Increased activity of Rho kinase contributes to hemoglobin-induced early disruption of the blood-brain barrier in vivo after the occurrence of intracerebral hemorrhage

This study is to examine whether the activation of Rho kinase (ROCK) accounts for hemoglobin (Hb)-induced disruption of blood-brain barrier (BBB) after the occurrence of intracerebral hemorrhage. A model of intracerebral injection of Hb was established in rats. Changes in the levels of mRNA of RhoA, ROCK2 and matrix metalloproteinase-9 (MMP-9) were measured using quantitative real-time polymerase chain reaction. Protein expression of RhoA, ROCK2, claudin-5 and MMP-9, as well as ROCK activity, were determined using Western blotting. Immunohistochemical assay was performed to visualize the expression of RhoA, ROCK2, claudin-5 and MMP-9 in endothelial cells. Hb injection produced a significant increase in BBB permeability and water content in the brain. Significant reduction of claudin-5 expression was detected by Western blotting and immunofluorescence in Hb group. The levels of RhoA and ROCK2 were significantly up-regulated from 6 h to 12 h after Hb injection and were concomitant with the increase in ROCK activity. Immunofluorescence double staining showed enhanced p-myosin light chain immunoreactivity but diminished claudin-5 staining in endothelial cells. Significant up-regulation of MMP-9 expression was detected after Hb injection, and statistical analyses further confirmed a positive correlation of MMP-9 expression with ROCK activity. The results showed that ROCK was activated in endothelial cells by Hb. This may account for the early disruption of the BBB via up-regulation of p-myosin light chain expression and aggravation of injuries to TJ proteins. The activation of ROCK may also increase MMP-9 expression, thereby leading to further BBB disruption.     

3′3-Diindolylmethane inhibits migration,invasion and metastasis of hepatocellular carcinoma by suppressing FAK signaling

Late stage hepatocellular carcinoma (HCC) usually has a low survival rate because it has high potential of metastases and there is no effective cure. 3′3-Diindolylmethane (DIM) is the major product of the acid-catalyzed oligomerization of indole-3-carbinol present in cruciferous vegetables. DIM has been proved to exhibit anticancer properties. In this study, we explored the effects and molecular mechanisms of anti-metastasis of DIM on HCC cells both in vitro and in vivo. We chose two HCC cell lines SMMC-7721 and MHCC-97H that have high potential of invasion. The results showed that DIM inhibited the proliferation, migration and invasion of these two cell lines in vitro. In addition, in vivo study demonstrated that DIM significantly decreased the volumes of SMMC-7721 orthotopic liver tumor and suppressed lung metastasis in nude mice. Focal Adhesion Kinase (FAK) is found over activated in HCC cells. We found that DIM decreased the level of phospho-FAK (Tyr397) both in vitro and in vivo. DIM inhibition of phospho-FAK (Tyr397) led to down-regulation of MMP2/9 and decreased potential of metastasis. DIM also repressed the migration and invasion induced by vitronectin through inactivation of FAK pathway and down-regulation of MMP2/9 in vitro. We also found that pTEN plays a role in down-regulation of FAK by DIM. These results demonstrated that DIM blocks HCC cell metastasis by suppressing tumor cell migration and invasion. The anti-metastasis effect of DIM could be explained to be its down-regulated expression and activation of MMP2/9 partly induced by up-regulation of pTEN and inhibition of phospho-FAK (Tyr397).     

