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.     

水通道蛋白在卵巢上皮癌中的表达及意义

目的探讨水通道蛋白1、3(AQP1、AQP3)在上皮性卵巢癌中的表达及其意义。方法采用免疫组织化学方法检测上皮性卵巢良性、交界性、恶性肿瘤中肿瘤及腹膜组织中AQP1和AQP3的表达,同时检测血管内皮生长因子(VEGF)表达并以CD105标记各组织微血管内皮细胞测定微血管密度(MVD)。结果卵巢癌患者腹膜中AQP1、AQP3表达明显高于良性肿瘤者(P<0.05)。与卵巢肿瘤性质有关;卵巢癌患者中,有腹水(尤其是腹水量≥1000ml)和淋巴结转移者AQP1、AQP3的表达明显高于无腹水者(P<0.05)。AQP3的表达与卵巢癌病理分期分级有关。结论 AQP1、AQP3参与卵巢癌发生发展、侵袭转移和腹水形成。腹膜组织上AQP1、AQP3的高表达与肿瘤盆腔扩散和腹水生成有密切关系。     

Curcumin attenuates brain edema in mice with intracerebral hemorrhage through inhibition of AQP4 and AQP9 expression

Aim:

Aquaporins (AQPs) are the water-channels that play important roles in brain water homeostasis and in cerebral edema induced by brain injury. In this study we investigated the relationship between AQPs and a neuroprotective agent curcumin that was effective in the treatment of brain edema in mice with intracerebral hemorrhage (ICH).

Methods:

ICH was induced in mice by autologous blood infusion. The mice immediately received curcumin (75, 150, 300 mg/kg, ip). The Rotarod test scores, brain water content and brain expression of AQPs were measured post ICH. Cultured primary mouse astrocytes were used for in vitro experiments. The expression of AQP1, AQP4 and AQP9 and NF-κB p65 were detected using Western blotting or immunochemistry staining.

Results:

Curcumin administration dose-dependently reduced the cerebral edema at d 3 post ICH, and significantly attenuated the neurological deficits at d 5 post ICH. Furthermore, curcumin dose-dependently decreased the gene and protein expression of AQP4 and AQP9, but not AQP1 post ICH. Treatment of the cultured astrocytes with Fe²⁺ (10–100 μmol/L) dose-dependently increased the expression and nuclear translocation of NF-κB p65 and the expression of AQP4 and AQP9, which were partly blocked by co-treatment with curcumin (20 μmol/L) or the NF-κB inhibitor PDTC (10 μmol/L).

Conclusion:

Curcumin effectively attenuates brain edema in mice with ICH through inhibition of the NF-κB pathway and subsequently the expression of AQP4 and AQP9. Curcumin may serve as a potential therapeutic agent for ICH.     

AQP4与星形胶质细胞胀亡之间的关系

细胞胀亡是不同于细胞凋亡的一种死亡方式,主要是因为细胞内ATP合成障碍,胞膜离子泵功能丧失,导致水及离子进入细胞内,引起细胞器及细胞肿胀,最终细胞死亡。有研究发现缺血缺氧状态下星形胶质细胞的死亡属于胀亡。AQP4是大脑内重要的水通道蛋白之一,主要在星形胶质细胞的足突表达,其主要作用为通过对水的转运．维持细胞内外水平衡。细胞胀亡与水通道的功能有关,星形胶质细胞的胀亡也与AQP4有密切关系,在缺血缺氧的情况下,星形胶质细胞的AQP4表达会出现上调或下调,从而导致星形胶质细胞内外水平衡被破坏．引起星形胶质细胞肿胀,最终出现细胞胀亡。     

