内皮素双受体拮抗剂在实验性急性肺栓塞中的应用研究

目的观察静脉内皮素双受体拮抗剂RO2610612在急性肺栓塞中的应用效果,研究内皮素与急性肺栓塞的关系。方法12只健康杂种犬随机分为对照组和实验组,每组6只。两组所有动物给予戊巴比妥静脉麻醉后分别于颈静脉插入热稀释漂浮导管连接生理监护仪;气管内插入气管插管连接呼吸描记器。采集栓塞前数据后于颈静脉注射自体血栓建立急性肺栓塞动物模型。实验组建立模型后1h开始持续静滴RO26106120.2mg/(kg.h),对照组同时输入等量生理盐水。对照组及实验组所有动物于栓塞前、栓塞后1,2,4,6h记录体、肺循环血流动力学指标、呼吸生理指标;取股动脉血3ml,采用放射免疫法测定动脉血内皮素-1水平,并测动脉血气。结果两组动物栓塞后较栓塞前血浆内皮素-1水平、肺动脉平均压、肺血管阻力、呼吸频率、动脉血二氧化碳分压和肺气道阻力显著性升高(P<0.05),心输出量、动脉血氧分压和肺动态顺应性显著性下降(P<0.05)。实验组于栓塞2h后肺动脉平均压、肺血管阻力、呼吸频率、肺气道阻力升高水平显著性低于同时期对照组(P<0.05),血浆内皮素-1水平、心输出量、和肺顺应性显著性高于对照组(P<0.05)。结论在急性肺栓塞病理过程中,内皮素参与了肺循环阻力升高与肺气道阻力升高和肺顺应性下降的形成。内皮素受体拮抗剂拮抗内皮素与其受体的结合,在急性肺栓塞中,可以减缓肺循环阻力、肺气道阻力的上升,因而可能成为急性肺栓塞临床治疗的新手段。     

Effect of Zataria multiflora Extract on Total and Differential White Blood Cell Count and Endothelin Level in Blood of Ovalbumin Sensitized Guinea Pigs

Objective: To investigate the effect of hydro-ethanolic extract of Zataria multiflora (Z. multiflora) on endothelin level, total and differential white blood cells (WBC) count of sensitized guinea pigs. Methods: Five groups of guinea pigs sensitized to ovalbumin (OA) were given drinking water alone (group S), drinking water containing three concentrations of Z. multiflora (0.2, 0.4 and 0.8 mg/mL as groups S+Z1, S+Z2 and S+Z3) and dexamethasone (group S+D), n=6 for each group. The endothelin levels as well as total and differential WBC count in blood of sensitized and control guinea pigs were evaluated using enzyme linked immunosorbent assay method, and hemocytometer and Wright-Giemsa''s staining of blood sample smear respectively. Results: Blood endothelin levels, total and most differential WBC count were increased but lymphocytes decreased in sensitized animals compared to controls (all P<0.01). In groups S+D, S+Z2 and S+Z3 endothelin level, total and differential WBC counts were significantly improved compared with group S (P<0.01). Although, all measured parameters in group S+Z1 was lower than group S+D (P<0.01), some parameters in group S+Z3 were greater than in group S+D (P<0.05 to P<0.01). Conclusion: The results showed an anti-inflammatory effect of Z. multiflora extract in sensitized guinea pigs, which may suggest a therapeutic potential for the plant on asthma.     

疏血通注射液联合纳洛酮治疗慢性肺心病急性加重期的临床研究

目的 探讨疏血通注射液联合纳洛酮治疗慢性肺心病急性加重期对血浆内皮素（ET-1）的影响。方法 采用回顾性与便利抽样研究方法,病例收治时间为2010年2月—2016年12月,选择在此期间在西安市五环集团职工医院诊治的慢性肺心病急性加重期患者122例,按照治疗方法的区别分为观察组与对照组各61例,对照组给予纳洛酮治疗,2 mg加入0.9% NaCl注射液100 mL中静滴,1次/d。观察组在对照组治疗的基础上给予疏血通注射6 mL加入0.9% NaCl注射液100 mL中静滴,1次/d。两组都治疗观察28 d。比较两组疗效、三尖瓣区最大返流速度（V_max）和肺动脉压（PAPs）及ET-1水平,同时比较两组凝血功能指标。结果 对照组总有效率为86.9%,观察组为98.4%,观察组显著高于对照组（P<0.05）。治疗后两组的V_max值、PAPs值与血浆ET-1含量均显著低于治疗前（P<0.05）,且观察组显著低于对照组（P<0.05）。治疗后两组的凝血酶原时间（PT）、活化部分凝血活酶时间（APTT）与凝血酶时间（TT）值都显著高于治疗前（P<0.05）,且治疗后观察组显著高于对照组（P<0.05）。结论 疏血通注射液联合纳洛酮治疗慢性肺心病急性加重期能抑制血浆ET-1的表达,改善肺动脉压力,具有抗凝的作用,从而有利于提高治疗疗效。     

