Aquaporin-1蛋白在缺血心肌中表达变化及其与心肌水肿的关系

目的在动物心肌梗塞模型中探讨心肌缺血状态下心肌细胞膜水通道蛋白Aquaporin-1的变化趋势及其与心肌水肿的联系。方法采用抗AQP1特异性免疫组化考查AQP1在心肌组织中的分布特点;以Real-timePCR及Westernblotting了解在缺血状态下心肌组织AQP1基因转录及蛋白表达的动态变化趋势;采用Evan’blue定量测定法了解缺血心肌组织中毛细血管通透性的变化及用心肌水含量测定法了解缺血心肌的水肿程度。结果AQP1在心肌中存在着广泛分布,并主要分布于心肌的血管及心肌细胞膜。缺血造成AQP1在心肌的转录及蛋白表达呈现先减后增的趋势;缺血后心肌水肿程度存在着双峰样变化趋势而且AQP1蛋白的表达增加的时间特征上与缺血后的第二个水肿高峰的变化趋势相一致。结论AQP1蛋白在心肌缺血后存在着先减后增的动态变化,AQP1蛋白的表达增加可能与部分地联系到缺血后的心肌水肿的形成。

缺血预适应后心肌水肿与水通道Aquaporin-1蛋白表达变化的关系

目的通过对缺血预适应动物模型与单纯心肌缺血动物模型之间的对照研究,探讨预适应对心肌细胞膜水通道蛋白Aquaporin-1表达的影响及与心肌水肿的关系。方法以Real-time PCR及Western blotting了解在心肌组织AQP1基因转录及蛋白表达的动态变化趋势;采用Evan blue定量测定法了解心肌组织中微血管通透性的变化及用心肌水含量测定法了解缺血心肌的水肿程度。结果与缺血对照组相似,缺血预适应后心肌AQP1基因及其蛋白的表达呈现先减后增的趋势,但增加的程度较对照组为高。预适应后组织血管通透性增高明显受到抑制,但仅伴随着缺血后早期阶段心肌水肿的减轻。与微血管通透性的持续降低形成对比的同时,心肌水肿状态在之后一段时期内保持较高水平与AQP1蛋白表达增高在时间关系上相对应。结论预适应早期心肌水肿的减轻与血管通透性的抑制有关,而AQP1蛋白表达的增加可能是心肌水肿状态维持的因素。

Aquaporin-1在大鼠睾丸输出小管的免疫组织化学定位研究

目的 研究正常大鼠睾丸输出小管上皮细胞上Aquaporin 1(AQP 1)的定位分布以期了解其在水重吸收上的作用机制。方法 对正常Wistar大鼠睾丸输出小管进行常规免疫组织化学方法染色观察。结果 在睾丸输出小管非纤毛上皮细胞的刷状缘及基侧部AQP 1阳性表达强烈 ,核上区的胞内体的质膜上也有阳性表达 ;纤毛上皮细胞的纤毛亦呈阳性反应。结论 Aquaporin 1可能与睾丸输出小管非纤毛上皮细胞水重吸收功能有密切关系。

Aquaporin-1 mRNA在人非小细胞肺癌组织中的表达和意义

目的:探讨Aquaporin-1(AQP1)mRNA在人非小细胞肺癌组织中的表达和意义。方法:以相应患者手术切除的正常肺组织为对照,采用半定量RT-PCR技术检测人非小细胞肺癌(17例鳞癌和18例腺癌)组织中AQP1 mRNA的表达水平。结果:人肺腺癌组织中AQP1 mRNA表达水平为0.419±0.135,高于正常肺组织中的表达水平(0.250±0.124),二者之间差异有统计学意义(P<0.05);人肺鳞癌组织中AQP1 mRNA表达水平为0.341±0.099,与正常肺组织中AQP1 mRNA表达水平(0.264±0.114)之间无统计学差异(P>0.05)。结论:AQP1mRNA在人肺腺癌组织中表达增加,提示AQP1可能与人肺腺癌的发生和发展有关系。

Effect of Dexamethasone and Aquaporin-1 Antisense Oligonucleotides on the Aquaporin-1 Expression in Cultured Human Trabecular Meshwork Cells

The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group （0 μg/mL）. In the 4 μg/ mL and 40 μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.

