腹腔注射β-七叶皂甙钠对缺血-再灌注小鼠脑IL-1β、AQP4表达的影响

目的探讨β-七叶皂甙钠对缺血-再灌注(I-R)小鼠脑内白细胞介素1β(IL-1β)和水通道蛋白4(AQP4)表达的影响。方法健康雄性昆明小白鼠随机分为四组,每组57只:I-R组(A组),β-七叶皂苷钠组(B组),生理盐水(NS)对照组(C组)和假手术组(D组)。建立大脑中动脉闭塞(MCAO)模型;2 h后,B、C组分别经腹腔给予β-七叶皂苷钠和同体积NS,然后每隔24小时重复1次。在MCAO2 h后再灌注不同时间点分别进行脑含水量、脑梗死体积、IL-1β及AQP4表达水平检测。结果 A组不同时间点病变侧脑梗死体积、脑含水量增高,AQP4及IL-1β表达均被上调,以48 h最为显著;与A组比较,B组不同时间点脑梗死体积明显减小,脑含水量降低,AQP4及IL-1β表达下调(P<0.05)。结论β-七叶皂甙钠能有效保护小鼠缺血性脑损伤;其机制与抑制脑内IL-1β生成和降低AQP4表达有关。     

膀胱癌患者PF4水平及AQP1、AQP3表达与疾病进展、预后的关系

目的 分析膀胱癌患者血小板因子4(PF4)水平及水通道蛋白1(AQP1)、水通道蛋白3(AQP3)表达与疾病进展、预后的关系。方法 将2016年1月—2018年1月我院92例膀胱癌作为观察组,同期体检健康者50例作为对照组。比较2组血清PF4水平及观察组癌、癌旁组织AQP1、AQP3表达情况;分析观察组临床病理特征与血清PF4及癌组织AQP1、AQP3的关系;比较不同预后患者血清PF4与癌组织AQP1、AQP3表达情况;采用Pearson相关性分析探讨血清PF4与癌组织AQP1、AQP3表达相关性;绘制受试者工作特征(ROC)曲线分析血清PF4及癌组织AQP1、AQP3表达对膀胱癌不良预后的预测效能。结果 观察组血清PF4水平高于对照组(P<0.01)。观察组癌组织AQP1、AQP3表达均高于癌旁组织(P<0.01)。TNM分期Ⅲ～Ⅳ期、肿瘤最大径>2 cm、有淋巴结转移患者血清PF4水平与癌组织AQP1、AQP3表达分别高于TNM分期Ⅰ～Ⅱ期、肿瘤最大径≤2 cm、无淋巴结转移患者(P<0.01)。膀胱癌患者血清PF4水平与癌组织AQP1、AQP3表达呈正相关...     

AQP1、AQP2及AQP3在人睾丸肿瘤组织中表达的检测

目的 研究水通道蛋白(Aquaporin,AQP)1、2、3在人睾丸肿瘤组织的分布及表达情况.方法 选取经手术切取的睾丸肿瘤组织10例,应用免疫组织化学S-P法检测AQP1、AQP2、AQP3的分布及表达.结果 在睾丸肿瘤组织中AQP1主要表达在微血管内皮细胞;AQP2及AQP3主要表达在睾丸肿瘤细胞的胞膜及胞质内.结论 AQP1、AQP2及AQP3表达于睾丸肿瘤组织,说明睾丸肿瘤组织水通道蛋白表达增加.     

乌司他丁对大鼠缺血性脑水肿脑组织AQP4表达的影响

目的探讨乌司他丁对大鼠缺血性脑水肿脑组织AQP4表达的影响及意义。方法采用线栓法制备大鼠大脑中动脉栓塞(MCAO)模型,随机分为3组:对照组、治疗1组、治疗2组,取24h时相点用免疫组化方法观察大鼠模型缺血区AQP4的表达。结果治疗1组与对照组比较,24h后AQP4表达水平减轻(P<0.05),治疗2组与对照组相比,24h后AQP4表达水平明显减轻(P<0.01)。结论乌司他丁能通过下调脑缺血组织AQP4的表达,起到脑保护作用。     

神经生长因子与正畸疼痛关系的初步研究

目的 探讨神经生长因子(Nerve growth factor,NGF)与正畸期间机体反应之间的关系,为深入了解正畸牙移动机制提供参考.方法 选择60只雄性成年SD大鼠,随机分为空白组,加力对照组与加力组,各组又分为1、3、5、7 d组,每组各5只.模拟临床牙受力过程,采用免疫组织化学方法检测三叉神经节内NGF的表达,进行统计学分析.结果 大鼠牙齿受力后,第1天即观察到三叉神经节内NGF表达明显增强,至第3天达到最高,以后逐渐下降,但仍高于两对照组.结论 大鼠牙受力初期,NGF表达显著增高,以后逐渐下降,可能与矫治初期疼痛有关.     

