内毒素作用下肺动脉内皮细胞ET—1基因表达与释放

应用斑点杂交、放免测定等方法检测分析了内毒素不同浓度同时相(A组)和内毒素同浓度不同时相(B组)作用下体外培养的新生牛肺动脉内皮细胞ET—1基因表达和释放的作用,且该作用有明显的剂量依赖关系和时间效应关系,两组ET—1基因,表达与释放。结果表明,内毒素具有促进内皮细胞ET—1基因表达的强弱与释放量的增减均呈显著的正相关。提示内毒素血症时血浆ET水平升高与ET合成增加和基础释放量增高有密切关系。

CHANGES OF ENDOTHELIN－1 GENE EXPRESSION IN RAT BRAINS DURING ISCHEMIA AND ISCHEMIC REPERFUSION

Objective. The experiment was designed to study the association of cerebral ischemia and reperfusion with endothelin- 1 (ET- 1) gene expression of rat brains and time-dependent changes of ET- 1 gene expression during cerebral ischemia.Materials and methods. Thirty- three male SD rats were divided into dot blot hybridization(n = 27) and in silu hybridization groups(n= 6). The focal cerebral ischemia and reperfusion models were made with suture embolism of middle cerebral artery. Dot blot hybridization groups were redivided into control and ischemic subgroups (ischemia for 0. 5 , 1 , 1. 5 , 3 , 6 , 12 , 24 , 48 and 72 h respectively). In situ hybridization groups were redivided into ischemia and reperfusion groups. After 24 h ischemia and 24 h reperfusion,ET1 gene expressions were investigated with in situ hybridization and the resuhs were analyzed with IBAS 2000 Image Analysis System.Results. Dot blot hybridization showed that ET-1 mRNA of cerebral cortex and caudate- putamen was increased at 6 h of ischemia and reached peak at 24 h (3. 9 and 3. 7 fold respectively) ,and at 72 h of ischemia it remained at high levels(3. 5 and 2. 1 fold respectively). In silu hybridization showed that the levels of ET- 1 mRNA of cerebral cortex and caudate-putamen were also markedly increased both in 24 h ischemia and 24 h reperfusion groups (P<0. 01 , P<0. 05 respectively) .Conclusions. ET-1 gene expression in focal ischemic brain tissue were markedly and progressively increased during cerebral ischemia and reperfusion and downregulation of ET- 1 gene expression may be a new approach to the treatment of ischemic cerebrovascular diseases.

Associations of big endothelin-1 and C-reactive protein in atrial fibrillation

Background Atrial fibrillation （AF） is associated with inflammation and endothelial dysfunction. However, the association between inflammation （as indexed by high-sensitivity C-reactive protein, hs-CRP） and endothelial function [as indexed by big endothelin-1 （ET-1）] in AF patients remains unclear. Methods We enrolled 128 patients with lone AF, among which 83 had paroxysmal AF, and 45 had persistent AF. Eighty-two age- and gender-matched controls of paroxysmal supraventricular tachycardia without AF history were evaluated. Plasma hs-CRP, big ET-1 levels and other clinical characteristics were compared among the groups. Results Patients with persistent AF had higher hs-CRP concentrations than those with paroxysmal AF （P 〈 0.05）, both groups had higher hs-CRP level than controls （P 〈 0.05）. Patients with persistent AF had higher big ET-1 level than those with paroxysmal AF, although the difference did not reach the statistical significance （P 〉 0.05）, and both groups had higher big ET-1 levels than controls （P 〈 0.05）. Multiple regression analyses revealed hs-CRP as an inde- pendent determinant of AF （P 〈 0.001）. Further adjusted for big ET-1, both big ET-1 and hs-CRP were independent predictors for AF （P 〈 0.001）, but the odds ratio for hs-CRP in predicting AF attenuated from 8.043 to 3.241. There was a positive relation between hs-CRP level and big ET-1 level in paroxysmal AF patients （r = 0.563, P 〈 0.05）, however, the relationship in persistent AF patients was poor （r = 0.094, P 〈 0.05）. Conclusions Both plasma hs-CRP and big ET-1 levels are elevated in lone AF patients, and are associated with AF. In paroxysmal lone AF patients, there were significant positive correlations between plasma hs-CRP level and big ET- 1 level.

