gene expression, increased endothelial cell binding with monocytes and activated the NF-kappaB pathway, which was inhibited by pretreatment with losartan or L-arginine. These data suggest that ADMA is a potential proinflammatory factor and may be involved in the inflammatory reaction induced by Ang II.

[Relevant factors of microalbuminuria in aged patients with essential hypertension].

OBJECTIVE: To investigate the relationship between microalbuminuria and endothelial-dependent relaxing function and atherosclerosis of common carotid artery (CCA) in aged patients with essential hypertension (EH). METHODS: Sixty-four aged EH patients were recruited. According to the albumin excretion rate (AER) in the urine measured by immunoturbidimetry, patients were divided into 2 groups: normoalbuminuria group (NAU group) and microalbuminuria group (MAU group). Thirty aged persons without EH were served as the control group. The endothelium-dependent relaxing function of blood vessels, intima-media thickness (IMT) and the plaque of CCA were measured by color Doppler ultrasound. RESULTS: The flow-mediated dilation in the MAU group [(4.98+/-1.35)%] and that in the NAU group [(6.31+/-1.14)%] were significantly lower than that in the control group [(9.09+/-1.83)%, P<0.05, respectively], especially lower in the MAU group. The IMT of CCA in the MAU group [(0.97+/-0.19)mm] and that in the NAU group [(0.86+/-0.10)mm] were significantly thicker than that in the control group [(0.78+/-0.13)mm] (P<0.05, respectively), especially thicker in the MAU group. The analysis of multiple stepwise regression showed that the microalbuminuria was successively related to EDF, the IMT of CCA, the plaque index of CCA, systolic blood pressure, etc. CONCLUSION: EDF is impaired, and there is the atherosclerosis of CCA in aged patients with EH. Microalbuminuria correlates with the decrease of endothelium-dependent relaxing function and the IMT of CCA in aged patients with EH.

[Effects of fenofibrate on the proliferation and apoptosis and nitric oxide synthase expression of cultured human umbilical vein endothelial cells induced by lysophosphatidylcholine].

OBJECTIVE: To investigate the effects of fenofibrate on the proliferation and apoptosis and endothelial nitric oxide synthase (eNOS) mRNA expression of cultured human umbilical vein endothelial cells (HUVECs) induced by lysophosphatidylcholine (LPC). METHODS: HUVECs were cultured in vitro. The study was designated to 5 groups according to fenofibrate concentration: control group, LPC group, LPC + low-concentration fenofibrate (10 micromol/L), LPC + middle-concentration fenofibrate (50 micromol/L), and LPC + high-concentration fenofibrate (100 micromol/L). The study was designated to 6 groups according to the intervention time: control group, LPC group, LPC + fenofibrate (50 micromol/L) 6 h, LPC + fenofibrate 12 h, LPC + fenofibrate 24 h, and LPC + fenofibrate 48 h. The proliferation and apoptosis of HUVECs were evaluated by MTT assay, flow cytometry and fluorescence microscopy, respectively. eNOS mRNA were assayed by real time-PCR. RESULTS: Compared with the control group, LPC could inhibit the proliferation and induce apoptosis, and downregulate eNOS mRNA expression and decrease NO production of HUVECs. Fenofibrate could increase the proliferation and decrease the apoptosis, and up-regulate eNOS mRNA expression and enhance NO production in HUVECs. CONCLUSION: Fenofibrate could improve the proliferation and inhibit the apoptosis, and up-regulate eNOS mRNA expression of HUVECs induced by LPC, which may be responsible for fenofibrate to prevent and treat atherosclerosis.

[Effects of advanced glycation end products on the activity of NF-kappaB and the expression of fibronectin mRNA in the endothelial cells in aged rats].

