Vitamin D prevents high glucose-induced human umbilical vein endothe-lial cell apoptosis by inhibition of prolyl isomerase 1-mediated mitochon-drial oxidative stress / 中国病理生理杂志

AIM:To observe the effects of vitamin D on the apoptosis ,prolyl isomerase 1(Pin1)protein ex-pression and activity ,mitochondrial translocation of p 66Shc,and reactive oxygen species(ROS)production in high glu-cose-cultured human umbilical vein endothelial cells(HUVECs),and to explore the role of vitamin D receptor(VDR)in these processes.METHODS:HUVECs were treated with high glucose(33 mmol/L)in the presence or absence of vita-min D or Pin1 inhibitor juglone.The cell apoptosis was measured by flow cytometry and TUNEL staining.Intracellular ROS levels were examined by flow cytometry and fluorescence microscopy.The protein levels of Pin1,p66Shc,p-p66Shc,mito-chondria to cytoplasm ratio of p66Shc,and caspase-3 in HUVECs were measured by Western blot.Pin1 activity in HU-VECs lysate was assessed by a commercial kit.Knockdown of VDR by siRNA was conducted to evaluate the role of VDR in the regulatory effects of vitamin D on Pin 1 protein expression and activity in HUVECs under high-glucose condition.RE-SULTS:Vitamin D suppressed the apoptosis and intracellular ROS generation of HUVECs induced by high glucose(P<0.05).Vitamin D inhibited high glucose-induced upregulation of Pin1 protein expression and activity(P<0.05).Vita-min D inhibited the phosphorylation and mitochondrial translocation of p 66Shc and caspase-3 protein expression induced by high glucose(P<0.05).Knockdown of VDR by siRNA abolished the inhibitory effects of vitamin D on high glucose-in-duced upregulation of Pin 1 protein expression and activity.CONCLUSION:Vitamin D alleviates high glucose-induced endothelial cell apoptosis by inhibition of Pin 1 protein expression and activity ,and attenuation of p66Shc-mediated mito-chondrial oxidative stress ,which are dependent on VDR activation.

The stability of the atherosclerotic plaque depends on the extent of injured endothelium:results from a novel model of ischemia/reperfusion induced atherosclerosis in carotid artery of rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To observe the atherogenic lesion progress in a novel ischemia/reperfusion induced atherosclerosis model in the carotid artery of rats.</p><p><b>METHODS</b>Rats were divided into normal control, sham-operated control and ischemia-reperfusion injury (IRI) groups (n = 10 each). IRI was induced by 30 min carotid artery occlusion with a 2 cm long artery clips in anesthetized rats. Four weeks later, hematoxylin and eosin (HE) and immunohistochemical stain were performed on carotid arteries of various groups. The ratio of neointima area/media area (I/M) and expression of platelet endothelial cell adhesion molecule (PECAM-1/CD31) were compared among groups.</p><p><b>RESULTS</b>(1) Neointimal hyperplasia was detected in carotid artery of IRI group and the I/M ratio was significantly higher than in normal control and sham-operated groups (1.328 ± 0.301 vs. 0.011 ± 0.004 and 0.017 ± 0.008, all P < 0.01). (2) Small to large-sized neointima were found in the IRI group and the small sized intima was stable while large sized intima which covered the whole cavity was instable and underwent spontaneous rupture and thrombosis formation. (3) CD31 expression was significantly upregulated in carotid artery of IRI group corresponding to the instability of neointima in this group.</p><p><b>CONCLUSION</b>Ischemia-reperfusion injury of carotid artery could result in atheroma in rats, this model could be used for future research on the pathogenesis of atherosclerosis. Our results show that endothelium injury of the arteries is the key factor to trigger atheroma and responsible for the disruption of the plaque.</p>

