ABSTRACT
ObjectiveTo investigate the effects of Huashi Runzao prescription （HRP） on the histopathological injury and function of submandibular gland in naive non-obese diabetic （NOD/Ltj） mouse model of Sjögren's syndrome （SS） and its regulatory effect on aquaporin 5 （AQP5） expression in submandibular gland cells. MethodThe SS model was induced in NOD/Ltj mice. The NOD/Ltj female mice aged nine weeks were selected and randomly assigned into model group，HRP group （7.15 g·kg-1·d-1），and hydroxychloroquine （HCQ） group （1.30 g·kg-1·d-1）， and female BALB/c mice in the same age were selected and assigned into the normal group， with six mice in each group. Drug intervention lasted eight weeks. The water consumption and salivary flow rate （SFR） of each group were recorded. The pathological staining results of the submandibular gland of mice in each group were observed and scored. AQP5 expression was determined by immunohistochemistry （IHC） and Western blot. ResultCompared with the normal group， the model group showed increased water consumption （P<0.05） and reduced SFR （P<0.05）. Compared with the model group， the HRP group showed decreased water consumption （P<0.05） and increased SFR （P<0.05）， and the HCQ group showed increased SFR （P<0.05）. In terms of histopathological results of the submandibular gland，compared with the normal group，the model group showed increased pathological score， number of lymphocyte infiltration foci，and percentage of lymphatic infiltration area （P<0.05）. Compared with the model group， the HRP group showed reduced pathological scores and number of lymphocyte infiltration foci （P<0.05）， and the HRP group and the HCQ group showed reduced percentage of lymphatic infiltration area（P<0.05）. The results of IHC and Western blot showed that compared with the normal group，the model group showed down-regulated expression level of AQP5 protein （P<0.05）， and compared with the model group and the HCQ group，the HRP group showed up-regulated expression level of AQP5 protein （P<0.05）. ConclusionHRP can improve the secretion function of submandibular gland acinous cells and glandular structure injury in SS model mice， and its mechanism may be related to the up-regulation of AQP5 protein expression level in submandibular gland cells.
ABSTRACT
Sj9gren's syndrome(SS) is an autoimmune disease with glandular dysfunction caused by the massive infiltration of the exocrine glands by lymphocytes. The pathogenesis of this disease is related to the chronic inflammatory response of the exocrine glands due to excessive activation of B cells and T cells. In addition to dry mouth and eyes, SS can also cause damage to other organs and systems in the human body, seriously affecting the quality of life of patients. Traditional Chinese medicine(TCM) has definite clinical efficacy in the treatment of SS as it can alleviate symptoms and regulate immune disorders without causing adverse reactions, demonstrating high safety. This paper reviews the current status of preclinical and clinical trials about the TCM treatment of SS in the past decade. TCM mainly mitigates SS symptoms such as dry mouth, dry eyes, dry skin, and joint pain and improves the prognosis and quality of life of patients by regulating the abnormally activated B cells and T cells, inhibiting the autoimmune response, restoring the balance between pro-inflammatory and anti-inflammatory cytokines, and reducing the pathological damage caused by immune complexes to exocrine glands and joints in SS patients.
Subject(s)
Humans , Sjogren's Syndrome/drug therapy , Medicine, Chinese Traditional , Quality of Life , Xerostomia , Autoimmune DiseasesABSTRACT
Objective@#To investigate the effects of sinomenine on brain edema and the expression of aquaporin 4 (AQP-4) and aquaporin 5 (AQP-5) in rats with acute brain injury.@*Methods@#According to random number table method, rats were divided into sham operation group, model group, low-dose sinomenine group, high-dose sinomenine group, 20 in each group. In addition to the sham operation group, the rat brain injury model was established by Feeney free falling impact method. Rats in low and high dose of sinomenine group were given sinomenine 30, 60 mg/kg by intraperitoneal injection, and the sham operation group and model group were injected intraperitoneally with the same volume of normal saline, once per day for 7 days. Histopathological changes in each group were observed by HE staining and electron microscope. Western blot and RT-PCR were used to detect AQP-4 and AQP-5 protein and gene expression in brain tissue.@*Results@#The pathological results showed that the nerve cells in the model group were loosely arranged, disordered in hierarchy, some cells appear to have degenerated. The degeneration and necrosis of nerve cells in the low and high dose sinomenine group were less than those in the model group. Compared to the model group, the expression of AQP-4 (0.74 ± 0.13, 0.49 ± 0.11 vs. 1.30 ± 0.21), AQP-5 (0.98 ± 0.07, 0.45 ± 0.10 vs. 1.47 ± 0.18) in the low dose and high dose sinomenine group significantly decreased (P<0.05). The expression of AQP-4 (1.48 ± 0.18, 1.26 ± 0.12 vs. 2.04 ± 0.14), AQP-5 (1.31 ± 0.17, 1.20 ± 0.11 vs. 1.87 ± 0.15) mRNA in the low dose and high dose sinomenine group significantly decreased (P<0.05).@*Conclusions@#Sinomenine could alleviate cerebral injury by lowering the expression of AQP-4 and AQP-5.
