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SUMMARY: This study assessed the effects of Acacia Senegal (AS) combined with insulin on Na+/K+-ATPase (NKA) activity and mRNA expression, serum glucose, renal function, and oxidative stress in a rat model of diabetic nephropathy (DN). Sixty rats were equally divided into six groups: normal control, normal+AS, diabetic (DM), DM+insulin, DM+AS, and DM+insulin+AS groups. Diabetes mellitus (type 1) was induced by a single injection of streptozotocin (65 mg/kg), and insulin and AS treatments were carried until rats were culled at the end of week 12. Serum glucose and creatinine levels, hemoglobin A1c (HbA1c) were measured. Renal homogenate levels of NKA activity and gene expression, malondialdehyde, superoxide dismutase (SOD), catalase and reduced glutathione (GSH) were evaluated as well as kidney tissue histology and ultrastructure. Diabetes caused glomerular damage and modulation of blood and tissue levels of creatinine, glucose, HbA1c, malondialdehyde, NKA activity and gene expression, SOD, catalase and GSH, which were significantly (p<0.05) treated with AS, insulin, and insulin plus AS. However, AS+insulin treatments were more effective. In conclusion, combined administration of AS with insulin to rats with DN decreased NKA activity and gene expression as well as oxidative stress, and improved glycemic state and renal structure and function.
Este estudio evaluó los efectos de Acacia senegal (AS) combinada con insulina sobre la actividad Na+/K+- ATPasa (NKA) y la expresión de ARNm, la glucosa sérica, la función renal y el estrés oxidativo en un modelo de nefropatía diabética (ND) en ratas. Sesenta ratas se dividieron equitativamente en seis grupos: control normal, normal+AS, diabética (DM), DM+insulina, DM+AS y DM+insulina+AS. La diabetes mellitus (tipo 1) se indujo mediante una única inyección de estreptozotocina (65 mg/kg), y los tratamientos con insulina y AS se llevaron a cabo hasta que las ratas fueron sacrificadas al final de la semana 12. Se midieron niveles séricos de glucosa y creatinina, hemoglobina A1c (HbA1c). Se evaluaron los niveles de homogeneizado renal de actividad NKA y expresión génica, malondialdehído, superóxido dismutasa (SOD), catalasa y glutatión reducido (GSH), así como la histología y ultraestructura del tejido renal. La diabetes causó daño glomerular y modulación de los niveles sanguíneos y tisulares de creatinina, glucosa, HbA1c, malondialdehído, actividad y expresión génica de NKA, SOD, catalasa y GSH, los cuales fueron tratados significativamente (p<0,05) con AS, insulina e insulina más AS. Sin embargo, los tratamientos con AS+insulina fueron más efectivos. En conclusión, la administración combinada de AS con insulina a ratas con DN disminuyó la actividad de NKA y la expresión genética, así como el estrés oxidativo, y mejoró el estado glucémico y la estructura y función renal.
Subject(s)
Animals , Male , Rats , Plant Extracts/administration & dosage , Sodium-Potassium-Exchanging ATPase/drug effects , Diabetic Nephropathies/drug therapy , Acacia/chemistry , Superoxide Dismutase , Glycated Hemoglobin/analysis , Plant Extracts/pharmacology , Gene Expression , Rats, Sprague-Dawley , Sodium-Potassium-Exchanging ATPase/genetics , Oxidative Stress , Microscopy, Electron, Transmission , Disease Models, Animal , Drug Therapy, Combination , Glycemic Control , Insulin/administration & dosage , Kidney/drug effects , MalondialdehydeABSTRACT
OBJECTIVE Na+/K+-ATPase(NKA)is a large membrane protein expressed universally in all cells.It is indispensable for the maintenance of ionic gradient.We previously reported that the dysfunction of this pump in neurons and astrocytes contributes to stroke and neurodegenerative diseases,respectively.However,its roles in the microglia and stress-related diseases are still unclear.METHODS Two classical models,chronic restraint stress(CRS)model and electronic foot shock(ES)model,were used to study the pathogenesis of anxi-ety in either NKAα1 global knockout(NKAα1 GKO)mice or NKA α1 conditional knockout(NKAα1 CKO)mice.Behavioral tests like open-field test,elevated plus maze,Morris water maze,novel object recognition test and gait imaging test were performed.A variety of molecular bio-logical methods were employed,including RNA sequenc-ing(RNA-seq)analyses,immunofluorescence and elec-trophysiological recordings etc.RESULTS NKAα1 defi-ciency had a broad impact on physical stress-induced anxiety-like behavior,but failed to exacerbate CRS induced memory deficits.Electrophysiology experiment showed that NKAα1 GKO and NKAα1 CKO mice exhibit-ed neuronal hyperexcitability under chronic stress.The underlying mechanisms may involve neuroinflammation,as NKAα1 deficiency exacerbated stress-induced microg-lia activation in vivo.Similarly,inhibition or downregula-tion of NKA α 1 aggravated LPS + ATP-induced inflam-mation in vitro.DR5-12D,a monoclonal antibody against the DR-region of NKAa1,improved stress-induced anxiety-like behavior through amelioration of the neuronal hyper-excitability and neurogenesis deficit in the ventral hippo-campus of mice.CONCLUSION NKA is closely related to neuroinflammation in microglia and DR-region of NKA a1 subunit may serve as a novel target to treat stress-induced anxiety.