Angiotensin-converting enzyme 2 gene transfer attenuates neointimal formation after carotid artery ischemia-reperfusion injury in rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the effects of lentiviral recombinant angiotensin-converting enzyme 2 (LV-ACE2) gene transfer on the neointimal formation after carotid artery ischemia-reperfusion injury (IRI) and related mechanisms.</p><p><b>METHODS</b>IRI was induced in SD rats through the carotid artery clipping and rats were divided into IRI, IRI+LV-GFP, IRI+LV-ACE2, IRI+ paclitaxel groups (n = 10 each). Sham operated rats serve as normal control. Four weeks later, neointimal formation was observed on HE stained carotid artery sections. The protein expression of ACE2, α-SM-actin, CD31, AT1R and P-ERK were detected by immunohistochemistry.</p><p><b>RESULTS</b>(1) Carotid artery neointimal hyperplasia was readily shown in IRI group [I/M: 1.517 ± 0.151 (4 weeks later) vs. 0.011 ± 0.004 (Sham), P < 0.01], which was significantly reduced in IRI+LV-ACE2 (0.71 ± 0.17) and IRI+ paclitaxel (0.89 ± 0.21) groups. (2) The growth of vascular smooth muscle cells and neovascularization were also significantly inhibited in IRI+LV-ACE2 group and the expression of α-SM-actin (5 843 ± 839 vs. 12 648 ± 1 760, P < 0.01) and CD31 [(12.40 ± 4.01)/mm(2) vs. (96.20 ± 17.79)/mm(2), P < 0.01], AT1R (1 219 ± 175 vs. 4 861 ± 545, P < 0.01) and P-ERK1/2 phosphorylation (1 040 ± 215 vs. 2 938 ± 286, P < 0.01) in the neointimal of the injury arteries in IRI+LV-ACE2 group were significantly downregulated compared to IRI group.</p><p><b>CONCLUSION</b>This data suggest that ACE2 gene overexpression is able to attenuate neointimal formation after ischemia-reperfusion injury possibly through downregulating AT1 receptor expression and signal pathway of ERK1/2 phosphorylation.</p>

Overexpression of angiotensin-converting enzyme 2 inhibits angiotensin II type 1 receptor expression and signal transducer and activator of transcription 3 phosphorylation in cultured smooth muscle cells / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the effects of recombinated lentiviral angiotensin-converting enzyme 2 (ACE2) vector transfer on the expression of angiotensin II type 1 (AT1) receptor in cultured vascular smooth muscle cells (VSMCs).</p><p><b>METHODS</b>VSMCs were divided into 7 groups: (1) CONTROL: serum-free culture medium; (2) Lentiviral-GFP vector group: Lentiviral-GFP vector (MOI = 10); (3) Ang II group (10(-7) mol/L); (4) Ang II (10(-7) mol/L) + Lentiviral-ACE2 (MOI = 10) group; (5) Ang II (10(-7) mol/L) + Irbesartan (10(-7) mol/L) group ; (6) Ang II (10(-7) mol/L) + irbesartan (10(-7) mol/L) + Lentiviral-ACE2 (MOI = 10) group ; (7) Lentiviral-ACE2 (MOI = 10) group. Ninety-six hours later, the proliferation of VSMCs was determined with CCK-8 Kit. AT1 receptor mRNA and protein expressions were detected with quantitative real-time PCR and Western blot, the signaling pathway of signal transducer and activator of transcription 3 (STAT3) was also detected.</p><p><b>RESULTS</b>ACE2 gene transfer significantly inhibited the VSMCs proliferation in the absence or presence of Ang II. AT1 receptor mRNA and protein expressions were also significantly downregulated in the absence or presence of Ang II. Similar to AT1 receptor mRNA and protein expression changes, STAT3 phosphorylation was also significantly inhibited by ACE2 overexpression.</p><p><b>CONCLUSION</b>Our results suggest that overexpression of ACE2 gene could inhibit the VSMCs proliferation by downregulating AT1 receptor expression and STAT3 phosphorylation. ACE2 could also directly inhibit AT1 receptor in cultured VSMCs.</p>