ABSTRACT
In many ways, cancer cells are different from healthy cells. A lot of tactical nano-based drug delivery systems are based on the difference between cancer and healthy cells. Currently, nanotechnology-based delivery systems are the most promising tool to deliver DNA-based products to cancer cells. This review aims to highlight the latest development in the lipids and polymeric nanocarrier for siRNA delivery to the cancer cells. It also provides the necessary information about siRNA development and its mechanism of action. Overall, this review gives us a clear picture of lipid and polymer-based drug delivery systems, which in the future could form the base to translate the basic siRNA biology into siRNA-based cancer therapies.
ABSTRACT
Objective::To observe the effect of Bimin decoction(BMD) on nuclear factor-kappa B(NF-κB) signaling pathway and aquaporin 5(AQP5) expression in allergic rhinitis (AR) rats with lung and spleen Qi deficiency syndrome(LSQDS), in order to study the mechanism in treating AR. Method::Fifty-six SD rats were randomly divided into seven groups: control group, AR group, LSQDS AR group, BMD low dose two weeks group and four weeks group, BMD high dose two weeks group and four weeks group. The control group did not intervened, the AR group established the AR disease model with ovalbumin (OVA) as the allergen, the other five groups established the LSDQS model with smoke and senna gavage, and also established the AR disease model with OVA sensitization at the same time as the AR group. After the model was established successfully, four BMD intervention groups were separately given low dose BMD (11.3 g·kg-1) for 2 weeks and 4 weeks, and high dose BMD (22.6 g·kg-1) for 2 weeks and 4 weeks. To observe the general situation of the rats, hematoxylin eosin (HE) staining was used to observe the pathological changes of the nasal mucosa, immunohistochemistry and Western blot were used to detect the expression levels of NF-κB and AQP5, real-time fluorescent quantitative polymerase chain reaction technique (Real-time PCR)was used to detect mRNA levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6). Result::The typical AR symptoms were found in AR rats, the AR symptoms and lung and spleen Qi deficiency symptoms were found in AR rats with LSQDS at the same time, and the AR symptoms and lung and spleen Qi deficiency symptoms were significantly improved after the intervention of BMD. Compared with the control group, the typical histopathological changes of nasal mucosa were found in AR group and LSQDS AR group, with a higher behavioral score (P<0.05), and the expression of NF-κB and AQP5 protein increased (P<0.05), the expression of IL-1β, TNF-α, IL-6 and AQP5 mRNA increased (P<0.05). Compared with AR group, the pathological changes of nasal mucosa in LSQDS AR group were more serious, and the expression of NF-κB protein in nucleus increased (P<0.05), the expression of TNF-α and AQP5 mRNA increased (P<0.05). Compared with LSQDS AR group, the pathological changes of nasal mucosa in the groups which interfered by BMD were improved, and the expression of NF-κB and AQP5 protein decreased (P<0.05), the expression of IL-1β, TNF-α, IL-6 and AQP5 mRNA decreased (P<0.05). Compared with BMD low-dose two-week group, the expression of NF-κB protein in nucleus decreased (P<0.05) in BMD high dose four week group. Conclusion::Compared with AR group, the AR condition of the rats with LSQDS is more serious under the same allergen stimulation, BMD can treat AR and reduce the over secretion of glands, which may be related to inhibit the expression of AQP5 by inhibiting the NF-κB signaling pathway.
