Clinical Hemorheology and Microcirculation - Volume 34, issue 1-2

Authors: Xue, Meilang | Thompson, Patrick | Sambrook, Philip N. | March, Lyn | Jackson, Christopher J.

Article Type: Research Article

Abstract: Activated protein C (APC) is a serine protease that plays a central role in physiological anticoagulation, and has more recently been shown to be a potent anti-inflammatory mediator. We show here that APC upregulates the angiogenic promoters, vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8) or matrix metalloproteinase-2 (MMP-2) in cultured human skin fibroblasts (HF), keratinocytes (HK) or umbilical vein endothelial cells (HUVE). In the chick embryo chorio-allantoic membrane assay, APC promoted angiogenesis. In a full-thickness rat skin healing model, a single topical application of APC enhanced wound healing compared to saline control. In summary, our results …demonstrate that APC promotes cutaneous wound healing at least partly by stimulation of angiogenesis. Show more

Keywords: Activated protein C, angiogenic factors, wound healing, keratinocytes, umbilical vein endothelial cells

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 153-161, 2006

Authors: Itoh, Yoshiaki | Takaoka, Rie | Ohira, Masayuki | Abe, Takato | Tanahashi, Norio | Suzuki, Norihiro

Article Type: Research Article

Abstract: Generation of reactive oxygen species (ROS) and their detrimental effects on the brain after transient ischemia are widely recognized. We studied ROS production from mitochondria in human brain microvessel endothelial cells (HBEC) under chemical hypoxia. HBEC in confluent conditions were incubated for 30 min with 10 μM 5-(and-6)-carboxy-2′,7′-dichlorodihydrofluorescein (DCF) diacetate, which was hydrolyzed and trapped inside the cells. ROS were measured with a fluorescent microscope, a CCD camera and an image analyzing system. Injury to mitochondrial respiratory chain was induced either with rotenone (an inhibitor of mitochondrial complex I) or with m-chlorocarbonyl cyanide phenylhydrazone (CCCP) (an uncoupler of ATP synthetase). …Shortly after application of 10 μM rotenone, fluorescent intensity started to increase and the gradual increase continued for 10 min. Similarly, CCCP (10, 50 and 100 μM) dose-dependently increased the fluorescent intensity (p<0.01). Edaravone, a free radical scavenger widely used for treatment of cerebral infarction in Japan, at 100 μM successfully suppressed this ROS production (p<0.05). These data show that chemical hypoxia with normal concentration of oxygen in the medium induced free radicals generation in HBEC. Importance of endothelial mitochondria as a source of free radicals after reperfusion is suggested. Show more

Keywords: Human brain, microvessel, endothelial cell, reactive oxygen species (ROS), dichlorodihydrofluorescein

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 163-168, 2006

Authors: Fu, Panfeng | Wang, Chunling | Xiu, Ruijuan

Article Type: Research Article

Abstract: Serum C-reactive protein (CRP) and soluble angiopoietin receptor Tie-2 (sTie) were detected in patients with acute myocardial infarction (AMI). The results indicated that mean serum CRP and Tie-2 levels were significantly higher in the patients with AMI than the control group. Increasing CRP was related to an increased infarction area and adverse prognosis. Levels of sTie-2 increased in the AMI patients and the maximum level of sTie-2 appeared at day 2 after onset of AMI. The feasibility of using to detect serum CRP and Tie-2 was also presented in this study. Measurement for CRP by optical proteinchip with imaging ellipsometry …(OPC-IE) and immunobidimetric analyzer showed no obvious difference (p<0.01). Also, the measurement for Tie-2 by ELISA and OPC-IE showed no obvious difference (p<0.01). Show more

Keywords: C-reactive protein (CRP), angiopoietin receptor Tie-2, optical proteinchip, acute myocardial infarction

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 169-175, 2006

Authors: Yada, Toyotaka | Shimokawa, Hiroaki | Kajiya, Fumihiko

Article Type: Research Article

Abstract: Rho-kinase modulates calcium sensitivity of the myosin light chain in smooth muscle cells and has been implicated as playing a pathogenetic role in cardiovascular disorders. This paper was aimed to determine whether hydroxyfasudil (a specific Rho-kinase inhibitor) exerts cardioprotective effect on coronary ischemia–reperfusion (I/R) injury, and if so, whether NO is involved. Canine subepicardial small arteries (diameter≥100 μm) and arterioles (diameter<100 μm) were observed by a CCD intravital microscope during coronary I/R. Coronary vascular responses to endothelium-dependent (acetylcholine) and -independent (papaverine) vasodilators were examined after I/R under three conditions: control, preconditioning, and hydroxyfasudil. Coronary I/R significantly impaired coronary vasodilation to …acetylcholine, whereas hydroxyfasudil completely preserved the responses, as did preconditioning. Hydroxyfasudil also significantly reduced myocardial infarct size. These results indicated that hydroxyfasudil exerts cardioprotective effects on coronary I/R injury in vivo, for which NO-mediated mechanism may be involved. Show more

