OBJECTIVETo determine the effect of AQP1 gene on facial nerve edema following injury through investigation of the relationship between the expression of AQP1 gene and Schwann cells swelling.

METHODSThe AQP1 expression in Schwann cells of mouse facial nerve tissues was detected by immunofluorescent staining. The transgenic protocol by lentivirus transduction was used to specifically upregulate AQP1 expression in Schwann cells. Lenti-AQP1 and CTRL (empty vector) transduced cells were observed during gene overexpression every 24 h for 6 days by using phase contrast microscopy. Cell volume of CTRL and Lenti-AQP1 treated cells was measured daily from the day of treatment, through day 6.

RESULTSSchwann cell primary cultures maintained a high level of AQP1 water channels, representing an ideal cell model to study the role of AQP1 in the facial nerve. The expression of AQP1 mRNA and protein in Schwann cells infected with the Lenti-AQP1 was increased significantly compared with CTRL lentivirus (P < 0.05). Lenti-AQP1 caused cell swelling in cultured Schwann cells, as validated by cell volume determinations (P < 0.01).

CONCLUSIONSAQP1 is an important factor responsible for the fast water transport of cultured Schwann cells. It plays an important role in facial nerve edema.

Animals ; Aquaporin 1 ; genetics ; metabolism ; Cell Size ; Edema ; etiology ; Facial Nerve ; metabolism ; Facial Nerve Diseases ; etiology ; Lentivirus ; Mice ; RNA, Messenger ; metabolism ; Schwann Cells ; cytology ; metabolism ; virology ; Time Factors ; Transduction, Genetic ; methods ; Up-Regulation

This study aims to elucidate the mechanisms by which dexmedetomidine alleviates pulmonary edema in rats with acute lung injury induced by lipopolysaccharide (LPS). Male Wistar rats were randomly divided into five groups: normal saline control (NS) group, receiving intravenous 0.9% normal saline (5 mL/kg); LPS group, receiving intravenous LPS (10 mg/kg); small-dose dexmedetomidine (S) group, treated with a small dose of dexmedetomidine (0.5 μg · kg(-1) · h(-1)); medium-dose dexmedetomidine (M) group, treated with a medium dose of dexmedetomidine (2.5 μg · kg(-1) · h(-1)); high-dose dexmedetomidine (H) group, treated with a high dose of dexmedetomidine (5 μg · kg(-1) · h(-1)). The rats were sacrificed 6 h after intravenous injection of LPS or NS, and the lungs were removed for evaluating histological characteristics and determining the lung wet/dry weight ratio (W/D). The levels of tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) in the lung tissues were assessed by enzyme- linked immunosorbent assay (ELISA). The mRNA and protein expression levels of aquaporin-1 (AQP1) and aquaporin-5 (AQP5) were detected by RT-PCR, immunohistochemistry, and Western blotting. The lung tissues from the LPS groups were significantly damaged, which were less pronounced in the H group but not in the small-dose dexmedetomidine group or medium-dose dexmedetomidine group. The W/D and the concentrations of TNF-α and IL-1β in the pulmonary tissues were increased in the LPS group as compared with those in NS group, which were reduced in the H group but not in S group or M group (P<0.01). The expression of AQP1 and AQP5 was lower in the LPS group than in the NS group, and significantly increased in the H group but not in the S group or M group (P<0.01). Our findings suggest that dexmedetomidine may alleviate pulmonary edema by increasing the expression of AQP-1 and AQP-5.
Acute Lung Injury ; chemically induced ; drug therapy ; genetics ; pathology ; Adrenergic alpha-2 Receptor Agonists ; pharmacology ; Animals ; Aquaporin 1 ; agonists ; genetics ; immunology ; Aquaporin 5 ; agonists ; genetics ; immunology ; Dexmedetomidine ; pharmacology ; Dose-Response Relationship, Drug ; Drug Administration Schedule ; Gene Expression Regulation ; Injections, Intravenous ; Interleukin-1beta ; antagonists & inhibitors ; genetics ; immunology ; Lipopolysaccharides ; Lung ; drug effects ; immunology ; pathology ; Male ; Organ Size ; drug effects ; Pulmonary Edema ; chemically induced ; drug therapy ; genetics ; pathology ; Rats ; Rats, Wistar ; Signal Transduction ; Transcription, Genetic ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; genetics ; immunology

OBJECTIVETo observe the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer, and study its anti-tumor mechanism.