水通道蛋白3在银屑病皮损中的表达及意义

目的:探讨水通道蛋白3(AQP3)mRNA在寻常型银屑病皮损中的表达及意义。方法:选取18例寻常型银屑病患者(寻常型银屑病组),取材部位为非曝光部位典型银屑病皮损处;另外选择18例整形外科手术患者(正常对照组),取材部位为非曝光部位正常皮肤。先用无创性皮肤生理功能测试仪检测角质层含水量、皮脂含量及经表皮水分流失(TEWL);再用RT-PCR的方法分别检测皮肤中AQP3 mRNA的表达量。结果:寻常型银屑病组角质层含水量、皮脂含量均较正常对照组降低(P<0.05),经表皮水分流失较正常对照组增加(P<0.05)。寻常型银屑病组AQP3 mRNA的表达量为(0.154±0.091),正常对照组为(0.401±0.121),寻常型银屑病组明显低于正常对照组(P<0.05)。结论:AQP3表达下降可能是银屑病皮损干燥、皮肤屏障破坏的原因之一。     

Emodin ameliorates acute lung injury induced by severe acute pancreatitis through the up‐regulated expressions of AQP1 and AQP5 in lung

The present study investigates the ameliorating effects of emodin on acute lung injury (ALI) induced by severe acute pancreatitis (SAP). An ALI rat model was constructed by sodium ursodeoxycholate and they were divided into four groups: SHAM, ALI, emodin and dexamethasone (DEX) (n=24 per group). Blood samples and lung tissues were collected 6, 12 and 24 hours after the induction of SAP‐associated ALI. Lung wet/dry ratio, blood gases, serum amylase and tumor necrosis factor‐α (TNF‐α) were measured at each time point. The expressions of AQP1 and AQP5 in lung tissue were detected by immunohistochemical staining, western blotting and real‐time PCR. As the results show, there were no statistical differences in the levels of serum amylase, lung wet/dry ratio, blood gases indexes, serum TNF‐α and pathological changes between emodin and DEX groups. However, significant differences were observed when compared with the ALI group. AQP1 and AQP5 expressions were significantly increased and lung oedemas were alleviated with the treatment of emodin and DEX. The expressions of AQP1 and AQP5 were significantly decreased in SAP‐associated ALI rats. Emodin up‐regulated the expression of AQP1 and AQP5, it could reduce pulmonary oedema and ameliorate SAP‐induced ALI. Regulations on AQP1 and AQP5 expression had a great value in clinical application.     

AQP4参与雌激素对单胺类神经递质的调节作用

目的：探讨AQP4对雌激素调节神经递质作用的影响。方法：应用雄性AQP4基因敲除型CD1小鼠与野生型CD1小鼠，给予不同剂量雌激素后测定不同脑区中单胺类神经递质的含量。结果：AQP4基因敲除型雄性小鼠纹状体多巴胺（DA）及5-羟色胺（5-HT）、皮层去甲肾上腺素（NE）含量增加；海马NE、下丘脑DA水平下降。同时，给予雌激素后，野生型雄性CD1小鼠纹状体DA、5-HT含量及海马5-HT含量升高，皮层DA、5-HT水平及下丘脑NE、DA水平下降；而雌激素对AQP4敲除型雄性CD1小鼠单胺类递质无显著影响。结论：AQP4敲除可改变雌激素对单胺类递质水平的调节作用，对纹状体内DA水平的影响尤为显著。AQP4参与了雌激素对单胺类神经递质的调节。     