Aquaporin-1 down regulation associated with inhibiting cell viability and inducing apoptosis of human lens epithelial cells

AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparency maintenance.Herein,AQP-1 expression in LECs was investigated to evaluate its influence on cell survival in association with its potential role in cataract formation.·M ETHODS：LECs were transfected with lentivirus carrying AQP-1 small interfering RNA（si RNA）.Real-time polymerase chain reaction（PCR）and Western blotting were conducted to detect AQP-1 expression in LECs from different groups.Meanwhile,cell counting kit-8（CCK-8）assay and flow cytometry were performed to measure LEC proliferation and apoptosis,respectively.·RESULTS：AQP-1 expression was significantly reduced in LECs,both at m RNA and protein levels（〈0.05）,after si RNA treatment.Decreased cell viability was detected by CCK-8 assay in LECs with si RNA interference,compared to control cells（〈0.05）.The apoptosis rate significantly increased in cells after si RNA interference（〈0.05）.·CONCLUSION：The decreased cell viability following AQP-1 down regulation is largely due to its induction of apoptosis of LECs.AQP-1 reduction might lead to changes of physiological functions in LECs,which might be associated with the occurrence and development of cataracts.

Increased aquaporin-1 levels in lens epithelial cells with primary angle-closure glaucoma

AIM： To determine the levels of aquaporin-1（AQP-1） in the lens epithelial cells（LECs） of primary glaucoma and to clarify its correlation with lens thickness.METHODS： This study comprised 64 eyes of 64 patients with primary glaucoma, who were divided into 3 groups： 25 eyes of 25 patients with acute primary angle-closure glaucoma（APACG）, 19 eyes of 19 patients with chronic primary angle-closure glaucoma（CPACG） and 20 eyes of 20 patients with primary open angle glaucoma（POAG）. This study also included 12 eyes of 12 patients with senile cataract as controls. The levels of AQP-1 in LECs were examined by real-time quantitative polymerase chain reaction（RT-q PCR） and immunohistochemistry. The lens thickness was measured by A-scan ultrasonography. RESULTS： The AQP-1 m RNA levels of LECs were 0.84±0.27, 0.69±0.34, 0.44±0.19 and 0.51±0.21 in APACG, CPACG, POAG and senile cataract group, respectively. The levels of AQP-1m RNA were significantly higher in PACG groups compared with those in senile cataract and POAG group（all P〈0.05）. The immunohistochemistry showed the AQP-1 expression were strong-positive in PACG groups, but weak-positive in senile cataract and POAG group. A positive correlation was found between AQP-1 m RNA levels and the lens thickness（r=0.645, P〈0.001）. CONCLUSION： These findings show that the higher expression of AQP-1 in LECs may contribute to increased lens thickness, which might be associated with the occurrence and development of PACG.

Regulatory Effect of Dexamethasone on Aquaporin-1 Expression in Cultured Bovine Trabecular Meshwork Cells

To evaluate the effect of dexamethasone on the expression of aquaporin-1 （AQP-1） in cultured bovine trabeeular meshwork cells, bovine trabeeular meshwork cells were cultured in vitro and reproduced to the third and the fourth generation, then treated with dexamethasone at the concentrations of 5, 25, 50, 250μg/L respectively for 7 days. Immunohistochemical technique-supervision method was employed to measure, and image analysis system to analyze the expression of AQP-1 in normal cultured bovine trahecular meshwork cells and those treated with dexamethasone. In normal bovine trabeeular meshwork cells, the grayseale of AQP-1 positive staining was 167.94± 1.18, while it was 168.92±0.91, 176.72±1.80, 180. 64±1.31, 185.64±1.58 in cells treated with 5, 25, 50, 250μg/L concentrations of dexamethasone. When the concentration of dexamethasone was higher than 25 μg/L, the expression of AQP-1 was significantly inhibited （P〈0.05）. The regulation of AQP-1 expression by dexamethasone in cultured bovine trahecular meshwork cells in vitro may be one of causes that retard the aqueous outflow in glueoeorticoid-induced glaucoma.

Lung injury and aquaporin-1mRNA expression during cardiopulmonary bypass

Objective To testify the lung injury induced by cardiopulmonary bypass(CPB) in canine model and observe the influence of CPB on the aquaporin 1 (AQP1) mRNA expression in canine lung. Methods 8 mongrel dogs were used to perform the cardiopulmonary bypass. The hearts arrested for 90 minutes with mild hypothermia and rebeated for 6 hours. The hemodynamics,the ratio of lung dry weight and wet weight,the plasmic

Efficacy of the nucleotide-binding oligomerization domain 1 inhibitor Nodinhibit-1 on corneal alkali burns in rats