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@#Objective To investigate the mechanism of insulin alleviating pulmonary edema in mice with acute lung injury(ALI) by serine/threonine protein kinase-1(SGK1).Methods 32 male adult C3H/HeN mice were randomly divided into control group(only pumped with the same amount of normal saline as the treatment group),ALI group(continuously pumped with the same amount of normal saline as the treatment group after modeling),treatment group [continuously pumped with 0.1 U/(kg·h) of insulin through jugular vein after establishing ALI model] and SGK1 siRNA group[continuously pumped with 0.1 U/(kg·h) of insulin and given SGK1 siRNA(75 μg SGK1 siRNA diluted in 100 μL saline) simultaneously after establishing ALI model] with 8 mice in each group.After 8 h,the mice were killed for arterial blood gas analysis(1 h after establishment of the model) and the changes of plasma glucose levels were detected(0,1,4and 8 h);The bronchoalveolar lavage fluid(BALF) was collected to detect the content of total protein,and the alveolar epithelial permeability and lung water content were measured;The pathological changes of lung tissue and apoptosis of lung epithelial cells were observed;The protein expressions of alveolar epithelial sodium channel(ENaC) and α_1-Na~+,K~+-ATPase and the phosphorylation level of SGK1 were determined by Western blot.Results There was no significant difference in plasma glucose level of ALI and treatment group at 0,1,4 and 8 h after insulin infusion(t=1.330 0,0.986 0,0.565 7 and 0.724 3,P=0.204 7,0.340 7,0.580 6 and 0.480 8,respectively).Compared with ALI group,the partial pressure of oxygen in arterial blood in treatment group increased significantly(t=6.026,P <0.000 1),while the BALF protein content,alveolar epithelial permeability,lung water content and lung epithelial cells apoptosis decreased significantly(t=7.39,5.286,5.651 and 3.312,P <0.000 1,=0.000 4,=0.000 2 and=0.007 8,respectively),and the expression of α-ENaC and α_1-Na~+,K~+-ATPase and the phosphorylation level of SGK1 in lung tissue significantly increased(t=26,18.67 and 8.547,P <0.000 1,<0.000 1 and=0.000 1,respectively);Compared with the treatment group,the BALF protein content,alveolar epithelial permeability,lung water content and lung epithelial cells apoptosis increased significantly in SGK1 siRNA group(t=5.964,3.449,3.148 and 3.520,P=0.000 2,0.006 2,0.010 4 and0.016 9,respectively),while α-ENaC and α_1-Na~+,K~+-ATPase protein expression and SGK1 phosphorylation level decreased significantly(t=13,9.874 and 7.741,P <0.000 1,<0.000 1 and=0.001 5,respectively).Conclusion Exogenous insulin can alleviate the pulmonary edema in ALI mice,which might be mediated via up-regulation of the expressions of α-ENaC and α_1-Na~+,K~+-ATPase by SGK1.
