蛛网膜下腔出血后大鼠基底动脉p38MAPK信号传导途径研究

目的探讨蛛网膜下腔出血(SAH)后大鼠基底动脉中p38丝裂原活化蛋白激酶(p38MAPK)信号传导通路的活化情况以及与脑血管痉挛(CVS)的关系。方法通过枕大池二次注血方法制作大鼠SAH模型,以免疫组化方法和逆转录酶-多聚酶链反应分析,分别从蛋白、基因水平分析SAH后基底动脉中p38MAPK信号传导通路的活化情况。结果 SAH后大鼠基底动脉逐渐出现痉挛。基底动脉磷酸化p38MAPK表达逐渐增加,3 d时达高峰并持续至第5 d,14 d时恢复正常。p38MAPK基因表达在注血后1 d明显增加,逐渐增加,于5 d时达高峰,14 d仍维持较高水平。结论SAH后大鼠基底动脉中p38MAPK信号传导通路激活,可能诱导CVS的发生。     

蛛网膜下腔出血后脑血管痉挛的肿瘤坏死因子-α基因启动子相关遗传易感性分析

目的 探讨蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)与肿瘤坏死因子-α基因(TNF-α)启动子相关遗传易感性。方法 经临床确诊SAH患者168例,采用TCD检测和评定SAH患者CVS情况,分为CVS(-)组84例和CVS(+)组84例,另选84名正常成年人作为对照组,应用聚合酶链反应和序列特异性引物-PCR(SSP-PCR)方法对TNF-α基因启动子区5个位点进行基因分型, 用EPI 和EH 等软件分析各位点等位基因、基因型及其组间差异。结果 CVS(+)组与CVS(-)组Hunt-Hess分级构成比有明显差异(P<0.05)。38GG基因型和-863CC基因型是SAH后发生CVS的易感因素(P均<0.05)。多因素Logistic回归分析发现-238G/A、-863C/A基因型和Hunt-Hess分级为与脑血管痉挛有关的危险因素(P<0.05)。CVS(+)组与CVS(-)组年龄、性别、高血压病史及CT Fisher分级构成比均无明显差异(P均>0.05)。结论 TNF-α基因启动子区多态性可能是影响SAH患者发生CVS的高危因素。     

蛛网膜下腔出血后脑血管痉挛的治疗

自发性蛛网膜下腔出血(spontaneous subarachnoid hemor- rhage,sSAH)是临床常见的一种神经外科急症,约80%～95%的sSAH患者是颅内动脉瘤源性蛛网膜下腔出血(aneu- rysmal SAH,aSAH)。aSAH患者常易并发脑血管痉挛(cerebral vasospasm,CV),严重影响预后。CV是一种持续性的对血管舒张药物无反应的血管收缩状态。依据患者神经系统症状的有无可将其分为无症状性脑     

蛛网膜下腔出血后脑血管痉挛的研究进展

脑血管痉挛（ＣＶＳ）也称颅内动脉痉挛,为脑底大动脉的一支或多支由于动脉壁平滑肌的收缩或血管损伤引起其管腔形态学变化,从而在动脉造影时表现的管腔狭窄。严重者可造成脑缺血和脑梗塞,引起迟发性神经功能障碍。动脉瘤破裂性蛛网膜下腔出血（ＳＡＨ）常引起ＣＶＳ,...     

刺五加防治蛛网膜下腔出血后脑血管痉挛

目的 观察刺五加注射液防治蛛网膜下腔出血（ＳＡＨ）后脑血管痉挛（ＣＶＳ）的疗效。方法 将ＳＡＨ患者４４例,随机分为刺五加组（２０例）和ＳＡＨ对照组（２４例）,２组均于发病４８小时内接受治疗,ＳＡＨ对照组用脱不、止血等常规疗法,刺五加组在常规疗法基础上加用刺五加注射液１００毫升,静脉滴注,每日一次,共７日。结果 一个月内,刺五加组的ＣＶＳ发生率、死亡率明显低于ＳＡＨ对照组（Ｐ〈０．０５）,再出血发生率２组比较无明显差异（Ｐ〉０．０５）。发病第１４天时,刺五加组患者的病情级别明显低于ＳＡＨ对照组（Ｐ〈０．０５）。结论 刺五加注射液防治ＳＡＨ后ＣＶＳ疗效确切,并能促进神经功能的尽快恢复,且无增加再出血的危险。     

