1.Effect of rhubarb on expressions of aquaporin-2 and -4 in rat's kidney.
Jun-qiang BAO ; Feng LI ; Wen-sheng ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2008;28(12):1108-1111
OBJECTIVETo investigate the effect of rhubarb on expressions of aquaporin-2 and 4 (AQP2 and AQP4) in rat's kidney.
METHODSThirty-two SD rats were randomly divided into 4 groups, the normal control group, and the three rhubarb groups medicated via gastrogavage with low, mid and high dose of rhubarb extract (total anthraquinone) respectively. The 6 h and 24 h urine volume were measured, and the protein and mRNA expressions of AQP2 and AQP4 in renal tissue were determined with immunohistochemistry, Western blot and RT-PCR.
RESULTSNo significant difference between the control group and the low dose rhubarb treated group was found in urine volume, as well as in AQP2 and AQP4 protein and mRNA expressions. But the urine volume was obviously higher, the protein and mRNA expressions of AQP2 and AQP4 were markedly lower in rats after mid/high dose rhubarb medication respectively when compared with those in the normal controls (all P < 0.01).
CONCLUSIONRhubarb can inhibit the protein and mRNA expressions of AQP2 and AQP4 in rats' kidney, which probably is one of the mechanisms of rhubarb for diuresis.
Animals ; Aquaporin 2 ; genetics ; metabolism ; Aquaporin 4 ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Kidney ; drug effects ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry
2.Clinical characteristics and molecular genetics of seven neonates with congenital nephrogenic diabetes insipidus.
Xiaoyun DONG ; Huiru DONG ; Wenqing KANG ; Hong XIONG ; Bangli XU ; Fatao LIN ; Xuan ZHENG
Chinese Journal of Medical Genetics 2021;38(12):1185-1189
OBJECTIVE:
To explore the clinical characteristics, genetic basis and clinical treatment of seven neonates with congenital nephrogenic diabetes insipidus (NDI).
METHODS:
Clinical data of the patients were collected. High-throughput sequencing was carried out to detect potential variants. Sanger sequencing was used to verify the results.
RESULTS:
The patients were all males, with the age of onset being 10 to 21 days. All patients were admitted to the hospital for intermittent fever as the first symptom during the neonatal period. Additional symptoms had included polydipsia and polyuria. After the treatment, 5 patients had recovered, the remainders still had NDI symptoms and developmental retardation. Five children were found to harbor pathogenic variants of the AVPR2/AQP2 gene, which included one in-frame mutation of c.645_646insGCACCTACCCTGGGTATCGCC, two missense mutations of c.541C>T and c.419C>A, and two hemizygous deletions of the AVPR2/AQP2 gene. Among these, two were unreported previously. Cases 6 and 7 were a pair of twins. Both had carried homozygous missense variants of c.538G>A of the AVPR2/AQP2 gene, which was known to be pathogenic.
CONCLUSION
AVPR2/AQP2 is the main pathogenic gene for congenital NDI, for which two novel pathogenic variants have been discovered in this study. Above results have provided a basis for clinical diagnosis and genetic counseling for the affected pedigrees.
Aquaporin 2/genetics*
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Child
;
Diabetes Insipidus, Nephrogenic/genetics*
;
Diabetes Mellitus
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Humans
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Infant, Newborn
;
Male
;
Molecular Biology
;
Mutation
;
Pedigree
;
Receptors, Vasopressin/genetics*
3.Effect and mechanism of emodin for regulating aquaporin-2 expression in cultured NRK cells.
Qing LIU ; Feng LI ; Qin-you REN
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(8):871-874
OBJECTIVETo investigate the effect and mechanism of emodin for regulating aquapoin-2 (AQP2) in NRK cells cultured in vitro.
METHODSExperiments on NRK cells cultured with alpha-DMEM medium in vitro were conducted in two steps. (1) Cells were randomly divided into 4 groups: the control group, and the three emodin treated groups treated with different dosages of emodin (5, 10 and 20 mg/L) respectively. After 24 h treatment, the location of AQP2 was decided by indirect immunofluorescene, and the AQP2 protein and mRNA expression levels were detected by Western blot and semiquantive RT-PCR. (2) Cells were randomly divided into 4 groups, the control group, and the three treated groups treated respectively with 10 mg/L 8-Bromo-cAMP, 20 mg/L emodin, and 20 mg/L emodin +10 mg/L 8-Bromo-cAMP. The activity of protein kinase A (PKA) in NRK cells after 24 h treatment was determined with non-radioactive detecting method.
