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aquaporin-1 and aquaporin-3 expressions in the temporo-mandibular joint condylar cartilage after an experimentally induced osteoarthritis

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Chinese Medical Journal 2007年第24期120卷 2191-2194页

作者： MENG Juan-hong MA Xu-chen LI Zhi-min WU Deng-cheng Center for TMD ＆ Orofacial Pain Peking University School of Stomatology Beijing 100081 China Department of Oral and Maxillolacial Surgery Peking University School of Stomatology Beijing 100081 China Laboratory of Oral and Maxillofacial Surgery PekingUniversity School of Stomatology Beijing 100081 China

Background Over 70% of the total tissue weight in the cartilage matrix consists of water, and the early-stage osteoarthritic cartilage is characterized by swelling. Water transport in the cartilage matrix and across the membranes of chondrocytes may be important in normal and pathological conditions of cartilage. The purpose of this study was to identify aquaporin-1 （AQP1） and aquaporin-3 （AQP3） expressions in the mandibular condylar cartilage after experimentally induced osteoarthritis （OA） in rats. Methods An experimental temporomandibular joint OA was induced by partial discectomy in rats. The pathological characteristics of the normal, early-stage, and late-stage osteoarthritic TMJ cartilages were verified by histological techniques. The AQP1 and AQP3 gene expressions in the normal and osteoarthritic cartilages were measured using quantitative real-time reverse-transcription PCR analysis. The cartilage sections were incubated in primary polyclonal antibodies to AQP3; immunofluorescent microscopy was used to examine the AQP3 expression shown by its protein level. Results The mRNA expression levels of AQP1 and AQP3, analyzed using quantitative PCR, revealed that AQP3 mRNA was highly up-regulated in the OA cartilage, which was considered significant. There was no notable difference in the expression of AQP1 mRNA between OA and normal controls. With the progressing of the OA, the localization of the AQP3 protein was quite different from that of the normal cartilage. Compared to the normal cartilage, the expressions of AQP3 protein were observed mainly in the proliferative zone and the upper mid-zone chondrocytes at the early-stage of OA, and were observed to appear frequently throughout the mid- and deep zone during the late-stage of OA. Conclusions The high expression of AQP3 mRNA in the OA cartilage and the different localization of the AQP3 protein suggest that it may play a particular role in OA pathogenesis. Further study of AQP3 function may provide new insight into the understanding of the molecular mech

关键词： aquaporin osteoarthritis temporomandibular joint

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Modulatory effect of aquaporin 5 on estrogen-induced epithelial-mesenchymal transition in prostate epithelial cells

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Chinese Medical Journal 2021年第4期134卷 448-455页

作者： Yu Fan Tu-Run Song Qiang Wei Lu Yang Tao Lin Xiao-Bing Feng Xian-Ding Wang Zhong-Li Huang Shi-Jian Feng Department of Urology Institute of UrologyOrgan Transplantation CenterWest China Hospital of Sichuan UniversityChengduSichuan 610041China