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ObjectiveTo observe the regulatory effect of Xiao Qinglongtang and its ingredients on lung water transport-related proteins， and to explain the biological connotation of lung governing water movement， based on which the regulatory mechanism of Xiao Qinglongtang will be explored. MethodAccording to the composition rules of classical formula， Xiao Qinglongtang （11.22 g·kg-1）， Guizhi Gancao （2.70 g·kg-1）， Shaoyao Gancao （2.70 g·kg-1）， Jiangxinwei （3.90 g·kg-1）and Banxia Muahuang （0.032 7 g·kg-1） were prepared. The pathological model of syndrome of cold fluid accumulated in lung of rats was established by the "coldness of body + drinking cold + cold bath" method， and Xiao Qinglongtang and its ingredients were administrated to intervene with the model rats. Lung function parameters of forced vital capacity （FVC）， functional residual capacity （FRC）， mean mid-expiratory flow （MMEF）， inspiratory time （tI）， and inspiratory time （tE） were determined by lung function analyzer. Hematoxylin and eosin （HE） staining was used to observe the changes in pathological morphology. The expression of aquaporin （AQP）1， AQP5， epithelial sodium channel α subunit（α-ENaC） and Na+-K+-ATPase in lung tissues of rats， the content of tumor necrosis factor -α（TNF-α）， the mRNA expression of cyclic adenosine monophosphate （cAMP）， protein kinase A （PKA） and cAMP-response element binding protein （CREB）， and the protein expression of cAMP， PKA， CREB， and phosphorylated-CREB （p-CREB） were detected by immunohistochemistry （IHC）， enzyme-linked immunosorbent assay （ELISA）， Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and Western blot， respectively. ResultCompared with normal group， functions of FVC， FRC and MMEF in model group were significantly decreased (P<0.01)， and the time of tI and tE was significantly prolonged （P<0.05，P<0.01）. The content of TNF-α in lung tissue was significantly increased （P<0.01）. The mRNA and protein expressions of cAMP， PKA and CREB in lung tissue were significantly decreased （P<0.01）. The expression of AQP5 and α-ENAC in lung tissue decreased significantly. The alveolar cavity of rats was filled with edema fluid， surrounding tissue hyperemia， inflammatory cell infiltration, bronchial mucosa epithelial adhesion. Compared with model group， Xiao Qinglongtang and its fangyuan group could significantly enhance the FVC, FRC and MMEF functions of model rats （P<0.05，P<0.01）， and tI and tE time were shortened （P<0.05，P<0.01）. The content of TNF-α in lung tissues of Xiao Qinglongtang group， Guizhi Gancao group and Banxia Mahuang group was significantly decreased （P<0.01）. The mRNA expressions of cAMP， PKA and CREB in Xiao Qinglongtang group were significantly up-regulated （P<0.01）， and the mRNA expressions of cAMP and PKA in Guizhi Gancao， Jiangxinwei and Banxia Mahuang groups were significantly up-regulated （P<0.01）. The protein expressions of cAMP， PKA and CREB in Xiao Qinglongtang group， Guizhi Gancao group， Jiangxinwei group and Banxia Mahuang group were significantly up-regulated （P<0.01）， and the protein expression of CREB in Shaoyao Gancao group was significantly up-regulated（P<0.05）. Xiao Qinglongtang could up-regulate the positive expression of AQP5 and α-ENAC， and Guizhi Gancao group could up-regulate the positive expression of α-ENAC. Xiao Qinglongtang and its fangyuan can reduce the lung edema， inflammatory cell infiltration and bronchial mucosal adhesion of model rats. ConclusionXiao Qinglongtang and its ingredients can reduce lung edema and inhibit inflammation by improving the expression of lung water transport-related proteins AQP1， AQP5， and α-ENaC through cAMP/PKA pathway， thereby restoring the lung functions in rats with syndrome of cold fluid accumulated in lung. Na+-K+-ATPase may play an auxiliary role in the regulation of lung water transport. This provides a certain objective basis for preliminarily elucidating the connotation of lung governing water movement from the perspective of lung water transport-related proteins.
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Objective:To investigate the expression of RASAL3 in intrahepatic cholangiocarcinoma (iCCA) and the mechanism of promoting iCCA development.Methods:Tumor and paracancerous tissues were collected from 185 iCCA patients, the expression of RASAL3 was detected by immunohistochemistry, RT-qPCR and Western blot. The expression of RASAL3 and FXYD6 mRNA and protein in human cholangiocarcinoma cell line and human bile duct epithelial cells were detected with RT-qPCR and Western blot, the cell proliferation was detected with CCK-8 assay, and the activity of Na +-K +-ATPase was also detected. Results:RASAL3 was highly expressed in cholangiocarcinoma tissues and cell lines; Survival analysis showed that RASAL3 overexpression was associated with poor prognosis of cholangiocarcinoma( P<0.05) and knockdown of RASAL3 inhibits the proliferation of cholangiocarcinoma cells; Silencing RASAL3 decreases the expression of FXYD6 inhibiting the activity of Na +-K +-ATPase. Conclusion:RASAL3 is up-regulated in human cholangiocarcinoma, which can promote the occurrence and development of cholangiocarcinoma by activating FXYD6 and affecting Na +-K +-ATPase activity.
