he effects of constant magnetic field on apoptosis,adhesion molecule expression in endothelial cells and their adhesion rates with monocytes

AIM: To investigate the effects of constant magnetic field on apoptosis, secretion and expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVEC), and their adhesion rates with THP-1 monocytes induced by angiotensin Ⅱ (AngⅡ).METHODS: The third passage of cultured HUVEC was used.There were six groups: control group, Ang Ⅱ (10^-6 mol/L) group, Ang Ⅱ with 1, 5, 10 or 20 gausses of constant magnetic field group.Samples were collected 24 h after incubation with or without magnetic field.Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and propidinm iodide staining with flow cytometry.Secretion and expression of ICAM-1 and VCAM-1 were detected by ELISA and immunocytochemistry, respectively.Adhesion rate between HUVEC and THP-1 was measured by counting method.RESULTS: Ang Ⅱ at concentration of 10^-6mol/L induced apoptosis in HUVECs (P<0.05 vs control), whereas in 1, 5, 10 and 20 gausses group, apoptosis of HUVECs was significantly lower than that in Ang Ⅱ group (P<0.05).Ang Ⅱ at concentration of 10-6 mol/L significantly increased secretion and expression of ICAM-1 and VCAM-1 (P<0.05 vs control), whereas secretion and expression of ICAM-1 and VCAM-1 in 1, 5, 10 and 20 gausses group significantly decreased, compared with Ang Ⅱ group (P<0.05).The adhesion rates between HUVEC and THP-1 significantly increased 24 h after incubation of HUVEC with Ang Ⅱ (P<0.05 vs control), in contrast, the adhesion rates between HUVEC and THP-1 in 1, 5, 10 and 20 gausses group significantly decreased, compaed with Ang Ⅱ group (P<0.05).CONCLUSIONS: One gauss to 20 gausses of constant magnetic field antagonizes the effects of Ang Ⅱ on HUVEC, decreases apoptosis and expression of ICAM-1 and VCAM-1 in HUVEC, and also decreases the adhesion rates between HUVEC and monocytes induced by Ang Ⅱ.     

Proliferation and apoptosis of neuroendocrine cells in ovarian epithelial tumors

AIM: The purpose of this study was to observe the morphological features of neuroendocrine cells (NECs)， their proliferation and apoptosis in ovarian epithelial tumors, and to discuss their biological and clinical significance. METHODS: 79 specimens of ovarian epithelial tumor samples were collected, of them 20 benign, 18 boderline, 41 milignant tumors, and 22 normal ovaries were investigated immunohistochemically. Chromogranin A was used to detect NECs and their proliferation and apoptosis were examined by double-label staining of chromogranin A and Ki67 or TUNEL. RESULTS: The positive rate of CgA, distribution and staining intensity in ovarian epithelial tumors were higher than those in normal ovary. NECs showed various shapes with neuronoid protuberances stretching to the neighboring cells or basement membrane. Occasionally, they might touch together. No TUNEL positive coexpression in all NECs was observed by double-label staining, but some NECs were coexpressed with Ki67. CONCLUSION: NECs of ovarian epithelial tumors like cancer cells showed a proliferation, but no apoptosis. Their secretion might promote their neighboring non-NECs to proliferate and prevent them from apoptosis.     

The protective effect of composite salviae dropping pills on human umbilical vein endothelial cells injured by lipopolysaccharide

AIM: To observe the function and morphological change of human umbilical vein endothelial cells (HUVECs) treated with lipopolysaccharide (LPS) and composite salviae dropping pills (DSP). METHODS: HUVECs were cultured and incubated within 10 mg/L LPS for 12 hours. Different final concentrations of composite salviae dropping pills (1 g/L, 0.5 g/L, 0.25 g/L, 0.1 g/L) were added before and after LPS treatment. Cell viability, NO, NOS, ET-1 and intracellular calcium were measured. The cells were observed under inverted microscope, inverted phase contrast microscope, laser confocal scanning microscopy and transmission electron microscope. RESULTS: When given after LPS treatment, different final concentrations of composite salviae dropping pills played a protective role (P<0.05), and the concentration of 0.5 g/L was the most effective (P<0.01). When given before LPS treatment, 0.5 g/L and 0.25 g/L of composite salviae dropping pills protected the human vascular endothelial cells (P<0.05), but 1 g/L and 0.1 g/L didn't play a protective role (P>0.05). The HUVECs injured by LPS underwent apparent morphological change after treatment with composite salviae dropping pills. CONCLUSION: Composite salviae dropping pills have an evident protective effect on human umbilical vein endothelial cells exposed to LPS.     

Chronic cigarette smoking induces alteration of FIZZ1/RELMα expression in rat lung

AIM: To investigate the expression of FIZZ1/RELMα in lung tissue of chronic cigarette smoking rat, and to determine the relationship between airway inflammation and airway hyperresponsiveness. METHODS: Made rat model of chronic cigarette smoking was used. The expression of FIZZ1/RELMα in lung tissue was determined by immuno-histochemistry and in situ hybridization. RESULTS: In control rats, FIZZ1/RELMα protein and mRNA expressions were observed at low levels. In cigarette smoking rats, FIZZ1/RELMα expression increased in all the cells especially in bronchial smooth muscle cells, vascular wall cells and alveolar epithelial cells. CONCLUSION: FIZZ1/RELMα is a secreted peptide specifically expressed in lung. Cigarette smoking induces its upregulation, which possibly contributes to cigarette smoking-induced airway hyperresponsiveness.     