AIM: To evaluate the therapeutic effect of Nodinhibit-1 on alkali-burn-induced corneal neovascularization (CNV) and inflammation. The nucleotide-binding oligomerzation domain 1 (NOD1) is a potent angiogenic gene. METHODS: The alkali -burned rat corneas (32 right eyes) were treated with eye drops containing Nodinhibit-1 or phosphate buffered solution (PBS, pH 7.4) only, four times per day. CNV and inflammation were monitored using slit lamp microscopy, and the area of CNV was measured by formula. Vascular endothelial growth factor (VEGF) and pigment epithelium -derived factor (PEDF) was determined by Western blot analysis. The TUNEL assay was used to assess the corneal apoptosis cells. RESULTS: Alkali-burn-induced progressive CNV and inflammation in the cornea. After treatment for 7d and 14d, there were statistically significant differences in the CNV areas and inflammatory index on that between two group(P<0.05, respectively). Epithelial defect quantification showed a significant difference between the two groups at days 4 and 7 after the alkali burns (P<0.05). The apoptotic. cells on days 1, 4, and 7 between the two groups showed significant differences at all time points (P<0.05, respectively). Compared to that in control group, the protein level of VEGF expression was significantly reduced whereas the PEDF expression was increase in the Nodinhibit-1 groups on day 14(P<0.05, respectively). CONCLUSION: Topical application of 10.0 mu g/mL Nodinhibit -1 may have potential effect for the alkali burn -induced CNV and inflammation. The effect of Nodinhibit -1 on CNV may be by regulation the equilibrium of VEGF and PEDF in the wounded cornea.

LRG1 promotes corneal angiogenesis and lymphangiogenesis in a corneal alkali burn mouse model

AIM:To investigate the potential effect and mechanism of leucine-richα-2-glycoprotein-1(LRG1)on corneal angiogenesis and lymphangiogenesis.METHODS:Corneal neovascularization and lymphatics were induced by establishing alkali burn mouse model.Immunofluorescence staining was performed to detect the location of LRG1 in cornea tissues and to verify the source of LRG1-positive cells.Corneal whole-mount staining for CD31(a panendothelial cell marker)and lymphatic endothelial hyluronan receptor-1(LYVE-1;lymphatic marker)was performed to detect the growth of blood and lymphatic vessels after local application of exogenous LRG1 protein or LRG1 si RNA.In addition,expressions of the proangiogenic vascular endothelial growth factor(VEGF)related proteins were detected using Western blot analysis.RESULTS:LRG1 was dramatically increased in alkali burned corneal stroma in both the limbal and central areas.LRG1-positive cells in the corneal stroma were mainly derived from Vimentin-positive cells.Local application ofexogenous LRG1 protein not only aggravated angiogenesis but also lymphangiogenesis significantly(P<0.01).LRG1 group upregulated the levels of VEGF and the vascular endothelial growth factor receptor(VEGFR)family when compared with the phosphate-buffered saline(PBS)control group.We also found that LRG1-specific si RNA could suppress corneal angiogenesis and lymphangiogenesis when compared with the scramble si RNA-treated group(P<0.01).CONCLUSION:LRG1 can facilitate corneal angiogenesis and lymphangiogenesis through heightening the stromal expression of VEGF-A,B,C,D and VEGFR-1,2,3;LRG1-specific si RNA can suppress corneal angiogenesis and lymphangiogenesis in corneal alkali burn mice.

Topical dihydroartemisinin inhibits suture-induced neovascularization in rat corneas through ERK1/2 and p38 pathways

AIM: To determine if topical instillation of dihydroartemisinin (DHA) inhibits corneal neovascularization (NV) in rats and to investigate the role of the extracellular regulated kinases (ERK) 1/2 and p38 pathways in this process. O METHODS: Suture-induced corneal NV was produced in rats and the eyes were topically treated with different concentrations of DHA (20mg/L, 10mg/L or 5mg/L) or normal saline 4 times a day for 7 days. The corneal NV was quantified as the proportion of NV area to the whole cornea. Western blot was used to determine the expressions of vascular endothelial growth factor (VEGF) and the phosphorylation status of VEGF receptor-2, ERK1/2 and p38 in the corneas. Immunofluorescent staining was used to determine the expressions of phospho-ERK1/2 and phospho-p38 in the corneal tissues from the eyes treated with 20 mg/L DHA (DHA group) or normal saline (control group). RESULTS: The proportion of corneal NV area in the eyes treated with normal saline or DHA at dosages of 20mg/L, 10mg/L or 5mg/L was (23.74 +/- 3.00)%, (15.73 +/- 2.88)%, (19.53 +/- 2.42)%, and (23.38 +/- 2.79)%, respectively. In the eyes treated with 20mg/L or 10mg/L DHA, the corneal NV area was significantly reduced when compared to that in eyes with normal saline (P < 0.05). Western blot analyses revealed that 20mg/L DHA significantly inhibited the expressions of VEGF and phospho-VEGFR-2. Both 20mg/L and 10mg/L DHA inhibited the expressions of phospho-ERK1/2 and phospho-p38. Immunofluorescent staining further demonstrated that 20mg/L DHA lowered the Expression levels of phospho-ERK1/2 and phospho-p38 in the corneas with suture-induced NV. O CONCLUSION: Suture-induced NV in rat corneas was significantly inhibited by topical treatment with 20mg/L and 10mg/L DHA. The results suggest that the effects could be partially dependent on the DHA-mediated inhibitions of the ERK1/2 and p38 pathways.

