scholarly journals Gene expression and enzyme activities of carbonic anhydrase and glutaminase in rat kidneys induced by chronic systemic hypoxia

2015 ◽  
Vol 24 (3) ◽  
pp. 139-45
Author(s):  
Andi N.K. Syarifin ◽  
Sri W.A. Jusman ◽  
Mohamad Sadikin

Background: Hypoxia can cause acidosis. Kidney plays an essential role in maintaining acid-base balance, which involves the activities of carbonic anhydrase (CA) and glutaminase (GLS). This study is aimed to determine the expression and activities of the CA9 and GLS1 enzymes in relation to hypoxia inducible factor-1α (HIF-1α), a transcription factor protein which is a marker of hypoxia.Methods: This study was an in vivo experimental study with coupled paralel design. used 25 male Sprague-Dawley rats weighing 150-200 g. Rats were divided into 5 groups: the control group (normoxic condition) and 4 treatment groups. The latter were kept in a hypoxic chamber (10% O2: 90% N2) for 1, 3, 5 and 7 days. All rats were euthanized after treatment, kidneys excised, tissues homogenized and investigated for gene expression of CA9, GLS1 and HIF-1α. On protein level, total enzymatic activities of CA and GLS and protein of HIF-1α were also investigated. Data were analyzed statistically using ANOVA for significance, and as its alternative, used Mann-Whitney and Kruskal-Wallis test.Results: Results showed that HIF-1α mRNA increased during hypoxia, but not HIF-1α protein. It seemed that acidosis occurs in kidney tissue, indicated by increased CA9 and GLS1 mRNA expression and specific activity of total CA and GLS1. Expression of CA9 and GLS1 mRNA both showed strong positive correlation with HIF-1α mRNA, but not with HIF-1α protein.Conclusion: It is suggested that during chronic systemic hypoxia, gene expression of CA9 and GLS1 and their enzyme activities were increased as a response to acidosis and related with the expression of HIF-1α mRNA.

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
H R Helmi ◽  
A P Sunjaya ◽  
D Limanan ◽  
A R Prijanti ◽  
S W A Jusman ◽  
...  

Abstract Background Apelin, an adipokine peptide and its receptor has recently emerged as a key signaling pathway in maintaining cardiac performance at chronic pressure loads. Apelin has been linked to ventricular dysfunction and therefore maybe of pathophysiologic relevance as a candidate biomarker in HF patients. Purpose This study aims to investigate Apelin-13 gene expression and level, and Apelin receptor (APJ) level in a rat model of heart failure induced by chronic systemic hypoxia and their correlation to BNP-45 gene expression and level, the current gold standard biomarker for heart failure, and to cardiac histopathologic changes. The effect of chronic systemic hypoxia on cardiac hypertrophy, remodeling and heart failure parameters is also of interest. Methods Twenty-eight male Sprague-Dawley rats (8–12 weeks of age) were placed in special hypoxic chambers divided into 7 groups – a control group provided with normoxia (atmospheric O2 levels) and 6 exposure groups exposed to hypoxia (8% O2) for 6 hours, 1, 3, 5, 7 and 14 days respectively prior to measurement. Changes in the expression of Apelin and BNP-45 were measured using quantitative real-time PCR, whereas changes in Apelin-13, APJ and BNP-45 levels were measured using ELISA. Histopathology staining using Hematoxylin and Eosin was performed on cardiac tissues post-termination. Results Compared to control, BNP-45 mRNA expression in the hypoxic heart was only significantly different in day 14, whereas, Apelin mRNA expression had showed significantly higher values starting from day 7 onward. This is in line with the evidence of cardiac hypertrophy based on histopathologic examination present from day 7 onwards. BNP-45 and Apelin-13 levels were significantly higher compared to control from day 5 onwards with a peak on day 7. Although significantly higher than control, Apelin-13 and BNP-45 level decreases in day 14 as compared to day 7. Mean APJ levels showed a similar profile with Apelin-13 and BNP-45 levels with a peak in day 7 (4.619 ng/mL). The cardiac Apelin-13 level shows strong significant correlation with BNP-45 levels (r 0.823, p-value 0.0001). There was also a strong significant correlation between APJ receptor levels with Apelin-13 (r 0.9029, p-value 0.001) and BNP-45 (r 0.9062, p-value 0.0009) levels. Apelin-13, APJ and BNP-45 levels also showed strong significant positive correlation to the duration of hypoxia exposure. Conclusion Chronic (≥5 days) and not acute systemic hypoxia in an experimental rat model leads to increase in Apelin-13, APJ and BNP-45 levels. Apelin-13 and BNP-45 were found to significantly increase from 5 days onwards. Apelin mRNA expression was found to show significant increase earlier compared to BNP-45 mRNA expression. Hence, Apelin may serve as a new candidate biomarker for detection of HF due to oxidative stress compared to BNP-45. Exposure to chronic systemic hypoxia can serve as an easily replicable rat model for heart failure. Acknowledgement/Funding Department of Biochemistry and Molecular Biology, Faculty of Medicine, Tarumanagara University, Jakarta, Indonesia