OBJECTIVE: To explore the effects of advanced glycation end products (AGEs) on the activity of NF-kappaB and fibronectin (Fn) synthesis in the endothelial cells in aged rats. METHODS: Endothelial cells were cultured in M199 from the aorta of 24 month old rats and divided into 3 groups: Group A (5 mmol/L glucose) as controls, Group B (25 mg/L AGEs for 48 h), and Group C (50 mg/L AGEs for 48 h). The activity of NF-kappaB was evaluated by immunofluorescence and the expression of Fn mRNA was measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Compared with the controls, AGEs induced the activity of NF-kappaB and increased the Fn mRNA expression in a concentration-dependent manner (P<0.05). CONCLUSION: The activity of NF-kappaB and up-regulated expression of Fn mRNA induced by AGEs may contribute to chronic complications of diabetes.

[Effect of atorvastatin on the nuclear factor-kappaB and soluble inter-cellular adhesion molecules-1 in patients with acute coronary syndrome].

OBJECTIVE: To determine the effect of atorvastatin on the activity of peripheral blood lymocyte nuclear factor-kappaB (NF-kappaB) and plasma soluble inter-cellular adhesion molecules-1 (sICAM-1) in acute coronary syndromes. METHODS: Sixty-eight patients with acute coronary syndrome were randomly divided into atorvastatin therapeutic group (n = 37) and conventional therapeutic group (n = 31). Enzyme-linked immunosorbent assay was used to measure the plasma sICAM-1, and immunohistochemical method was used to measure the activity of NF-kappaB in the peripheral blood lymphocyte before and 12 weeks after the therapy in the two groups. RESULTS: Before the therapy, the level of NF-kappaB activity in the peripheral blood lymphocytes [(45.2 +/- 8.4)% vs (40.8 +/- 9.2)%, P > 0.05] and the plasma sICAM-1 [(357.2 +/- 84.5) ng/ml vs (365.5 +/- 91.3) ng/ml, P > 0.05] in the two groups had no significant difference. After 12 weeks of the therapy, in the conventional group the level of NF-kappaB activity in the peripheral blood lymphocytes [(40.8 +/- 9.2)% vs (38.7 +/- 8.9)%, P > 0.05] and the plasma sICAM-1 [(357.2 +/- 84.5) ng/ml vs (365.5 +/- 91.3) ng/ml, P > 0.05] still had no significant difference. But in the atorvastatin group the level of NF-kappaB activity in the peripheral blood lymphocyte [(45.2 +/- 8.4)% vs (25.6 +/- 7.9)%, P < 0.05 and the plasma soluble inter-cellular adhesion molecule-1 [(357.2 +/- 84.5) ng/ml vs (175.8 +/- 75.6) ng/ml, P < 0.05] did show a significant difference. CONCLUSION: The cholesterol-lowering therapy with atorvastatin can reduce the inflammation and stabilize the plaque in the acute coronary syndrome, which may be related to the inhibition of NF-KB and sICAM-1 activities.

[Related inflammation markers among different types of coronary heart disease].

OBJECTIVE: To study the relationship between levels of activity of NF-kappaB p65, plasma soluble inter-cellular adhesion molecule-1, C-reactive protein on plaque stability, and different types of coronary heart disease. METHODS: We measured the levels of plasma soluble inter-cellular adhesion molecule-1 and C-reactive protein by enzyme-linked immunosorbant assay and the activity of NF-kappaB p65 in peripheral blood lymocytes immunohistochemically. RESULTS: Compared with the stable angina and the control group, the baseline activity of NF-kappaB p65, sICAM-1 and C-reactive protein was significantly elevated in the acute myocardial infarction and the unstable angina (P <0.01). After 3 month follow-up, the levels of activity of NF-kappaB p65 and sICAM-1 were unchanged (P > 0.05). In all groups, C-reactive proteins were lowered at the review (P <0.01). CONCLUSION: The levels of activity of NF-kappaB p65, sICAM-1 and C-reactive protein are related to the plaque stability among different types of coronary heart disease. NF-kappaB p65, and sICAM-1 are not affected by the acute event. These plasma markers may be important risk factors for the development of the acute coronary syndrome.