Keywords: Rho-kinase inhibitor, coronary microcirculation, ischemia–reperfusion, nitric oxide

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 177-183, 2006

Authors: Luo, Zhaohua | Li, Hongwei | Zhang, Jing | Zhang, Honggang | Xiu, Ruijuan

Article Type: Research Article

Abstract: Connective tissue growth factor (CTGF) is a 38-kDa cysteine-rich protein and an important regulator of angiogenesis. In order to study the role CTGF gene playing in angiogenesis, the eukaryotic expression vector of CTGF gene was constructed in this study, and the role of endogenous CTGF on migration of human umbilical vein endothelial cell (HUVECs) was investigated. According to human CTGF cDNA sequence, a pair of specific primers containing digestion sites of Xba I and Hind III on the 5′ end respectively were designed. Reverse transcript polymerase chain reaction (RT-PCR) was used to amplify CTGF cDNA from HUVECs. The eukaryotic expression …vector pcDNA3.1(−)/CTGF containing the entire coding region was constructed successfully. Compared with CTGF sequence of GenBank, DNA sequence analysis showed that this reformed plasmid contained the same full length of CTGF cDNA. The results of Western blotting demonstrated that CTGF was over-expressed at 48 h after transfection. Migration of HUVECs transfected with CTGF vector increased significantly compared with those transfected with vector control. In conclusion, the eukaryotic expression vector pcDNA3.1(−)/CTGF was constructed successfully and the endogenous CTGF promoted the migration of HUVECs. This study lays a foundation for further study on the role CTGF gene playing in angiogenesis. Show more

Keywords: Connective tissue growth factor (CTGF), cell migration, angiogenesis, tumor

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 185-192, 2006

Authors: Nakamura, Masahiko | Asada, Maki | Matsui, Hidenori | Hibi, Norifumi | Tsuchimoto, Kanji | Inoue, Jun | Oda, Masaya

Article Type: Research Article

Abstract: To clarify the microvascular changes and the effector sites of lansoprazole during the formation of colitis, the dextran sulfate sodium (DSS)-induced colitis was induced by the oral administration for 3 and 7 days. The alteration of the microvascular permeability was estimated by the intraaortic infusion of FITC-dextran. The effector sites of 3 H-lansoprazole were examined by the intraaortic infusion of the radiolabelled compound and the autoradiographic procedure of water-soluble compounds. As a result, marked increase of the microvascular permeability was detected three days after DSS treatment near the inflammatory cells in the tip portion of the colonic mucosa. 3 H-lansoprazole …in the control rat colon was localized in the goblet cells, while in DSS-treated rats, 3 H-lansoprazole was accumulated in the cytoplasm of the mesenchymal cells, and most of them coincided with polymorphonuclear leucocytes and macrophages. Show more

Keywords: Dextran sulfate sodium, FITC-dextran, lansoprazole, autoradiography, colitis, neutrophil, macrophage

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 193-199, 2006

Authors: Su, Yan | Li, Hongwei | Zhang, Jing | Xiu, Ruijuan

Article Type: Research Article

Abstract: Hypoxia is generally considered to represent a fundamental stimulus for angiogenesis. Most angiogenic factors were up-regulated by hypoxia, but as a strong angiogenic and antiapoptotic factor, hepatocyte growth factor (HGF) was down-regulated by hypoxia. In order to investigate whether hypoxia inducible factor 1a (HIF-1α) participate in the transcriptional regulation of HGF under hypoxia, rat cardiac myocytes were treated with cobalt chloride (CoCl2 ), the specific inducer of HIF-1α, for different times, and total RNA and protein were isolated to perform RT-PCR and Western blot. Results show the expression of HGF mRNA in cardiac myocytes decreased distinctively after treating with CoCl2 …for 12 hours. However, at the same time, the expression of HIF-1α protein was up-regulated. HIF-1α may be participate in the transcriptional regulation of HGF indirectly. Show more

Keywords: Hepatocyte growth factor, hypoxia inducible factor 1a, hypoxia, cobalt chloride