RESULTSAQP2 was located at the cell membrane of NRK cells. Western blot and semiquantitive RT-PCR found that AQP2 protein and mRNA expressions were significantly decreased in NRK cells of groups treated by 10 mg/L and 20 mg/L emodin (P < 0.05). PKA activity determination showed significantly decreased phosphorylation level of PKA in NRK cells of groups treated with 20 mg/L emodin group (P < 0.05).
CONCLUSIONEmodin can inhibit the genetic transcription and the translation of AQP2 gene in NRK cells, which demonstrates that the change of AQP2 expression regulated by emodin may be correlated with the diuresis effect of rhubarb, and it is likely that the regulation is going through PKA signal pathway.
Animals ; Aquaporin 2 ; genetics ; metabolism ; Cell Line ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; Emodin ; pharmacology ; Kidney ; cytology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Signal Transduction ; drug effects
4.Effect of Yixintai Granule on mRNA and protein expression levels of AQP, in renal medulla of chronic heart failure rabbits.
Yun TANG ; Zhi-Hua GUO ; Ya LI ; Tao SUN ; Gang-Qiang WU ; Li LIU ; Jie DENG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(3):333-337
OBJECTIVETo explore the effect of Yixintai Granule (YG) on mRNA and protein expression levels of AQP2 in renal medulla of chronic heart failure (CHF) rabbits.
METHODSCHF rat model was established by ear marginal vein injection of adriamycin. Successfully modeled rabbits were divided into the model group, the high (8.4 g/kg), middle (4.2 g/kg), and low dose (2.1 g/kg) YG group, and the Furosemide group (2 mg/kg). Besides, a normal control group was set up. Equal volume of physiological saline was administered to rabbits of the model group and the normal control group by gastrogavage. YG at different doses was administered to rabbits of the 3 YG groups by gastrogavage. The intervention lasted for 4 weeks, once per day. After treatment the urine volume and pathomorphological changes of renal medulla tissue were observed. mRNA and its protein expression levels of AQP2 were detected.
RESULTSCompared with the normal control group, the urine volume decreased significantly, mRNA and protein expression levels of renal medulla AQP2 increased significantly in the model group (all P < 0.01). Compared with the model group, the urine volume increased significantly, and mRNA and protein expression levels of renal medulla AQP2 decreased significantly in all medicated groups (all P < 0.01). Compared with the low dose YG group, the urine volume significantly increased and the mRNA expression level of renal medulla AQP2 significantly decreased in the middle and high dose YG groups (all P < 0.01). The expression level of AQP2 protein significantly decreased in the high dose YG group (P < 0.01). Pathological changes of the renal medulla was the most obviously seen in the model group. But they were alleviated to various degrees in all medicated groups. They were more obviously attenuated in the middle and high dose YG groups.
CONCLUSIONYG could improve CHF possibly through down-regulating mRNA and protein expression levels of AQP2 in renal medulla, and elevating the urine volume.
Animals ; Aquaporin 2 ; genetics ; metabolism ; Chronic Disease ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Heart Failure ; drug therapy ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Rats, Sprague-Dawley
5.Two Novel Mutations in the Aquaporin 2 Gene in a Girl with Congenital Nephrogenic Diabetes Insipidus.