Background:Estrogen is involved in the pathophysiological process of benign prostatic hyperplasia(BPH),in which epithelial-mesenchymal transition(EMT)plays an important role.Upregulation of aquaporin(AQP)5,which is directly activated by estrogen,has been reported to promote EMT in multiple cells.This study aimed to examine the effects of AQP5 on estrogen-induced EMT in the prostate.Methods:Normal prostate(NP)tissue samples without any histopathological changes and BPH tissue samples with pathologically confirmed hyperplasia were obtained.An EMT cell model was subsequently established by adding estradiol(E2)to RWPE-1 cells,after which AQP5 knockdown was performed.Tissue morphological and immunohistochemical features were examined using hematoxylin-eosin and immunohistochemical staining.Western blot analysis was performed to determine the expression of AQPs,estrogen receptors,and EMT-related proteins.Cell proliferation was assessed and supernatants were collected for enzyme-linked immunosorbent assay to determine transforming growth factor-β1(TGF-β1)concentrations.Immunofluorescence staining was performed to assess protein expressions in RWPE-1 cells.Results:BPH tissues exhibited greater EMT(TGF-β1:1.362±0.196 vs.0.107±0.067,P=0.003;vimentin:1.581±0.508 vs.0.221±0.047,P<0.001;E-cadherin:0.197±0.188 vs.1.344±0.088,P<0.001),higher AQP5(1.268±0.136 vs.0.227±0.055,P<0.001)and estrogen receptor(ER)α(1.250±0.117 vs.0.329±0.134,P<0.001)expression but lower ERβ(0.271±0.184 vs.1.564±0.130,P<0.001)expression than NP tissues.E2-stimulated cells had higher AQP5 expression(1.298±0.058 vs.1.085±0.104,P=0.049),increased cell proliferation(1.510±0.089 vs.1.000±0.038,P<0.001),and EMT(TGF-β1 concentration:0.352±0.021 ng/mL vs.0.125±0.014 ng/mL,P<0.001;vimentin:1.641±0.120 vs.0.188±0.020,P=0.002;E-cadherin:0.075±0.030 vs.0.843±0.046,P<0.001)than controls.E2-stimulated cells with AQP5 knockdown exhibited decreased EMT(TGF-β1 concentration:0.223±0.041 ng/mL vs.0.352±0.021 ng/mL,P=0.016;vimentin:0.675±0.056 vs.1.641±0.120,P=0.001;E-cadh

关键词： aquaporin Benign prostatic hyperplasia Epithelial-mesenchymal transition Estrogen Prostate epithelial cells

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Interactions between Transmembrane Helices within Monomers of the aquaporin AtPIP2;1 Play a Crucial Role in Tetramer Formation

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Molecular Plant 2016年第7期9卷 1004-1017页

作者： Yun-Joo Yoo Hyun Kyung Lee Wonhee Han Dae Heon Kim Myoung Hui Lee Jouhyun Jeon Dong Wook Lee Junho Lee Yongjik Lee Juhun Lee Jin Seok Kim Yunje Cho Jin-Kwan Han Inhwan Hwang School of Interdisciplinary Bioscience and Bioengineering Pohang University of Science and Technology Pohang 790-784 Korea Department of Life Sciences Pohang University of Science and Technology Pohang 790-784 Korea Department of Biology Sunchon National University Sunchon 57922 Korea Division of Integrative Biosciences and Biotechnology Pohang University of Science and Technology Pohang 790-784 Korea

aquaporin （AQP） is a water channel protein found in various subcellular membranes of both prokaryotic and eukaryotic cells. The physiological functions of AQPs have been elucidated in many organisms. However, understanding their biogenesis remains elusive, particularly regarding how they assemble into tetramers. Here, we investigated the amino acid residues involved in the tetramer formation of the Arabidopsis plasma membrane AQP AtPIP2;1 using extensive amino acid substitution mutagenesis. The mutant proteins V41A/ E44A, F51A/L52A, F87A/191A, F92A/193A, V95A/Y96A, and H216A/L217A, harboring alanine substitutions in the transmembrane （TM） helices of AtPIP2;1 polymerized into multiple oligomeric complexes with a vari- able number of subunits greater than four. Moreover, these mutant proteins failed to traffic to the plasma membrane, instead of accumulating in the endoplasmic reticulum （ER）. Structure-based modeling revealed that these residues are largely involved in interactions between TM helices within monomers. These results suggest that inter-TM interactions occurring both within and between monomers play crucial roles in tetramer formation in the AtPIP2;1 complex. Moreover, the assembly of AtPIP2;1 tetramers is critical for their trafficking from the ER to the plasma membrane, as well as water permeability.

关键词： aquaporin AtPIP2 tetramer formation interaction between transmembrane helices

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Overexpression of the tonoplast aquaporin AtTIP5;1 conferred tolerance to boron toxicity in Arabidopsis

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Journal of Genetics and Genomics 2010年第6期37卷 389-397页

作者： Yongqi Pang, Lijuan Li, Fei Ren, Pingli Lu, Pengcheng Wei, Jinghui Cai, Lingguo Xin, Juan Zhang, Jia Chen, Xuechen Wang State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100193, China State Key Laboratory of Plant Physiology and Biochemistry College of Biological Sciences China Agricultural University Beijing 100193 China