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SUMMARY: Six Lonchura striata and six Copsychus saularis birds were selected in this study, morphological index of the small intestine was measured by quantitative biology and image analysis. The changes of goblet cells and Na+/K+ATPase were detected by AB-PAS staining and ELISA to inform the different mechanisms of the digestion and absorption of nutrients between the Lonchura striata and Copsychus saularis. The villus height, crypt depth and muscle thickness of each segment of small intestine of Lonchura striata were smaller than those of Copsychus saularis, and the difference of ileum muscle thickness was significant. In addition, the ileum villus height/crypt depth (VH/CD) value of Lonchura striata was significantly less than that of Copsychus saularis. The number of goblet cells in duodenum and jejunum of Lonchura striata and Copsychus saularis had no significant difference, but the number of goblet cells in ileum of Copsychus saularis was significantly larger than that of Lonchura striata. The vitality of Na+/K+-ATPase in different intestinal segments of the Lonchura striata and the Copsychus saularis was different. The vitality of Na+/K+-ATPase in the Lonchura striata was significantly higher than that of the Copsychus saularis. It can be concluded that the digestion and absorption capacity of Copsychus saularis and Lonchura striata are significantly different, and the reason may be due to their different diets and intestinal floras.
RESUMEN: En este estudio se seleccionaron seis aves Lonchura striata y seis Copsychus saularis, a las cuales se midió mediante biología cuantitativa y análisis de imágenes el índice morfológico del intestino delgado. Los cambios de las células caliciformes y Na+/K+ATPasa se detectaron mediante tinción AB- PAS y ELISA para informar los diferentes mecanismos de digestión y absorción de nutrientes entre Lonchura striata y Copsychus saularis. La altura de las vellosidades, la profundidad de las criptas y el grosor del músculo de cada segmento del intestino delgado de Lonchura striata fueron menores que los de Copsychus saularis, y se observó una diferencia significativa en el grosor de la músculatura del íleon. Además, el valor de la altura de la vellosidad del íleon/profundidad de la cripta (VH/CD) de Lonchura striata fue significativamente menor que el de Copsychus saularis. En el número de células caliciformes del duodeno y del yeyuno de Lonchura striata y Copsychus saularis no hubo una diferencia significativa, pero el número de células caliciformes en el íleon de Copsychus saularis fue significativamente mayor que el de Lonchura striata. Hubo diferencias en la vitalidad de Na+/K+-ATPasa en diferentes segmentos intestinales de Lonchura striata y Copsychus saularis. La vitalidad de Na+/K+-ATPasa en Lonchura striata fue significativamente mayor que la de Copsychus saularis. Se puede concluir que la capacidad de digestión y absorción de Copsychus saularis y Lonchura striata son significativamente diferentes, posiblemente debido a sus distintas dietas y floras intestinales.
Subject(s)
Animals , Birds/anatomy & histology , Intestine, Small/anatomy & histology , Passeriformes/anatomy & histologyABSTRACT
Eccrine sweat glands (ESGs) perform critical functions in temperature regulation in humans. Foxa1 plays an important role in ESG maturation and sweat secretion. Its molecular mechanism, however, remains unknown. This study investigated the expression of Foxa1 and Na-K-ATPase (NKA) in rat footpads at different development stages using immunofluorescence staining, qRT-PCR, and immunoblotting. Also, bioinformatics analysis and Foxa1 overexpression and silencing were employed to evaluate Foxa1 regulation of NKA. The results demonstrated that Foxa1 was consistently expressed during the late stages of ESGs and had a significant role in secretory coil maturation during sweat secretion. Furthermore, the mRNA abundance and protein expression of NKA had similar accumulation trends to those of Foxa1, confirming their underlying connections. Bioinformatics analysis revealed that Foxa1 may interact with these two proteins via binding to conserved motifs in their promoter regions. Foxa1 gain-of-function and loss-of-function experiments in Foxa1-modified cells demonstrated that the activities of NKA were dependent on the presence of Foxa1. Collectively, these data provided evidence that Foxa1 may influence ESG development through transcriptional regulation of NKA expression.
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Acute respiratory distress syndrome (ARDS) is a critical disease with high mortality, and currently there is no specific treatment. ARDS is characterized by refractory hypoxemia secondary to pulmonary edema, but the underlying mechanism is not yet fully understood. Alveolar edema fluid is mainly actively transported and reabsorbed by sodium-water transport system. The sodium pump (Na +-K +-ATPase-mediated Na + transport) on the basal side of type Ⅱ alveolar epithelial cells (ATⅡ) is the main driving force for pulmonary edema clearance. Na +-K +-ATPase regulation is affected by many regulatory factors through a variety of ways, among which "long-term regulation" mechanism plays an important role, including positively regulating the gene transcription and protein expression of Na +-K +-ATPase. Na +-K +-ATPase can also be degraded by ubiquitin-proteasome pathway (UPP) and autophagy lysosome pathway to affect its abundance and enzyme activity, meanwhile, Na +-K +-ATPase α1 plays a key role in sodium water transport. We review the "long-term regulation" mechanism of Na +-K +-ATPase related pathways in pulmonary edema clearance and explore the possibility of new therapies for ARDS based on this mechanism, so as to provide new targets for the treatment of ARDS.