Effect of atorvastatin on advanced glycation end products induced monocyte chemoattractant protein-1 expression in cultured human endothelial cells / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of atorvastatin on advanced glycation end products (AGE) induced monocyte chemoattractant protein-1 (MCP-1) expression in human umbilical vein endothelial cells (HUVECs) and whether this effect could be linked to peroxisome proliferator-activated receptor-γ (PPAR-γ) and nuclear factor-κB (NF-κB).</p><p><b>METHODS</b>Grouping: (1) Blank control group; (2) BSA group; (3) AGE group: cells were incubated with different concentrations of AGE (10(-4), 10(-3), 10(-2) and 10(-1) g/L) for 24 hours; (4) AGE + Atorvastatin group: cells were incubated with different concentrations of atorvastatin (0.1, 1, 10 µmol/L) for 1 hour, then incubated with AGE (10(-1) g/L) for 24 hours; (5) PPAR-γ agonist (15 d-PGJ2) group: cells were incubated with 15 d-PGJ2 (10 µmol/L) for 1 hour, then incubated with AGE (10(-1) g/L) for 24 hours; (6) PPAR-γ inhibitor (GW9662) group: cells were incubated with GW9662 (5000 nmol/L) for 1 hour, then incubated with atorvastatin (1 µmol/L) and AGE (10(-1) g/L) for 24 hours. Collagenase was used to isolate the endothelial cell from human umbilical vein; RT-PCR was performed to examine the mRNA expression of MCP-1 and PPAR-γ; Western blot was performed to detect NF-κB p65 protein.</p><p><b>RESULTS</b>(1) The expression of MCP-1 mRNA was increased in proportion with increasing concentrations of AGEs which could be blocked by atorvastatin in a dose-dependent manner. (2) AGE (10(-1) g/L) significantly downregulated the expression of PPAR-γ mRNA (0.22 ± 0.08 vs. 0.69 ± 0.09, P < 0.01) while upregulated the expression of phospho-NF-κB p65 protein (0.78 ± 0.06 vs. 0.31 ± 0.01, P < 0.01) and nonphospho-NF-κB p65 protein (1.61 ± 0.16 vs. 0.59 ± 0.14, P < 0.01) compared with the control group which could be significantly attenuated by atorvastatin. (3) PPAR-γ agonist decreased the expression of phospho-NF-κB p65 protein (0.21 ± 0.01 vs. 0.78 ± 0.06, P < 0.01), nonphospho-NF-κB p65 protein (0.67 ± 0.14 vs. 1.61 ± 0.16, P < 0.01) and MCP-1 mRNA (0.17 ± 0.02 vs. 0.93 ± 0.12, P < 0.01) compared with AGE (10(-1) g/L) group. (4) PPAR-γ inhibitor antagonized the effect of atorvastatin on the expression of phospho-NF-κB p65 protein, nonphospho-NF-κB p65 protein and MCP-1 mRNA stimulated by AGE in HUVECs (P < 0.01).</p><p><b>CONCLUSION</b>The anti-inflammatory properties of atorvastatin in AGE stimulated HUVECs may partly be attributed to the effect on upregulation of PPAR-γ and downregulation of NF-κB signaling pathway.</p>

Effects of human tissue kallikrein 1 gene delivery on carotid artery neointima formation after balloon angioplasty in spontaneously hypertensive rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of human tissue kallikrein 1(Ad-hKLK1) gene delivery on the neointima formation in carotid arteries of spontaneously hypertensive rats (SHRs).</p><p><b>METHODS</b>Carotid artery restenosis was induced in male SHR rats by balloon-injury. Rats were randomly assigned into 4 groups: Sham-operated (n = 6); Angioplasty (phosphate buffered solution 50 microl, n = 8); Vector virus (control virus, 1 x 10(9) IU in 50 microl, n = 8) and Ad-hKLK1(Ad-hKLK1, 1 x 10(9) IU in 50 microl, n = 8). Rats were sacrificed 4 weeks later. The wall-to-lumen area ratio and intima/media ratio in carotid artery were assessed by image analysis in HE stained sections. The mRNA bradykinin receptor (B1R and B2R) expressions were detected by RT-PCR. The protein expression of the cycle-independent kinase inhibitors p27Kip1 and p2lCip1 were determined by Western blot analysis.</p><p><b>RESULTS</b>Wall-to-lumen area ratio reduced 35.6% and intima/media ratio reduced 38.8%in Ad-hKLK1 treated SHRs compared to angioplasty group (all P < 0.001). The expression of p27Kip1 and p2lCip1 increased significantly in Ad-hKLK1 treated SHRs compared with angioplasty rats (all P < 0.001). The mRNA expression of B2R was significantly upregulated in angioplasty rats compared with sham-operated rats (P < 0.05) while mRNA expression of B1R was similar between the 2 groups.</p><p><b>CONCLUSION</b>hKLK1 gene delivery may effectively reduce neointimal formation via downregulating bradykinin B2R and up-regulating the expressions of p27Kip1, p2lCip1 signaling pathways in carotid arteries of SHRs after balloon injury.</p>