ABSTRACT
Xylitol is well-known to have an anti-caries effect by inhibiting the replication of cariogenic bacteria. In addition, xylitol enhances saliva secretion. However, the precise molecular mechanism of xylitol on saliva secretion is yet to be elucidated. Thus, in this study, we aimed to investigate the stimulatory effect of xylitol on saliva secretion and to further evaluate the involvement of xylitol in muscarinic type 3 receptor (M3R) signaling. For determining these effects, we measured the saliva flow rate following xylitol treatment in healthy individuals and patients with dry mouth. We further tested the effects of xylitol on M3R signaling in human salivary gland (HSG) cells using real-time quantitative reverse-transcriptase polymerase chain reaction, immunoblotting, and immunostaining. Xylitol candy significantly increased the salivary flow rate and intracellular calcium release in HSG cells via the M3R signaling pathway. In addition, the expressions of M3R and aquaporin 5 were induced by xylitol treatment. Lastly, we investigated the distribution of M3R and aquaporin 5 in HSG cells. Xylitol was found to activate M3R, thereby inducing increases in Ca²⁺ concentration. Stimulation of the muscarinic receptor induced by xylitol activated the internalization of M3R and subsequent trafficking of aquaporin 5. Taken together, these findings suggest a molecular mechanism for secretory effects of xylitol on salivary epithelial cells.
Subject(s)
Humans , Aquaporin 5 , Bacteria , Calcium , Calcium Signaling , Candy , Epithelial Cells , Immunoblotting , Mouth , Polymerase Chain Reaction , Receptors, Muscarinic , Saliva , Salivary Glands , XylitolABSTRACT
Objective To evaluate the effect of ulinastatin (UT1) on the expression of aquaporin 1 (AQP1) and AQP5 in rats with acute lung injury induced by cardiopulmonary bypass (CPB).Methods Forty-eight clean-grade healthy adult male Sprague-Dawley rats,weighing 200-250 g,were divided into 3 groups (n=16 each) using a random number table method:sham operation group (Sham group),CPB group and UTI group.UTI 200 000 U/kg was injected intravenously at 10 min prior to CPB in UTI group.The model of CPB was established in CPB and UTI groups.The equal volume of normal saline was intravenously injected at 10 min prior to puncture or at 10 min prior to CPB in Sham and CPB groups.Rats were sacrificed,and lung tissues were excised for determination of weight to dry weight ratio (W/D ratio),expression of AQP1 and AQP5 (by immunohistochemistry),expression of AQP1 and AQP5 protein and mRNA (by real-time polymerase chain reaction or Western blot) and for examination of morphological structure (with a light microscope) and ultrastructure of lung tissues (with an electron microscope).Injured alveolar rate (IAR) and rates of AQP1 and AQP5 positive cells were calculated.Results Compared with Sham group,W/D ratio and IAR were significantly increased,rates of AQP1 and AQP5 positive ceils were decreased,and the expression of AQP1 and AQP5 protein and mRNA was down-regulated in CPB and UTI groups (P＜0.05).Compared with CPB group,W/D ratio and IAR were significantly decreased,rates of AQP1 and AQP5 positive cells were increased,and the expression of AQP1 and AQP5 protein and mRNA was up-regulated in UTI group (P＜0.05).The injury to morphological structure and ultrastructure was significantly attenuated in UTI group when compared with CPB group.Conclusion The mechanism by which UTI pretreatment reduces CPB-induced acute lung injury is related to up-regulating the expression of AQP1 and AQP5 in rats.