七氟烷后处理减轻兔肺缺血再灌注损伤中AQP1的表达及意义研究

目的 探讨水通道蛋白1 (aquaporin1,AQP1) mRNA在七氟烷后处理中对兔肺组织的作用.方法 选择新西兰大白兔12只,雌雄不拘,随机分为两组,每组6只:Ⅰ组为肺缺血再灌注损伤模型组;S组为七氟烷后处理组.所有实验动物于阻断左肺门前(T1)、阻断左肺门60 min(T2)、实验结束时(T3)3个时间点分别检测肺组织AQP1 mRNA表达及病理改变,实验结束时检测肺湿/干重比.结果 两组肺组织AQP1 mRNA表达随时间推移逐渐下降;T3时点,S组AQP1mRNA表达高于Ⅰ组,肺湿/干重比低于Ⅰ组,差异有统计学意义(P＜0.05).结论 七氟烷后处理对肺缺血再灌注损伤有保护作用,可能与AQP1 mRNA表达上调有关.     

AQP1基因沉默对前列腺癌PC-3细胞的影响

目的:探讨水通道蛋白1(AQP1)基因沉默对在前列腺癌PC-3细胞的作用.方法:培养PC-3细胞至对数生长期,随机分为质粒转染组和质粒空白组.针对AQP1基因设计合成小片段RNA,并构建高效表达载体.利用转染试剂Lipo-fectamineTM2000用脂质体介导的基因转染技术将质粒转染入PC-3细胞.RT-PCR检测PC-3细胞AQP1mRNA的表达情况及激光共聚焦观察室观察、拍照AQP1表达的情况,转染后72h.将鼠龄为6周的60只BALB/C-nu/nu雄裸鼠随机分成两组,其中正常对照组30只,转染组30只.转染组把转染后的前列腺癌PC-3细胞注射裸鼠腋窝处皮下;对照组把未经转染的前列腺癌PC-3细胞注射到裸鼠腋窝处皮下.每日观察肿瘤生长及体重情况,裸鼠饲养6周处死,完整取出移植瘤瘤体测量体积和重量.结果:与质粒空白组相比,转染组应用RNA干扰技术后AQP-1在前列腺癌PC-3细胞系中mRNA的水平降低,与对照组相比,转染组裸鼠移植瘤体积为(573.39±175.24) mm3明显小于对照组(1482.50±327.86) mm3,(P＜0.05).转染组肿瘤重量为(0.55±0.11)g明显小于对照组(1.31±0.29)g,差别具有统计学意义(P＜0.05).结论:AQP-1广泛的表达在正常前列腺组织和前列腺癌组织中,对内环境稳态起着重要的作用.更多的AQP-1促进了前列腺癌细胞的增长,抑制AQP-1的表达可以使前列腺癌移植瘤生长减慢,体积减小.     

白三烯受体拮抗剂对哮喘小鼠肺组织HMGB1表达的影响

目的 探讨白三烯受体拮抗剂对哮喘小鼠肺组织高迁移率族蛋白B1(HMGB1)表达的影响.方法 72只SPF级Balb/c小鼠随机分为哮喘组(A组)、孟鲁司特治疗组(B组)和生理盐水对照组(C组),每组24只.建守哮喘模型.于第1、2、3、4周4个时段,HE染色观察肺组织病理改变,RT-PCR法检测肺组织HMGB1 mRNA表达,免疫组化标记分析HMGB1在肺组织的分布.结果 HE染色B组肺组织炎症较A组明显减轻.第2周时A、B组肺组织HMGB1的表达明显高于C组[(1.58±0.47)、(1.42士0.30)vs.(0.65±0.15)](P<0.01);B组肺组织HMGB1的表达随治疗时间的延长而减少.结论 白三烯受体拮抗剂能抑制哮喘小鼠气道炎症,减少HMGB1的生成.     