METHODIn vitro, MTT assay and scratch assay were adopted to detect the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer cells. The cell autophagy was detected by the acridine orange staining. The gap-junction intercellular communication (GJIC) was investigated by the fluorescent yellow transfer. The expression of aquaporin 1 (AQP1) was analyzed by the Western blotting. In vivo, the subcutaneous implant model and the experimental pulmonary metastasis model of Lewis lung cancer in mice were established to evaluate the anti-tumor and anti-metastasis effects of alcohol extract from Pharbitidis Semen. The serum carcinoembryonic antigen (CEA) and beta2 microglobulin (beta2-MG) of mice bearing Lewis lung cancer were detected by the electrochemiluminesence immunoassay. The expressions of lung AQP1 and Connexin 43 (Cx43) were examined by the immunohistochemical method.

RESULTIn vitro, alcohol extracts from Pharbitidis Semen inhibited the cell proliferation in a dose-dependent matter, significantly prevented the cell migration, down-regulated AQP1 proteins of cells, promoted GJIC, and decreased the serum-free autophagy of tumor cells. In vivo, compared with untreated model mice, alcohol extracts from Pharbitidis Semen inhibited the tumor growth in a dose-dependent matter, prevented the tumor metastasis and prolonged the life span of mice bearing Lewis lung cancer, while decreasing serum CEA and beta2-MG of mice bearing Lewis lung cancer, enhancing the immumohistochemical staining intensity of Cx43 and weakening aquaporins AQP1 positive intensity.

CONCLUSIONAlcohol extracts from Pharbitidis Semen could prevent the proliferation and metastasis in Lewis lung cancer cells. Its mechanism may be related to the promotion of GJIC and the down-regulation of AQP1.

Animals ; Antineoplastic Agents ; administration & dosage ; Aquaporin 1 ; genetics ; metabolism ; Carcinoma, Lewis Lung ; drug therapy ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Connexin 43 ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Ipomoea ; chemistry ; Lung Neoplasms ; drug therapy ; genetics ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Metastasis ; Seeds ; chemistry

BACKGROUNDThe extract of Ginkgo biloba leaves tablets, ginaton, is widely used in treating ischemic cerebrovascular disease in the clinic. This study aimed to investigate the expression of aquaporin-1 (AQP-1) in rat lung with ischemia/reperfusion injury after pretreatment with ginaton, and whether the pretreatment with ginaton reduces the acute lung injury caused by ischemia/reperfusion injury.

METHODSAdult Wistar rats were divided into two groups. Some rats were used as donors (n = 20), the others as recipients (n = 20). Left lungs of donor rats were used for the isolated lung reperfusion model, which perfused only with low potassium dextran (LPD) solution as group A (n = 10); the others were pretreated with ginaton before reperfusion as group C (n = 10). Right lung of donor rat without any treatment was used as a control group (group B and group D, n = 10 for each group). After the model was established, the expression of AQP-1 in the lung tissues was examined by immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction.

RESULTSImmunohistochemical examination revealed that AQP-1 was expressed in endothelia. Immunoblotting demonstrated that the relative gray values of AQP-1 protein in groups A and C were 0.65±0.06, 0.88±0.11, respectively. The relative gray values of the mRNA expression in groups A and C were 0.30±0.08, 0.49±0.11, respectively. The expression of AQP-1 protein and mRNA in group C was significantly higher than in group A (P < 0. 05).

CONCLUSIONThe pretreatment with ginaton can reduce the acute lung injury caused by ischemia/reperfusion.

Animals ; Aquaporin 1 ; genetics ; metabolism ; Ginkgo biloba ; chemistry ; Immunohistochemistry ; Lung ; drug effects ; metabolism ; Plant Extracts ; therapeutic use ; Plant Leaves ; chemistry ; Rats ; Rats, Wistar ; Reperfusion Injury ; drug therapy ; metabolism ; Tablets

OBJECTIVETo explore the role of aquaporin-1 (AQP1) gene in erythroid differentiation of erythroleukemia K562 cells induced by retinoic acid (RA).