高氧液对脑缺血大鼠内皮素及降钙素基因相关肽含量的影响

目的 :探讨高氧液对缺血大鼠脑细胞的保护作用。方法 :40只 Wistar大鼠 (雌雄各半 )随机分成高氧液组、对照组、模型组和假手术组 ,每组各 10只。阻断 3条动脉造成急性不完全性脑缺血 ,制备动物模型。制模成功后 ,高氧液组立即以 10 ml· kg- 1 · h- 1 的速率经尾静脉输入高氧液 ,对照组以同种方法输入等量生理盐水 ,模型组和假手术组不治疗。制模后 1小时各组动物经腹主静脉取血 ,然后立即断头取脑 ,测定内皮素 (ET)和降钙素基因相关肽 (CGRP)含量。结果 :高氧液可明显降低脑缺氧后 ET水平 ,而升高 CGRP水平 ,纠正 ET、CGRP失衡。结论 :高氧液对缺血大鼠脑细胞有明显保护作用。     

阿托伐他汀联合坎地沙坦酯治疗原发性高血压的临床研究

目的探究阿托伐他汀联合坎地沙坦酯治疗原发性高血压的临床疗效。方法选取2012年1月—2014年11月义马煤业集团股份有限公司收治的原发性高血压患者300例,随机分为对照组和治疗组,每组150例。对照组口服坎地沙坦酯片,2片/次,1次/d。治疗组口服阿托伐他汀钙片1片/次,1次/d,坎地沙坦酯片的用法用量同对照组。两组患者均连续治疗10周。观察两组的临床疗效,同时比较治疗前后两组患者收缩压、舒张压、超敏C反应蛋白(hs-CPR)、内皮素(ET)、一氧化氮(NO)的变化。结果治疗后,两组总有效率分别为76.67%、90.67%,两组比较差异有统计学意义(P0.05)。治疗后,两组患者收缩压、舒张压、hs-CRP、ET均较治疗前显著降低,NO显著升高,同组治疗前后比较差异有统计学意义(P0.05);且治疗组这些观察指标的改善程度优于对照组,两组比较差异有统计学意义(P0.05)。结论阿托伐他汀联合坎地沙坦酯治疗原发性高血压具有较好的临床疗效,可改善患者的血管内皮功能和炎症反应,值得在临床上进一步推广和应用。     

In vivo imaging reveals an essential role of vasoconstriction in rupture of the ovarian follicle at ovulation