一氧化氮在蛛网膜下腔出血后脑血管痉挛中的作用

脑血管痉挛(cerebral vasospasm,CVS)是动脉瘤性蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后高致死率、致残率的主要原因之一。SAH后CVS的发病机制尚不明确,目前比较肯定的机制是蛛网膜下腔血液和红细胞崩解产物氧合血红蛋白(OxyHb)是CVS发生的第一推动力,起关键作用。血管     

蛛网膜下腔出血后脑血管痉挛危险因素探讨

目的 通过对临床观测指标与蛛网膜下腔出血(subaraehnoid hemorrhage,SAH)后脑血管痉挛(cerebral vasospasm,CVS)关系的回顾性分析,筛选出相关危险因素,为指导临床SAH后CVS治疗提供依据.方法 收集符合研究标准的99例SAH患者临床资料,按是否发生CVS分为两组,分析入院时年龄、发病意识、发病时体温、血钙水平及钙离子拮抗剂干预、Hunt-Hess分级、白细胞计数等相关指标.结果 两组患者体温、白细胞计数、血钙、钙离子拮抗剂干预、Hunt-Hess分级、发病意识均有统计学差异(P＜0.05).中年、高Hunt-Hess分级、白细胞计数升高均与CVS的发生呈正相关;钙离子拮抗剂干预为CVS发生的负相关因素.结论 年龄、Hunt-Hess分级、白细胞计数为SAH后发生CVS的独立危险因素,钙离子拮抗剂的使用为保护因素;发病意识为SAH后发生CVS正相关因素;血钙为SAH后发生CVS负相关因素.     

Trapidil对蛛网膜下腔出血后脑血管痉挛作用的实验研究

目的 探讨蛛网膜下腔出血（SAH）后脑血管痉挛的发生机制及其可能的治疗方法。方法 利用家兔枕大池内注血构建SAH模型，观察血小板衍生生长因子（PDGF）拮抗剂trapidil对脑基底动脉的影响。结果 脑基底动脉于SAH后48h明显变细；静脉或动脉内持续灌注trapidil 15min（1．5mg／min）后，数字减影脑血管造影（DSA）显示痉挛血管已明显扩张变粗，30min时达高峰。结论 PDGF可能参与脑血管痉挛发生的病理过程，PDGF拮抗剂trapidil可有效缓解实验性SAH后脑血管痉挛，有望成为脑血管痉挛的治疗药物。     

尼莫地平防治蛛网膜下腔出血后脑血管痉挛疗效观察

脑血管痉挛(CVS)是蛛网膜下腔出血(SAH)最常见、最严重的并发症之一,发生率高达70%,是SAH患者病残和死亡的主要原因,SAH后积极防治CVS的发生有重要临床意义。1资料与方法1.1病例选择2008-06-2011-06我院神经内科收治SAH     

Role of P38 mitogen-activated protein kinase on Cx43 phosphorylation in cerebral vasospasm after subarachnoid hemorrhage

Objectives: Cx43 phosphorylation is involved in the pathogenesis of cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH). However, the exact phosphorylation mechanism of Cx43 in CVS is not fully elucidated. Thus, we examined the role of P38MAPK in CVS and Cx43 phosphorylation, using a double hemorrhage rat model.

Methods: Sprague–Dawley rats weighing 300–350?g were grouped into sham, SAH, vehicle, and SAH?+?SB203580. SAH was induced by double injecting blood into the prechiasmatic cisterns. Neurological score was measured with the Garcia scoring system, and the diameters of basilar arteries and the expression of pCx43, pP38MAPK, and P38MAPK proteins were measured through pressure myograph measurement and Western blot analysis, respectively.