2015 ◽  
Vol 24 (3) ◽  
pp. 133-8
Author(s):  
Ulfah Lutfiah ◽  
Sri W.A. Jusman ◽  
Mochammad Sadikin

Background: Hypoxia may induce gastric ulcer associated with excessive hidrogen chloride (HCl) secretion. Synthesis of HCl involves 2 enzymes, H,K-ATPase and carbonic anhydrase (CA). This study aimed to clarify the underlying cause of gastric ulcer in chronic hypoxic condition, by investigating the H,K-ATPase and CA9 response in rats.Methods: This study was an in vivo experiment, to know the relationship between hypoxia to expression of H,K-ATPase and CA9 mRNA, and H,K-ATPase and total CA specific activity of chronic systemic rat gastric hypoxia. The result was compared to control. Data was analyzed by SPSS. If the data distribution was normal and homogeneous, ANOVA and LSD post-hoc test were used. However, if the distribution was not normal and not homogeneous, and still as such after transformation, data was treated in non-parametric using Kruskal-Wallis and Mann Whitney test. Twenty five male Sprague-Dawley rats were divided into 5 groups: rats undergoing hypoxia for 1, 3, 5, and 7 days placed in hypoxia chamber (10% O2, 90% N2), and one control group. Following this treatment, stomach of the rats was extracted and homogenized. Expression of H,K-ATPase and CA9 mRNA was measured using real time RT-PCR. Specific activity of H,K-ATPase was measured using phosphate standard solution, and specific activity of total CA was measured using p-nitrophenol solution.Results: The expression of H,K-ATPase mRNA was higher in the first day (2.159), and drastically lowered from the third to seventh day (0.289; 0.108; 0.062). Specific activities of H,K-ATPase was slightly higher in the first day (0.765), then was lowered in the third (0.685) and fifth day (0.655), and was higher in the seventh day (0.884). The expression of CA9 mRNA was lowered progressively from the first to seventh day (0.84; 0.766; 0.736; 0.343). Specific activities of total CA was low in the first day (0.083), and was higher from the third to seventh day (0.111; 0.136; 0.144).Conclusion: In hypoxia condition, expression of H,K-ATPase and CA9 mRNA were decreased, but the specific activity of H,K-ATPase and total CA were increased.


Animals ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 2585
Author(s):  
Fang Jiang ◽  
Yan Lin ◽  
Linghong Miao ◽  
Jingyuan Hao

The ability of bamboo charcoal to reduce the negative effects of high dietary selenium (Se) concentrations was assessed by feeding juvenile blunt snout bream (Megalobrama amblycephala) one of five Se-rich diets (1.5 mg/kg Se; 36% protein, 8.7% lipid) containing graded levels (0–4 g/kg) of bamboo charcoal powder for eight weeks. There were four tanks (350 L) of fish (initial weight 16.0 ± 0.5 g) for each treatment, and the fish were fed to satiation four times each day. At the end of the feeding trial, all of the fish from each tank were weighed to calculate the growth performance. Blood samples were firstly obtained to collect plasma for the biochemical indexes determination. Liver tissues were then collected to determine the antioxidant enzyme activities and gene expression. Dorsal muscles were also collected to determine the nutrient composition. The results show that when the bamboo charcoal content in the Se-rich feed ranged between 0 and 3 g/kg, the weight growth rate (WGR) and specific growth rate (SGR) values increased with the higher dietary bamboo charcoal content, and the maximum WGR and SGR values were achieved when the bamboo charcoal content in the Se-rich feed was 2–3 g/kg (p < 0.05). The Se content in muscle tissues decreased significantly with the increased bamboo charcoal content (p < 0.05) in the Se-rich feed, which ranged from 0 to 4 g/kg. When the bamboo charcoal content in the Se-rich feed was 2–3 g/kg, the levels of glucose (GLU) and albumin (ALB) in plasma reached a maximum (p < 0.05), whereas the level of alkaline phosphatase (ALP) reached a minimum (p < 0.05). Additionally, the activities of catalase (CAT), total superoxide dismutase (T-SOD), total antioxidative capacity (T-AOC), and glutathione peroxidase (GSH-Px) were significantly enhanced (p < 0.05) when the bamboo charcoal content was 3 g/kg. In contrast, the malondialdehyde (MDA) level increased sharply when the bamboo charcoal content in the Se-rich feed was 1 g/kg, compared to the control group and the groups supplemented with 2–3 g/kg bamboo charcoal (p < 0.05). Regarding mRNA-level gene expression, the results show that dietary supplementation with 0 to 3 g/kg of bamboo charcoal increased the expression of keap1 and nrf2, whereas nfkb expression was inhibited (p < 0.05). The mRNA expression of the antioxidant enzymes cat, gpx, and mn-sod was consistently enhanced in the group fed with the 3 g/kg bamboo charcoal diet (p < 0.05). The expression of the pro-inflammatory cytokines tnfα and tgfβ was inhibited in the groups supplemented with 2–3 g/kg bamboo charcoal, whereas the expression of anti-inflammatory cytokines (il10) increased in the bamboo charcoal supplementation groups compared to the control group (p < 0.05). Generally, supplementation with 2–3 g/kg of bamboo charcoal in Se-rich feed improved the growth performance, physiological status, and antioxidant enzyme activities of blunt snout bream. Moreover, bamboo charcoal supplementation in Se-rich diets stimulated the antioxidant system and inhibited the inflammatory response by activating Nrf2-Keap1 and suppressing NF-κB.