AB021.Discovery of a new role for the WNK1 kinase in corneal wound healing

Background:Damage to the corneal epithelium triggers important changes in the composition of the extracellular matrix(ECM)to which the basal human corneal epithelial cells(hCECs)attach.These changes are perceived by integrins,a family of trans-membrane receptors that activate different intracellular signalling pathways,ultimately leading to re-epithelialization of the injured epithelium.Our goal is to study the impact of the pharmacological inhibition/activation of specific signal transduction mediators on corneal wound healing using both monolayers of hCECs and tissue-engineered human corneas(hTECs)as in vitro models.Methods:hTECs were produced by the self-assembly approach and wounded with a 8-mm biopsy punch.Total RNA and proteins were isolated from the wounded and unwounded hTECs to conduct gene profiling analyses and protein kinase arrays.The wounded tissues were then incubated either with the lysine deficient protein kinase 1(WNK1)inhibitor WNK463,the WNK1 indirect agonist AM1241,or with the vehicle alone(DMSO;negative control)and wound healing was monitored for 6 days.The impact of WNK1 inhibition/activation on hCECs monolayers was determined using scratch wound assays.Results:Gene profiling analyses and protein kinases arrays revealed that expression and activity of several mediators from the integrin-dependent signalling pathways were altered in response to the ECM changes taking place during corneal wound healing.Phosphorylation of the WNK1 kinase turned out to be the most striking activation event occurring during wound healing.Since the pharmacological inhibition of WNK1 by WNK463 significantly reduced the rate of corneal wound closure in our hTECs and hCECs monolayers compared to their respective negative controls,we believe that the pharmacological activation of WNK1 could turn out to be an interesting avenue to accelerate corneal wound closure.Conclusions:These results will contribute to a better understanding of the cellular and molecular mechanisms involved in corneal wound healing.Furthermore,they identified a new function for the WNK1 kinase in corneal wound healing and might lead to the identification of a new therapeutic target in the field of corneal wounds.

Molecular mechanism of the inhibition effect of Lipoxin A4 on corneal dissolving pathology process

AIM:Excessive dissolve of corneal tissue induced by MMPs which were activated by cytokins and chemokines will lead to corneal ulcer. The molecular mechanism of Lipoxin A4 (LXA4) on corneal collagen degradation in three dimensions was investigated. ·METHODS:Rabbit corneal fibroblasts were harvested and suspended in serum -free MEM. Type I collagen, DMEM, collagen reconstitution buffer and corneal fibroblast suspension were mixed on ice. The resultant mixture solidified in an incubator, after which test reagents and plasminogen was overlaid and the cultures were returned to the incubator. The supernatants from collagen gel incubations were collected and the amount of hydroxyproline in the hydrolysate was measured. Immunoblot analysis of MMP-1,-3 and TMMP-1,-2 was performed. MMP-2, -9 was detected by the method of Gelatin zymography. Cytotoxicity assay was measured. RESULTS:LXA4 inhibited corneal collagen degradation in a dose and time manner. LXA4 inhibited the IL -1β induced increases in the pro-MMP-1, -2, -3, -9 and active MMP -1,-2,-3,-9 in a concentration dependent manner. LXA4 also inhibited the IL-1β induced increases in TIMP-1, -2. CONCLUSION:As a potent anti-inflammation reagent, LXA4 can inhibit corneal collagen degradation induced by IL-1β in corneal fibroblasts thus inhibiting corneal dissolving pathology process.