Effects of human tissue kallikrein gene transfer on the migration of vascular smooth muscule cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of adenovirus-mediated human tissue kallikrein (Ad-hKLK1) gene transfer on platelet-derived growth factor-BB (PDGF-BB)-induced migration of vascular smooth muscle cells from spontaneously hypertensive rats (VSMC(SHR)).</p><p><b>METHODS</b>A bicistronic recombinant adenovirus vector (Ad-hKLK1) carrying the target hKLK1 gene and the reporter gene EGFP was constructed. VSMCs isolated from the thoracic aorta of male SHR were passaged, and the quiescent VSMC(SHR) in passages 3-6 seeded in 6-well plates were treated with Ad-hKLK1 and control virus. Human PDGF-BB or icatibant Hoe140, a BK B2 antagonistat, was used as the chemoattractant and placed in the bottom chamber of the Boyden chamber. The mRNA expressions of bradykinin B1 receptor and B2 receptor were detected by RT-PCR in VSMC(SHR).</p><p><b>RESULTS</b>hKLK1 gene transfer significantly inhibited PDGF-BB-induced migration of VSMC(SHR), with the peak inhibition rate of 34.6% (P<0.001). PDGF-BB significantly increased the mRNA expression of B2 receptor but not B1 receptor in VSMC(SHR).</p><p><b>CONCLUSIONS</b>hKLK1 gene transfer can inhibit the migration of VSMC(SHR) induced by PDGF-BB, and the inhibitory effects may be not mediated by bradykinin B2 receptor.</p>

Effects of human tissue kallikrein gene delivery on the proliferation of vascular smooth muscle cells / 中华心血管病杂志

<p><b>OBJECTIVE</b>Tissue kallikrein cleaves kininogen substrate to produce vasoactive kinin peptides that have been implicated in the proliferation of vascular smooth muscle cells. We investigated the effects of adenovirus-mediated human tissue kallikrein (Ad-hKLK1) gene delivery on the proliferation of vascular smooth muscle cells of SHR (VSMCs(SHR)) induced by platelet derived growth factor-BB (PDGF-BB).</p><p><b>METHODS</b>Primary VSMCs(SHR) were isolated and cultured from thoracic aorta of male SHR. The VSMCs(SHR) proliferation induced by PDGF-BB was accessed by cell counting and methyl thiazolyl tetrazolium (MTT). Western blot was used to determine the protein expression of hKLK1, the cycle-independent kinase inhibitors p27(Kip1) and p21(Cip1). The mRNA expressions of bradykinin B1 receptor and B2 receptor were detected by RT-PCR in VSMCs(SHR).</p><p><b>RESULTS</b>Proliferation of VSMCs(SHR) induced by PDGF-BB was significantly inhibited post transfection of Ad-hKLK1 (20-100 MOI) in a MOI-dependent manner. The peak inhibition titer of Ad-hKLK1 was 100 MOI with peak inhibition rate of 39.3% (cell counting, n = 3, P < 0.01), 30.2% (MTT, n = 3, P < 0.01) and 36.4% (peak stunning rate of cell-cycle in phase G(0)/G(1)). The inhibitory effects of proliferation and cell-cycle caused by hKLK1 gene delivery could be abolished by Hoe140, a bradykinin B2 receptor antagonist. The protein expression of p27(Kip1) and p21(Cip1) increased significantly after the hKLK1 gene delivery, whereas Hoe140 nearly completely blocked these effects (n = 3, P < 0.001, respectively). PDGF-BB also significantly upregulated the mRNA expression of B2 receptor but not B1 receptor in VSMCs(SHR).</p><p><b>CONCLUSION</b>The hKLK1 gene delivery could inhibit PDGF-BB induced proliferation in VSMCs(SHR) through Bradykinin B2 receptor and up-regulate expression of p27(Kip1) and p2l(Cip1).</p>