ABSTRACT
Objective To evaluate the effects of dexmedetomidine on the expression of aquaporins 1 (AQP1) and AQP5 during lung ischemia/reperfusion (I/R) in rats in an in vitro experiment.Methods SPF healthy male Sprague-Dawley rats,aged 3-4 months,weighing 250-320 g,were used in this study.Forty-five isolated rat lungs in which the model of isolated lung perfusion was successfully established were divided into 3 groups (n=15 each) using a random number table:sham operation group (S group),I/R group and dexmedetomidine group (D group).The isolated rat lungs were continuously perfused for 150 min in group S.After 15 min of perfusion,the isolated rat lungs were subjected to 60 min of ischemia and apnea followed by 75 min of ventilation and reperfusion to establish the model of isolated lung I/R injury in group I/R.In group D,the isolated rat lungs were perfused for 15 min with K-H perfusion fluid containing 10 nmol/L dexmedetomidine,and then subjected to 60 min of ischemia and apnea followed by 75 min of ventilation and reperfusion with K-H perfusion fluid containing 10 nmol/L dexmedetotnidine.The lung compliance,airway resistance,perfusion flow and partial pressure of arterial oxygen (PaO2) were recorded at 10 min of perfusion and 15,45 and 75 min after restoration of perfusion.Lung tissues were obtained at 75 min after restoration of perfusion for determination of wet/dry weight ratio (W/D ratio) and for examination of the pathological changes and changes in the uhrastructure.The expression of AQP1 and AQP5 protein and mRNA in lung tissues was detected by Western blot and real-time polymerase chain reaction,respectively.Results Compared with S group,the airway resistance was significantly increased and lung compliance,perfusion flow and PaO2 were decreased during reperfusion,and W/D ratio was increased in I/R and D groups (P＜0.05),the expression of AQP1 and AQP5 protein and mRNA in lung tissues was significantly down-regulated in group I/R,and the expression of AQP 1 and AQP5 protein and mRNA in lung tissues was significantly up-regulated in group D (P＜0.05).Compared with I/R group,the airway resistance was significantly decreased and lung compliance,perfusion flow and PaO2 were increased during reperfusion,W/D ratio was decreased,the expression of AQP1 and AQP5 protein and mRNA in lung tissues was up-regulated (P＜0.05),and the pathological changes of lung tissues was significantly attenuated in group D.Conclusion The mechanism by which dexmedetomidine reduces I/R injury may be related to up-regulation of the expression of AQP1 and AQP5 in rats in an in vitro experiment.
ABSTRACT
Objective To observe the expression of protein AQP5 and CC16 in lung after hemorrhagic shock resuscitation in rats in order to explore the mechanism of acute lung injury.Methods Thirty-two healthy and clean male SD rats were randomly (random number) divided into two groups:control group and hemorrhagic shock resuscitation group (n =16 in each).Besides,each group was further divided into two subgroups according to the experiment done at 12 h and 24 h after hemorrhagic shock resuscitation (n =8).The hemorrhagic shock model was made by using Wiggers' modified method.Resuscitation was done by transfusing the autologous blood and the equal volume of Ringer's solution.Blood samples were obtained from abdominal aorta at each given interval to measure the level of plasma endotoxin,and assay the CC16 and AQP5 by using ELISA.After the rats were sacrificed,the left lung tissue was taken to measure lung water content and the dry/wet ratio,and to examine the levels of CC16 and AQP5 in lung tissue by immunohistochemical method.Results ①The level of plasma endotoxin in the experimental group was significantly higher than that in the control group (P ＜ 0.01).②The content of plasma CC16 in the experimental group was higher than that in the control group (P ＜ 0.05).③Compared with the control group,the content of pulmonary homogenate AQP5 in the experimental group was significantly lower (P ＜0.05).④The lung water content (the dry/wet ratio) of the experimental group was obviously higher than that of the control group (P ＜ 0.05).⑤Hislogogical observation with HE staining showed in the control group,the alveolar structure was complete,the alveolar sacs were clear,and the alveolar septum was intact;but in the experimental group,the alveolar septum was widened,and there were obvious hemorrhage and neutrophil infiltration in the alveolar space.⑥ The level of lung tissue CC16 in control group was significantly higher compared with experimental group (P ＜ 0.05).⑦ The level of lung tissue AQP5 was significantly higher in control group compared with experimental group (P ＜ 0.05).Conclusions The proteins of AQP5 and CC16 were involved in the process of acute lung injury after hemorrhagic shock resuscitation in rats,and their levels were positively correlated with length of time after hemorrhagic shock.
ABSTRACT
Objective To determine if aquaporin1 ( AQP1) and aquaporin5 ( AQP5) are expressed in the alveolar-capillary membrane in rats, and to investigate the changes of AQP1 and AQP5 expression in the rat with acute lung injury.Methods The distribution of AQP1 and AQP5 in alveolar capillary membrane was investigated by immunohistochemistry and immunoelectron microscopy with affinity-purified antibodies to human AQP1 and AQP5.The possibility that alveolar capillary membrane AQP1 and AQP5 undergo altered regulation was studied by a rat model established using intra-tracheal instillation of lipopolysaccharide (LPS).Results Immunolabelling showed that AQP1 was stained primarily in the microvascular endothelium of normal lungs, while AQP5 was expressed in type I pneumocytes. Immunohistochemical analysis showed a significant decrease in the expression of AQP1 and AQP5 in injured lungs at 4 -48 h after LPS instillation.AQP1 protein was resumed partly at 24 h after LPS instillation and steroid administration, whereas AQP5 was unchanged.Conclusions The decreased expressions of AQP1 and AQP5 in injured lungs suggest that both of them may play a role in abnormal fluid transportation.