METHODSK562 cells were cultured in the presence of 1 µmol/L RA for varying lengths of time, and γ-globin mRNA expression and hemoglobin content in the cells were detected by real-time PCR (RT-PCR) and ultraviolet spectrophotometry, respectively, to evaluate the erythroid differentiation of K562 cells. RT-PCR and Western blotting were used to examine AQP1 expression in the cells following RA treatment. A retroviral expression vector of AQP1 small interfering RNA (pSUPER-retro-puro-shAQP1) was constructed and transfected into K562 cells to establish a K562 cell line with stable AQP1 down-regulation (K562-shAQP1), in which the changes in γ-globin and hemoglobin expressions after RA treatment were detected.

RESULTSRA treatment significantly increased γ-globin and hemoglobin expressions in K562 cells (P<0.01), causing also significantly enhanced AQP1 mRNA and protein expressions over time (P<0.01). Transfection with the recombinant plasmids pSuper-retro-puro-shAQP1 resulted in stable AQP1 suppression in K562 cells (P<0.01), which showed markedly reduced γ-globin and hemoglobin expressions after RA induction as compared to the control K562 cells (P<0.01).

CONCLUSIONK562 cells show a significant increase of AQP1 expression after RA-induced erythroid differentiation, and suppression of AQP1 expression can partially block the effect of RA, suggesting the important role of AQP1 in RA-induced erythroid differentiation of K562 cells.

Aquaporin 1 ; antagonists & inhibitors ; metabolism ; Cell Differentiation ; drug effects ; Humans ; K562 Cells ; Leukemia, Erythroblastic, Acute ; metabolism ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Tretinoin ; pharmacology

OBJECTIVETo investigate the expressions of aquaporins (AQPs) in the mouse prostatic tissue and their significance, and to provide some evidence for a deeper insight into the physiological function and regulation of AQP expressions in normal and diseased prostatic tissues.

METHODSThe mRNA expressions of AQP0 - 4 in the mouse prostatic tissue were determined by RT-PCR, and the expressions and localizations of AQP1 and AQP3 proteins were characterized by Western blot and immunohistochemistry.

RESULTSRT-PCR exhibited the mRNA expressions of AQP1 and AQP3, but not those of AQP0, AQP2 and AQP4 in the prostate tissue, while Western blot showed the expression of the AQP1 protein with the relative molecular mass (RMM) of 28 000 and those of the glycosylated and non-glycosylated AQP3 proteins with the RMM of 35 000 and 27 000, respectively. Immunohistochemistry indicated the strong expression of AQP1 in the cyst and plasma membrane of the secretary cells and that of AQP3 in the stroma cells of the prostate.

CONCLUSIONThe AQP1 and AQP3 genes were expressed in the secretary epithelia of the mouse prostate tissue, which suggests that AQP1 and AQP3 may play an important role in the secretion of prostatic fluid.

Animals ; Aquaporin 1 ; genetics ; metabolism ; Aquaporin 3 ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Prostate ; metabolism ; pathology

OBJECTIVETo investigate the impact of energy metabolism at the cellular level on the expression of the water channel protein aquaporin 1 (AQP1).

METHODSBalb/c mouse fibroblasts were incubated with iodoacetamide (IA) in vitro, and the changes in AQP1 expression were detected by immunoblotting and immunohistochemistry at 0, 4, and 6 h.

RESULTSIA induced the expression of AQP1 at 4 and 6 h accompanied with cell death. Reverse transcription PCR showed an increased expression of AQP1 mRNA in the cells. AQP1 expression was also upregulated by the inhibitor of microtubule and cytochrome C oxidase.

CONCLUSIONA pretranslational regulation occurs in IA-induced AQP1 expression in mouse fibroblasts, and the up-regulated AQP1 accumulation is characterized by mitochondria-related energy dependence.

Animals ; Aquaporin 1 ; genetics ; metabolism ; Cells, Cultured ; Energy Metabolism ; Fibroblasts ; cytology ; metabolism ; Iodoacetamide ; pharmacology ; Mice ; Mice, Inbred BALB C ; Mitochondria ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Up-Regulation ; drug effects

BACKGROUNDAquaporin-1 (AQP1) has involved in fluid transport in diverse pulmonary edema diseases. Our study aimed to explore the dynamic changes of AQP1 in pulmonary water metabolism in rats following traumatic brain injury (TBI) and the protective effect provided by shenmai injection.

METHODSSixty male Sprague Dawley rats weighting 280 - 300 g were randomly divided into three groups: the normal control group, the model group and the shenmai injection (SMI) group. One piece skull was taken away without injuring cerebral tissue in normal control group, while rats in model group and SMI group were subject to free fall injury in the cerebral hemisphere. Rats in model group received intraperitoneal normal sodium (15 ml/kg) at one hour post-injury and the same dose of shenmai injection instead in SMI group, respectively. The expression of AQP1 was detected by immunohistochemical analysis and semi-quantitative RT-PCR at 0 hour, 10 hours, 72 hours and 120 hours after TBI. Arterial blood gas analysis and lung wet to dry were also measured.

RESULTSAQP1 was mainly presented in the capillary endothelium and slightly alveolar epithelial cells in three groups, but the expression of AQP1 in the normal control group was positive and tenuous, weakly positive in the model and SMI groups, respectively. Compared with normal control group, AQP1 mRNA levels were down regulated in the model and SMI groups at 10 hours, 72 hours and 120 hours (P < 0.05). While AQP1 mRNA levels in the SMI group was up-regulated than that in the model group (P < 0.05). Lung wet to dry weight ratio (W/D) in the model and SMI groups at 10 hours were higher than that in normal control group (P < 0.05). Compared with normal control group, PaO2 was markedly lower in the model and SMI groups (P < 0.05), but there were no statistically significant differences between model and SMI groups (P > 0.05).

CONCLUSIONSThe decreased AQP1 expression may be involved in the increased lung water content and dysfunction of pulmonary water metabolism following TBI. The treatment with SMI could improve water metabolism by promoting AQP1 expression.

Animals ; Aquaporin 1 ; genetics ; metabolism ; Brain Injuries ; drug therapy ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Immunohistochemistry ; Injections ; Lung ; drug effects ; metabolism ; Male ; Random Allocation ; Rats ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the changes of vascular endothelial growth factor(VEGF), aquaporin (AQP) gene and protein expression during hypoxic encephaledema so as to provide the basis for elucidating the brain injury caused by acute hypoxic exposure and pathogenesis of the encephaledema.

METHODSWistar rats were randomly divided into 4 groups, i.e. control group, hypoxia 4 000 m group, hypoxia 6 000 m group and hypoxia 8 000 m group. Rats in hypoxic groups were exposed to hypoxia at simulated altitude of 4 000 m, 6 000 m and 8 000 m above sea level for 8 hours respectively in order to establish hypoxic encephaledema model. The water content in brain was determined by dry-weight method. The changes in morphology of brains were observed under optical microscope. The changes in expression of VEGF, AQP1 and AQP4 genes and protein were determined by RT-PCR and immunohistochemistry.

RESULTS(1) The results determined by dry-weight method indicated that water content of rats brain increased markedly after rats were exposed to a simulated altitude at 6 000 m, 8 000 m. (2) The results determined by microscopy indicated that during the rats exposed to hypoxia, nerve cells, vascular endothelial cells and astrocyte foot processes swelled lightly, transudate occurred in tissues at 4 000 m. The swelling of vascular endothelial cell (VEC) and astrocyte foot processes aggravated, interspace between vessels and tissues enlarged, and transudate in tissue increased at 6 000 m. The swelling of VEC and astrocyte foot processes went from bad to worse, interspace between vessels and tissues enlarged further, and transudate in tissue increased evidently at 8 000 m. (3) During hypoxic encephaledema, the expression of VEGF, AQP1 and AQP4 mRNA increased, AQP1 was abnormally expressed on the surface of VEC, and the expressive level of VEGF and AQP1 on VEC and AQP4 on astrocyte foot processes increased.

CONCLUSIONThe changes in expression and distribution of VEGF, AQP1 and AQP4 during encephaledema caused by hypoxic exposure may induce blood-brain barrier injury, and may be one of the pathogenesis of hypoxic encephaledema.

Animals ; Aquaporin 1 ; genetics ; metabolism ; Aquaporin 4 ; genetics ; metabolism ; Blood-Brain Barrier ; pathology ; Brain ; metabolism ; Brain Edema ; etiology ; metabolism ; physiopathology ; Hypoxia ; complications ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Animals ; Aquaporin 1 ; analysis ; genetics ; physiology ; Aquaporin 3 ; analysis ; genetics ; physiology ; Disease Models, Animal ; Immunohistochemistry ; Lung ; blood supply ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Reperfusion Injury ; etiology ; metabolism