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 201-206, 2006

Authors: Xia, Zuo-Li | Sun, Bao-Liang | Zheng, Yan-Hong | Yang, Ming-Feng | Song, Xi-Jun | Hao, Fang | Zhao, Xiao-Min | Xu, Chang-Qing

Article Type: Research Article

Abstract: The model of lymphatostatic encephalopathy was established by occluding and removing profound cervical nodes in rats, and the kinetic alteration of nitric oxide (NO), maleic dialdehyde (MDA), free radical scavenger (CuZn-SOD) and arterial systolic blood pressure were determined on different days after the blockage. The results showed that the level of NO significantly decreased at 1 day (P<0.05) and further decreased at 3, 5 and 7 day (P<0.01). The levels of MDA at 1, 3, 5 and 7 day significantly increased, but the contents of CuZn-SOD significantly decreased compared with the control (P<0.01). There was negative correlation between the levels …of MDA and CuZn-SOD, but there was no relationship between MDA an NO. Arterial systolic blood pressure decreased progressively after cervical lymphatic blockage. The results showed that NO, oxide free radicals and the disturbances of the cardiovascular regulation may play important roles in lymphatostatic encephalopathy. Show more

Keywords: Lymphatostatic encephalopathy, nitric oxide, free radical, arterial systolic blood pressure

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 207-211, 2006

Authors: Li, Yuan | Zhou, Zong-Guang | Zhang, Jie | Chen, You-Dai | Li, Hong-Guang | Gao, Hong-Kai | Wang, Rong | Hu, Ting-Ze

Article Type: Research Article

Abstract: This paper was aimed to detect Toll-like receptor 4 (TLR4) microcirculatory expression and localization in rat pancreas and intestine. Acute pancreatitis (AP) was induced by twice injections of cerulein (20 μg in total) and acute necrotizing pancreatitis (ANP) was induced by intraductal injection of 5% taurocholate (1 ml/kg.bw). Reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) were used to detect and localize TLR4 in the pancreas and intestine. Results showed that RT-PCR of RNA isolated from pancreatic and intestinal tissue yielded the predicted amplicon for TLR4; IHC analysis localized TLR4 expression to the endothelium of pancreatic arteriole, venule, acinar …capillary network and sinusoidal capillary of endocrine islet; TLR4 expression in intestine was principally in the microvascular endothelium and leucocytes within the mucosa lamina propria. TLR4 staining in intestine was more intense in taurocholate-induced pancreatitis (TIP) than that in cerulein-induced pancreatitis (CIP). In conclusion, TLR4 could be detected in the pancreatic and intestinal microcirculation, suggesting TLR4 involved in the microcirculatory impairment in AP; the more intense intestinal TLR4 expression in TIP suggests a potential risk for secondary infection. Show more

Keywords: Acute pancreatitis, microcirculation, Toll-like receptor 4

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 213-219, 2006

Authors: Osada, Takashi | Tomita, Minoru | Tanahashi, Norio | Takeda, Hidetaka | Nagai, Toshihiro | Suzuki, Norihiro

Article Type: Research Article

Abstract: The source and target of edema fluid for ischemic brain swelling clinically often observed in “malignant infarction” was examined in ex vivo. Wister rat brain hemispheres were removed and incubated air-tightly in a deoxygenated artificial cerebrospinal fluid at 37° for 30 min. Ionic movement into the brain tissue was calculated from their concentration changes in the incubation fluid. We found a weight increase by 11.3±2.5% (p<0.01) and a decrease in Na+ from 148.0 to 139.0±8.2 mEq/l (p<0.01) and an increase in K+ from 4.3 to 11.2±1.2 mEq/l. Video tape recording revealed that the brain swelling started immediately upon …the incubation, and the electronmicroscopical investigation of the swollen cortical tissue revealed that the fluid moved mainly into astroglial cells. The astroglial swelling was quite similar to that of specimen taken from clinical cases at autopsy. The driving force of the water shift can be explained by discharge of thermodynamic potential, i.e., a coupled transport of water with Na+ across the cell membrane (anomalous osmosis). The swelling was not affected by addition of aquaporin blocker, mercuric chloride. It is concluded that cerebrospinal fluid bathing the brain in situ can be the source of edema fluid for ischemic brain swelling. Show more

Keywords: Edema fluid, anomalous osmosis, membrane depolization, ischemic brain swelling

Citation: Clinical Hemorheology and Microcirculation, vol. 34, no. 1-2, pp. 223-226, 2006