ABSTRACT
OBJECTIVE: To observe whether bone marrow mesenchymal stem cell( BMMSC) could be induced by alveolar epithelial cell( AEC) of rats exposed to silica dust or not. METHODS: BMMSCs were isolated and cultivated from 6specific pathogen free healthy male SD rats through bone marrow adherent method. The AECs from other 6 rats randomly selected from the same batch were cultivated by immune adherent purification method. Three rats were treated with 1. 0 m L( 40 g/L mass concentration) of silicosis dust suspension by one time intratracheal injection as silicosis dust exposure model,and the other 3 rats were given 0. 9% sodium chloride solution as normal. Experimental group was the co-culture of BMMSCs and AECs from silicosis dust exposure rats. Control group A was the co-culture of BMMSCs and AECs from normal rats. Control group B was the culture of BMMSCs alone. The morphology changes were observed by the inverted phase contrast microscope at the time points of the 4th and the 8th day. Double immunofluorescence staining using aquaporin 5( AQP5) and surfactant protein C( SP-C) was performed on the treated BMMSCs. The fluorescence staining was observed using the inverted fluorescence microscope( IFM) and laser scanning confocal microscope( LSCM). Integral optical density( IOD) analysis was conducted on fluorescence of 2 kinds of proteins by Image-pro plus 6. 0 graphic analysis software. RESULTS: After the co-culture,the BMMSCs in experimental group and control group A changed from long spindle shape to cubic and polygonal shape,the variation of morphology on day 8 was more obvious than that on day 4,and the change in control group A was less obvious than that of experimental group. There was no obvious morphology change in BMMSCs of control group B. By IFM and LSCM,on day 4 and day 8,the expression of green fluorescence AQP5 and red fluorescence SP-C were all observed in BMMSCs of experimental group and control group A. The BMMSCs of control group B only showed a little green fluorescence expression of AQP5,no expression of red SP-C fluorescence was seen. Both by IFM and LSCM,on day 4 and day 8,the 2 kinds of IOD of BMMSCs in experiment group were higher than those of control group A and B at the same time points( P < 0. 01); the IOD of control group A was higher than that of control group B at the same time point( P < 0. 01). The IOD of experiment group and control group A on day 8 were higher than those on day4 in the same group( P < 0. 01). CONCLUSION: AEC of rats exposed to silica dust can effectively induce BMMSC to be differentiated into AEC.
ABSTRACT
BACKGROUND: Aquaporins are water channel proteins that play a major role in the movement of water in various human tissues. Recently, it has been found that aquaporins have influence in the carcinogenesis of human malignancies. We analyzed the prognostic impact of aquaporin 5 (AQP5) in non-small lung cancer (NSCLC). METHODS: Seventy-six cases of NSCLC were studied, including 44 cases of adenocarcinoma (ADC) and 32 cases of squamous cell carcinoma (SQCC). Tissue microarray was constructed and immunohistochemical staining for AQP5 was performed. RESULTS: AQP5 was positive in 59.2% of the total enrolled NSCLCs (63.7% in ADC and 53.1% in SQCC). The difference in expression of AQP5 according to the histologic grade of the tumor was significant (p<.047), but not in a serial order. When ADC and SQCC were separately evaluated, no significant difference was observed according to the histologic grade of the tumor (p=.076 in ADC and p=.631 in SQCC). No difference was observed between AQP5 expression and other demographic data and tumor characteristics. Disease-free survival (DFS) was higher in AQP5 negative cases than positive cases in ADC (p=.047), but no significance was found in SQCC (p=.068). We were unable to find a significance between AQP5 overexpression and overall survival in either ADC (p=.210) or SQCC (p=.533). CONCLUSIONS: AQP5 expression is associated with DFS in ADC of the lung and tumor grade of NSCLC. The present study suggests that AQP5 can be a prognostic factor of NSCLC.