Hae Il CHEONG ; Su Jin CHO ; Shou Huan ZHENG ; Hee Yeon CHO ; Il Soo HA ; Yong CHOI
Journal of Korean Medical Science 2005;20(6):1076-1078
Congenital nephrogenic diabetes insipidus (CNDI) is a rare inherited disorder characterized by insensitivity of the kidney to the antidiuretic effect of vasopressin. There are three inheritance patterns of CNDI: the X-linked recessive form associated with vasopressin V2 receptor gene mutations, and the autosomal recessive and dominant forms associated with aquaporin-2 gene (AQP2) mutations. The evaluation for polyuria and polydipsia in a one-month-old Korean girl revealed no response to vasopressin and confirmed the diagnosis of CNDI. Because the child was female without family history of CNDI, her disease was thought to be an autosomal recessive form. We analyzed the AQP2 gene and detected a compound heterozygous missense point mutation: (70)Ala (GCC) to Asp (GAC) in exon 1 inherited from her father and (187)Arg (CGC) to His (CAC) in exon 3 inherited from her mother. The first mutation is located within the first NPA motif of the AQP2 molecule and the second one right after the second NPA motif. This is the first report to characterize AQP2 mutations in Korean patients with autosomal recessive CNDI, and expands the spectrum of AQP2 mutations by reporting two novel mutation, (70)Ala (GCC) to Asp (GAC) and (187)Arg (CGC) to His (CAC).
Aquaporin 2/*genetics
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Base Sequence
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Child, Preschool
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DNA/genetics
;
DNA Mutational Analysis
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Diabetes Insipidus, Nephrogenic/congenital/*genetics
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Female
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Genes, Recessive
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Heterozygote
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Humans
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Infant
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Male
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Mutation, Missense
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Point Mutation
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Research Support, Non-U.S. Gov't
6.Effect of Euphorbia kansui on urination and kidney AQP2, IL-1beta and TNF-alpha mRNA expression of mice injected with normal saline.
Huiyu LI ; Fan LEI ; Yugang WANG ; Xinyue XIAO ; Jun HU ; Xianlong CHENG ; Dongming XING ; Lei HUA ; Ruichao LIN ; Lijun DU
China Journal of Chinese Materia Medica 2012;37(5):606-610
OBJECTIVETo observe the effect of Euphorbia kansui (E. KS) alcohol extracts on urination and kidney-related expressions of mice injected with normal saline and to discuss its impact on kidney.
METHODMice intraperitoneally injected with normal saline were observed for urination and changes in kidney-related histiocytic factors of after intragastrical administration of E. KS and compared with normal mice.
RESULTE. KS alcohol extracts can promote urination of mice injected with normal saline and enhance peripheral serum creatinine, with no obvious pathological change showed in tissue sections. It had a certain effect on reducing AQP2 expression and enhancing TNF-alpha expression.
CONCLUSIONEuphorbia kansui in large dose has a remarkable effect on kidney but may be accompanied with pathological reactions to some extent, especially the dose of 1.2 g x kg(-1). The pathological reactions may be related with increased serum creatinine and TNF-alpha expression.
Animals ; Aquaporin 2 ; genetics ; Euphorbia ; Interleukin-1beta ; genetics ; Kidney ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Plant Extracts ; pharmacology ; RNA, Messenger ; analysis ; Tumor Necrosis Factor-alpha ; genetics ; Urination ; drug effects
7.Relationship between regulation effect of salvia miltiorrhiza on AQP2 in kidney and promoting blood circulation and diuresis.
Xiao-Jing DONG ; Liang-Feng GUO ; Rui YAO ; Song-Yan XUE ; Feng LI
China Journal of Chinese Materia Medica 2014;39(16):3162-3165
Partial nature of "promoting blood circulation and dieresis" of Salvia Miltiorrhizain was initially demonstrated by investigating the regulation effect of AQP2 expression in kidney of trauma blood stasis model rats with the Salvia Miltiorrhizain so as to provide guidance for its clinical deployment of administration. Random allocation was taken to averagely divide 30 SD rats into two groups: 10 rats in normal group and 20 rats in blood stasis syndrome group. Trauma blood stasis rat model was established by quantitatively beating. Then the rat model group was divided into model group and salvia group. After 7 days of treatment, the rat kidney AQP2 expression was detected, the content of urine AQP2 was compared and the damaged local muscle and kidney pathological changes were observed by immunohistochemical method and western blot method. Compared with that of the normal group, rats in model group had inflammatory cells infiltration, blood stasis and edema of the injured local muscles and up-regulated AQP2 expression, decreasing urinary output, and kidney tissues blood stasis and edema (P < 0.05). On the other hand, compared with that of the model group, those parameters of rats in salvia group were all decreasing except urine output (P < 0.05). Such result indicated that Salvia Miltiorrhiza can reduce trauma blood stasis rat content of urine AQP2 and down-regulated AQP2 expression in kidney tissue, so as to reduce the reabsorption of water by renal tubular and increase urine output. The promoting blood circulation effect of Salvia Miltiorrhizain can alleviate the degree of the damaged tissue edema and encourage urine drainage. This therapy is closely related to the effect of regulating AQP2 in kidney by salvia, so the purpose of this study by verifying "promoting blood circulation and diuresis" as the mechanism for the regulation effect of the salvia on AQP2 expression.