Boron （B） toxicity to plants is responsible for low crop productivity in many regions of the world. Here we report a novel and effective means to alleviate the B toxicity to plants under high B circumstance. Functional characterization of AtTIP5;1, an aquaporin gene, re- vealed that overexpression ofAtTIP5;1 （OxAtT1P5;1） in Arabidopsis significantly increased its tolerance to high B toxicity. Compared to wild-type plants, OxAtTIP5;1 plants exhibited longer hypocotyls, accelerated development, increased silique production under high B treatments. GUS staining and quantitative RT-PCR （qRT-PCR） results demonstrated that the expression of AtTIP5;1 was induced by high B concentration treatment. Subcellular localization analysis revealed that the AtTIP5;1-GFP fusion protein was localized on the tonoplast membrane, which was consistent with the prediction based on bioinformatics. Taken together, our results suggest that AtTIP5; 1 is in- volved in B transport pathway possibly via vacuolar compartmentation for B, and that overexpression of AtTIP5;1 in plants may provide an effective way to overcome the problem resulting from high B concentration toxicity.

关键词： aquaporin boron toxicity tolerance

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Effect of emodin on aquaporin 5 expression in rats with sepsis-induced acute lung injury

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Journal of Traditional Chinese Medicine 2015年第6期35卷 679-684页

作者： Sun Yanni Sun Lijing Liu Shuqing Song Juan Cheng Jiejing Liu Jun Department of Emergency Medicine Putuo HospitalShanghai University of Traditional Chinese Medicine Department of Nephrology Medicine Xinhua Hospital Affiliated to Shanghai Jiaotong University

OBJECTIVE: To investigate the effects of emodin on aquaporin 5(AQP5) expression in rats with sepsis-induced acute lung injury.METHODS: We divided 60 adult male Sprague-Dawley rats, weighing 200-230 g, into four groups: control, sham surgery, model and emodin groups(n = 15 for each). We created a sepsis model with cecal ligation and puncture; the sham surgery group had their cecums replaced after exposure outside the abdominal cavity. Each group was further divided into three subgroups(n = 5 for each)and expressions of AQP5 m RNA and proteins in lung tissue were measured by real-time fluorescence polymerase chain reaction and western blot at 6, 12 and 24 h after surgery.RESULTS: AQP5 expression did not change over time in the control group and sham surgery group,but decreased over time in the model group. The lowest expression was found in 12-h subgroup,which significantly differed from the 6-h subgroup(P < 0.01). Compared with the model group, AQP5 expression in the emodin group was significantly higher in all the subgroups(all P <0.01). Expressions in the 12-h subgroup were the highest, and significantly differed from the other subgroups. We found that lung tissue damage,such as pulmonary edema, alveolar damage and the exudation of red blood cells in pulmonary interstitium and alveolar, was significantly milder in the emodin group under light microscope than the model group.CONCLUSION: AQP5 expression was significantly down-regulated in rats with sepsis-induced acute lung injury induced by cecal ligation and puncture.Early prophylactic use of emodin can significantly enhance the AQP5 expression, thus effectively reducing the degree of pulmonary edema in septic rats.

关键词： Emodin Sepsis Acute lung injury aquaporin 5

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Regulatory effect of miRNA 320a on expression of aquaporin 4 in brain tissue of epileptic rats

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Asian Pacific Journal of Tropical Medicine 2015年第10期8卷 791-796页

作者： Yu-Cheng Song Wen-Juan Li Liu-Zhi Li Department of Neurology Shandong Ankang Hospital Institute of Forensic Medicine and Laboratory Medicine Jining Medical University

Objective:To study the expression of miRNA 320 a in the brain tissue of epileptic rats and analyze its effect on the expression of aquaporin 4(AQP4).Methods:All rats were performed with the intraperitoneal injection of lithium chloride(3 mmol/kg) and then the intraperitoneal injection of pilocarpine(30 mg/kg) 24 h later(injected twice) to prepare the epileptic model of Wistar rats.Rats in the control group were injected with the equal volume of normal saline.According to the Racine scale,rats with over stage 3 of epilepsy were chosen and the brain tissue was separated quickly and then stored at-80 ℃.The immunohistochemistry was used to detect the expression of aquaporin in the brain tissue of epileptic model and the Real-time PCR was employed to determine the difference in the expression of miRNA 320 a and AQP4 in the brain tissue of rats between the epileptic model group and control one.Five 5-day neonatal Wistar rats were chosen to collect the cerebral cortex and their primary astrocytes were separated and cultured.They were transfected with miRNA mimic and imitated to the endogenous miRNA 320 a to up-regulate the expression of miRNA 320 a.Results:In the model group,the expression of AQP4 was significantly higher than the control group(P<0.01).However,the expression of miRNA 320 a in the model group was lower than control group(P<0.05),which was negatively correlated to AQP4.In the primary astrocytes,the transfection of miRNA 320 a mimic could significantly reduce the expression of AQP4,while its inhibitor could up-regulate the expression of AQP4,which indicated that miRNA 320 a could reduce the expression of AQP4.Conclusions:In the primary astrocytes of rats,the miRNA 320 a could inhibit the expression of AQP4 and after adding the inhibitor of miRNA 320 a,the expression of AQP4 was up-regulated.

关键词： miRNA 320a Epilepsy aquaporin AQP4

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Fasudil alleviates LPS-induced lung injury by restoring aquaporin 5 expression and inhibiting inflammation in lungs

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The Journal of Biomedical Research 2019年第3期33卷 156-163页

作者： Jingjing Wang Hui Kong Jian Xu Yanli Wang Hong Wang Weiping Xie Department of Respiratory & Critical Care Medicine the First Affiliated Hospital of Nanjing Medical University

Fasudil, a selective rho kinase(ROCK) inhibitor, has been reported to play a beneficial role in systemic inflammation in acute lung injury, but its mechanism for ameliorating pulmonary edema and inflammation remains unclear. Using hematoxylin-and-eosin(H&E) staining, immunohistochemistry, enzyme-linked immunosorbent assay,quantitative real time PCR and Western blotting, we found that fasudil attenuated LPS-induced lung injury, decreased lung edema, and suppressed inflammatory responses including leukocyte infiltration and IL-6 production. Further,fasudil upregulated LPS-induced aquaporin 5 reduction and inhibited NF-κB activation in the lungs of mice. Our results suggest that fasudil could restore the expression of aquaporin 5 to eliminate LPS-induced lung edema and prevent LPS-induced pulmonary inflammation by blocking the inflammatory pathway. Collectively, blockade of the ROCK pathway by fasudil may be a potential strategy for the treatment of acute lung injury.

关键词： acute lung injury aquaporin 5 fasudil fliud transport NF-κB

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A cluster of mutagenesis revealed an osmotic regulatory role of the OsPIP1 genes in enhancing rice salt tolerance

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The Crop Journal 2023年第4期11卷 1204-1217页

作者： Leyuan Tao Bing Wang Shichao Xin Wei Li Shengcai Huang Laihua Liu Jing Cui Qianru Zhang Xianguo Cheng State Key Laboratory of Efficient Utilization of Arid and Semi-arid Arable Land in Northern China Institute of Agricultural Resources and Regional PlanningChinese Academy of Agricultural SciencesBeijing 100081China Department of Plant Nutrition College of Resources and Environmental SciencesChina Agricultural UniversityBeijing 100193China Key Laboratory of Rubber Biology and Genetic Resources of the Ministry of Agriculture Rubber Research InstituteChinese Academy of Tropical Agricultural SciencesHaikou 571011HainanChina International Joint Research Center of Technology Innovation and Achievement Transformation on Palatability of Rice Tianjin Agricultural UniversityTianjin 300384China

aquaporins play important regulatory roles in improving plant abiotic stress tolerance.To better understand whether the Os PIP1 genes collectively dominate the osmotic regulation in rice under salt stress,a cluster editing of the Os PIP1;1,Os PIP1;2 and Os PIP1;3 genes in rice was performed by CRISPR/Cas9 system.Sequencing showed that two mutants with Cas9-free,line 14 and line 18 were successfully edited.Briefly,line 14 deleted a single C base in both the Os PIP1;1 and Os PIP1;3 genes,and inserted a single T base in the Os PIP1;2 gene,respectively.While line 18 demonstrated an insertion of a single A base in the Os PIP1;1gene and a single T base in both the Os PIP1;2 and Os PIP1;3 genes,respectively.Multiplex editing of the Os PIP1 genes significantly inhibited photosynthetic rate and accumulation of compatible metabolites,but increased MDA contents and osmotic potentials in the mutants,thus delaying rice growth under salt stress.Functional loss of the Os PIP1 genes obviously suppressed the expressions of the Os PIP1,Os SOS1,Os CIPK24 and Os CBL4 genes,and increased the influxes of Na+and effluxes of K^(+)/H^(+)in the roots,thus accumulating more Na+in rice mutants under salt stress.This study suggests that the Os PIP1 genes are essential modulators collectively contributing to the enhancement of rice salt stress tolerance,and multiplex editing of the Os PIP1 genes provides insight into the osmotic regulation of the PIP genes.

关键词： aquaporin Multiplex gene editing CRISPR/Cas9 OsPIP1 genes Rice(Oryza sativa L.) Salt tolerance

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Reduced Bone Mineral Density and Bone Metabolism in aquaporin-1 Knockout Mice

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Chemical Research in Chinese Universities 2007年第3期23卷 297-299页

作者： WU Qing-tian MA Qing-jie HE Cheng-yan WANG Cai-xia GAO Shi HOU Xia MA Tong-hui Membrane Channel Research Laboratory Northeast Normal University Changchun 130024 P. R. China China-Japan Union Hospital Jilin University Changchun 130033 P. R. China School of Basic Medicine Jamasi University Jamasi 154002 P. R. China

An overt phenotype of aquaporin-1 knockout（AQP1 ko） mice is growth retardation, suggesting possible defects in bone development and metabolism. In the present study, we analyzed the bone mineral density（ BMD）, bone calcium and phosphorus contents, and bone metabolism in an AQP1 ko mouse model. The BMD of femurs in AQP1 ko mice was significantly lower than that of litter-matched wildtype mice as measured by dual energy X-ray absorptiometry. Consistently, the contents of bone total calcium and phosphorus were also significantly lower in AQP1 ko mice. The reduced BMD caused by AQP1 deficiency mainly affect male mice. Bone metabolic activity, as indicated by 99m^Tc-MDP absorption measurements, was remarkably reduced in AQP1 ko mice. These results provide the first evidence that AQP1 play an important role in bone structure and metabolism.

关键词： aquaporin Gene knockout Bone mineral density Bone metabolism

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Molecular Cloning of a Glycyrrhiza uralensis F.aquaporin GuPIP1 Up-regulated in Response to Drought,Salt and ABA Stress

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Chemical Research in Chinese Universities 2007年第1期23卷 52-57页

作者： WANG Fang JIANG Yong FENG Xue-chao XU Li-na LI Ming-tang LIANG Hai-tao LI Yong-ming ZHU Na LIU Yan-li MA Tong-hui Membrane Channel Research Laboratory Northeast Normal University Changchun 130024 P. R. China Department of Biology Quanzhou Normal University Quanzhou 362000 P. R. China

A PCR-bosed homologous cloning strategy was used to identify aquaporin genes from the roots of Chinese licorice （ Glycyrrhiza uralertsis F. ）. A 1236 bp cDNA with 870 bp open reading frame encoding a 290 amino acids aquaporin ortholog, GuPIP1, was successfully cloned and characterized. The deduced GuPIP1 protein contains six putative transmembrane domains; two conserved NPA motifs as well as the MIP and PIP family signature sequences. A rabbit polyelonal antibody against N-terminal peptide of GuPIP1 corresponded to a 31 kDa GuPIP1 protein on Western blot of plasma membrane preparation of root tissue. RT-PCR and Western blot analysis indicated the expression of GuPIP1 in the root, leaf, and stem tissues. Thus far, GuPIP1 is the first Glycyrrhiza uralensis F. aquaporin that has been identified at a molecular level. Quantitative real-time PCR analysis showed that the expression of GuPIP1 was up-regulated in response to drought, ABA, and salt stress.

关键词： aquaporin Cloning Gtyeyrrhiza uralensis F. PIP C.ene regulation

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