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This study aimed to assess the effects of low and high water hardness in interaction with different water pH in pacu (Piaractus mesopotamicus). Pacu juveniles were subjected to low (50 mg CaCO3 L-1 - LWH) or high water hardness (120 mg CaCO3 L-1 - HWH) at water pH of 5.5 (acidic), 7.5 (circumneutral) or 9.0 (alkaline) for 15 days. Gills and kidneys were collected (days 1, 5 and 15). Gill Na+/K+-ATPase (NKA) and vacuolar-type H+-ATPase (V-ATPase) activities were higher in alkaline pH with HWH on day 1. Gill and kidney NKA and V-ATPase activities were higher in acidic pH with LWH on day 15. Gill NKA activity of pacus under alkaline pH with LWH was higher than those exposed to HWH. Reduced antioxidant capacity in the gills and kidney and enhanced thiobarbituric acid reactive substances (TBARS) levels were demonstrated in fish exposed to acidic or alkaline pH, mainly with LWH. HWH increased glutathione-S-transferase (GST) activity and reduced TBARS levels in the gills and kidney. On day 15, GST activity was increased at acidic pH with LWH. In conclusion, circumneutral pH presents less oxidative stress and fewer variations in ATPases and HWH reduced deleterious effects in fish exposed to acidic or alkaline pH.(AU)
Este estudo objetivou analisar o efeito de baixa e alta dureza da água em interação com diferentes pH da água em pacu (Piaractus mesopotamicus). Juvenis de pacu foram submetidos a baixa (50 mg CaCO3 L-1 - BDA) ou alta dureza da água (120 mg CaCO3 L-1 - ADA) em pH da água de 5.5 ácido), 7.5 (circum-neutro) ou 9.0 (alcalino) por 15 dias. Foram coletados brânquias e rim (dias 1, 5 e 15). Atividade de Na+/K+-ATPase (NKA) e H+-ATPase do tipo vacuolar (V-ATPase) branquial foram maiores em pH alcalino com ADA no dia 1. Atividade de NKA e V-ATPase branquial e renal foram maiores em pH ácido com BDA no dia 15. Atividade de NKA branquial de pacus submetidos a pH alcalino com BDA foi maior que aqueles expostos para ADA. Em peixes expostos a pH ácido ou alcalino com BDA houve redução da capacidade antioxidante nas brânquias e rim e aumento dos níveis de "substâncias reativas ao ácido tiobarbitúrico" (TBARS). Em ADA aumentou a atividade da "glutationa-S-transferase" (GST) e reduziu níveis de TBARS nas brânquias e rim. No dia 15, a atividade da GST foi maior em pH ácido com BDA. Em conclusão, pH circum-neutro apresentou menor estresse oxidativo e poucas variações na atividade de ATPases e ADA reduziu efeitos deletérios em peixes expostos a pH ácido ou alcalino.(AU)
Subject(s)
Animals , Water/chemistry , Characiformes/anatomy & histology , Characiformes/physiology , Oxidative Stress , Hydrogen-Ion ConcentrationABSTRACT
OBJECTIVE@#This study investigated the ameliorative potential of Zingiber officinale Roscoe extract against lead-induced brain damage in rats.@*METHODS@#Thirty male rats were divided into 5 groups of 6 rats each. Lead-acetate toxicity was induced by intraperitoneal injection (10 mg/kg body weight (b.w.)) in Groups B-E. Group A (control) and Group B (lead-acetate) were left untreated; vitamin C (200 mg/kg b.w.) was administered to Group C; ethyl acetate fraction from Z. officinale extract (200 and 100 mg/kg b.w.) was administered to Group D and E by oral gavage once daily for 7 days. Changes in the content of some key marker enzymes such as acetylcholinesterase (AChE), butyrylcholinesterase (BChE), monoamine oxidase (MAO), epinephrine, dopamine, Na/K-ATPase, catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) as well as malonaldehyde (MDA) levels were determined in serum.@*RESULTS@#Exposure to lead acetate resulted in a significant decrease (P < 0.05) in the activities of BChE, AChE, Na/K-ATPase, SOD, CAT and GPx with a corresponding increase in the levels of MDA, xanthine oxidase, epinephrine, dopamine and MAO relative to the control group. Levels of all disrupted parameters were alleviated by co-administration of Z. officinale fraction and by the standard drug, vitamin C.@*CONCLUSION@#These results suggest that ethyl acetate fraction of Z. officinale extract attenuates lead-induced brain damage and might have therapeutic potential as a supplement that can be applied in lead poisoning.