2021 ◽  
Author(s):  
Atta Mohammad Dost ◽  
Mehmet Gunata ◽  
Onural Ozhan ◽  
Azibe Yildiz ◽  
Nigar Vardi ◽  
...  

Abstract Amikacin (AK) is frequently used in the treatment of gram-negative and some gram-positive infections. However, its use is limited due to nephrotoxicity due to the increase in reactive oxygen radicals. The aim of this study was to investigate the role of carvacrol (CAR) against AK-induced nephrotoxicity in rats. Thirty-two Sprague Dawley rats were randomly divided into four groups as control (Vehicle), AK (400 mg/kg), CAR + AK (80 mg/kg CAR + 400 mg/kg AK), and AK + CAR (400 mg/kg AK + 80 mg/kg CAR) groups. AK and CAR were administered via intramuscular and per-oral for 7 days, respectively. Blood and kidney tissue samples were taken at the end of the experiment. Renal function and histopathological changes were compared, and the relevant parameters of oxidative stress and inflammation were detected. Histopathological findings (necrotic changes and dilatation and inflammatory cell infiltration) significantly increased in the AK group compared to the control group. Also, the rats in the AK group lost weight significantly. It was found that CAR treatment before and after AK significantly improved nephrotoxicity histopathologically (p < 0.05). However, this improvement was not detected biochemically. These results show that CAR treatment before and after AK improves nephrotoxicity in the histopathological level.


2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Chang Ho Lee ◽  
Kyung Woon Kim ◽  
So Min Lee ◽  
So Young Kim

Abstract Background This study aimed to investigate the changes in the expression of hippocampal genes upon acute noise exposure. Methods Three-week-old Sprague–Dawley rats were assigned to control (n = 15) and noise (n = 15) groups. White noise (2–20 kHz, 115 dB sound pressure level [SPL]) was delivered for 4 h per day for 3 days to the noise group. All rats were sacrificed on the last day of noise exposure, and gene expression in the hippocampus was analyzed using a microarray. Pathway analyses were conducted for genes that showed differential expression ≥ 1.5-fold and P ≤ 0.05 compared to the control group. The genes included in the putative pathways were measured using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Results Thirty-eight upregulated genes and 81 downregulated genes were identified. The pathway analyses revealed that upregulated genes were involved in the cellular responses to external stimuli and immune system pathways. qRT-PCR confirmed the upregulation of the involved genes. The downregulated genes were involved in neuronal systems and synapse-related pathways, and qRT-PCR confirmed the downregulation of the involved genes. Conclusions Acute noise exposure upregulated the expression of immune-related genes and downregulated the expression of neurotransmission-related genes in the hippocampus.


Author(s):  
Koffi Kouame ◽  
Aniekan Imo Peter ◽  
Edidiong Nnamso Akang ◽  
Roshila Moodley ◽  
Edwin Coleridge Naidu ◽  
...  

This study investigated the antidiabetic activity of Cinnamomum cassia (C. cassia, Cc) silver nanoparticles (CcAgNPS) and effects of C. cassia on the kidneys of rats with induced type 2 diabetes. Twenty-four Sprague-Dawley rats weighing 250 ± 20 g were induced with diabetes by intraperitoneal injection of streptozotocin (STZ, 60 mg/kg). Animals were randomly assigned to one of four groups (n = 6) and treated for eight weeks with normal saline (control, group A), 5 mg/kg of CcAgNPs (group B), 10 mg/kg of CcAgNPs (group C), or 200 mg/kg of Cc (group D). Body weight and fasting blood glucose (FBG) was measured weekly and fortnightly, respectively. At the end of experiments, animals were euthanized, blood and kidney tissue samples were collected for biochemistry (oxidative stress markers and renal function parameters) and kidneys were harvested for histology (PAS and HE staining). Body weight was significantly higher in group B and C vs. control (p < 0.05), while no significant differences were observed in the kidney-to-body weight ratio between groups. FBG, glutathione, malondialdehyde, alanine aminotransferase, aspartate aminotransferase, serum urea and creatinine were significantly lower in group B, C and/or D vs. control (all p < 0.05). In group A, severe distortion of the glomerular network was observed, marked by the loss of capsular integrity, thickened basement membrane, tubular cells with pyknotic nuclei, vacuolization, and interstitial space with infiltrations. These adverse effects were mitigated by 5 mg/kg and 10 mg/kg of CcAgNPs. Our study confirms structural and functional damage to kidneys caused by diabetes. CcAgNPs have a regenerative potential in diabetes-induced kidney damage and may be used as an antidiabetic agent.