Rupture of the ovarian follicle releases the oocyte at ovulation, a timed event that is critical for fertilization. It is not understood how the protease activity required for rupture is directed with precise timing and localization to the outer surface, or apex, of the follicle. We hypothesized that vasoconstriction at the apex is essential for rupture. The diameter and blood flow of individual vessels and the thickness of the apical follicle wall were examined over time to expected ovulation using intravital multiphoton microscopy. Vasoconstriction of apical vessels occurred within hours preceding follicle rupture in wild-type mice, but vasoconstriction and rupture were absent in Amhr2^cre/+SmoM2 mice in which follicle vessels lack the normal association with vascular smooth muscle. Vasoconstriction is not simply a response to reduced thickness of the follicle wall; vasoconstriction persisted in wild-type mice when thinning of the follicle wall was prevented by infusion of protease inhibitors into the ovarian bursa. Ovulation was inhibited by preventing the periovulatory rise in the expression of the vasoconstrictor endothelin 2 by follicle cells of wild-type mice. In these mice, infusion of vasoconstrictors (either endothelin 2 or angiotensin 2) into the bursa restored the vasoconstriction of apical vessels and ovulation. Additionally, infusion of endothelin receptor antagonists into the bursa of wild-type mice prevented vasoconstriction and follicle rupture. Processing tissue to allow imaging at increased depth through the follicle and transabdominal ultrasonography in vivo showed that decreased blood flow is restricted to the apex. These results demonstrate that vasoconstriction at the apex of the follicle is essential for ovulation.During ovulation in typically mono-ovulatory species such as humans, as well as in poly-ovulatory species such as rodents, the oocyte is released from the preovulatory follicle by extrusion through a rupture site on the outer surface, or apex, of the follicle, which protrudes from the surface of the ovary (1). Precise timing and accurate spatial localization of rupture at the apex are essential to allow capture of the oocyte by a hormonally primed oviduct where fertilization occurs, but the mechanisms involved are not yet understood. The rupture site breaches multiple layers of cells and their associated extracellular matrix and basement membranes (2). These include the single layer of epithelial cells that covers the surface of the ovary, the basement membrane that supports it, and the multiple cell layers comprising the wall of the preovulatory follicle. The outer wall of the ovarian follicle contains androgen-secreting theca cells and extensive vasculature. This vasculature consists of an inner and an outer plexus of capillaries with associated arterioles and venules that supply nutrients to the entire follicle (3–5). Underlying the theca and separated from it by a basement membrane is the avascular granulosa cell layer that serves as the major source of estrogen. The oocyte resides in the center of the follicle surrounded by multiple layers of specialized granulosa cells known as “cumulus cells.” In a mature preovulatory follicle, formation of a fluid-filled antral cavity separates the oocyte–cumulus complex from the mural granulosa cells that form the wall of the follicle except at a region known as the “stalk,” which connects the oocyte–cumulus complex to the antral granulosa cells of the follicle wall. At ovulation the oocyte is released from the follicle in association with attached cumulus cells.The preovulatory release of surge levels of luteinizing hormone (LH) from the anterior pituitary acts on receptors in the follicle to trigger events critical for the rupture and remodeling of the follicle and differentiation of granulosa and theca cells into progesterone-producing cells of the corpus luteum. The cumulus cells are induced to secrete a mucoelastic extracellular matrix which causes loosening of contacts between granulosa cells and between granulosa cells and the oocyte, a process known as “cumulus expansion,” which is essential for ovulation (1). Expression of proteases belonging to several major families, including the matrix metalloproteinase, plasminogen activator/plasmin, and ADAMTS (a disintegrin and metalloproteinase with thrombospondin-like motifs) families, increases. Simultaneously, follicle cells express protease inhibitors such as tissue inhibitors of metalloproteinases (TIMPs 1–4) and plasminogen activator inhibitors (PAI 1–3) (6, 7). The increase in protease activity is essential for rupture of the follicle and for the breakdown of the basement membrane separating theca and granulosa cells to allow the ingrowth of blood vessels to establish the corpus luteum. The mechanisms that regulate the balance of protease and protease inhibitor activity in the follicle to allow precise rupture at the apex while protecting most of the follicle structure from protease activity are not understood (1, 6, 7).We postulated that vasoconstriction of vessels within the theca at the apex of the follicle is required to promote follicle rupture. Our first approach was to examine mice with conditional expression of a dominant active allele of smoothened (SMO), the transmembrane protein that relays signaling by the hedgehog (HH) pathway. In these Amhr2^cre/+SmoM2 mice, preovulatory follicles develop normally in many respects, including changes in the expression of critical genes in response to the preovulatory LH surge (8, 9). However, follicles fail to rupture, and oocytes remain trapped as the follicles luteinize. The major ovarian phenotype in these mice is a pronounced deficiency of vascular smooth muscle (VSM) surrounding vessels in the theca cell layer, whereas other vessels that are present throughout the stroma of the ovary have normal maturation with VSM. Because VSM is required for vasoconstriction, the mice provided a model to test whether failure of vasoconstriction contributes to anovulation. In additional experiments with wild-type mice, we blocked the increase in the expression of endothelin 2 (Edn2) by granulosa cells that normally occurs within hours before follicle rupture (10, 11). Because EDN2 is a potent vasoconstrictor, this approach allowed us to test the effect on follicle rupture of inhibiting vasoconstriction versus treatment with exogenous compounds to restore vasoconstriction. In addition, treatment of wild-type mice with EDN2 receptor antagonists was used to test the role of EDN2 in vasoconstriction and rupture. Vasoconstriction and changes in the follicle wall were monitored repeatedly relative to the time of ovulation using intravital multiphoton microscopy.     