Subject(s)
Humans , Adenocarcinoma , Aquaporin 5 , Aquaporins , Carcinogenesis , Carcinoma, Non-Small-Cell Lung , Carcinoma, Squamous Cell , Disease-Free Survival , Lung , Lung Neoplasms , WaterABSTRACT
Objective To investigate the effect of all-trans retinoic acid (at-RA) on fetal alveolar epithelial type Ⅱ cells (fAEC Ⅱ s) proliferation and the expression of pulmonary surfactant C (SPC) as well as aquaporin 5 (AQP5).Methods fAEC Ⅱ s were isolated and purified from fetal lung of pregnant SD rats (19 days).After being cultured for 1 day,and the fAEC Ⅱ s were interfered by at-RA for 1,2 and 3 days.Cell proliferation,viability as well as growth state,expressions of SPC mRNA as well as AQP5 mRNA and expressions of protein SPC as well as AQP5 were respectively detected by using 4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT),inverted microscope,real-time fluorescence quantitative PCR (RT-PCR) and Western blot.Results (1) When fAEC Ⅱ s were treated with at-RA for 1 day,and the cell proliferation and viability did not change (P ＞ 0.05),while the proliferation and viability were significantly improved in 2 days (P ＜ 0.05),and the difference was the most obvious (P ＜ 0.05) at 3 days.(2)Compared with the control group,the cell growth state was better,and the cell adherence was tighter and the refraction was higher in at-RA group.(3) Compared with the control group,at-RA up-regulated the expressions of AQP5 mRNA and AQP5 protein(t =-19.58,-10.44,-16.01,-46.25,-12.79,-27.96,all P ＜ 0.05),and the percentages of control group were 281.07％,766.67％,1 163.33％ and 792.65％,1 310.52％,1 561.56％ in 1,2 and 3 days respectively.(4) Compared with control group,the expressions of SP-C mRNA and SPC protein were up regulated when cells were exposed to at-RA for 1 and 3 d,but while they were down-regulated at 2 days(protein:the percentages of control group were 615.480％,369.450％ and 11.269％,respectively ; mRNA:728.33 ％,400.83 ％,66.57％,respectively)(t=-26.34,-25.26,13.80,-25.25,-31.71,9.12,all P＜0.05).Conclusions at-RA can promote the proliferation and differentiation of fAEC Ⅱs,enhance the fAEC Ⅱ s viability,and improve the expression of SPC and AQP5.
ABSTRACT
Objective To evaluate the effect of long-term use of simvastatin on aquaporin 5 (AQP5) expression in a rat model of ventilator-induced lung injury.Methods Thirty-two adult male Sprague-Dawley rats, weighing 250-280 g, were randomly divided into 4 groups (n=8 each) using a random number table: control group (group C), simvastatin group (group Sim), mechanical ventilation group (group MV) and simvastatin + mechanical ventilation group (group SMV).In C and Sim groups, normal saline 1 ml/d and simvastatin 10 mg · kg-1 · d-1 were injected, respectively, through a gastric tube into stomach for 4 weeks, and then the rats were tracheostomized, and mechanically ventilated, and the animals kept spontaneous breathing for 4 h.In MV and SMV groups, normal saline 1 ml/d and simvastatin 10 mg · kg-1 · d-1were injected, respectively, through a gastric tube into stomach for 4 weeks, and then the rats were tracheostomized, and mechanically ventilated (tidal volume 50 ml/kg) for 4 h.The rats were then sacrificed, and lungs were removed for determination of wet to dry lung weight ratio (W/D ratio), activities of myeloperoxidase (MPO) and superoxide dismutase (SOD), malondialdehyde (MDA) content,and expression of AQP5 protein and mRNA in lung tissues, and for microscopic examination of pathological changes.Results Compared with group C, the W/D ratio, MPO activity, and MDA content were significantly increased, the SOD activity was decreased, and the expression of AQP5 protein and mRNA was down-regulated in group MV (P＜0.01).Compared with group MV, the W/D ratio, MPO activity, and MDA content were significantly decreased, the SOD activity was increased, and the expression of AQP5 protein and mRNA was up-regulated in group SMV (P＜0.01).The pathological changes of lungs were significantly mitigated in group SMV as compared with group MV.Conclusion Long-term use of simvastatin alleviates ventilator-induced lung injury, and the mechanism is related to down-regulated expression of AQP5 in rats.