大鼠脑缺血/再灌注后早期AQP4的动态表达及其与脑水肿关系的研究

目的研究局灶性脑缺血/再灌注损伤大鼠脑组织病理变化及脑组织水通道蛋白4(AQP4)和相关蛋白表达变化,以探索AQP4与脑水肿的关系。方法♂成年SD大鼠150只,体质量250~300 g,随机分为假手术(Sham)组和脑缺血/再灌注损伤模型(I/R)组,其中I/R组又分为I/R-6 h、I/R-12 h、I/R-24 h、I/R-48 h 4个时间点组别。采用线栓法阻塞大脑中动脉建立局灶性脑缺血/再灌注模型,于相应时间点进行神经症状评分,EB染色检测脑组织通透性,TTC染色观察脑梗死体积,干湿重法检测脑含水量的变化,免疫组化(IHC)检测大鼠脑缺血/再灌注后不同时间点梗死灶周围AQP4的表达,Western blot及RT-PCR检测AQP4及相关蛋白表达情况。结果与Sham组相比较,随着再灌注时间增加,I/R模型组大鼠神经功能评分、脑梗死体积、脑组织通透性,脑组织含水量均持续增加,IHC显示AQP4的表达逐渐上调,分布愈加广泛;Western blot及RT-PCR结果验证了AQP4表达水平的增高趋势,同时对相关蛋白表达检测显示,MMP-9表达增高,Occludin及JAM-1表达显示降低趋势,在I/R-48h模型组表达差异性均最明显(均P<0.01 vs Sham)。结论脑缺血/再灌注后,伴随脑组织损伤的加剧,AQP4与MMP-9表达共同被激活,导致Occludin及JAM-1等TJPs蛋白降解增加,共同参与了脑水肿的形成。     

结肠癌组织中AQP1表达与临床病理特征的关系与预后研究

目的探析结肠癌组织中AQP1表达与临床病理特征之间的关系与预后。方法选择2010年1月至2013年1月期间我院收治的55例结肠癌患者为研究对象,对AQP1表达情况进行检测,对结肠癌病理特征与AQP1之间的关系进行分析。结果癌旁组织与结肠癌的AQP1表达比较有统计学意义(P<0.05)。结论结肠癌组织中AQP1表达明显升高,与预后和临床病理特征有关,是比较重要的一个指标。     

Effects of lysine aspirin on lung AQP5 expression and lymphocyte apoptosis in paraquat-poisoned rats

Objective: This study explored the effects of lysine aspirin on lung aquaporin 5 (AQP5) expression and lymphocyte apoptosis in paraquat-poisoned rats. Methods: Thirty healthy male Wales rats were randomly divided into three groups (n?=?10): the control group received 0.9% sodium chloride (0.4?mL, intragastric administration; 0.8?mL, intraperitoneal injection); the paraquat group received 40?mg/kg paraquat (intragastric administration) and 0.9% sodium chloride (intraperitoneal injection); and the paraquat + lysine aspirin group received 40?mg/kg paraquat (intragastric administration) and 20?mg/kg lysine aspirin (intraperitoneal injection). Rats were killed at 24 and 48?h. RT-PCR and immunohistochemical staining were performed in lung tissue to determine the AQP5 mRNA and protein expression. Blood from the arterial vein was used to evaluate lymphocyte apoptosis. Results: The lung tissue of paraquat-treated rats displayed pulmonary hemorrhage, interstitial edema and inflammatory cell infiltration. AQP5 mRNA and protein expression in the paraquat-treated group decreased after 24 and 48?h, whereas the peripheral blood lymphocyte apoptosis ratio significantly increased. In contrast, paraquat + lysine aspirin treatment ameliorated these changes. Conclusion: Paraquat decreases AQP5 expression in rat lungs and increases peripheral blood lymphocyte apoptosis. Lysine aspirin can reduce these alterations.     

Effect of D-serine on AQP4 knockout-induced changes in behavior alteration to cocaine

OBJECTIVE AQP4 is widely expressed in brain astrocytes,and its function is mainly to maintain the steady-state of brain water.D-serine as an important neurotransmitter secreted by glial cells is an endogenous co-agonist of NMDA receptor,and is essential for the induction of long-term potentiation.The present study is to explore whether D-serine is mediated the change of synaptic plasticity and cocaine-induced addictive behavior induced by AQP4 knockout.METHODS The effects of D-serine on NAc LTD and behavior of AQP4 knockout mice were investigated by electrophysiological recording and behavioral tests.RESULTS 10 μmol·L-1 D-serine significantly reversed NMDA receptor-dependent LTD in AQP4 KO mice NAc slices.D-serine concentration-dependently antagonized the effects of AQP4 knockout on cocaine addiction behavior.500 mg·kg-1 D-serine significantly increased cocaine-induced locomotor activity in AQP4 KO mice.D-serine(500 mg·kg-1) significantly restored cocaine-induced conditioned place preferences in AQP4 KO mice.CONCLUSION AQP4 knockout changed the behavior of cocaine addiction through affecting the release of D-serine in glial cells.     