Gelatinases expression and their activity in rat alveolar macrophages induced by cigarette smoke medium

AIM: To study the effect of cigarette smoke medium (CSM) on the gene expression and activity of gelatinases from alveolar macrophages (AMs) in the rat, and then to explore their role in the pathogenesis of chronic obstructive pulmonary disease (COPD). METHODS: AMs were obtained from BALF of the rats that had smoked for 12 weeks. CSM was produced following the method of Wirtz and colleagues, and the cultured AMs were respectively stimulated for 24 h by 0%, 1%, 3%, 5%, 10%, 15% CSM. The mRNA levels of MMP-9 and MMP-2 were detected by semi-quantitative RT-PCR, and the enzyme activity was measured by Zymography. RESULTS: When the concentration of CSM was below 5%, the expression and activity of MMP-9 and MMP-2 signficantly increased with the concentration of CSM in a dose-depended manner (P<0.05). While the concentration of CSM exceeded 5%, the expression and activity of MMP-9 and MMP-2 correspondingly decreased with the increase in CSM concentrations (P<0.05), which possibly were related with the cytotoxicity of CSM. CONCLUSION: The expression and activity of MMP-9 and MMP-2 are induced by CSM. The gelatinases induced by smoking from AM may play an important role in the pathogenesis of COPD.     

Expression of nerve growth inhibiting factor Nogo-A mRNA and protein in the brain ischemic infarction rats

AIM: To investigate the changes of expression of Nogo-A at different time points in brain ischemic infarct rats. METHODS: The model of 80 cases of middle cerebral artery occlusion (MCAO) in rats was established. The expression of Nogo-A mRNA and protein were determined by Western blotting and hybridization, and the relationship between functional scoring and Nogo-A was also evaluated. RESULTS: In the brain of MCAO rats, Nogo-A mRNA expression was decreased on day 3 and increased significantly on day 7. The highest level was observed at the 21th d, keeping the same level at the 28th d. Nogo-A protein expression showed the same results. These results were correlated with the brain function scoring. CONCLUSION: Expression of Nogo-A does not increase in the early stage, but increases significantly in the late stage of MCAO, suggesting that Nogo-A expression may play an important role in the nerve regeneration of brain ischemic injury.     

Influence of ovariectomy and estrogen replacement on gene expression profile of myocardium in rats

AIM:To investigate the influence of ovariectomy and estrogen replacement treatment on profile of gene expression in myocardium by cDNA microarray,and to characterize the targeting genes of estrogen.METHODS:cDNA microarray containing 1 400 rat cDNAs was used to study the genes differentially expressed in myocardium between sham (Ⅰ),ovariectomy (Ⅱ,OVX) and estrogen replacement treatment (Ⅲ,OVX+E2) group.Then down-regulated genes in myocardium of OVX rats were further confirmed by RT-PCR.RESULTS:177 genes were differentially expressed in myocardium between sham and OVX rats,with 91 genes up-regulated and 86 genes down-regulated in OVX rats.164 genes were differentially expressed in myocardium between OVX and OVX+E2 rats,with 113 genes up-regulated and 54 genes down-regulated in OVX rats.There were 54 genes differentially expressed in OVX compared to sham and OVX+E2.They are involved in membrane channels and transporters (18),cell receptors (9),intracellular transducers/effectors/modulator (7) and metabolism (6).Most of the genes (45) were down-regulated in OVX rats and up-regulated in OVX+E2 rats.RT-PCR test confirmed the results of cDNA microarray.CONCLUSIONS:Long-term estrogen replacement may influence the expression of genes involved in membrane channels and transporters,cell receptors,intracellular transducers/effectors/ modulator and metabolism.Long-term estrogen replacement has some beneficial effects on ionic concentration and cardiac function which partially comes from the results of influence of expression on Na+,K+-ATPase and Na+/H+ exchanger.Estrogen has an inhibitory effect on the expression of dopamine receptor,which partially clarify the myocardial protection of estrogen.     

Protective effects of ［Gly14］-humanin on cultured neural stem cells treated with β-amyloid protein in vitro

目的：研究［Gly14］-humanin对β淀粉样蛋白1-42引起的神经干细胞毒性的作用。方法：用［Gly14］-humanin和β淀粉样蛋白1-42处理神经干细胞，观察其对神经干细胞增殖、分化的影响。结果：较低浓度的［Gly14］-humanin加入有β淀粉样蛋白1-42处理的神经干细胞培养基，经过琼脂糖凝胶电泳分析及流式细胞测定DNA含量，神经干细胞显示出对β淀粉样蛋白1-42的耐受，而对照组则显示神经干细胞出现凋亡现象。高浓度的［Gly14］-humanin加入培养基，经台盼蓝拒染法计数细胞，神经干细胞死亡率明显低于对照组，对照组神经干细胞出现大量死亡。结论：［Gly14］-humanin 不但具有抑制β淀粉样蛋白1-42对神经干细胞的毒性作用，而且在低浓度的情况下，可以促进神经干细胞的增殖和分化为神经元。