Influence of ox-LDL on migration function of THP-1 macrophages and microRNA21 expression and MAPK pathway phosphorylation / 心血管康复医学杂志

Objective :To explore influence of oxidized low density lipoprotein (ox-LDL ) on migration function of THP-1 macrophages ,expression of microRNA 21 (miR-21) and mitogen-activated protein kinase (MAPK) path-way.Methods :Phorbol myristate acetate (PMA) of 160nmol/L was used to induce THP-1 cells to differentiate into macrophages.According to application of ox-LDL treatment and liposomes-mediated miR-21 inhibitor transfecting THP-1 macrophages (transfection for short) or not ,THP-1 macrophages were divided into blank control group (re-ceived neither ox-LDL treatment nor transfection ) ,ox-LDL group (received 50mg/L ox-LDL treatment without transfection) ,miR-21 inhibitor group (received 50mg/L ox-LDL treatment after transfection ) and miR-21 inhibitor negative-control group (received 50mg/L ox-LDL treatment after negative-control transfection ).THP-1 macro-phage migration number was measured by transwell method ,miR-21 expression was measured by real-time quantita-tive PCR ,and expression of dual specific phosphate 8 (DUSP-8) and phosphorylation level of MAPK pathway were measured by Western-blot method .Results :Compared with blank control group ,there were significant rise in mi-gration number of THP-1 macrophages [(74.10 ± 15.10) vs.(184.10 ± 26.28)] ,miR-21 expression [(1.00 ± 0.21) vs.(2.02 ± 0.27)] and phosphorylation levels of JNK and P38 protein ,and significant reduction in expression of DUSP-8 protein in ox-LDL group ,P＜0.01 all.Compared with ox-LDL group ,there were significant reductions in migration number of THP-1 macrophages [ (184.10 ± 26.28) vs.(58.50 ± 10.24)] ,miR-21 expression [ (2.02 ± 0.27) vs.(0.66 ± 0.16)] and phosphorylation levels of JNK and P38 protein ,and significant rise in expression of DUSP-8 protein in ox-LDL group , P＜0. 01 all .Conclusion : Ox-LDL enhances migration function of macrophages , which may be related to its effects of upregulating miR-21 expression ,enhancing phosphorylation of JNK and P38 protein of MAPK pathway and reducing DUSP-8 expression .

Lipid-lowering effect of seven traditional Chinese medicine monomers in zebrafish system / 生理学报

The present study aimed to study lipid-lowering effect of seven traditional Chinese medicine monomers in zebrafish system. Zebrafish were fed with high fat diet to establish a hyperlipemia model, then fasted and bathed with seven traditional Chinese medicine monomers stigmasterol, triacontanol, chrysophanol, vanillic acid, shikimic acid, polydatin and oleanolic acid respectively. The oil red O staining was used to detect the blood lipids of zebrafish. Serum total cholesterol and triglyceride levels were detected to validate the lipid-lowering effect. The result showed that a zebrafish model of hyperlipemia could be established by feeding larvae zebrafish with high fat diet. Among the seven traditional Chinese medicine monomers, chrysophanol had lipid-lowering effect. Chrysophanol significantly reduced serum total cholesterol and triglyceride levels in adult zebrafish fed with high fat diet. Chrysophanol accelerated peristalsis frequency of zebrafish intestine and the excretion of high fat food. It is concluded that chrysophanol has lipid- lowering effect in zebrafish, and the mechanism of the effect may be due to the roles of chrysophanol in reducing lipid absorption from gastrointestinal tract and accelerating the excretion of food.

Overexpression of hypoxia inducible factor-1alpha (HIF-1alpha) promotes the differentiation of endothelial progenitor cell ex vivo / 中国实验血液学杂志

To investigate the influence of HIF-1alpha overexpression on the differentiation of endothelial progenitor cells (EPCs) ex vivo, EPCs were isolated from human peripheral blood by density gradient centrifugation, overexpressed HIF-1alpha was transfected to EPCs by electroporation; HIF1alpha, HIF1beta, vascular endothelial growth factor (VEGF) mRNA level were measured with RT-PCR; HIF-1alpha protein was detected with immunohistochemistry in a time course. CD31(+) cells were measured with flow cytometry. Cell morphology was observed after transfection. The results showed that the transfection efficiency of HIF-1alpha to EPCs was about 20%. HIF-1alpha and its controlled target gene VEGF were markedly induced by HIF-1alpha vector (P < 0.05). HIF1beta had its same level as it before interference (P > 0.05). HIF-1alpha protein was induced by HIF-1alpha transfection after 12 hours but was undetectable at 24 hours. After 7 - 14 days cultured in 21% oxygen pressure, fluorescence-trace experiments revealed that CD31 + EPCs/EC could be generated more efficiently from overexpressed HIF-1alpha than that from pEGFP transfected group (P > 0.05). EPC morphology was observed by light microscopy. HIF-1alpha-transfected cells under normoxia sprouted more rapidly from the EPC colonies than the untransfected cells or cells transfected with an GFP vector, which essentially maintained the original colony formation. HIF-1alpha transfected cells took on an array-like arrangement rather than random dispersal, suggesting that they were in an advanced state of differentiation. It is concluded that the utility of overexpression of HIF-1alpha can induce target genes which have influence on cell differentiation. HIF-1alpha transfection was found to give a prospected way to do the insight research on ischemic treatment in vivo.