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Aim To investigate the Na+,k +-ATPase, cyclc-AMP (cAMP), protein kinase A (PKA), cAMP-responsive element binding protein (CREB) and aquaporin 2 (AQP2) of the inner medullary col-lecting duct cell (IMCD3) induced by adramycin (ADR) , and to study on the protection mechanism of Danggui-Shaoyao-San (DSS) from the perspective of water-liquid balance. Methods IMCD3 cells were used as the research object. The effects of different concentrations of ADR on the proliferation of IMCD3 cells were determined by MIT assay. There were six groups in the cell experiment, namely, control group, model group, low-, medium-, and high-dose DSS extract (concentrations of 0. 8, 1.6, 3.2 g L"1) and H-89 inhibitor group. ELISA was used to detect the content of cAMP in cells. Changes of Na+ , K +-ATPase activity in cells were detected by Na+,k +-ATPase assay kit. The level of PKA, CREB and AQP2 mRNA in cells were detected by real-time PCR. The protein expression of PKA, CREB and AQP2 in IM-CD3 cells was assessed by Western blot. Results 1 x 10~8 mol L"1 ADR was the optimum concentration in IMCD3 cells. Different concentrations of DSS extract could effectively inhibit the injuiy of IMCD3 cells induced by ADR, and increase the activity of Na+,k +-ATPase and the content of cAMP in cell supernatant. DSS (1.6, 3.2 g L"1) extract could up-regulate the expressions of PKA, CREB and AQP2 mRNA and pro-tein expression (P < 0. 05 , P < 0. 01). Conclusion DSS extract can increase Na+,k +-ATPase activity and activate the cAMP-PKA-CREB pathway, thus inhibiting IMCD3 cell contraction mediated by ADR.
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The aim of this research is to investigate the effects of paeoniflorin and menthol on the physiological function of Calu-3 cell membrane during the transport of puerarin. Calu-3 cell was used as the cell model to simulate nasal mucosa tissues, and the cell membrane fluidity, Na⁺-K⁺-ATPase activity and Ca²⁺-ATPase activity were detected by fluorescence recovery after photobleaching(FRAP) and ultramicro enzyme activity testing, in order to explore the mechanism of compatible drugs on promoting puerarin transport. The results showed that when puerarin associated with low, middle and high concentration of menthol or both paeoniflorin and menthol, the fluorescence recovery rate was increased significantly, while Na⁺-K⁺-ATPase activity had no significant change and Ca²⁺-ATPase activity was enhanced significantly as compared with puerarin alone. Therefore, it was concluded that menthol had the abilit of promoting the transport and the mechanism might be related to increasing membrane fluidity and activating Ca²⁺-ATPase.
Subject(s)
Humans , Calcium-Transporting ATPases , Metabolism , Cell Line, Tumor , Cell Membrane , Glucosides , Chemistry , Isoflavones , Metabolism , Membrane Fluidity , Menthol , Chemistry , Monoterpenes , Chemistry , Sodium-Potassium-Exchanging ATPase , MetabolismABSTRACT
Objective:To evaluate the neuroprotective effect of ashwagandla extract against aluminum chloride-induced neurotoxicity in rats.Methods:Rats were divided into control,aluminumintoxicated rats treated daily with aluminum trichloride (AlCl3) (100 mg/kg,orally)for 30 d and aluminum-intoxicated animals protected by receiving daily ashwagandha extract (200 mg/kg,orally) one hour before AlCl3 administration for 30 d.Levels of lipid peroxidation,nitric oxide,reduced glutathione and tumor necrosis factor-α were measured in the cortex,hippocampus and striatum.In addition,the activities of Na+,K+,ATPase and acetylcholinesterase were determined in the three studied brain regions.Results:Aluminum increased the levels of lipid peroxidation and nitric oxide in the cortex,hippocampus and striatum and decreased the reduced glutathione level in the hippocampus and striatum.In rats protected with ashwagandha extract,non significant changes were observed in lipid peroxidation,nitric oxide and reduced glutathione.In addition,ashwagandha extracts prevented the increased activity of acetylcholinesterase and Na+,K+,ATPase induced by AlCl3 in the cortex,hippocampus and striatum.The present findings also showed that the significant increase in tumor necrosis factor-or induced by AlCl3 in the cortex and hippocampus was prevented by ashwagandha extract.Conclusions:The present results suggest that ashwagandha extract possesses antioxidant and anti-inflammatory effects against aluminum neurotoxicity.In addition,ashwagandha extract could prevent the decline in cholinergic activity by maintaining normal acetylcholinesterase activity.The later effect could recommend the use of ashwagandha as a memory enhancer.