间歇低氧小鼠血清氧化应激指标的变化及N-乙酰半胱氨酸的治疗作用

目的观察间歇低氧(I H)小鼠血清氧化应激指标的变化情况,并探讨N-乙酰半胱氨酸(NAC)的治疗作用。方法随机将50只小鼠分为对照组(A组,20只),IH组(B组,30只),分别造模3d、1周、2周、3周、4周;另设IH4周+NAC组(C组,6只)。检测各组小鼠血清超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-PX)表达水平。结果与A组同期比较,B组MDA表达均升高(P<0.05);造模1周后,SOD、GSH-PX表达均降低(P<0.05)。随低氧时间的延长,B组MDA逐渐升高,SOD、GSH-PX逐渐降低(P<0.05)。与B组比较,C组MDA降低,SOD、GSH-PX升高(P<0.05)。结论 IH可导致机体氧化应激水平增加,而NAC治疗能够改善IH导致的氧化应激反应。     

他克莫司滴眼液对损伤角膜上皮AQP3、5表达的影响

目的通过研究他克莫司(tacrolimus)滴眼液(0.1%FK506)在损伤角膜上皮愈合过程中水通道蛋白(AQP)3、5的表达及其作用机制,为研制角膜上皮创伤修复的有效治疗药物提供理论依据。方法采用机械法刮除小鼠角膜中央上皮的方法建立模型,观察角膜上皮修复情况。取损伤若干小时用药后大鼠角膜进行AQP3、5免疫组化染色,观察AQP3、5表达情况。结果角膜上皮基底细胞层可见AQP3表达,并且实验组表达比对照组更高,表达高峰出现在损伤后18 h。角膜上皮损伤后用0.1%FK506滴眼,伤后6 h实验组愈合面积与对照组比较,差异无统计学意义,而损伤后12、18、24 h比较,差异有统计学意义(P<0.05)。上皮全层可见AQP5表达,随着角膜的愈合,其表达量降低。实验组6、12、18 h表达量与对照组比较,差异有统计学意义(P<0.05)。结论应用0.1%FK506滴眼,可通过增加AQP3表达而加快小鼠角膜上皮损伤的修复;而AQP5表达量则随角膜愈合时间延长而降低。     

Applications of aquaporin inhibitors

The aquaporins are a family of small, integral membrane proteins that function as plasma membrane transporters of water and in some cases small polar solutes such as glycerol. There are at least 10 distinct aquaporins in mammals with specific patterns of expression in epithelial, endothelial and other tissues. Recent studies in aquaporin-null mice have indicated key roles for certain aquaporins in the urinary concentrating mechanism, fluid secretion by glands, brain swelling, skin moisture, hearing and vision, and gastrointestinal absorption. The only known inhibitors of some aquaporins are mercurial sulfhydryl-reactive compounds, which are too toxic and nonspecific for use in vivo. Small-molecule or peptide aquaporin blockers have potential applications in the treatment of disorders of fluid/pressure homeostasis such as heart failure, hypertension, brain swelling and glaucoma.     

Brain edema and aquaporin]

Aquaporin is a membrane-bound water channel and highly related to the development and resolution of brain edema. We have detected mRNA and protein expression of aquaporin-4 in ischemic and traumatic brain injury. The aquaporin-4 expression is related to the resolution of brain edema rather than the evolution of the edema. We also detected the relation of aquaporin-1 expression and the evolution of hydrocephalus. Regulation of aquaporin function may be a new venue of treatment in brain edema.     