老年全麻患者血清脂联素、基质金属蛋白酶-9水平与术后认知功能障碍的关系

目的 评价老年全身麻醉患者血清脂联素(adiponectin,ADP)及基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)水平与术后认知功能障碍(postoperation cognitive dysfunction,POCD)的关系.方法 择期全身麻醉下行全髋关节置换术老年患者98例,年龄65岁～83岁,美国麻醉医师协会(ASA)分级Ⅰ～Ⅱ级,性别不限.所有患者分别于手术前3d及术后1、2、3、7d采用蒙特利尔认知评估表(montreal cognitie assessment,MoCA)评估认知功能,并采集空腹静脉血标本测定血清ADP及MMP-9水平.根据术后3d是否发生POCD分为POCD组和无POCD组.结果 28例患者发生POCD,发生率为28.5％,POCD组患者术后1、2、3d及7d血清MMP-9水平[(537±68)、(481±64)、(432±64)、(393±48) μg/L]较术前[(293±50) μg/L]明显升高(P＜0.05),血清ADP水平[(4.3±1.6)、(4.7±1.2)、(5.1±1.6)、(6.0±1.1) mg/L]较术前[(9.39± 1.36) mg/L]明显下降(P＜0.05).无POCD组术后1、2d血清MMP-9水平较术前明显升高(P＜0.05),血清ADP水平较术前明显下降,术后3d均恢复至术前水平.组间比较,POCD组术后各时间血清MMP-9水平明显高于无POCD组[(439±53)、(387±66)、(301±67)、(296±54)μg/L](P＜0.05);而ADP水平均明显低于无POCD组[(5.7±1.0)、(6.7±1.4)、(9.1±1.0)、(9.4±1.2)mg/L] (P＜0.05).直线相关分析:POCD组患者血清MMP-9水平与MoCA评分呈负相关(r=-0.833,P＜0.01),ADP水平与MoCA评分呈正相关(r=0.513,P＜0.01).结论 老年全麻患者术后血清ADP水平下降与MMP-9水平升高可能参与了POCD发生的病理生理过程.     

基质金属蛋白酶-9和骨桥蛋白在甲状腺癌和腺瘤组织中的表达及其临床意义

目的 探讨基质金属蛋白酶-9 (MMP-9)和骨桥蛋白(OPN)在甲状腺癌及腺瘤组织中的表达和临床意义.方法 应用免疫组织化学法检测MMP-9和OPN在85例甲状腺癌组织、26例甲状腺瘤和15例正常甲状腺组织中的表达水平.结果 MMP-9在正常甲状腺、甲状腺瘤和甲状腺癌组织中的表达率分别为20.00％ (3/15)、53.85％ (14/26)和84.71％ (72/85),两两比较差异有统计学意义(P＜0.05),MMP-9表达水平与甲状腺癌美国肿瘤联合委员会(AJCC)分期、组织分化、颈部淋巴结转移、包膜浸润明显相关(P＜0.05);OPN在正常甲状腺、甲状腺瘤和甲状腺癌组织中的表达率分别为13.33％ (2/15)、46.15％ (12/26)和74.12％ (63/85),两两比较差异有统计学意义(P＜0.05),OPN表达水平与甲状腺癌AJCC分期、颈部淋巴结转移、包膜浸润明显相关(P＜0.05).结论 检测MMP-9和OPN有助于判断甲状腺癌浸润转移潜能.     

阿托伐他汀对心肌梗死后大鼠基质金属蛋白酶-2及微小RNA-21的作用

目的 观察阿托伐他汀对心肌梗死后大鼠基质金属蛋白酶(MMP)-2、微小RNA(miRNA,miR)-21表达的影响.方法 选取140只Wistar大鼠建立急性心肌梗死模型,分为假手术组、对照组、阿托伐他汀低剂量组和高剂量组,进行左室重量指数、心肌胶原容积分数、miRNA-21及MMP-2 mRNA表达测定.结果 阿托伐他汀治疗组左室重量指数、心肌胶原容积分数、梗死周边区miR-21 、MMP-2 mRNA的表达均降低(P＜0.05);各组梗死周边区miR-21表达量与MMP-2 mRNA水平呈正相关(r对照组=0.611,P＜0.05;r阿托伐他汀低剂量组=0.502,P＜0.05;r阿托伐他汀低剂量组=0.541,P＜0.05).结论 阿托伐他汀能降低梗死周边区MMP-2和miR-21的表达,发挥减轻心室重构的作用.     

异氟醚和七氟醚对肺癌根治术患者血清E-selectin、ICAM-1、MMP-2、MMP-9表达的影响

目的评价异氟醚和七氟醚吸入麻醉对肺癌根治术患者血清E-选择素(E-selectin)、内皮细胞间黏附分子(ICAM)-1、基质金属蛋白酶(MMP)-2、MMP-9浓度变化的影响。方法择期行肺癌根治术患者60例,年龄43~68岁,ASAⅠ或Ⅱ级。采用随机数字表法,将患者随机均分为两组:异氟醚(I组)和七氟醚组(S组)。两组患者气管插管后吸入1.7%~2.3%异氟醚或2.5%~3.4%七氟醚,维持I组异氟醚呼气末浓度为1.5~2.0 MAC、S组七氟醚呼气末浓度维持在1.5~2.0 MAC至术毕。于麻醉诱导前5min(T0)、手术开始后1h(T1)、手术开始后2h(T2)和术后1h(T3)时采集静脉血样,酶联免疫吸附法测定血清E-selectin、ICAM-1、MMP-2、MMP-9表达。结果与T0时比较,T1~T3时I组血清E-selectin、ICAM-1、MMP-2、MMP-9表达明显增加(P0.05),T2、T3时S组血清E-selectin、ICAM-1、MMP-2、MMP-9表达明显降低(P0.05)。T1~T3时S组血清E-selectin、ICAM-1、MMP-2、MMP-9表达明显低于I组(P0.05)。结论七氟醚能抑制肺癌根治术患者血清E-selectin、ICAM-1、MMP-2、MMP-9的表达。     