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Objective: To evaluate the neuroprotective effect of ashwagandha extract against aluminum chloride-induced neurotoxicity in rats. Methods: Rats were divided into control, aluminum-intoxicated rats treated daily with aluminum trichloride (AlCl
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ABSTRACT It is widely known that high fat diet (HFD) can contribute to the advent of health problems. Recent studies have indicated that obesity imposes a hemodynamic overload to the kidneys. In order to further investigate such injuries, two groups of six Swiss mice each were fed with a controlled AIN93G diet or a high fat (AIN93G modified) diet for eight weeks. Blood samples were collected to determine the hormonal, lipid profile, glucose, urea, and creatinine levels. Histopathological and immunohistochemical analysis were carried out to analysis the kidney damage. Fractions of renal membranes were prepared to assess the Na,K-ATPase activity, lipid peroxidation, total cholesterol, and phospholipid content. The results indicated that the blood lipid profile, urea and creatinine was not altered by the HFD. On the other hand, it was observed in HFD diet mice elevated glucose blood levels along with an augment on insulin and a decrease on corticosterone release. HFD provoked a reduction in the diameter of the convoluted tubules and cell volume in Bowman's capsule and an increased number of positive cells with Na,K-ATPase, but reduced the Na,K-ATPase activity and the cholesterol content in the kidney cell membrane but favored the lipid peroxidation.
Subject(s)
Animals , Male , Mice , Sodium/analysis , H(+)-K(+)-Exchanging ATPase/analysis , Diet, High-Fat/adverse effects , Immunohistochemistry/methods , Cell Membrane , Data Interpretation, Statistical , Renal Insufficiency/physiopathology , Kidney/physiopathologyABSTRACT
ABSTRACT Uliginosin B, a phloroglucinol isolated from Hypericum polyanthemum Klotzsch ex Reichardt, Hypericaceae, has antidepressant-like effect in the forced swimming test in rodents and inhibits monoamines neuronal reuptake without binding to their neuronal carriers. Studies showed the involvement of Na+,K+-ATPase brain activity in depressive disorders, as well as the dependence of neuronal monoamine transport from Na+ gradient generated by Na+,K+-ATPase. This study aimed at evaluating the effect of uliginosin B on Na+,K+-ATPase activity in mice cerebral cortex and hippocampus (1 and 3 h after the last administration) as well as the influence of veratrine, a Na+ channel opener, on the antidepressant-like effect of uliginosin B. Mice were treated (p.o.) with uliginosin B single (10 mg/kg) or repeated doses (10 mg/kg/day, 3 days). Acute administration reduced the immobility in the forced swimming test and tail suspension test and increased Na+,K+-ATPase activity in cerebral cortex 1 h after treating, whereas the repeated treatment induced the antidepressant-like effect and increased the Na+,K+-ATPase activity at both times evaluated. None treatment affected the hippocampus enzyme activity. Veratrine pretreatment prevented uliginosin B antidepressant-like effect in the forced swimming test, suggesting the involvement of Na+ balance regulation on this effect. Altogether, these data indicate that uliginosin B reduces the monoamine uptake by altering Na+ gradient.
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Aim To explore the relationship between Mre11/Rad50/Nbs1 ( MRN ) complex focus formation and DNA double-strand breaks( DSBs) caused by cinob-ufagin in human hepatocellular carcinoma HepG2 cells. Methods The Na+,K+-ATPaseα1 subunit expression level in liver cancer tissues was detected by immunohis-tochemistry. After HepG2 cells were treated with 5μmol·L-1 cinobufagin for 6, 12 and 24 h, the drug-in-duced DSBs were assessed by single cell gel electro-phroesis ( SCGE ) , the gene transcription and protein levels of Mrel1, Nbs1, Rad50 and p53 were evaluated by Real time-PCR and Western blot. The cell cycle in parallel was analyzed by flow cytometry. Results The Na+, K+-ATPase α1 subunit expression level in liver cancer tissues was significantly increased compared with the tissue adjacent to carcinoma ( P <0. 05 ) . The 5μmol · L-1 cinobufagin could induce the DSBs in a time-dependent manner (P <0. 05), and it could up-regulate the gene expression levels of Mre11, Nbs1, Rad50 and p53 in HepG2 cells ( P<0. 05 ) . The pro-portions of HepG2 cells in S phase were ( 21. 32 ± 4. 21) % in the control group, and (33. 25 ± 5. 72) %, (56. 72 ± 6. 29) % and (67. 32 ± 9. 42) % in HepG2 cells treated with 5 μmol · L-1 cinobufagin for 6, 12 and 24 h, respectively. The proportions of cells in S phase in cinobufagin groups were significantly increased compared with the control group ( P<0. 05 ) . Conclu-sion Cinobufagin could induce the cell cycle arrest in liver cancer HepG2 cells by activation of Mre11/Rad50/Nbs1 Complex.