Results: The neurological scores remarkably decreased after SAH but remarkably improved after SB203580 was used. The results of pressure myograph analysis on the SAH and vehicle groups showed the considerable narrowing of the basilar arteries in comparison with that of the sham group. By contrast, the arterial diameters in the SAH?+?SB203580 group were much larger than those observed in the SAH and vehicle groups. Moreover, the P38MAPK expression in the sham group had no substantial change in contrast to the SAH and vehicle groups, and pCx43 and pP38MAPK increased in the SAH and vehicle groups. Meanwhile, the SAH?+?SB203580 group showed marked decrease in Cx43 and P38MAPK phosphorylation levels relative to the SAH and vehicle groups.

Conclusions: P38MAPK pathway facilitates the development of CVS through the upregulation of Cx43 phosphorylation.     

p38丝裂原活化蛋白激酶在实验性蛛网膜下腔出血后早期脑损伤中的作用

目的探讨p38丝裂原活化蛋白激酶(p38MAPK)在蛛网膜下腔出血(SAH)后早期脑损伤(EBI)中的作用。方法成年雄性SD大鼠随机分配至对照组、SAH组及p38MAPK干预组,每组18只。采用血管内穿刺法制作SAH模型,干预组于术前30 min经侧脑室注射p38MAPK特异性抑制剂SB203580,造模后24 h处死。观察各组大鼠脑含水量和神经功能评分,RT-PCR及免疫组化检测脑组织p38MAPK表达。结果与对照组相比,SAH组大鼠脑含水量(t=-196.35,P0.01)及p38 MAPK的mRNA水平(t=-24.75,P0.01)均明显升高,神经功能评分明显减低(t=201.08,P0.01)。与SAH组相比,干预组脑含水量(t=75.67,P0.01)及p38 MAPK的mRNA水平(t=9.43,P0.01)均明显下降,神经功能评分明显升高(t=-81.68,P0.01)。免疫组化示SAH组及干预组均有p38MAPK表达,但干预组较SAH组表达水平明显下降(t=-3.37,P0.01)。结论 p38 MAPK在EBI形成机制中起重要作用,有望成为防治EBI的药物作用新靶点。     

Activation of p38 mitogen-activated protein kinase in spinal microglia mediates morphine antinociceptive tolerance

Compelling evidence has suggested that spinal glial cells were activated by chronic morphine treatment and involved in the development of morphine tolerance. However, the mechanisms of glial activation were still largely unknown in morphine tolerance. In present study, we investigated the role of p38 mitogen-activated protein kinase (p38 MAPK) in the spinal cord in the development of chronic morphine antinociceptive tolerance. We found that intrathecal administration of morphine (15 microg) daily for 7 consecutive days significantly induced an increase in number of phospho-p38 (p-p38) immunoreactive cells in the spinal cord compared with chronic saline or acute morphine treated rats. Double immunofluorescence staining revealed that p-p38 immunoreactivity was exclusively restricted in the activated spinal microglia, not in astrocytes or neurons. Repeated intrathecal administration of 4-(4-fluorophenyl)-2-(4-methylsulfonylphenyl)-5-(4-pyridyl)-1H-imidazole (SB203580) (10 microg or 2 microg), a specific p38 inhibitor, 30 min before each morphine injection for 7 consecutive days significantly attenuated tolerance to morphine analgesia assessed by tail flick test. However, a single intrathecal administration of SB203580 (10 microg) did not antagonize the established tolerance to morphine analgesia. Taken together, these findings suggested that p38 MAPK activation in the spinal microglia was involved in the development of morphine antinociceptive tolerance. Inhibition of p38 MAPK by SB203580 in the spinal cord attenuated but not reversed the tolerance to morphine analgesia. The present study provides the first evidence that p38 activation in spinal microglia played an important role in the development of tolerance to morphine analgesia.     