Animals
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Aquaporin 2
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genetics
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metabolism
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Blood Circulation
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drug effects
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Diuresis
;
drug effects
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Drugs, Chinese Herbal
;
administration & dosage
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Humans
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Kidney
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blood supply
;
drug effects
;
metabolism
;
physiopathology
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Kidney Diseases
;
drug therapy
;
genetics
;
metabolism
;
physiopathology
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Male
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Rats
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Salvia miltiorrhiza
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chemistry
8.Dexmedetomidine alleviates pulmonary edema by upregulating AQP1 and AQP5 expression in rats with acute lung injury induced by lipopolysaccharide.
Yuan-xu JIANG ; Zhong-liang DAI ; Xue-ping ZHANG ; Wei ZHAO ; Qiang HUANG ; Li-kun GAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(5):684-688
This study aims to elucidate the mechanisms by which dexmedetomidine alleviates pulmonary edema in rats with acute lung injury induced by lipopolysaccharide (LPS). Male Wistar rats were randomly divided into five groups: normal saline control (NS) group, receiving intravenous 0.9% normal saline (5 mL/kg); LPS group, receiving intravenous LPS (10 mg/kg); small-dose dexmedetomidine (S) group, treated with a small dose of dexmedetomidine (0.5 μg · kg(-1) · h(-1)); medium-dose dexmedetomidine (M) group, treated with a medium dose of dexmedetomidine (2.5 μg · kg(-1) · h(-1)); high-dose dexmedetomidine (H) group, treated with a high dose of dexmedetomidine (5 μg · kg(-1) · h(-1)). The rats were sacrificed 6 h after intravenous injection of LPS or NS, and the lungs were removed for evaluating histological characteristics and determining the lung wet/dry weight ratio (W/D). The levels of tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) in the lung tissues were assessed by enzyme- linked immunosorbent assay (ELISA). The mRNA and protein expression levels of aquaporin-1 (AQP1) and aquaporin-5 (AQP5) were detected by RT-PCR, immunohistochemistry, and Western blotting. The lung tissues from the LPS groups were significantly damaged, which were less pronounced in the H group but not in the small-dose dexmedetomidine group or medium-dose dexmedetomidine group. The W/D and the concentrations of TNF-α and IL-1β in the pulmonary tissues were increased in the LPS group as compared with those in NS group, which were reduced in the H group but not in S group or M group (P<0.01). The expression of AQP1 and AQP5 was lower in the LPS group than in the NS group, and significantly increased in the H group but not in the S group or M group (P<0.01). Our findings suggest that dexmedetomidine may alleviate pulmonary edema by increasing the expression of AQP-1 and AQP-5.
Acute Lung Injury
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chemically induced
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drug therapy
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genetics
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pathology
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Adrenergic alpha-2 Receptor Agonists
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pharmacology
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Animals
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Aquaporin 1
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agonists
;
genetics
;
immunology
;
Aquaporin 5
;
agonists
;
genetics
;
immunology
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Dexmedetomidine
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pharmacology
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Dose-Response Relationship, Drug
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Drug Administration Schedule
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Gene Expression Regulation
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Injections, Intravenous
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Interleukin-1beta
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antagonists & inhibitors
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genetics
;
immunology
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Lipopolysaccharides
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Lung
;
drug effects
;
immunology
;
pathology
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Male
;
Organ Size
;
drug effects
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Pulmonary Edema
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chemically induced
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drug therapy
;
genetics
;
pathology
;
Rats
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Rats, Wistar
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Signal Transduction
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Transcription, Genetic
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Tumor Necrosis Factor-alpha
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antagonists & inhibitors
;
genetics
;
immunology
9.Effect of total anthraquinone in rheum on aquaporin 2 expression in rat distal colon.