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Objective: To study the antipyretic effect of Shufeng Jiedu Capsules (SJC) and its mechanism. Methods: Using the yeast-reduced fever model of rats to study the antipyretic effects of SJC and its mechanism, collecting blood plasma and hypothalamus to measure the indicators related to body temperature such as IL-1α, IL-1β, TNF-α, IFN-γ, PGE2, cAMP, cGMP, Na-K-ATPase, IL-6, and AVP. Results: SJC can significantly decline the contents of TNF-α, IL-6, IL-1α, and IL-1β, and it can also decline the content of pyrogenic medium-cAMP, ratio of cAMP/cGMP, reduce thermogenesis and levels of Na+, K+-ATPase, and significantly enhance the content of endogenous antipyretic medium-AVP. Conclusion: SJC has a significant antipyretic effect which could decline pyrogenic medium and reduce thermogenesis through reducing the content of inflammatory cytokines-PGE2 and cytokines, it also enhances the content of endogenous antipyretic medium-AVP to play its antipyretic effect.
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OBJECTIVE: To observe whether low concentration (1×10-8 mol·L-1) of ouabain (OUA) can increase the contractility in rat cardiocytes and investigate the Na/K pump signal transduction pathways related to positive inotropic action following the low concentration of OUA. METHODS: On enzymatic isolation of rats ventricular myocytes, the Na+/K+ pump current (Ip) was by whole-cell patch-clamp, in order to observe the low concentration of OUA on Ip. The contraction of a single myocyte was assessed by a video-based motion edge-detection system. (1) To detect and compare the potentiations of 1×10-8-1×10-3 mol·L-1 OUA on the contractility in rat cardiocytes. (2) The cardiocytes were pre-treated with PP2 (1 μmol·L-1), NAC (100 μmol·L-1), PD98059(50 μmol·L-1) for 5 min, and the effects of the signals transduction inhibitors on the positive inotropic effect of 1×10-8 mol·L-1 OUA was recorded. RESULTS: The 1×10-8-1×10-3 mol·L-1 OUA increased the contractility of rat cardiocytes (P0.05). CONCLUSION: The 1×10-8-1×10-3 mol·L-1 OUA could increase the contraction amplitude of cardiocytes in rats in concentration-dependent manner. Positive inotropic effect of OUA in low concentration is related to Na/K pump signal transduction. Multiple signal pathways regulate the positive inotropic effect of 1×10-8 mol·L-1 of OUA, including the Src/ROS signal pathway.
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OBJECTIVE: To study the effects of dimethyltin chloride( DMT) on the activity of renal H~+K~+-ATPase( HKA)and Na~+K~+-ATPase( NKA) in SD rats. METHODS: i) In vitro experiment. Five specific pathogen free( SPF) healthy female SD rats were used. The kidney homogenates made with 0. 90% sodium chloride solution was added with DMT( mass concentration,1. 0 g/L) to make final concentrations of 0,1,25,125 and 625 mg/L respectively,then the HKA and NKA activities were detected by the enzyme-linked immunosorbent assay( ELISA). ii) In vivo experiment. Forty SPF healthy SD rats were divided into control group and exposure group,with 20 rats( 10 males and 10 females) in each group. The exposure group was given one-time intraperitoneal injection with DMT( 16. 000 mg / kg body weight),while the control group was given one-time intraperitoneal injection with same volume of 0. 90% sodium chloride solution. The rats were executed 1 and 24 hours after exposure. The kidney tissue was extracted to make kidney homogenates for determination of HKA and NKA activity by microplate reader. The blood from abdominal aorta was collected to measure the levels of serum K~+,Na~+and Cl-. RESULTS: i) In vitro experiment. The HKA activity was inhibited by DMT,and the effect of inhibition increased with the increase of DMT exposure dose( P < 0. 01),showing a dose-effect relationship. The DMT had no effect on NKA activity( P > 0. 05). ii) In vivo experiment. The body weight of rats at 24 hours time point in exposed group was lower than that in control group( P < 0. 01). The HKA activity of the kidney tissue in rats in exposed group was lower than that of control group( P < 0. 01). The NKA activity in kidney tissue of rats and the level serum K~+,Na~+and Cl-did not show statistical difference in main and interactive effects concerning treatment and exposure time( P > 0. 05). CONCLUSION: DMT could inhibit the HKA activity in kidney homogenates,but had no obvious effect on NKA activity.