Characteristic changes in astrocyte properties during astrocyte-to-neuron conversion induced by NeuroD1/Ascl1/Dlx2

Direct in vivo conversion of astrocytes into functional new neurons induced by neural transcription factors has been recognized as a potential new therapeutic intervention for neural injury and degenerative disorders. However, a few recent studies have claimed that neural transcription factors cannot convert astrocytes into neurons, attributing the converted neurons to pre-existing neurons mis-expressing transgenes. In this study, we overexpressed three distinct neural transcription factors––NeuroD1, Ascl1, and Dlx2––in reactive astrocytes in mouse cortices subjected to stab injury, resulting in a series of significant changes in astrocyte properties. Initially, the three neural transcription factors were exclusively expressed in the nuclei of astrocytes. Over time, however, these astrocytes gradually adopted neuronal morphology, and the neural transcription factors was gradually observed in the nuclei of neuron-like cells instead of astrocytes. Furthermore,we noted that transcription factor-infected astrocytes showed a progressive decrease in the expression of astrocytic markers AQP4(astrocyte endfeet signal), CX43(gap junction signal), and S100β. Importantly, none of these changes could be attributed to transgene leakage into preexisting neurons. Therefore, our findings suggest that neural transcription factors such as NeuroD1, Ascl1, and Dlx2 can effectively convert reactive astrocytes into neurons in the adult mammalian brain.

Ischemic postconditioning alleviates lung injury and maintains a better expression of aquaporin-1 during cardiopulmonary bypass

Background It has found that ischemic postconditioning （IPO） might decrease pulmonary ischemia/reperfusion （I/ R） injury,which is one of the main reasons of lung injury caused by cardiopulmonary bypass （CPB）.It was found that aquaporins （AQPs） play a role in the maintenance of fluid homeostasis.But it is still unclear whether IPO influences the expression of aquaporin-1 （AQP1）.This study was designed to investigate whether IPO can reduce CPB-related lung injury and affect the expression of AQP1 of lungs.Methods Twelve healthy dogs were divided into control group （C group） and ischemia postconditioning group （IPO group）.CPB procedures were implemented.Ten minutes later,the left pulmonary artery was separated and blocked.Postconditioning consisted of two cycles of 5-minute pulmonary artery reperfusion/5-minute reocclusion starting at the beginning of reperfusion.The 2×4 cm tissues of both sides of pulmonary apex,superior,middle and inferior lobe were taken before CPB （T1）,before occlusion and reopening of left pulmonary artery （T2,T3）,and 2 hours after CPB （T4）.Samples were used to evaluate lung injury degrees and to detect the expression of AQP1.At T1 and T4,blood was collected from femoral artery to calculate pulmonary function.Results At T4,each pulmonary function showed significant deterioration compared with T1.Lung injury could be found at the onset of CPB.However,the expression of AQP1 decreased and wet to dry weight ratio （W/D） increased after T2.In the left lung of C group,the worst pulmonary function and structures were detected.The slightest changes were discovered in the right lung of C group.A close relationship between W/D and lung injury score was found.The lung injury score was negatively related with the expression of AQP1.It was found that the expression of AQP1 was negatively connected with W/D.Conclusions In dog CPB models,lung injury induced by CPB was related with down regulated expression of AQP1.AQP1 is believed to be involved in the mechanisms of lung ischemia/reperfusion （I/R） injury caused by CPB.IPO increases the expression of AQP1,provides a protective effect on lung suffering from CPB,and alleviates CPB-related lung injury.

Expression of aquaporin-1 in rat pleural mesothelial cells and its specific inhibition by RNA interference in vitro