盐酸法舒地尔治疗蛛网膜下腔出血所致脑血管痉挛的临床Ⅱ期试验研究

目的通过Ⅱ期临床试验评价盐酸法舒地尔(FSD)注射液治疗蛛网膜下腔出血 (SAH)所致脑血管痉挛(CVS)的临床疗效和安全性。方法采用随机双盲双模拟、阳性药物对照方法。观察64例颅内动脉瘤破所致SAH。分为A、8两组各32例。治疗方式:手术夹闭47例,介入治疗17例。治疗方法:A组FSD 30mg+生理盐水40mL／次、3次／d,B组尼莫地平8mg(40ml)／次、3次／d。比较用药前后两组临床表现、意识变化以GCS、病情以Hunt&Hess分级、预后以GOS 为指标,CT、TCD,生命体征,肝、肾功能、血K、Na、Cl及血、尿、便常规。结果(1)总体病情两组均明显好转,但两组间差异无统计学意义(P=0．3028)。(2)头颅CT:用药前无低密度区者用药后亦未见到改变。(3)TCD:治疗前MCA平均流速(单位:cm／s)A组97．15±51．01,治疗后第 7、14天分别下降为90．64±40．93,78．52±25．62;B组91．06±22．20,治疗后第7、14天分别为84．53± 26．17,85．88±35．80,两组比较无统计学意义(P>0．06)。(4)预后:总有效率A组93．94％(PP)、 93．94％(ITT)和93．94%(GOS);B组相应为96．88％、94．12％和96．88％。两组间差异无统计学意义。(5)对于肝、肾功能、电解质和血、尿、便常规没有任何影响。结论此药具有安全、可靠和有效性．无毒副作用及不良反应。是一种新型抗CVS药物。     

Cilostazol attenuates cerebral vasospasm after experimental subarachnoid hemorrhage

Abstract

Background and purpose: Cerebral vasospasm is a major cause of morbidity and mortality in patients with subarachnoid hemorrhage (SAH). Cilostazol, a selective inhibitor of phosphodiesterase 3, is a peripheral vasodilator, an anti-inflammatory, and causes antiplatelet aggregation. We investigated these effects on cerebral vasospasm after rat SAH.

Methods: Thirty-eight Sprague–Dawley rats were randomly divided into three groups: SAH + normal feed (SAH group; n=14), SAH + feed containing 0·1% cilostazol (cilostazol group; n=12) and sham-operated rats (sham group; n=12). The basilar arteries (BA) of all groups were analysed by measuring wall thickness, internal luminal perimeter and cross-sectional area on day 7. Immunohistochemical study with RM-4, an anti-rat macrophage/dendritic cells monoclonal antibody and ultrastructural study with transmission electron microscopy were performed.

Results: Although most animals in the SAH group presented with typical vasospasm, the means of inner perimeter and cross-section area of the BA in the cilostazol group were significantly greater than the SAH group (836 ± 134 μm versus 771 ± 125 μm and 39177 ± 15405 μm² versus 33098 ± 13871 μm², respectively). Wall thickness of the BA in the cilostazol group demonstrated significant decrease, compared with the SAH group (17·4 ± 2·3 versus 21·0 ± 2·7 μm). In immunohistological study, SAH induced an obvious increase in mean perivascular RM-4-positive cell count, whereas cilostazol significantly reduced it by 59%. Ultrastructural study depicted cilostazol markedly attenuating structural deterioration of the vascular wall due to SAH.

Conclusions: This work demonstrates that cilostazol attenuates cerebral vasospasm after SAH in rat, possibly in part due to the anti-inflammatory effect.     