HYPOXIA AND ENDOTHELIN－1 STIMULATE DNA SYNTHESIS OF PULMONARY ARTERY SMOOTH MUSCLE CELLS

Hypoxia and endothelin-1 (ET-1) are associated with constriction of pulmonary vasculature both in vivo and in vitro. However, the role of hypoxia and ET-1 in the vascular remodelling during the development of pulmonary hypertension is unclear. This study demonstrated that ET-1(0. 1 nmol/L to 100 nmol/L) increased the [￣3H]thymidine uptake in a dose-dependent manner in cultured bovine pulmonary artery smooth muscle cells(PASMC). which was enhanced by exposing PASMC to hypoxia (2%O2, 93%N2,5%CO2 ). BQ123, the specific antagonist of endothelin receptor subtype A,eliminated the ET-1 medicated proliferation of PASMC and the cooperative effect of hypoxia. Some dilatory drugs could inhibite the mitogenic effect of ET-1. We also observed that hypoxia significantly increased [￣3H] thymidine uptake in PASMC without ET-1 and BQ123 could inhibite this effect. Radioimmunoassay suggested that there was an autocrine of ET-1 in cultured PASMC which was enhanced by hypoxia significantly.

Expression of mucosal addressin cell adhesion molecule 1 on vascular endothelium of gastric mucosa in patients with nodular gastritis

AIM:The interaction of mucosal addressin cell adhesion molecule 1 (MAdCAM-1) with integrin α4β7 mediates lymphocyte recruitment into mucosa-associated lymphoid tissue (MALT).Nodular gastritis is characterized by a unique military pattern on endoscopy representing increased numbers of lymphoid follicles with germinal center,strongly associated with H pylori infection.The purpose of this study was to address the implication of the MAdCAM-1/integrin β7 pathway in NG. METHODS:We studied 17 patients with NG and H pylori infection and 19 H pylori-positive and 14 H pylori-negative controls.A biopsy sample was taken from the antrum and snap-frozen for immunohistochemical analysis of MAdCAM- 1 and integrin β7.In simultaneous viewing of serial sections, the percentage of MAdCAM-1-positive to von Willebrand factor-positive vessels was calculated.We also performed immunostaining with anti-CD20,CD4,CD8 and CD68 antibodies to determine the lymphocyte subsets co- expressing integrin β7. RESULTS:Vascular endothelial MAdCAM-1 expression was more enhanced in gastric mucosa with than without H pylori infection.Of note,the percentages of MAdCAM-1-positive vessels were significantly higher in the lamina propria of NG patients than in H pylori-positive controls.Strong expression of MAdCAM-1 was identified adjacent to lymphoid follicles and dense lymphoid aggregates.Integrin β7-expressing mononuclear cells,mainly composed of CD20 and CD4 lymphocytes,were associated with vessels lined with MAdCAM-1-expressing endothelium.CONCLUSION: Our results suggest that the MAdCAM一1/ integrin a4p7 homing system may participate in gastric inflammation in response to H py/o}i-infection and contributes to MALT formation, typically leading to the development of NG.

INTRACELLULAR REDISTRIBUTION OF CARDIAC ENDOTHELIN- 1 RECEPTOR IN RAT DURING MYOCARDIAL HYPERTROPHY

Objective. In a model of rat cardiac hypertrophy, the changes in the distribution of ET- 1 receptors in two subcellular fractions, the sarcolemma and the light vesicles during myocardial hypertrophy were studied. Methods. Cardiac hypertrophy was produced by placing a constricting clip around the suprarenal abdominal aorta of rats, and ET- 1 receptor was assayed with radioactive analysis method. Results. It was found that plasma and ventricular ET- 1 levels increased significantly on week 2 and week 4 of pressure overload. ET- 1 binding studies showed that during myocardial hypertrophy, the maximum binding capacity (Bmax) was increased by 41％ (P< 0.01) and 65％ (P< 0.01) in sarcolemma in H- 2 week and H- 4 week groups, but was decreased by 24％ (P< 0.01) and 21％ (P< 0.01) in light vesicles. The sum of Bmax of sarcolemmal and light vesicle fractions was increased by 33％ (P< 0.01) and 57％ (P< 0.01) in group H- 2 week and H- 4 week, respectively. Conclusion. ET- 1 receptors in rat heart were externalized from light vesicles to sarcolemma, which may contribute to the development of myocardial hypertrophy.