ABSTRACT
Objective To investigate the expression patterns and significance of aquaporin 5 (AQP‐5) and multidrug‐resistance associated genes in human colon cancer with different differentiation degree and their correlation .Methods The expression of aqua‐porin 5 and resistance genes P‐gp ,GST‐π,TopoⅡ ,and TS in human 45 cases colon cancer tissues with different differentiation de‐gree and 36 cases of adjacent mucosa tissues as well as 58 cases of normal colonic epithelium were detected by quantitative RT‐PCR ,Western blot and immunohistochemistry .Results Immunohistochemistry results showed that AQP‐5 distributed mainly in the cell membrane and the cytoplasm .Fluorescence quantitative RT‐PCR and Western blot showed that AQP‐5 expression could not be detected in adjacent mucosa tissues and normal colonic epithelium tissues .The AQP‐5 expression level was higher in colon cancer tissues compared with adjacent mucosa tissues and normal colonic epithelium tissues (P0 .05) .Positive correlation was found between the expression of AQP‐5 and P‐gp ,GST‐π,Topo Ⅱ(P0 .05) .Conclusion The AQP‐5 and re‐sistance gene expression were increased in colon cancer tissues .The AQP‐5 expression level was higher in colon cancer compared with adjacent control or normal tissues ,which may promote the transfer and progress of colon cancer .
ABSTRACT
Objective To determine the relationship of the expression of aquaporin 5 (AQP5)with clinicopathology and prognosis of colorectal cancer.Methods We collected data from 45 patients with primary colorectal cancer without any adjuvant therapy before operation.The expression of AQP5 was measured by immunohistochemistry (IHC).Then we analyzed the correlation between AQP5 expression and clinicopathological parameters (including age,tumor size,clinical staging,tumor location,lymph node and pathological type)and the connection between AQP5 expression and prognosis based on follow-up data.Results Of the 45 tumor specimens,14 (31.1%)had a high level of AQP5 expression,29 (64.4%)exhibited a moderate level of staining,and 2 (4.4%)had an absence of AQP5 staining.AQP5 was only occasionally detected in para-neoplastic [3/45 (6.67%)]and normal tissues [3/45 (6.67%)].The overexpression of AQP5 was also positively associated with TNM stage (P =0.002),lymph node metastasis (P =0.01 6),and distant metastasis (P =0.000).However,it had no significant association with age, gender,histologic grade or tumor size (P > 0.05 ).Conclusion AQP5 may be used as a novel biomarker for predicting the prognosis of colorectal cancer.
ABSTRACT
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.
Subject(s)
Animals , Male , Rats , Acute Lung Injury , Drug Therapy , Genetics , Pathology , Adrenergic alpha-2 Receptor Agonists , Pharmacology , Aquaporin 1 , Genetics , Allergy and Immunology , Aquaporin 5 , Genetics , Allergy and Immunology , Dexmedetomidine , Pharmacology , Dose-Response Relationship, Drug , Drug Administration Schedule , Gene Expression Regulation , Injections, Intravenous , Interleukin-1beta , Genetics , Allergy and Immunology , Lipopolysaccharides , Lung , Allergy and Immunology , Pathology , Organ Size , Pulmonary Edema , Drug Therapy , Genetics , Pathology , Rats, Wistar , Signal Transduction , Transcription, Genetic , Tumor Necrosis Factor-alpha , Genetics , Allergy and ImmunologyABSTRACT
Objective To explore the effect of different doses of dobutamine on acute lung injury (ALI) in rabbits with septic shock and to clarify the possible mechanism.Methods The rabbits model of septic shock was made by cecal ligation and puncture combined with intravenous injection of endotoxin,70 male New Zealand white rabbits were randomly divided into five groups (14 rabbits in each groups):shamc operation group (group A),ALI group (group B),dobutamine low-dose group (group C),dobutamine medium-dose group (group D) and dobutamine high-dose group (group E),7 rabbits from each group were sacrificed 3 h and 6 h after septic shock.The level of cyclic adenosine monophosphate (cAMP) in lung tissue was detected by ELISA.The expression of aquaporin 5 (AQP5) protein was determined by western blotting.The wet to dry weight (W/D) ratio was measured.The pathological and ultrastructural changes of lung tissue were evaluated by optical microscopy and electron microscope,and lung injury score was assessed.The differences among the different groups were analyzed by one-way ANOVA (LSD test).Results The level of cAMP and expression of AQP5 protein in lung tissue at 3 h and 6 h were dramatically lower in group B than those in group A (3.53 ±0.43) pmol/mLvs.