Resveratrol inhibits expression of EMMPRIN from macrophages / 药学学报

<p><b>AIM</b>To investigate the effect of resveratrol on EMMPRIN expression of macrophages.</p><p><b>METHODS</b>Human monocytic cell line THP-1 cells were co-cultured with EMMPRIN-highly-expressed MCF-7 cells; MMP-9 production was assayed by zymography. THP-1 cells were induced by PMA, expression of EMMPRIN was assayed by Western blotting. Cells were treated with resveratrol or PPARgamma agonist--pioglitazone during differentiation, EMMPRIN expression and MMP-9 activity were assayed. U937 cells were co-transfected with PPARy expression and luciferase-coding reporter vector, then cultured with pioglitazone or resveratrol, the activating capability of resveratrol on PPARgamma was evaluated by measuring the luciferase activity. THP-1 cells were pretreated with PPARgamma antagonist--GW9662 before pioglitazone or resveratrol treatment, then assayed for EMMPRIN expression and MMP-9 production.</p><p><b>RESULTS</b>EMMPRIN expression was greatly increased during the differentiation from monocytes to macrophages; co-culturing with MCF-7 cells significantly increased MMP-9 production by monocytes. Both resveratrol and pioglitazone markedly inhibited EMMPRIN expression during monocytes differentiation. Resveratrol significantly activated PPARgamma and GW9662 greatly decreased the effect of resveratrol on EMMPRIN and MMP-9.</p><p><b>CONCLUSION</b>EMMPRIN expression is greatly up-regulated from monocytes to macrophages, which may play a role in inducing MMPs production by monocytes/macrophages. Resveratrol can significantly inhibit EMMPRIN expression via activating PPARgamma, which may be the underlying mechanism of its inhibitory effect on MMPs production by monocytes/macrophages.</p>

Effects of nocturnal hypoxemia on carotid artery atherosclerosis and plagues in patients with acute coronary syndrome / 上海第二医科大学学报

0.05).In group Ⅱand Ⅲ,the carotid artery IMT was thicker and the amount of plagues were larger than those in group Ⅰ(P

Effects of TIMP-2 gene transfer on atherosclerotic plaque in rabbits / 中华心血管病杂志

<p><b>OBJECTIVE</b>To evaluate the effects of TIMP-2 local gene transfer on atherosclerotic plaque.</p><p><b>METHODS</b>Atherosclerosis models were induced by denuding femoral artery endothelium plus high lipid diet in rabbits. TIMP-2 gene was transferred locally by balloons eluted with pcDNA3-TIMP-2. RT-PCR and Western blot were performed to verify exogenous genes transfer. MMPs activity in atherosclerotic plaque was evaluated by zymography. HE and VG staining and automatic image analysis system were used for pathological analysis of atherosclerotic femoral arteries. The lumen area of the vessel and the collagen contents in the atherosclerotic plaque were measured.</p><p><b>RESULTS</b>The expression of TIMP-2 gene in pcDNA3-TIMP-2 transferred group was significantly higher than control-vector transferred group at the end of week 2 after operation and reached the peak at the end of week 4. Comparing with the control group, the expression of TIMP-2 protein in treated group was also higher at the end of week 2, 4, and 8 after operation. Correspondingly, the MMP-2 and MMP-9 activities were lower in treated group. The thickness of fibrous cap of atherosclerotic plaque and the amount of collagen of the lesion were increased significantly in treated group compared with the control group, but there were no significant differences in vessel lumen area.</p><p><b>CONCLUSION</b>TIMP-2 gene transfer locally in atherosclerotic plaque could inhibit the activities of MMP-2 and MMP-9 in the lesion, increase the thickness of fibrous cap and the amount of collagen of the lesion, but may have no effect on the degree of the stenosis.</p>

Effect of danshensu, protocatechualdehyde and danshen injection on calcium ion concentration in cytoplasm of human erythrocytes / 中国中药杂志

<p><b>OBJECTIVE</b>To study the predominant calcium-antagonist components of Danshen injection.</p><p><b>METHOD</b>The effects of danshensu, protocatechualdehyde and Danshen injection on calcium concentration in cytoplasm of erythrocytes were examined in vitro by the fluorescent Ca+ -chelator fura-2.</p><p><b>RESULT</b>Either DS182 or PCAD can decrease in dose-dependent cytosolic free calcium concentration in human erythrocytes. They had additive effect when mixed, which was similar to Danshen injection.</p><p><b>CONCLUSION</b>DS182 and PCAD may be predominant calcium-antagonist components of Danshen injection.</p>