依达拉奉对心搏骤停大鼠心肌保护作用及水通道蛋白4、 水通道蛋白9的作用

目的 分析依达拉奉对水通道蛋白4(AQP4)、水通道蛋白9(AQP9)表达的影响,探讨依达拉奉对心搏骤停(CA)大鼠心肌保护作用机制。方法 选取96只健康SD大鼠,将其采用随机数字表法分为假手术组(28例),依达拉奉组(34例)、生理盐水组(34例),依达拉奉组及生理盐水组诱导建立CA模型,模型建立5 min后开始进行心肺复苏(CPR),待自主循环恢复后,依达拉奉组立即静脉注射3 mg/mL依达拉奉,生理盐水组给予等体积生理盐水,假手术组不诱导CA模型,仅进行动、静脉置管及气管插管。连续监测三组心电图及血流动力学1 h后缝合伤口,待模型大鼠苏醒后拔除插管,放入笼中常规饲养。分别于自主循环恢复后12、24、48、72 h处死大鼠,测定三组心肌组织中超氧化物歧化酶(SOD)、丙二醛(MDA)水平,光学显微镜下观察不同时间点心肌病理学变化、测定脑组织含水量,采用实时荧光定量聚合酶链式反应(RT-PCR)检测脑组织AQP4、AQP9 mRNA水平。结果 三组大鼠体质量、平均动脉压、舒张压、收缩压及心率等基础参数比较,均差异无统计学意义(P＞0.05);12、24、48、72 h处死大鼠,假手术组心肌组织中SOD水平均明显高于依达拉奉组、生理盐水组,差异有统计学意义(P＜0.05),MDA水平明显低于依达拉奉组及生理盐水组(P＜0.05);依达拉奉组12、24、48、72 h心肌组织中SOD水平分别为[(76.54±9.03)、(66.53±8.25)、(57.21±7.82)、(67.02±8.47)U/mg]均明显高于生理盐水组(P＜0.05),依达拉奉组12、24、48、72 h心肌组织中MDA水平[(11.36±1.15)、(18.29±2.96)、(23.07±2.37)、(20.83±1.82)nmol/mg]明显低于生理盐水组(P＜0.05);各时间点生理盐水组心肌病理学损伤程度均高于依达拉奉组;12、24、48、72 h处死大鼠,依达拉奉组脑组织含水量[(74.36±7.41)、(74.51±7.31)、(75.32±7.25)、(74.23±7.14)%]均明显低于生理盐水组(P＜0.05),依达拉奉组AQP4水平分别为[(1.21±0.11)、(1.13±0.12)、(0.87±0.10)、(0.79±0.11)]明显低于生理盐水组(P＜0.05);依达拉奉组AQP9 mRNA水平分别为[(0.82±0.14)、(0.70±0.11)、(0.62±0.12)、(0.50±0.11)]均明显低于生理盐水组(P＜0.05)。结论 依达拉奉可减轻CA大鼠CPR后心肌损伤,可通过下调AQP4、AQP9表达减轻大鼠脑水肿。     

水通道蛋白1与羊水量的关系

目的 研究水通道蛋白1(AQP1)与羊水量关系.方法 运用免疫组织化学法分别检测20例羊水过少(A组)、17例羊水过多(B组)、22例羊水正常(C组)的胎盘、胎膜组织中AQP1的表达;同时每组随机抽取10例用免疫印迹法分别检测胎盘、胎膜组织中AQP1的蛋白表达量.结果 AQP1主要在羊膜上皮细胞、平滑绒毛膜细胞滋养细胞、胎盘合体滋养细胞和血管内皮细胞的胞浆中表达.免疫组织化学法结果显示,AQP1在A、B和C组胎膜的表达强度分别为(5.82±1.33)、(7.94±1.85)和(6.75±1.25)(P<0.01);免疫印迹法结果显示,AQP1在胎膜的表达强度分别为(13.35±3.33)、(19.51±2.84)和(16.21±2.45)(P<0.01).胎膜AQP1蛋白表达最随着羊水量的增加相应上调.结论 胎膜中AQP1的表达随着羊水量的增多而增加.AQ21在胎膜的上调节可能促进了羊水的跨膜转运.     

Pregnant phenotype in aquaporin 8-deficient mice

Aim:

Aquaporin 8 (AQP8) is expressed within the female reproductive system but its physiological function reminds to be elucidated. This study investigates the role of AQP8 during pregnancy using AQP8-knockout (AQP8-KO) mice.

Methods:

Homozygous AQP8-KO mice were mated, and the conception rate was recorded. AQP8-KO pregnant mice or their offspring were divided into 5 subgroups according to fetal gestational day (7, 13, 16, 18 GD) and newborn. Wild type C57 pregnant mice served as the control group. The number of pregnant mice, total embryos and atrophic embryos, as well as fetal weight, placental weight and placental area were recorded for each subgroup. The amount of amniotic fluid in each sac at 13, 16, and 18 GD was calculated. Statistical significance was determined by analysis of variance of factorial design and chi-square tests.

Results:

Conception rates did not differ significantly between AQP8-KO and wild type mice. AQP8-KO pregnant mice had a significantly higher number of embryos compared to wild type controls. Fetal/neonatal weight was also significantly greater in the AQP8-KO group compared to age-matched wild type controls. The amount of amniotic fluid was greater in AQP8-KO pregnant mice than wild type controls, although the FM/AFA (fetal weight/amniotic fluid amount) did not differ. While AQP8-KO placental weight was significantly larger than wild type controls, there was no evidence of placental pathology in either group.

Conclusion:

The results suggest that AQP8 deficiency plays an important role in pregnancy outcome.     