川芎嗪干预转化生长因子β1诱导肝星状细胞结缔组织生长因子表达中p38丝裂酶原激活蛋白激酶的途径

背景：前期研究发现川芎嗪可通过抑制肝星状细胞的增殖和阻断Ⅰ,Ⅲ胶原的合成,下调结缔组织生长因子的表达等,发挥抗肝纤维化的作用,但具体机制尚不清楚。 目的：观察川芎嗪对体外培养肝星状细胞表达结缔组织生长因子的影响,以及p38丝裂酶原激活蛋白激酶(p38MAPK)信号通路在其中的作用。 方法：用5 μg/L转化生长因子β1诱导活化体外培养的肝星状细胞,用川芎嗪和p38MAPK特异阻断剂SB203580进行干预,以RT-PCR法检测结缔组织生长因子 mRNA和Ⅰ型胶原mRNA的表达,Western blot法检测磷酸化p38MAPK蛋白的表达。 结果与结论：经转化生长因子β1诱导后,肝星状细胞中结缔组织生长因子和Ⅰ型胶原mRNA表达显著增强(P < 0.01),用川芎嗪和SB203580干预后,结缔组织生长因子和Ⅰ型胶原mRNA的表达均出现不同程度的下降。但川芎嗪和川芎嗪+SB203580混合干预对这两者的基因表达抑制作用比单独的SB203580干预更强。川芎嗪和SB203580对磷酸化p38MAPK蛋白表达也都有明显的抑制作用(P < 0.01),但SB203580和川芎嗪+SB203580对其磷酸化蛋白表达抑制作用更明显,而SB203580与川芎嗪+SB203580无明显差异(P > 0.05)。因此,推测川芎嗪可能通过抑制转化生长因子β1诱导的结缔组织生长因子基因表达,阻断Ⅰ型胶原合成,其作用途径可能与抑制p38mapk信号通路有关,同时认为川芎嗪抗纤维化可能是多重作用靶点。     

Involvement of caspase-3 and p38 mitogen-activated protein kinase in cobalt chloride-induced apoptosis in PC12 cells

Our previous study showed that cobalt chloride (CoCl2) could induce PC12 cell apoptosis and that the CoCl2-treated PC12 cells may serve as a simple in vitro model for the study of the mechanism of hypoxia-linked neuronal disorders. The aim of this study is to elucidate the mechanism of CoCl2-induced apoptosis in PC12 cells. Caspases are known to be involved in the apoptosis induced by various stimuli in many cell types. To investigate the involvement of caspases in CoCl2-induced apoptosis in PC12 cells, we generated PC12 cells that stably express the viral caspases inhibitor gene p35 and analyzed the effect of p35 on the process of apoptosis induced by CoCl2. We also examined the effect of cell-permeable peptide inhibitors of caspases. The results showed that the baculovirus p35 gene and the general caspases inhibitor Z-VAD-FMK significantly block apoptosis induced by CoCl2, confirming that caspase is involved in CoCl2-induced apoptosis. Further investigation showed that in this process the caspase-3-like activity is increased, as indicated by the cells' ability to cleave the fluorogenic peptide substrate Ac-Asp-Glu-Val-Asp-7-AMC and to degrade the DNA-repairing enzyme poly-(ADP-ribose) polymerase (PARP), an endogenous caspase-3 substrate. At the same time, caspase-3-specific inhibitors, namely, the peptide Ac-DEVD-CHO, Ac-DEVD-FMK, partially inhibit CoCl2-induced apoptosis. These findings suggested that caspase-3 or caspase-3-like proteases are involved in the apoptosis induced by CoCl2 in PC12 cells. Additionally, we have observed that another apoptotic marker, p38 mitogen-activated protein kinase (MAPK), is significantly activated in this process in a time-dependent manner and that a selective p38 MAPK inhibitor, SB203580, partially inhibits this cell death. The addition of SB203580 also partially suppresses caspase-3-like activity. All these results confirm that the CoCl2-treated PC12 cell is a useful in vitro model with which to study hypoxia-linked neuronal disorders. Furthermore, the results showing that the baculovirus p35 gene and caspase inhibitors possess a remarkable ability to rescue PC12 cells from CoCl2-induced cell death may have implications for future neuroprotective therapeutic approaches for the hypoxia-associated disorders.