(21.18 ±0.62) pmol/mL; (0.44 ± 0.04) pmol/mLvs.(0.99±0.06)pmol/mL; (2.71±0.56)pmol/mLvs.(21.78±0.62)pmol/mL; (0.29 ±0.05) pmol/mLvs.(0.91 ±0.06) pmol/mL; all P ＜0.001,while the W/D ratio was obviously higher in group B than those in group A (all P ＜0.001).Compared with group B,the level of cAMP and AQP5 protein expression in lung tissue were significantly increased at 6 h in group C (8.48 ±0.61) pmol./ mLvs.(2.71±0.56) pmol/mL,P＜0.01; (0.49 ±0.04) pmol/mLvs.(0.29 ±0.05) pmol/mL,P=0.001 and at3 hand6 hin groupDandE (10.86±0.66) pmol/mLvs.(3.53±0.43) pmol/mL; (0.60±0.05) pmol/mLvs.(0.44±0.04) pmol/mL; (13.80±0.49) pmol/mLvs.(2.71±0.56) pmol/mL; (0.64 ± 0.03) pmol/mLvs.(0.29 ± 0.05) pmol/mL; (15.57 ± 0.60) pmol/mL vs.(3.53±0.43) pmol/mL; (0.91 ±0.05) pmol/mLvs.(0.44 ±0.04) pmol/mL; (19.30±0.42) pmol/mL vs.(2.71 ±0.56) pmol/mL; (0.89 ±0.08) pmol/mL vs.(0.29 ±0.05) pmol/mL; all P ＜ 0.01,while the W/D ratio in group E was decreased obviously (P =0.002; P =0.001).Compared with group C and D,the level of cAMP and the expression of AQP5 protein at 3 h and 6 h in group E increased significantly (all P ＜0.01.The pathological and ultrastructural changes of lung tissue were more intensive in group B than those in group A and the lung injury scores were obviously higher (P ＜0.01).The degree of lung pathological and ultrastructural lesion was ameliorated after administration of dobutanmine.Additionally,histological scores decreased significantly (P ＜ 0.01).Conclusions Our study demonstrated that dobutamine could improve ALI induced by endotoxin,the mechanism of protective effect may involve in increasing the level of cAMP and up-regulating the AQP5 protein expression,and high-dose dobutamine had better effects.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of H₂S on lower limb ischemia-reperfusion (LIR) induced lung injury and explore the underlying mechanism.</p><p><b>METHODS</b>Wistar rats were randomly divided into control group, IR group, IR+ Sodium Hydrosulphide (NaHS) group and IR+ DL-propargylglycine (PPG) group. IR group as lung injury model induced by LIR were given 4 h reperfusion following 4 h ischemia of bilateral hindlimbs with rubber bands. NaHS (0.78 mg/kg) as exogenous H₂S donor and PPG (60 mg/kg) which can suppress endogenous H₂S production were administrated before LIR, respectively. The lungs were removed for histologic analysis, the determination of wet-to-dry weight ratios and the measurement of mRNA and protein levels of aquaporin-1 (AQP₁), aquaporin-5 (AQP₅) as indexes of water transport abnormality, and mRNA and protein levels of Toll-like receptor 4 (TLR₄), myeloid differentiation primary-response gene 88 (MyD88) and p-NF-κB as indexes of inflammation.</p><p><b>RESULTS</b>LIR induced lung injury was accompanied with upregulation of TLR₄-Myd88-NF-κB pathway and downregulation of AQP1/AQP₅. NaHS pre-treatment reduced lung injury with increasing AQP₁/AQP₅ expression and inhibition of TLR₄-Myd88-NF-κB pathway, but PPG adjusted AQP₁/AQP₅ and TLR4 pathway to the opposite side and exacerbated lung injury.</p><p><b>CONCLUSION</b>Endogenous H₂S, TLR₄-Myd88-NF-κB pathway and AQP₁/AQP₅ were involved in LIR induced lung injury. Increased H₂S would alleviate lung injury and the effect is at least partially depend on the adjustment of TLR₄-Myd88-NF-κB pathway and AQP₁/AQP₅ expression to reduce inflammatory reaction and lessen pulmonary edema.</p>
Subject(s)
Animals , Male , Rats , Acute Lung Injury , Pathology , Aquaporins , Metabolism , Drug Evaluation, Preclinical , Edema , Pathology , Hydrogen Sulfide , Pharmacology , Therapeutic Uses , Inflammation , Lung , Pathology , Myeloid Differentiation Factor 88 , Metabolism , NF-kappa B , Metabolism , Random Allocation , Rats, Wistar , Reperfusion Injury , Pathology , Toll-Like Receptor 4 , Metabolism , Water , MetabolismABSTRACT
10.3969/j.issn.1007-3969.2013.04.007