Attenuation of opioid addiction in mice lacking aquaporin 4

OBJECTIVE To examine the effects of aquaporin 4(AQP4) on opioid addiction and underlie the mechanism behind it. METHODS(1) In the heroin-induced self-administration(SA) experiment,we explored the role of AQP4 on heroin-induced psychological addiction. After the mice were trained to learn heroin-induced SA under a fixedratio1(FR1) reinforcement program for 7 d,we randomly switched the heroin doses to 0.00625,0.0125,0.025,0.05 or 0.1 mg·kg~(-1)per infusion to counterbalance assignment design. In the end,all mice underwent extinction training and reinstatement testing.(2) In oral sucrose self-administration,5% sucrose solution was used for the mice and the procedures were similar to heroin SA.(3) In morphine-induced hyperactivity test,mice were habituated in the test apparatus for 30 min and then were given saline(10 mL·kg~(-1),sc) or morphine(10 or 20 mg·kg~(-1),sc) to record the locomotion for 1.5 h.(4) For the in vivo microdialysis experiment,mice were surgically implanted with intracranial guide cannula into nucleus accumbens(AP +1.4 mm,ML ±0.9 mm,DV-3.8 mm from bregma). After 5 d of recovery from surgery,the mice were challenged by saline(10 mL·kg~(-1),sc)or morphine(10 mg·kg~(-1),sc),and then samples were collected every 20 min. RESULTS We found that AQP4 deletion had no effects on sucrose-seeking and sucrose-taking,but it significantly attenuated heroin-taking and heroin-seeking behaviors in heroin self-administration. Besides these,AQP4 deletion had no effects on basal level of locomotion,but dramatically decreased morphine-induced hyperactivity.Furthermore,the in vivo microdialysis studies showed that AQP4 deficiency inhibited morphine(10mg · kg~(-1),sc)-induced elevation of extracellular dopamine levels in nucleus accumbens in mice.CONCLUSION Our present findings demonstrate that AQP4 was potentially involved in the properties of opioid rewarding by inhibiting dopamine release in nucleus accumbens(NAc).     

Down-regulation of aquaporin 4 in human placenta throughout pregnancy

BACKGROUND: The family of mammalian aquaporins (AQP) consists of 12 known members, each with a specific tissue distribution and membrane localization pattern. AQP4 is the first member of this family identified in biological membranes. This water channel protein is primarily expressed in astrocytes but is also localized in ependymocytes and endothelial cells, suggesting its involvement in the movement of water between the blood and brain, and between the brain and cerebrospinal fluid (CSF). To date, the regulation of AQP4 expression in the human placenta has not been studied. The purpose of this work was to investigate AQP4 localization and expression in the human placenta during gestation. MATERIALS AND METHODS: A total of 30 samples, 15 full-term placentae and 15 chorionic villous samples from first trimester, for the immunohistochemical analysis of AQP4 expression were used. The gestation period ranged from 5 to 40 weeks. RESULTS: A decrease of AQP4 expression in the syncytiotiophoblast from the first to the third trimester of gestation, in contrast with an increased expression shown by endothelial cells and stroma of placental villi was found. CONCLUSION: Our results may suggest that AQP4-mediated maternal-fetal fluid exchange could play an important role in the control of ion homeostasis and water balance in the human placenta throughout pregnancy.     

腹泻大鼠结肠水通道蛋白4表达与分布的研究

目的检测水通道蛋白4(AQP4)在腹泻大鼠结肠的表达和分布,探讨AQP4与肠道水代谢紊乱性疾病的关系。方法用20%的甘露醇灌胃制备大鼠渗透性腹泻模型,采用免疫组织化学方法对各组大鼠升、降结肠黏膜细胞AQP4进行定位、定量分析。结果①腹泻组和对照组大鼠升、降结肠黏膜都有AQP4表达,且升结肠表达量高于降结肠;②腹泻组大鼠升、降结肠黏膜AQP4表达均较对照组减少(P<0.01),9h时腹泻最严重,AQP4蛋白减少亦最显著(P<0.01);③AQP4主要表达于细胞膜,胞质仅有少量表达。结论①AQP4在升结肠表达高于降结肠,这与肠腔内水分主要是在近段吸收相一致;②腹泻状态下大鼠结肠黏膜AQP4表达下调,导致结肠对肠腔内水分的吸收减少,引起大便稀薄,含水量增加形成腹泻;③AQP4主要表达于细胞膜上,这与其功能密切相关。