Clinical features in 65 patients with acute myocardial infarction underwent successful thrombolytic therapy post cardiopulmonary resuscitation / 中华心血管病杂志

<p><b>OBJECTIVE</b>To analyze the clinical features of patients with acute myocardial infarction underwent successful thrombolytic therapy post cardiopulmonary resuscitation.</p><p><b>METHODS</b>This retrospective analysis included 65 patients with acute myocardial infarction underwent successful intravenous thrombolysis post cardiopulmonary resuscitation. The cases were collected from Chinese Journal Full-text Database from 1996 to 2006, only patients met the recanalization criteria of coronary artery were included.</p><p><b>RESULTS</b>Most of the patients were male (93.8%, 61/65) and aged less than 65 years (81.5%, 53/65). Cardiopulmonary resuscitation was performed within 5 min after cardiac arrest in 63 patients (96.9%). Defibrillation was performed 3.2 times per patient, chest compression in 52 patients (80.0%) and tracheal intubation in 21 patients (32.3%). The restoration time of spontaneous circulation were achieved within 10 min in 36 cases (55.4%), between 11 - 30 min in 19 cases (29.2%)and between 31 - 107 min in 10 cases (15.4%). Thrombolysis agents (urokinase, recombinant streptokinase or recombinant tissue-type plasminogen activator) were given intravenously at 172 +/- 92 min after acute myocardial infarction. Mild hemorrhage was seen in 12 cases (18.5%) and there was no report on severe hemorrhage event. The hemorrhage incidence tended to be higher than that of reported large Chinese thrombolysis trials (11.1% - 15.1%, P > 0.05).</p><p><b>CONCLUSION</b>Thrombolytic therapy was relatively safe and effective for those middle-aged male AMI patients received rapid cardiopulmonary resuscitation (< 5 min after cardiac arrest) and with shorter spontaneous circulation restoration time (<30 min).</p>

Meta-analysis of mesenteric arterial embolism or mesenteric arterial thrombosis / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To summarize the clinical characteristics of mesenteric arterial embolism (MAE) and mesenteric arterial thrombosis (MAT), and to clarify the diagnosis and treatment status of MAE and MAT in China.</p><p><b>METHODS</b>A retrospective analysis of 111 cases suffering from MAE or MAT was performed. Data of these cases were collected from Chinese Journal Full-text Database from 1994 to 2006.</p><p><b>RESULTS</b>There were 61 cases (54.9%) with MAE and 50 cases (45.1%) with MAT. Fifty-two patients (46.8%) had arterial fibrillation. Ninety-seven cases (87.4%) were diagnosed by exploratory laparotomy or autopsy, and 14 cases (12.6%) by imageology. Embolism or thrombosis in superior mesenteric artery (SMA) accounted for 92.8%, 4.5% in SMA plus inferior mesenteric artery. 15.2%(14/92) necrosis were located in jejunum or ileum, 39.1%(36/92) in jejunum and ileum, 38.0%(35/92) in jejunum, ileum and colon. Thrombolysis or anticoagulation in artery were operated in 7 cases(6.3%). Extraction of embolism or thrombosis in operation were implemented in 18 cases(16.2%). Intestinal resection were finished in 76 cases(68.5%). Sixty-eight patients (61.3%) were misdiagnosed. Sixty-three cases (60.6%) died.</p><p><b>CONCLUSION</b>The manifestation of MAE or MAT is quite complicated and changeable, so that many cases are misdiagnosed. The clinic and image characteristics of MAE and MAT have not been well known by doctors.</p>