Jun-qiang BAO ; Feng LI ; Wen-sheng ZHANG ; Yan-bo XU ; Qing LIU ; Xin WANG ; Yi-ling ZHAO ; Chang-hai WANG
China Journal of Chinese Materia Medica 2008;33(14):1732-1735
OBJECTIVETo investigate effect of total anthraquinone in rheum on aquaporin 2 expression in rat distal colon.
METHODSD rats were randomly divided into control group, low dose group, middle dose group and high dose group. Gavaged to control group, and treated group were administered saline and total anthraquinone in rheum with dosage of 0.14, 2.5, 4.5 g x kg(-1) x d(-1), respectively. All rats were put sacrificed after 5 days and stool in full length colon was gently collected to detect water content stool. Distal colon was removed to detect AQP2 expression with immunohistochemistry, western blot and RT-PCR.
RESULTNo diarrhea was found in low dose group and control group, there were not significant difference water content of stool and AQP2 expression between low dose group and control group. However, soft feces and loose stools occurred in diarrheic dose group, loose stools and watery stool appeared in high dose group. Stool water content increased in diarrheic dose group and High dose group, expression of AQP2 decreased evidently in these two groups (P < 0.01).
CONCLUSIONTotal anthraquinone in rheum can reduce the transcription and translation of AQP2 in rats' distal colon, increase fecal water content, which probably is one of the mechanisms of diarrhea caused by total anthraquinone in rheum.
Animals ; Anthraquinones ; chemistry ; pharmacology ; Aquaporin 2 ; genetics ; metabolism ; Colon ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Immunohistochemistry ; In Vitro Techniques ; Male ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Rheum ; chemistry
10.Effects of furosemide, antisterone and hydrochlorothiazide on expression of kidney aquaporin-2 gene and urine aquaporin-2 excretion in rats.
Sheng LIN ; Ding-li XU ; Wen-yan LAI ; Qian-bo SHEN ; Jing-xiong GAO ; Bin ZHANG ; Hao REN
Journal of Southern Medical University 2007;27(6):802-804
OBJECTIVETo investigate effects of the furosemide, antisterone and hydrochlorothiazide on expression of kidney aquaporin-2 (AQP(2)) gene and urine aquaporin-2 excretion in rats.
METHODSForty SD rats were randomized into 4 groups, namely the control group, furosemide group, antisterone group and hydrochlorothiazide group with corresponding treatment. Blood and urine samples were collected from the rats for measurement of serum Na(+), urine volume and urine osmolality during medication. Semi-quantitative RT-PCR was performed to measure kidney inner medullary AQP(2) and vasopressin V(2)-R mRNA. Western blotting was employed to detect kidney inner medullary AQP(2) protein expression. Urine AQP(2) concentration was measured by enzyme-linked immunosorbent assay (ELISA).
RESULTUrine volume and urinary AQP(2) excretion were both increased in rats treated with the 3 drugs as compared with that of the control group. However, urine osmolality was lower in furosemide group but higher inhydrochlorothiazide and antisterone groups than in the control group (P<0.05). The kidney inner medullary AQP(2) mRNA, V(2)-R mRNA and AQP(2) protein expression of furosemide group increased in comparison with that of the control group (Plt;0.05). In hydrochlorothiazide group, however, the above parameters were all decreased (Plt;0.05).
CONCLUSIONThe three classes of diuretics can all increase the excretion of the urinary AQP(2) but have different effects on the inner medullary AQP(2) mRNA and protein expression in normal rats. Hydrochlorothiazide reduces kidney AQP(2) mRNA and protein expression, while furosemide increased kidney AQP(2) gene expression.
Animals ; Aquaporin 2 ; genetics ; metabolism ; urine ; Blotting, Western ; Diuretics ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Furosemide ; pharmacology ; Gene Expression ; drug effects ; Hydrochlorothiazide ; pharmacology ; Kidney ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction