OBJECTIVE: To investigate the effect of aquaporin 5 (AQP5) on Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor κB (NF-κB) signaling pathway in Sjögren syndrome (SS) rats. METHODS: The SS gene expression data sets GSE406611 and GSE84844 were extracted from the Gene Expression Omnibus (GEO), and the AQP5 mRNA expression was analyzed by R software. The rat SS model was constructed. The successfully modeled rats were divided into SS group, SS+NC group, and SS+pc group, 10 rats in each group; and 10 rats were set as Normal group. The rats in the SS+NC group were injected with 10 μg of rno-pcDNA3.1-AQP5-NC at the submandibular gland, subcutaneously every day for 28 days. The rats in the SS+pc group were injected with 10 μg of rno-pcDNA3.1-AQP5 at the submandibular gland, subcutaneously every day for 28 days. The enzyme-linked immunosorbent assay (ELISA) kit was used to detect the content of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the serum. High-throughput sequencing was used to identify the target genes. Quantitative real-time PCR (qPCR) and Western blot were used to detect the mRNA and protein expressions of AQP5, TLR4, MyD88, and NF-κB in the rat submandibular gland tissue. RESULTS: In the SS dataset GSE406611 and GSE84844, the mRNA expression of AQP5 in SS was significantly reduced. Compared with the Normal group, the content of TNF-α and IL-1β in the serum, the mRNA and protein expressions of TLR4, MyD88, and NF-κB in the SS group were significantly increased, the mRNA and protein expressions of AQP5 were significantly decreased. After overexpression of AQP5, the content of TNF-α and IL-1β in the serum, the mRNA and protein expressions of TLR4, MyD88, and NF-κB in the SS+pc group were significantly decreased, the mRNA and protein expressions of AQP5 were significantly increased. The differences were statistically significant (all P < 0.05). CONCLUSION The expression of AQP5 is involved in the progression of SS. Increasing the expression of AQP5 can significantly inhibit inflammatory stress and reduce the pathological damage of submandibular gland tissue. This may be related to the inhibition of TLR4/MyD88/NF-κB conduction.
Animals ; Toll-Like Receptor 4/genetics* ; Myeloid Differentiation Factor 88/genetics* ; Aquaporin 5/metabolism* ; Sjogren's Syndrome/genetics* ; Signal Transduction ; NF-kappa B/metabolism* ; Rats ; Rats, Sprague-Dawley ; Interleukin-1beta/metabolism* ; Female

OBJECTIVETo explore the molecular mechanism of exocrine immune inflammatory injury of Sjögren's Syndrome and the intervention of Banxia Qinlian Decoction (BQD).

METHODSTotally 18 female NOD mice were randomly divided into the model group, the positive drug group, and the BQD group, 6 in each group. Six female BALB/c mice were recruited as a blank control group. Mice in the blank control group and the model group were gavaged with deionized water at the daily dose of 0.1 mL/10 g body weight. Tripterygium Tablet was administered by gastrogavage to mice in the positive group at the daily dose of 10 mg/kg. BQD was administered by gastrogavage to mice in the BQD group at the daily dose of 60 g crude drugs/kg. After 12 weeks of medication, mice were sacrificed. Their eyeballs were excised and blood collected. Tissues of bilateral parotids and submandibular glands were kept. mRNA transcriptional levels of IL-17, IL-6, type 3 muscarinic acetylcholine receptors (M3R), aquaporin protein-5 (AQP5) were detected by RT-PCR. Expression levels of M3R and AQP5 protein were detected by Western blot. Protein expression levels of IL-17 and IL-6 were detected by ELISA.

RESULTSCompared with the normal group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly up-regulated in the model group (P < 0.01). Compared with the model group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly down-regulated in the positive drug group and the BQD group with statistical difference (P < 0.01, P < 0.05). Compared with the BQD group, mRNA-transcriptional levels of IL-17, IL-6, and M3R, as well as M3R and AQP5 protein expression levels were significantly down-regulated in the positive drug group (all P < 0.01).

CONCLUSIONThe molecular mechanism of BQD in inhibiting SS exocrine neurotoxic injury might be possibly related to regulating Th17/IL-17 immune inflammatory way.

Animals ; Aquaporin 5 ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Interleukin-17 ; metabolism ; Interleukin-6 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Inbred NOD ; Sjogren's Syndrome ; drug therapy ; immunology ; Submandibular Gland ; Th17 Cells ; Up-Regulation

OBJECTIVETo investigate the effect of aquaporin 5(AQP5) on proliferation and migration of ectopic endometrial epithelial cells.

METHODSAQP5 shRNA interference fragments were designed and transfected into ectopic endometrial epithelial cells stably by lentivirus technology. Fluorescence quantitative RT-PCR and Western blotting were used to detect the AQP5 mRNA and protein expression, respectively. The cell proliferation and migration were determined by using MTT method and Transwell system, respectively. Levels of phosphorylated AKT(p-AKT) and total AKT were examined by Western blotting. The nude mice model of endometriosis was constructed and the endometrial cell nodule formation was observed.

RESULTSAQP5 shRNA transfection inhibited cell proliferation and migration compared with control group (both P<0.05). The activation of AKT in AQP5 shRNA transfected cells was lower than that in control cells (P<0.01). Compared to control group, the endometrial cells nodule formation was suppressed in mice inoculated with AQP5 shRNA-silencing ectopic endometrial epithelial cells.

CONCLUSIONDown-regulation of AQP5 expression can suppress the proliferation and migration of ectopic endometrial epithelial cells and endometrial cell nodule formation in nude mice, in which AKT pathway may be involved.

Animals ; Aquaporin 5 ; genetics ; Cell Movement ; Cell Proliferation ; Disease Models, Animal ; Down-Regulation ; Endometriosis ; pathology ; Epithelial Cells ; cytology ; Female ; Gene Silencing ; Mice ; Mice, Nude ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; RNA, Small Interfering ; Transfection

OBJECTIVETo study the estrogen-like action mechanism of Menoprogen on ovariectomized female rats.

METHODOvariectomized rat model (OVX) was established and estradiol (17beta-estradiol, E2) was used as positive control. The uterine coefficient and serum E2 level were determined after administration of Menoprogen for 2 weeks. The uterine vascular endothelial growth factor (VEGF), water channel protein (aquaporin, AQP), estrogen receptor (ER), progesterone receptor (PR) and the expression of proto-oncogenes (c-jun, c-fos) were observed by immunohistochemical method. Yeast two-hybrid assay was applied to detect the existence of components combining with ERalpha or ERbeta in Menoprogen.

RESULTBoth Menoprogen and E2 could significantly elevate the uterine coefficient of OVX rats, increase the level of serum E2 and up-regulate the expressions of VEGF, AQP2 as well as AQP5 in uterus. E2, not as E2 Menoprogen couldn't promote the expressions of ERalpha, PR, c-jun and c-fos in OVX rat uterus. And yeast two-hybrid assay showed no components combining with ERalpha or ERbeta in Menoprogen.

CONCLUSIONMenoprogen has estrogen-like effect, and can be used to treat menopause syndrome. The risk of estrogen-mediated endometrial cancer is low for this treatment because its mechanism is different from estrogen-like substances.

Animals ; Aquaporin 2 ; metabolism ; Aquaporin 5 ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Estradiol ; blood ; Estrogen Receptor alpha ; metabolism ; Estrogens ; pharmacology ; Female ; Ovariectomy ; adverse effects ; Rats ; Rats, Wistar ; Receptors, Progesterone ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo explore the protective mechanism of Nervilia fordii (NF) by observing the effect of its pretreatment on lung aquaporin 1 and 5 (AQP-1, AQP-5) expression in rats with endotoxin-induced acute lung injury (ALI).

METHODSTwenty-four SD rats were randomly divided into 3 groups, the normal group (A), the NF pre-intervention group (B) and the endotoxin model group (C). Rats in Group B and C were made into ALI by endotoxin (5 mg/kg) injection via sublingual vein, and NF pretreatment was applied to Group B. Animals were sacrificed at the 8 h after modeling, their lung were taken for observing the water permeability change by wet/dry weight ratio (W/D) measuring, pathological feature by HE staining, and the expression of AQP-1, AQP-5 was detected by immunohistochemistry and RT-PCR.

RESULTSThe W/D ratio of lung was higher in model rats than in normal rats, but as compared with Group C, it was significantly lower (P < 0.05) in Group B. The pulmonary edematous change was significantly mild and the AQP-1 and AQP-5 protein expressions were significantly higher in Group B than in Group C (P < 0.05).

CONCLUSIONNF pretreatment can promote lung AQP-1 and AQP-5 expression up-regulation, increase lung water clearance and transportation to improve the water balance and eliminate pulmonary edema, so as to effectively protect lung from acute injury.

Acute Lung Injury ; chemically induced ; drug therapy ; prevention & control ; Animals ; Aquaporin 1 ; metabolism ; Aquaporin 5 ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Endotoxins ; Female ; Lung ; metabolism ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Up-Regulation

OBJECTIVE: To detect the expression of AQP5, HIF-1alpha and VEGF in human nasal polyps, and to observe the relationship of AQP5 with HIF-1a and VEGF, and to investigate their role in the pathogenesis of nasal polyps. METHOD: Using the techniques of Real-time Quantitative Polymerase chain Reaction and Western blot, the expressions of AQP5, HIF-1alpha, VEGF mRNA and protein were examined. The specimens were obtained from patients underwent endoscopic surgery at the same time, including eighteen nasal polyps and ten inferior turbinate tissues. RESULT: (1) The result of RT-PCR showed that the expressions of the AQP5 mRNA were lower in nasal polyps than those in inferior turbinate tissue (P < 0.01); there was no significant difference between the expression of HIF-1alpha mRNA or VEGF mRNA in nasal polyps and inferior turbinate tissues (P > 0.05); (2) According to the Western blot, there was no significant difference between the expression of AQP5 in nasal polyps and inferior turbinate tissues (P > 0.05); the expression of HIF-la or VEGF were higher in nasal polyps than those in inferior turbinate tissues (P < 0.05); (3) According to the Western blot, there was a positive correlation between the expression of AQP5 and that of HIF-1alpha (r = 0.633, P < 0.05), and the correlation was also existed between AQP5 and that of VEGF (r = 0.611, P < 0.05). CONCLUSION AQP5, HIF-1alpha, VEGF are all involved in the edema formation of nasal polyps. The changes of AQP5's distribution can cause accumulation of water in nasal polyps, HIF-1alpha, VEGF can induce new blood vessels and increase vasopermeability in nasal polyps; they have separate regulatory mechanism in edema formation, but might have some relationships in some ways.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Aquaporin 5 ; metabolism ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Middle Aged ; Nasal Polyps ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

OBJECTIVEAlveolar epithelium impairment is one of pathological changes associated with chronic lung disease (CLD). Hoxb5 is one of the few homeobox genes strongly expressed in the developing lung. This study investigated the expression of HoxB5, SPC and AQP5 in rats with CLD in order to explore the role of Hoxb-5 in impairment and reparation of alveolar epithelium.

METHODSEighty neonatal rats were randomly exposed to hyperoxia (model group) or to room air (control group) (n=40 each). The CLD model was induced by hyperoxia exposure. The expression of HoxB5, SPC and AQP5 protein and mRNA in the lung tissue was detected by immunohistochemistry and RT-PCR 1, 3, 7, 14 and 21 days after exposure.

RESULTSIn the model group HoxB5 expression significantly decreased 7, 14 and 21 days after hyperoxia exposure. SPC expression decreased 3 days after hyperoxia exposure but increased significantly 7, 14 and 21 days after hyperoxia exposure as compared to the control group. AQP5 expression was progressively reduced with prolonged hyperoxia exposure.

CONCLUSIONSHyperoxia exposure may lead to alveolar epithelial cell (AEC) damage in neonatal rats. The increased SPC expression and decreased AQP5 expression suggested that the ability of differentiation and transformation of AECII into AECI decreased in neonatal rats with CLD. The decreased HoxB5 expression following hyperoxia exposure might contribute to a decreased ability of differentiation of AECII.

Animals ; Animals, Newborn ; Aquaporin 5 ; analysis ; genetics ; Chronic Disease ; Female ; Homeodomain Proteins ; analysis ; genetics ; Hyperoxia ; complications ; Immunohistochemistry ; Lung ; pathology ; Lung Diseases ; etiology ; metabolism ; Male ; Pulmonary Surfactant-Associated Protein C ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore a feasibility of engraftment of systemically transplanted bone marrow stromal cells (BMSCs) and differentiation into lung epithelial cells in lipopolysaccharides (LPS)-injured lungs.

METHODSBMSCs were isolated from bone marrow of transgenic green fluorescent protein (GFP) C57BL/6J mice and systemically administered to bone marrow-suppressed wild-type C57BL/6J mice. A mouse model of lung injury was prepared by intratracheal instillation of LPS. Recipients were assigned to four groups: intratracheal PBS + BMSCs transplantation (CM), intratracheal LPS + BMSCs transplantation (LM), intratracheal PBS + irradiation + BMSCs transplantation (CIM) and intratracheal LPS+ irradiation + BMSCs transplantation (LIM). BMSCs engraftment in recipient lungs was determined by immunofluorescent staining 14 days after BMSCs administration. Alveolar epithelial type II cells were isolated from recipient lungs and the rate of GFP positive cells was measured by flow cytometry. Expression of surfactant protein (SP)-A, SP-C and aquaporin (AQP)-5 mRNA in the lungs was evaluated by real-time PCR.

RESULTSGFP and cytokeratin positive cells were observed in lung parenchyma of the CIM and the LIM groups, but not in the CM and the LM groups. The LIM group had more positive cells than the CIM group. The rates of GFP positive cells were higher in the CIM (11.10+/- 3.19%) and the LIM groups (14.40+/- 2.40%) than those in the CM and the LM groups (2.82+/- 1.03% and 3.81+/- 0.93%, respectively; P< 0.05). The LIM group had higher mRNA expression of SP-C than the CM group (2.09+/- 0.18 vs 1.38+/- 0.30; P< 0.05).

CONCLUSIONSDonor derived BMSCs can engraft in LPS-injured lungs and differentiate into lung epithelial cells, suggesting BMSCs transplantation might contribute to lung repair.

Animals ; Aquaporin 5 ; genetics ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Female ; Lipopolysaccharides ; toxicity ; Lung ; drug effects ; metabolism ; pathology ; Lung Injury ; therapy ; Male ; Mice ; Mice, Inbred C57BL ; Peptides ; genetics ; Pulmonary Surfactant-Associated Protein A ; genetics ; RNA, Messenger ; analysis ; Stromal Cells ; cytology ; transplantation

OBJECTIVE: To investigate the expression and distribution of aquaporin 5 (AQP5) in allergic rhinitis (AR) treated by fluticasone propionate and levocabastine. METHOD: Forty Wistar rats were divided randomly into AR (n=30) and control groups (n=10). After AR models were established, the AR rats were divided evenly into F group, L group and AR control group. Three groups were treated respectively for 28 days, then the expression of AQP5 in nasal mucous membrane were detected by immunohistochemistry assay. RESULT: The distribution of AQP5 was consistent in all groups. The expression of AQP5 in F group was significantly different from L group and AR group (P<0.05), while there was no significant difference between that of AR group and L group (P>0.05). The expression of AQP5 in L group was significantly different from that in control group (P<0.05). CONCLUSION High expressions of AQP5 in rat with AR indicated the positive correlation between AQP5 and AR. AQP5 might be one of pathological factors of AR concerned with glands excessive secretion and tissue edema. Glucocorticoid can down-regulate the expression of AQP5, but H1-receptor antagonist can not reduce the expression of AQP5.
Androstadienes ; pharmacology ; Animals ; Aquaporin 5 ; metabolism ; Fluticasone ; Nasal Mucosa ; drug effects ; metabolism ; pathology ; Piperidines ; pharmacology ; Rats ; Rats, Wistar ; Rhinitis, Allergic, Perennial ; metabolism ; pathology

Animals ; Aquaporin 5 ; physiology ; Cytokines ; physiology ; Gene Expression Regulation ; Humans ; Intracellular Signaling Peptides and Proteins ; physiology ; Membrane Proteins ; physiology ; Mucins ; genetics ; Mucus ; secretion ; Myristoylated Alanine-Rich C Kinase Substrate ; Pulmonary Disease, Chronic Obstructive ; metabolism ; Respiratory Mucosa ; secretion ; Signal Transduction