Protective effects of simultaneous splenectomy on small‐for‐size liver graft injury in rat liver transplantation

Splenectomy is an effective technique in living donor liver transplantation (LDLT) with small‐for‐size (SFS) liver grafts for overcoming SFS liver graft injury. However, the protective mechanism of splenectomy is still unclear. The aim of this study was to investigate how splenectomy could attenuate SFS graft injury through the measurement of biochemical factors, particularly the expression of endothelin (ET)‐1, which is a key molecule of microcirculatory disorders by mediating sinusoidal vasoconstriction. We performed rat orthotopic liver transplantation using SFS liver grafts with or without splenectomy. We investigated intragraft expression of ET‐1 mRNA and hepatic protein levels of ET‐1. In addition, portal pressure, hepatic injury and morphological changes, and survival rate were evaluated. In result, intragraft ET‐1 mRNA expression after SFS liver transplantation was significantly downregulated by splenectomy, and hepatic expression of ET‐1 in SFS grafts was rarely observed. Splenectomy inhibited the increase in portal pressure, ameliorated SFS liver graft injury and improved the graft survival rate after SFS liver transplantation. In conclusion, splenectomy improved the SFS liver injury and decreased the expression of ET‐1 by attenuating portal hypertension on SFS liver transplantation. Downregulation of intragraft ET‐1 expression plays important roles in the protective mechanism of splenectomy in SFS liver transplantation.     

大鼠闭合性脑损伤后星形胶质细胞的形态及GAP-43、ET-1 mRNA表达的变化

目的:探讨闭合性脑损伤后脑组织星形胶质细胞(Ast)形态学变化和生长相关蛋白(GAP-43)、内皮素-1(ET-1)mRNA表达的变化.方法:78只大鼠随机分为12个损伤组和1个假损伤对照组,每组6只.损伤组制备Marmaruu脑损伤模型.各组分别于伤后0h、0.5 h、1 h、1.5 h、2 h、6 h、12 h、36 h、48 h、60 h、66 h、72 h断头取脑,在脑干损伤灶周围取部分脑组织,常规固定切片HE染色,胶质纤维酸性蛋白免疫组化染色结合计算机彩色图像分析技术观察分析Ast形态,RT-PCR法检测GAP-43及ET-1 mRNA.结果:闭合性脑损伤后0.5 h Ast反应性肿胀最明显,以后随着时间的延长Ast反应性肿胀程度降低、肿胀Ast的数目减少.伤后6～72 h脑组织GAP-43 mRNA及ET-1 mRNA均较假损伤对照组增高(P＜0.01).结论:GAP-43及ET-1在脑损伤后血脑屏障损害的病理生理过程烫中起重要作用.     

急性冠脉综合征患者血浆内皮素及外周血单个核细胞内皮素受体mRNA检测

目的:研究急性冠脉综合征(ACS)患者血浆内皮素(ET)水平及外周血单个核细胞ETA及ETB受体mRNA的表达.方法:采用放免法测定36例ACS患者(其中急性心肌梗死(AMI)16例,不稳定型心绞痛(UAP)20例)与20例健康人外周血浆ET水平,逆转录聚合酶链反应(RT-PCR)测定2组外周血单个核细胞ETA及ETB受体mRNA的表达.结果:ACS患者外周血浆ET((0.126±0.013)μg/L)明显高于对照组((0.050±0.006)μg/L)(P＜0.05),AMI亚组血浆ET((0.129±0.016)μg/L)与UAP亚组((0.124±0.011) μg/L)差异无统计学意义(P＞0.05).ACS组ETA受体mRNA表达水平(0.86±0.15)与对照组(0.53±0.16)相比差异有统计学意义(P＜0.05),亚组间ETA受体mRNA表达差异无统计学意义(P＞0.05).2组间ETB受体mRNA表达差异无统计学意义.结论:在ACS的发生过程中血浆ET水平升高,ETA受体mRNA的表达增加,促进了ACS的发生、发展.