MMP-2和MMP-9在肝内胆管细胞癌组织中的表达及意义

目的:探讨基质金属蛋白酶2(MMP-2)与基质金属蛋白酶9(MMP-9)在肝内胆管细胞癌(ICC)组织中的表达及其与ICC患者临床病理特征及预后的关系。方法:收集2011年1月—2014年12月收治的50例ICC患者的癌组织与癌旁组织手术标本,采用免疫组织化学方法及RT-PCR方法检测MMP-2和MMP-9在以上组织中的表达,分析MMP-2和MMP-9表达与ICC患者临床病理参数以及术后生存率的关系。结果:免疫组织化学结果显示,ICC组织中MMP-2和MMP-9蛋白阳性表达率分别为34.0%和32.0%,而两者在癌旁组织中无阳性表达;RT-PCR结果显示,ICC组织中MMP-2和MMP-9的相对表达均明显高于癌旁组织(均P0.05)。MMP-2和MMP-9表达与ICC患者是否存在淋巴结转移和肿瘤分化程度有关(均P0.05),而与患者性别、年龄和肿瘤分期无关(均P0.05);MMP-2和MMP-9阳性患者生存率明显低于各自阴性者,而两者均阳性患者的生存率最低(均P0.05)。结论:ICC组织中MMP-2和MMP-9表达明显上调,且两者的表达与ICC患者恶性临床病理特征及不良预后密切相关。     

参地补肾胶囊对肾小球硬化大鼠MMP-9、TIMP-1及其受体mRNA表达的影响

目的:观察参地补肾胶囊对肾小球硬化大鼠基质金属蛋白酶(MMP-9)、基质金属蛋白酶抑制物(TIMP-1)及其受体mRNA表达的影响,探讨其抗肾小球硬化的机制。方法:采用单侧肾脏切除同时尾静脉注射阿霉素的方法建立肾小球硬化大鼠模型,SD大鼠随机分为空白对照组、模型组、参地补肾胶囊组、贝那普利组、苏黄泄浊丸组,检测血肌酐(Scr)、尿素氮(BUN)、血浆蛋白(Alb);免疫组化法检测MMP-9、TIMP-1的表达,计算MMP-9/TIMP-1比值,RT-PCR检测受体TIMP-1mRNA的表达水平。结果:与模型组比较,参地补肾胶囊组能抑制TIMP-1及其受体mRNA的表达(P<0.01),上调MMP-9表达(P<0.01),提高MMP-9/TIMP-1的比值。结论:参地补肾胶囊可抑制TIMP-1及其受体mRNA的表达,改善肾脏病理形态,进而延缓肾小球硬化进展。     

The importance of temperature on the neurovascular unit

Therapeutic hypothermia is the only treatment that has been shown to be of benefit to infant's ≥ 36 weeks of gestation with hypoxic–ischemic encephalopathy. The evidence for the benefit is based on multiple, well-designed randomized clinical trials. Based on this data, the use of therapeutic hypothermia has been widely disseminated throughout the neonatal community. An important concept in hypoxic–ischemic brain injury is the functioning of the neurovascular unit which links neurons, non-neuronal cellular elements and the capillary endothelial cells to promote optimal barrier maintenance between the brain and systemic circulation, regulation of blood flow and neuro-immunologic functioning. Hypoxic–ischemic injury can trigger increased permeability of the blood‐brain-barrier via molecular events within the neurovascular unit and initiate pathways to brain injury. In addition, exposure of the brain to cellular elements from the systemic circulation can further propagate the neuro-inflammatory response. The influence of temperature on injury to the neurovascular unit has received relatively little attention. This review will focus on one component of the neurovascular unit, the blood‐brain barrier and its constituents. Specifically, this review will address the effects of hypoxia–ischemia and temperature on the neurovascular unit and potential knowledge gaps which may serve as areas for further investigation.