Background The discovery of water channel aquaporins （AQPs） has greatly expanded the understanding of the regulation of the water permeability of biological membranes. Aquaporin-1 （AQP1） may be involved in fluid transport in numerous pathological conditions. The objective of the present study was to examine whether AQP1 is present in cultured rat pleural mesothelial cells （PMCs） and to investigate the specific inhibitory effect of RNA interference （RNAi） on AQP1 expression in PMCs, which may provide a new method for the further studies on the relation between expression of AQP1 in PMCs and pleural fluid removal in vivo. Methods PMCs were isolated and cultured from rat pleura. The expression of AQP1 in PMCs was confirmed by immunocytochemical staining and reverse transcriptase-polymerase chain reaction （RT-PCR）. Two eukaryotic expression plasmid vectors of short hairpin RNA （shRNA） specific for the AQP1 gene of rat sapien were designed and constructed. The recombinant plasmid vectors were transfected into cultured rat PMCs by cation liposomes. Flow cytometry was used to screen the most effective shRNA at 48 hours after transfection. The expressions of AQP1 mRNA and protein were detected by RT-PCR and Western blotting method at 48 hours after transfection. Results RT-PCR and immunostaining revealed that AQP1 mRNA and protein were present in cultured rat PMCs. Two effective eukaryotic expression plasmid vectors of shRNA specific for the AQP1 gene were constructed successfully. The levels of the expression of AQP1 were inhibited by 83.45%, 90.93%, respectively, at mRNA level and 41.24%, 67.60%, respectively at protein level by two recombinant plasmids at 48 hours after transfection. The expression of AQP1 in PMCs transfected with plasmid was significantly lower than that of the cells transfected with the control plasmid HK and that of the untransfected cells （P〈0.01）. There was no significant difference in AQP1 expression between the control group and the group transfected with AQP1 nonspecific shRNAs （P〈0.05）. Conclusions The expression of AQP1 was present in rat PMCs. The application of shRNA-AQP1 could markedly inhibit the expression of AQP1 in cultured rat PMCs. The use of RNAi is a promising tool for future research into the mechanisms of pleural fluid in vivo.

THE ROLE OF CONNECTIVE TISSUE GROWTH FACTOR, TRANSFORMING GROWTH FACTOR Β1 AND SMAD SIGNALING PATHWAY IN CORNEA WOUND HEALING

The cornea is a highly specialized and unique organ in the human body. Its main function is to project light from the external environment onto the retina, and it has a specific transparency to perform its function properly. The transparency and integrity of the cornea is of vital importance. The corneal wound, especially laceration deep to Bowman＇s membrane and stroma, which will inevitably cause scar formation, may cause the degeneration or even loss of sight.

培养人眼小梁细胞水通道蛋白1与Na^+-K^+-ATP酶、Ca^(2+)-ATP酶的相关研究

目的探讨人眼小梁细胞上水通道蛋白1(aquaporin-1,AQP1)与Na+-K+-ATP酶、Ca2+-ATP酶的相关关系。方法培养的人眼小梁细胞分别给予地塞米松(dexamethasone,DEX)0.4μg/mL(B组)、DEX0.4μg/mL+AQP1反义寡核苷酸(antisense oligonucleotides,AS-ODN)0.625μg/mL(C组)、DEX0.4μg/mL+AS-ODN 1.25μg/mL(D组)、DEX0.4μg/mL+AS-ODN 2.5μg/mL(E组)、DEX0.4μg/mL+AS-ODN 5μg/mL(F组)处理,以不加药组为对照组(A组)。5d后测定各组Na+-K+-ATP酶、Ca2+-ATP酶活性。结果与对照组的相比,DEX0.4μg/mL组Na+-K+-ATP酶活性、Ca2+-ATP酶活性明显下降,差异有统计学意义(P<0.05)。在各DEX+AS-ODN组,随着AS-ODN浓度加大,小梁细胞Na+-K+-ATP酶的活性逐渐提升。在AS-ODN浓度加到5μg/mL时,Na+-K+-ATP酶的活性恢复到最大。B、C、D、E组Na+-K+-ATP酶的活性与对照组及F组差异均有统计学意义(P<0.05),F组与对照组差异无统计学意义(P>0.05)。B、C、D、E、F组Ca2+-ATP酶活性与对照组差异均有统计学意义(P<0.05),B、C、D、E、F组之间Ca2+-ATP酶活性差异无统计学意义(P>0.05)。结论人眼小梁细胞AQP1与Na+-K+-ATP酶关系密切,与Ca2+-ATP酶关系不显著,AQP1的正常表达是Na+-K+-ATP酶活性维持的必要条件之一。

角膜AQP_1含量变化与死亡时间关系的法医学研究

目的观察角膜AQP1含量变化与死亡时间的关系。方法50只家兔随机分为10组,以空气栓塞方法致死,于死后不同时间取出角膜。应用免疫组织化学方法观察角膜AQP1含量变化,并采用图像分析方法测定AQP1阳性面积、积分光密度、平均光密度、平均灰度等参数,对各参数与死亡时间的关系进行统计学分析。结果死后84h内,角膜AQP1呈阳性着色,死后96h未见着色。图像分析结果显示,随着死后时间的延长,角膜AQP1各参数呈逐渐下降的趋势,并获得了上述各参数与死亡时间变化关系的回归方程。结论家兔死亡后角膜AQP1的含量呈现出规律性变化,其参数阳性面积、积分光密度、平均光密度、平均灰度等可以作为推测死亡时间的参考指标。