内毒素血症大鼠肾脏水通道蛋白2等基因表达及肾脏结构功能的改变

目的:探讨内毒素血症大鼠肾脏水通道蛋白(AQP)2、细胞间黏附分子(ICAM)-1、肿瘤坏死因子(TNF)-α mRNA和蛋白表达及肾脏结构功能的改变。方法:Wistar大鼠24只随机分为空白对照1(C1)组,只注射生理盐水和脂肪乳的对照2(C2)组及注射内毒素和脂肪乳的内毒素(L)组,每组8只,在内毒素血症模型造模成功12h后,处死取材。观察肾组织病理学改变,RT-PCR、Western blot及免疫组化方法测定AQP2、ICAM-1和TNF-α的表达。结果:L组较C1组、C2组肾组织出现病理损伤,尿量减少,尿渗透压下降,血渗透压、血肌酐及尿素氮增高,AQP2 mRNA及蛋白表达减少,ICAM-1、TNF-α mRNA及蛋白表达增多,差异均有统计学意义(均P<0.001)。结论:大鼠内毒素血症可导致肾组织形态改变和功能损伤,AQP2表达减少,ICAM-1和TNF-α表达增多。     

AQP1、ALP在肺腺癌早期诊断中临床研究

目的：本研究旨在探讨AQP1、ALP在肺腺癌患者血清中的表达情况,为找到一种简便易行且敏感性较高的肺腺癌早期诊断方法提供依据。方法：抽取高度可疑肺腺癌住院患者血清163例（不含酗酒者）,随访留取最终确诊的31例患者血清;抽取30例无吸烟、酗酒等不良嗜好健康者的血清作为对照组,采用ELISA法分别检测两组中AQP1、ALP表达情况。结果：肺腺癌患者血清中的AQP1、ALP的表达水平明显高于健康对照组（P〈0.05）。结论：血清中AQP1、ALP的表达异常有助于肺腺癌早期的诊断。     

大鼠颅脑损伤急性期水通道蛋白4表达变化的研究

目的探讨水通道蛋白4(AQP4)在大鼠重型颅脑损伤时的表达变化及其与脑水肿间的关系。方法98只成年雄性Wistar大鼠,随机分为对照组及实验组(伤后0.5、2、6、12、24、48、72 h共7组,对照组不打击)。制作大鼠液压冲击颅脑损伤模型,分别于伤后0.5、2、6、12、24、48、72 h采用干湿比重法测脑组织含水量,半定量反转录聚合酶链反应(RT-PCR)检测脑组织AQP4 mRNA表达及其变化。结果脑组织AQP4 mRNA在伤后0.5 h开始表达上调,2、6、12 h依次增高,24 h达到峰值(P<0.05),72 h时仍维持较高水平。脑组织含水量与AQP4 mRNA表达变化一致。经相关性分析,AQP4 mRNA的表达与脑组织含水量呈正相关(r=0.095,P<0.05)。结论重型脑损伤急性期,AQP4 mRNA表达变化与颅脑损伤后脑水肿的形成和发展密切相关。AQP4可能参与重型脑损伤后脑水肿的形成并起重要作用。     

P2 Receptor-mediated Inhibition of Vasopressin-stimulated Fluid Transport and cAMP Responses in AQP2-transfected MDCK Cells

We cultured canine kidney (MDCK) cells stably expressing aquaporin-2 (AQP2) on collagen-coated permeable membrane filters and examined the effect of extracellular ATP on arginine vasopressin (AVP)-stimulated fluid transport and cAMP production. Exposure of cell monolayers to basolateral AVP resulted in stimulation of apical to basolateral net fluid transport driven by osmotic gradient which was formed by addition of 500 mM mannitol to basolateral bathing solution. Pre-exposure of the basolateral surface of cell monolayers to ATP (100 µM) for 30 min significantly inhibited the AVP-stimulated net fluid transport. In these cells, AVP-stimulated cAMP production was suppressed as well. Profile of the effects of different nucleotides suggested that the P2Y₂ receptor is involved in the action of ATP. ATP inhibited the effect of isoproterenol as well, but not that of forskolin to stimulate cAMP production. The inhibitory effect of ATP on AVP-stimulated fluid movement was attenuated by a protein kinase C inhibitor, calphostin C or pertussis toxin. These results suggest that prolonged activation of the P2 receptors inhibits AVP-stimulated fluid transport and cAMP responses in AQP2 transfected MDCK cells. Depressed responsiveness of the adenylyl cyclase by PKC-mediated modification of the pertussis-toxin sensitive G_i protein seems to be the underlyihng mechanism.     

颈椎舒颗粒对椎间盘退变大鼠模型AQP1的影响

目的:探讨颈椎舒颗粒治疗颈椎病作用的机制。方法:以动静力失衡方法建立颈椎间盘退变模型,测定椎间盘含水量了解颈椎间盘退变情况,用免疫印迹法检测大鼠椎间盘中AQP1含量,观察模型大鼠给药4周、8周后的变化。结果:其中用药4周后,与模型组相比,各组组均能增加椎间盘的含水量,差异有显著性意义(P<0.05)。各药物干预组,以高剂量组疗效最佳,高剂量组含水量最高,模型组最低。用药8周后,与模型组相比,各组组均能增加椎间盘的含水量,差异有显著性意义(P<0.05)。各药物干预组,以高剂量组疗效最佳,高剂量组含水量最高,模型组最低。用药4周后,与模型组相比,各组均能增加颈椎间盘AQP1的含量,差异有显著性意义(P<0.05)。各药物干预组,以高剂量组疗效最佳,高剂量组颈椎间盘AQP1的含量最高,模型组最低。用药8周后,与模型组相比,各组组均能增加颈椎间盘AQP1的含量,差异有显著性意义(P<0.05)。各药物干预组,以高剂量组疗效最佳,高剂量组AQP1最高,模型组最低,并且随着治疗时间的延长,效果有增加的趋势。结论:颈椎舒具有改善颈椎间盘退变的作用,对水通道蛋白1的调节作用是其疗效机制之一。     

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。     

尼莫地平对早期脑出血大鼠AQP4mRNA、MMP-9mRNA表达的影响

目的：探讨尼莫地平对早期脑出血周边组织AQP4mRNA、MMP-9mRNA表达及由脑出血引起的继发性脑水肿与AQP4、MMP-9的关系。方法：制做脑出血大鼠模型,分为假手术组,脑出血组及尼莫地平组,分别观察6h、1d、3d、5d、7d等5个时间点出血周边脑组织含水量、AQP4mRNA、MMP-9mRNA。结果：（1）脑出血组3d含水量达到高峰,第7天明显下降,但仍高于正常水平;尼莫地平组3d含水量达高峰,但低于脑出血组。（2）脑出血组AQP4mRNA、MMP-9mRNA的表达逐渐升高,3d达到高峰,7d明显下降,5个时间点明显高于假手术组。（3）尼莫地平组大鼠脑水肿程度、AQP4mRNA、MMP-9mRNA表达逐渐升高,3d达到高峰,明显低于脑出血组。（4）脑出血的水肿程度与AQP4mRNA、MMP-9mRNA呈正相关。结论：（1）AQP4、MMP-9参与了出血性脑水肿损伤的过程。（2）尼莫地平可以抑制AQP4、MMP-9的表达,进而减轻脑水肿的程度。