INFLUENCE OF GINKGO BILOBA EXTRACT ON NITRIC OXIDE AND ENDOTHELIN-1 DURING CEREBRAL VASOSPASM IN RATS

Objective.To investigate the effects of Ginkgo biloba extracton cer ebral vasospasmafter subarachnoid hemorrhageand its influence on n itric oxideand endothelin-l .Methods.Noncraniotomy models of SAH in Wistar rats were used and animals were divided into sham-operated group ,SAH group,saline-treated group and EGb-treated group.Diameter of basilar a rtery was measured.Regional cerebral blood flow,NO and ET-1levels in blood,and calcium content in brain tissue within24hours after SAH were dete cted.Pathological examination of hippocampus CA1sub-field was also performed .Results.Sham operation did not alter the above parameters.Induction of SAH l ed to a marked spasm of basilar artery.rCBF decreased obviously and consecutive ly within24hours after SAH.Meanwhile NO level in serum decreased,ET-1level in plasma and calcium content in brain tissue significantly in-creased.Pyra midal cells in hippocampus CA1subfield were severely damaged.EGb significantly antago-nized the pathological alterations of the above parameters.Conclusion .Alterations of NO,and ET-1play an important role in the development of CV S after SAH.EGb exerts its protective effects on CVS by inhibitng the above pat hological alterations.

Effects of the heparin-coated Wiktor-I stent on recovering the balance of ET-1 and CGRP after intracoronary stent implantation

Objective: To study the effects of heparin-coated stent (Wiktor-I) on recovering the balance between endothelin- 1(ET- 1) and calcitonin gene-related peptide (CGRP) both in the plasma of coronary sinus and peripheral vein after stent implantation. Methods: The patients with coronary artery disease (n=30) were divided into non-coated stent (NCS) group (n=16) and heparin-coated stent (HCS) group (n=14). Circulating ET-1 and CGRP levels were measured by radioimmunoassay (RIA) in the 2 groups at the paired sampling sites, namely the coronary sinus (Cs) and peripheral vein (Pv). The ratio of ET/CGRP was also calculated to reflect the balance of ET and CORP. Results: In NCS group plasma ET-1 concentrations exhibited 2 peaks at 0 mm and 12 h after stent implantation, CORP levels were highly consistent with that of ET-1, and the ratio of ET/CGRP increased markedly at post-PTCA 5 mm, persisting until post-stent 12 h. While in HCS group ET-1 levels and the ratio of ET/CGRP were decreased dramatically compared with that of NCS group at 5 min,10 mm and 12 h post-stent (P＜0.05, P＜0.01 and P＜0.05, respectively＝. There was no remarkable difference of CGRP levels between the 2 groups. Conclusions: Heparin-coated stent can decrease the post-stent circulating ET-1 levels and recover the balance between ET-1 and CGRP.

Response of blood endothelin-1 and nitric oxide activity in duodenal ulcer patients undergoing Helicobacter pylori eradication

AIM: To investigate the effect of Helicobacter pylori eradication on endothelin-1 (ET-1) and nitric oxide (NO) in duodenal ulcer (DU) patients. METHODS: Sixty-six Hpylori-infected active DU patients were consecutively enrolled to receive one-week triple therapy (rabeprazole, amoxicillin and metronidazole) and then one-month rabeprazole therapy. They were asked back to determine ulcer and Hpylori status using endoscopy one month later. Thirty-seven healthy controls (H pylori +/-:17/20) were enrolled for comparison. Blood samples were collected in each visit to measure plasma ET-1 and nitrate/nitrite levels using an enzyme immunoassay kit. RESULTS: Sixty DU patients finished trial per protocol. The ulcer healing and Hpylori-eradication rates were 86.7% and 83.3%, respectively. Plasma ET-1 level in DU patients was higher than that of Hpylori-negative and positive controls (3.59±0.96 vs0.89±0.54 vs0.3±0.2 pg/mL,P<0.01), while nitrate/nitrite levels among them were also significantly different (8.55±0.71 vs5.27±0.68 vs 6.39±0.92 μmol/L, P<0.05). H pylori eradication diminished ET-1 levels (3.64±0.55 vs2.64±0.55 pg/mL, P<0.01) but elevated nitrate/ nitrite level (8.16±0.84 vs11.41±1.42 umol/L,P<0.05). CONCLUSION: Both plasma ET-1 and nitrate/nitrite levels increase in active DU patients. After an effective H pylori eradication, DU healing is associated with diminished blood ET-1 level and elevated nitrate/nitrite level.

EXTERNALIZATION AND INTERNALIZATION OF CARDIAC ENDOTHELIN RECEPTORS DURING DIFFERENT PHASES OF SEPSIS IN RAT

To study the redistribution of endothelin- 1 (ET- 1) receptors in two subcellular organelles , the sarcolemmal membrane and the light vesicle, of rat heart during the progression of sepsis. Methods. Sepsis was induced by cecal ligation and puncture (CLP). ET1 receptor was assayed by using [125I]- ET1 binding. Marker enzyme activities, protein yield, and dry- to- wet weight ratio of cardiac membranes were measured. Results. Septic rat heart exhibited two distinct phases: an initial hyperdynamic phase( 9h after CLP; early stage of sepsis) followed by a hypodynamic (18h after CLP, late stage of sepsis) phase. [125I]- ET1 binding study showed that during early stage of sepsis, the Bmax of ET1 receptors was increased by 30％ in sarcolemma but decreased by 19％ in light vesicles, while during late stage of sepsis, the Bmax was decreased by 24％ in sarcolemma but increased by 38％ in light vesicles.The total binding of sarcolemma and light vesicles was increased by 25％ during early stage of sepsis but decreased by 17％ during late stage of sepsis. Conclusions. These data indicated that ET1 receptors in the rat heart were externalized from light vesicles to sarcolemmal membranes during early hyperdynamic phase while internalized from surface membranes to intracellular compartment during late hypodynamic phase of sepsis.

Gene expression profiling and endothelin in acute experimental pancreatitis

AIM:To analyze gene expression profiles in an experimental pancreatitis and provide functional reversal of hypersensitivity with candidate gene endothelin-1 antagonists.METHODS:Dibutyltin dichloride(DBTC) is a chemical used as a polyvinyl carbonate stabilizer/catalyzer,biocide in agriculture,antifouling agent in paint and fabric.DBTC induces an acute pancreatitis flare through generation of reactive oxygen species.Lewis-inbred rats received a single i.v.injection with either DBTC or vehicle.Spinal cord and dorsal root ganglia(DRG) were taken at the peak of inflammation and processed for transcriptional profiling with a cDNA microarray biased for rat brain-specific genes.In a second study,groups of animals with DBTC-induced pancreatitis were treated with endothelin(ET) receptor antagonists [ET-A(BQ123) and ET-B BQ788)].Spontaneous pain related mechanical and thermal hypersensitivity were measured.Immunohistochemical analysis was performed using anti-ET-A and ET-B antibodies on sections from pancreatic tissues and DRG of the T10-12 spinal segments.RESULTS:Animals developed acute pancreatic inflammation persisting 7-10 d as confirmed by pathological studies(edema in parenchyma,loss of pancreatic architecture and islets,infiltration of inflammatory cells,neutrophil and mononuclear cells,degeneration,vacuolization and necrosis of acinar cells) and the painrelated behaviors(cutaneous secondary mechanical and thermal hypersensitivity).Gene expression profile was different in the spinal cord from animals with pancreatitis compared to the vehicle control group.Over 260 up-regulated and 60 down-regulated unique genes could be classified into 8 functional gene families:circulatory/acute phase/immunomodulatory;extracellular matrix;structural;channel/receptor/transporter;signaling transduction;transcription/translation-related;antioxidants/chaperones/heat shock;pancreatic and other enzymes.ET-1 was among the 52 candidate genes upregulated greater than 2-fold in animals with pancreatic inflammation and visceral pain-related behavior.Treatments with the ET-A(BQ123) and ET-B(BQ-788) antagonists revealed significant protection against inflammatory pain related mechanical and thermal hypersensitivity behaviors in animals with pancreatitis(P < 0.05).Open field spontaneous behavioral activity(at baseline,day 6 and 30 min after drug treatments(BQ123,BQ788) showed overall stable activity levels indicating that the drugs produced no undesirable effects on normal exploratory behaviors,except for a trend toward reduction of the active time and increase in resting time at the highest dose(300 μmol/L).Immunocytochemical localization revealed that expression of ET-A and ET-B receptors increased in DRG from animals with pancreatitis.Endothelin receptor localization was combined in dual staining with neuronal marker NeuN,and glia marker,glial fibrillary acidic protein.ET-A was expressed in the cell bodies and occasional nuclei of DRG neurons in na ve animals.However,phenotypic expression of ET-A receptor was greatly increased in neurons of all sizes in animals with pancreatitis.Similarly,ET-B receptor was localized in neurons and in the satellite glia,as well as in the Schwann cell glial myelin sheaths surrounding the axons passing through the DRG.CONCLUSION:Endothelin-receptor antagonists protect against inflammatory pain responses without interfering with normal exploratory behaviors.Candidate genes can serve as future biomarkers for diagnosis and/or targeted gene therapy.

Endothelin-1, an important mitogen of smooth muscle cells of spontaneously hypertensive rats

OBJECTIVE: To study the features of vascular smooth muscle cell (VSMC) proliferation induced by endothelin-1 (ET-1). METHODS: VSMCs of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats were cultured and treated with ET-1. Basic fibroblast growth factor (bFGF) gene expression was measured using both Northern blot and an enzyme-linked immunoassay. RESULTS: ET-1 resulted in an increase in bFGF transcripts at 8 - 24 h; bFGF levels were significantly higher in VSMCs treated with ET-1 than in those not treated. However, VSMCs growth responses in SHR and WKY were different. Smooth muscle cells of SHR were hyper-responsive to ET-1. Maximal bFGF mRNA levels were elevated 3.5-fold at 4 h of stimulation in WKY and 8-fold at 8h in SHR4. Moreover, the proliferation of VSMCs induced by ET-1 was inhibited by antisense phosphorothioate oligodeoxynucleotides (10 micromol/L AS-bFGF) but not sense bFGF oligomers at the same concentrations, being reduced by 80% in SHR and 40% in WKY vs control, respectively. Furthermore, the effect of AS-bFGF oligomers on SHR SMC proliferation is significantly greater than on WKY SMC proliferation. CONCLUSION: ET-1 may be required for exaggerated vascular growth responses in SHR and bFGF may be involved.

Effects of Liandou Qingmai Recipe(连豆清脉方) on Endothelin-1,Nitric Oxide,Interleukin-6 and Interleukin-10 Levels in Patients with Coronary Heart Disease

Objective:To observe effects of Liandou Qingmai Recipe(连豆清脉方) on endothelin-1(ET-1),nitric oxide(NO),interleukin-6(IL-6) and IL-10 levels in patients with coronary heart disease.Methods:Total 101 cases with coronary heart disease were randomly divided into a treatment group(n=45) treated by Liandou Qingmai Recipe and a standard treatment group(control group,n=56),with a normal group of 16 health persons set up.Changes of ET-1,NO,IL-6 and IL-10 levels were measured before treatment and after treatment for two weeks.And the data were analyzed by SPSS 16.0 statistic software.Results:Before treatment,the levels of ET-1,IL-6 and IL-10 levels were significantly higher and NO was significantly lower in the patients with coronary heart disease than those in the normal group(90.7±12.7 ng/L vs 41.8±13.5 ng/L,9.17±0.18 ng/L vs 1.10±0.08 ng/L,1.94±0.26 ng/L vs 1.09±0.06 ng/L,and 92.2±17.7 μmol/L vs 124.5±27.2 μmol/L;all P<0.05),with no significant differences in the levels of ET-1,NO,IL-6 and IL-10 between the treatment group and the control group(P>0.05);After treatment,ET-1 and IL-6 significantly decreased in the treatment group and the control group,and NO increased in the treatment group;And IL-6 level was significantly lower and NO level was higher in the treatment group than those in the control group(4.48±1.22 ng/L vs 5.13±1.85 ng/L,117.4±22.3 μmol/L vs 92.4±17.1 μmol/L;both P<0.05);There was a positive correlation between IL-6 and IL-10,and a negative correlation between NO and IL-10(r=0.142,r=-0.152;both P<0.05).Conclusion:Liandou Qingmai Recipe can decline IL-6,IL-10 and ET-1 levels,and raise NO level in patients with coronary heart disease on the basis of standard treatment,so as to inhibit endothelial inflammatory response,improve vascular endothelial function,with stronger anti-AS action;And vascular endothelial lesion is related with inflammatory response.

Effect of Sancaijiangtang on plasma nitric oxide and endothelin-1 levels in patients with type 2 diabetes mellitus and vascular dementia:a single-blind randomized controlled trial

OBJECTIVE: To observe the effect of Sancaijiangtang powders on plasma nitric oxide and endothelin-1 levels. We sought to identify the common pathological link and mechanism of action for Traditional Chinese medicine in type 2 diabetes mellitus and vascular dementia,and to explicate the material basis for treating the different diseases with the same method in Traditional Chinese Medicine.METHODS: In total,168 patients with type 2 diabetes mellitus and vascular dementia were enrolled in the study,and randomly divided into two groups by simple randomization. Patients in the treatment group received oral Sancaijiangtang powders with pioglitazone hydrochloride three times daily,while patients in the control group received pioglitazone hydrochloride alone. The treatment course was for12 weeks. Mini-mental state examinations(Chinese version) and Montreal Cognitive Assessments(Beijing version) were performed,and fasting plasma glucose,fasting insulin,hemoglobin A1 c,homeostasis model assessment of insulin resistance,plasma nitric oxide and endothelin-1 levels were measured before and after the treatment.RESULTS: The post-treatment levels for all measurements in both groups were better than pre-treatment levels(P < 0.05). The post-treatment levels for all measurements in the treatment group were better than the levels measured in the control group(P < 0.05).CONCLUSION: Type 2 diabetes mellitus and vascular dementia have common pathological mechanisms for insulin resistance and endothelium dysfunction. Sancaijiangtang powders could improve the release of nitric oxide and inhibit the secretion of endothelin-1. Therefore,the material basis exists for treating the different diseases with the same method in Traditional Chinese Medicine.

Mild moxibustion at different intervention times on the levels of ET-1 and NO in the uterine tissues of rats with cold-damp coagulation and stagnation type dysmenorrhea

Objective： To observe the differences in analgesic effect of moxibustion at different intervention times on dysmenorrhea rats and explore its effect mechanism.Methods： Forty-five female Wistar rats were randomly divided into blank control group（group A）, model group（group B）, pre-moxibustion group（group C）, instant moxibustion group（group D） and pre-instant moxibustion group（group E）,with 9 rats in each group. Cold-damp coagulation and stagnation type dysmenorrhea models were established. In group C,mild moxibustion on ＂Shenque（神阙 CV 8） ＂ and＂Guanyuan（关元 CV 4）＂ was carried out from the time after modeling on the 8 th day for 3 consecutive days. In group D, mild moxibustion was given as the same methods with group C after injection with oxytocin on the 11 th day. In group E, mild moxibustion was given as the same methods from the time after modeling on the 8 th day to that after injection with oxytocin on the 11 th day for 4 consecutive days. The writhing behavior and the changes in levels of ET-1 and NO in uterine tissues of rats with dysmenorrhea in each group were observed.Results： Comparison of the latent period： compared with（4.38 ± 1.06） min in group B,the latent period of rats in group C（9.67 ± 1.32） min,group D（11.78 ± 1.30） min and group E（15.00 ± 1.22） min obviously prolonged（all p 0.01）. Compared with group C, the latent period of group E obviously prolonged（p 0.01）. Compared with group D, the latent period of group E obviously prolonged（p 0.01）.Comparison of the writhing times： compared with（4.38 ± 1.06） in group B,the writhing times of rats in group C（9.67 ± 1.32）,group D（11.78 ± 1.30） and group E（15.00 ± 1.22） reduced（all p 0.01）. Compared with group C,the writhing times of rats in group D and group E reduced（both p 0.01）. Compared with group D, the writhing times in group E reduced（p 0.05）. Comparison of the total writhing score：compared with（4.38 ± 1.06） in group B,the total writhing score of rats in group C（9.67±1.32）,group D（11.78 ± 1.30） and group E（15.00 ± 1.22） decreased（all p 0.01）. Compared with group C,the total writhing score of rats in group D and group E decreased（both p 0.01）. Compared with group D,the total writhing score of rats in group E decreased（p 0.05）. Comparison of ET-1 level： compared with（4.80 ± 0.47） in group A,the ET-1 level in uterine tissues of rats in group B（7.57±0.69） significantly increased（P 0.01）. Compared with group B, the ET-1 level in uterine tissues of rats in group C（6.20 ±0.50）,group D（5.67 ±0.29） and group E（5.16±0.33） obviously decreased（all p 0.01）. Compared with group C, the ET-1 level in uterine tissues of rats in group D and group E obviously decreased（p 0.05, p 0.01）. Compared with group D, the ET-1 level in uterine tissues of rats in group Eobviously decreased（p 0.05）. Comparison of NO level： compared with（6.63±1.83） in group A, the NO level in uterine tissues of rats in group B（1.62 ±0.58） significantly decreased（p 0.01）. Compared with group B, the NO level in uterine tissues of rats in group C（3.60±0.59）,group D（4.77 ±0.67） and group E（5.99±0.63） obviously increased（all p 0.01）. Compared with group C,the NO level in uterine tissues of rats in group Dand group E obviously increased（p 0.05, p 0.01）. Compared with group D, the NO level in uterine tissues of rats in group E obviously increased（p 0.01）.Conclusion： The analgesic effect of mild moxibustion at different intervention times on cold-damp coagulation and stagnation type dysmenorrhea rats was different, which was the most significant in pre-instant moxibustion group. One of the mechanisms of action may be related with the adjustment of abnormal levels of ET-1 and NO.

Influence of moxibustion temperatures on blood lipids, endothelin-1, and nitric oxide in hyperlipidemia patients

OBJECTIVE： To observe the influence of moxibustion temperature on blood lipids, endothelin-1（ET-1）, nitric oxide（NO）, and ET-1/NO in hyperlipidemia patients. METHODS： Forty-two primary hyperlipidemia patients were randomly divided into two groups of 21 and treated with moxibustion at different temperatures. Moxibustion was performed with the moxa roll 2.5-3.0 cm from the skin in the treatment group and 4 cm in the control group, 10 min per point, once every other day. Skin temperature was precisely measured with a thermometer during moxibustion. After a 12-week treatment, seven measurements of blood lipids, ET-1, and NO were recorded. RESULTS： Total cholesterol and triglyceride, were lower in the treatment group than in the control group（P0.05）. Serum ET-1 and ET-1/NO was obvi-ously lowered in the treatment group（P0.001）. Moxibustion regulated NO and ET-1/NO in the treatment group much better than in the control group. CONCLUSION： Moxibustion can regulate blood lipids and clear blood vessels. Moxibustion at 45℃has a better effect than moxibustion at 38℃ on regulating blood lipids and protecting vascular endothelial function, indicating that suitable temperature influences the curative effect of moxibustion.

Alleviation of hypoxic pulmonary vascular structural remodeling by L-arginine

Objective To explore the effect of L-arginine on hypoxic pulmonary vascular structural remodeling and its possible mechanisms.Methods Eighteen Wistar rats were randomly divided into three groups: the hypoxia group, the hypoxia with L-arginine group and the control group. Pulmonary artery mean pressure was evaluated with right cardiac catheterization. Pulmonary vascular structural changes were also observed. Plasma concentration of nitric oxide (NO) was measured via spectrophotometry, and endothelin-1 (ET-1) mRNA expression in pulmonary artery endothelial cells was detected using in situ hybridization.Results The pulmonary artery mean pressure was significantly high in hypoxic rats than in normal controls (20.33±2.18?mm?Hg vs 15.38±1.05?mm?Hg, P<0.05). Microstructural and ultrastructural analysis revealed the development of hypoxic pulmonary vascular structural remodeling in the hypoxic rats. Meanwhile, the plasma NO concentration was markedly lower in the hypoxic rats than in controls (P<0.05). The expression signals of ET-1 mRNA by pulmonary artery endothelial cells of hypoxic rats strengthened obviously. L-arginine ameliorated pulmonary hypertension (16.73±1.35?mm?Hg vs 20.33±2.18?mm?Hg, P<0.05) as well as pulmonary vascular structural remodeling in the hypoxic rats in association with an increase in plasma NO concentration (P<0.05) and inhibited ET-1 mRNA expression by the endothelial cells of pulmonary arteries.Conclusion L-arginine might play an important role in the regulation of hypoxic pulmonary vascular structural remodeling and hypoxic pulmonary hypertension. The mechanism is probably related to promoting NO production and, as a result, inhibiting ET-1 mRNA expression by pulmonary artery endothelial cells in hypoxic rats.

Modified cold storage of rat livers with self-made HYD solution

Objective To investigate the cold preservation effect on rat livers of a modified storage method with self-made HYD solution.Methods The vascular bed of rat livers was expanded with an additional 20 to 40?ml self-made HYD solution/100?g liver. After resection of the liver, the extra HYD solution (expressed as % liver weight) was entrapped via portal infusion by tying off the supra- and infra-hepatic inferior vena cava. Forty rats were randomly divided into four groups including control group with conventional storage method, and 20%, 30% and 40% groups according to the amount of extra HYD solution. We compared the preservation effect of the modified storage method with that of the conventional storage method using an isolated perfused rat liver model.Results Bile production and all the indices of hepatic microcirculation including portal perfusion pressure, endothelin-1 in the effluent, trypan blue distribution time and histology were significantly superior in the modified method groups compared to those in the control group (P<0.05). The contents of dihydroxybenzoic acid (DHBA) in the modified method groups were significantly lower than those in the control group (P<0.05). Liver enzymes activities in the 30% group were markedly lower than those in the control group (P<0.05). The preservation effect on rat liver in the 30% group was the best among the modified method groups.Conclusion The modified cold storage method is effective and may have potential for clinical application in liver preservation.

Clinical Study on Acupuncture and Ischemic Stroke

Objective： To study the clinical efficacy of acupuncture for ischemic stroke and its effect on ET-1 levels in IS cases. Method： The 63 cases were randomized into a treatment group （31 cases）, receiving acupuncture and routine method, and control group （32 cases）, receiving routine method alone. Seven days constitute a course of treatment, a 2-day interval between two courses. The clinical efficacy and ET-1 level in two groups were compared after four weeks. Result： The total effective rate in the treatment group and control group were 96.8% and 75% respectively, with a statistical significance （P〈0.05）; the ET-1 level in blood plasma also with a statistical significance after the treatment （P〈0.05）; the pre-treatment and post-treatment ET-1 level in the treatment group showed a statistical significance （P〈0.01）, whereas the control group didn＇t. Conclusion： Acupuncture is effective for ischemic stroke and can lower the ET-1 level in ischemic stroke cases.

Effect of salvianolic acid B on TNF-a induced cerebral microcirculatory changes in a micro-invasive mouse model

Purpose： To investigate the effects of salvianolic acid B （SAB） on tumor necrosis factor a （TNF-a） induced alterations of cerebral microcirculation with a bone-abrading model. Methods： The influences of craniotomy model and bone-abrading model on cerebral microcirculation were compared. The bone-abrading method was used to detect the effects of intracerebroventricular application of 40 μg/kg.bw TNF-a on cerebral venular leakage of fluorescein isothiocyanate （FITC）- albulmin and the rolling and adhesion of leukocytes on venules with fluorescence tracer rhodamine 6G, The therapeutical effects of SAB on TNF-a induced microcirculatory alteration were observed, with continuous intravenous injection of 5 mg/kg.h SAB starting at 20 min before or 20 min after TNF-a administration, respectively, The expressions of CDllb/CD18 and CD62L in leukocytes were measured with flow cytometry, Immunohistochemical staining was also used to detect E-selectin and ICAM-1 expression in endothelial cells, Results： Compared with craniotomy method, the bone-abrading method preserved a higher erythrocyte velocity in cerebral venules and more opening capillaries. TNF-a intervention only caused responses of vascular hyperpermeability and leukocyte roiling on venular walls, without leukocyte adhesion and other hemodynamic changes. Pre- or post-SAB treatment attenuated those responses and suppressed the enhanced expressions of CDII b/CDI8 and CD62L in leukocytes and E-selectin and ICAM-I in endothelial cells induced by TNF-a. Conclusions： The pre- and post-applications of SAB during TNF-a stimulation could suppress adhesive molecular expression and subsequently attenuate the increase of cerebral vascular permeability and leukocyte rolling.