2019 ◽  
Vol 8 (2) ◽  
pp. 147 ◽  
Author(s):  
Yi-Hsin Chen ◽  
Yun-Ching Fu ◽  
Ming-Ju Wu

Contrast is widely used in invasive image examinations such as computed tomography (CT) and angiography; however, the risk of contrast-induced nephropathy (CIN) is high. The aim of this study was to investigate the protective effect of resveratrol in a rat model of CIN. Sprague-Dawley rats were divided into four groups: the control group (0.9% saline infusion only); resveratrol group (RSV, resveratrol, 30 mg/kg); contrast media group (CIN); and resveratrol + contrast media group (RCIN, resveratrol 30 mg/kg 60 min before CIN). CIN was induced via an intravenous injection of a single dose of indomethacin (10 mg/kg), one dose of N-nitro-L-arginine methyl ester (10 mg/kg), and a single dose of contrast medium iopromide (2 g/kg). Blood urea nitrogen, creatinine, and neutrophil gelatinase-associated lipocalin (NGAL) were higher in the CIN group compared to the other groups. Histopathological tubule injury scores were also higher in the CIN group compared to the other groups (p < 0.01). NLPR3 in kidney tissue were higher in the CIN group compared to the other groups; however, these results were improved by resveratrol in the RCIN group compared with the CIN group. The expressions of IL-1β and the percentage of apoptotic cells were higher in the CIN group than in the control and RSV groups, but they were lower in the RCIN group than in the CIN group. The expression of cleaved caspase-3 was higher in the CIN group than in the control and RSV groups, but lower in the RCIN group than in the CIN group. Resveratrol treatment attenuated both injury processes and apoptosis and inhibited the inflammasome pathway in this rat CIN model.


2020 ◽  
Vol 27 (7) ◽  
pp. 614-622
Author(s):  
Ahmet Savcı ◽  
Enver Fehim Koçpınar ◽  
Harun Budak ◽  
Mehmet Çiftci ◽  
Melda Şişecioğlu

Background: Free radicals lead to destruction in various organs of the organism. The improper use of antibiotics increases the formation of free radicals and causes oxidative stress. Objective: In this study, it was aimed to determine the effects of gentamicin, amoxicillin, and cefazolin antibiotics on the mouse heart. Methods: 20 male mice were divided into 4 groups (1st control, 2nd amoxicillin, 3rd cefazolin, and 4th gentamicin groups). The mice in the experimental groups were administered antibiotics intraperitoneally at a dose of 100 mg / kg for 6 days. The control group received normal saline in the same way. The gene expression levels and enzyme activities of SOD, CAT, GPx, GR, GST, and G6PD antioxidant enzymes were investigated. Results : GSH levels decreased in both the amoxicillin and cefazolin groups, while GR, CAT, and SOD enzyme activities increased. In the amoxicillin group, Gr, Gst, Cat, and Sod gene expression levels increased. Conclusion: As a result, it was concluded that amoxicillin and cefazolin caused oxidative stress in the heart, however, gentamicin did not cause any effects.


Author(s):  
SARI BEMA RAMDIKA ◽  
ENDANG MAHATI ◽  
ANANG MOHAMAD LEGOWO ◽  
MUFLIHATUL MUNIROH ◽  
YORA NINDITA ◽  
...  

Objective: This study aims to determine the effect of fortified dadih with vitamin D3 on IL-6 expression level and the concentration of caecum SCFA in obese rats. Methods: A total of 30 male Sprague Dawley rats were divided into five equal groups: healthy-control-(K-), obese-control-(K+), obese-intervention-(X1, X2, and X3). K(+), X1, X2, and X3 were in obesity conditions, which was induced by a high-fat sucrose diet (HFSD) and K(-) as a healthy-control-group. Furthermore, vitamin D3-fortified dadih at doses of 4 g/200 g-body-weight/d, dadih only at doses of 4 g/200 g-body-weight/d, and vitamin D3 only at 36 IU/200 g-body-weight/d was administered to X1, X2, and X3 groups, respectively. Results: Treatment using fortified dadih with vitamin D3 showed significantly reduce weight gain (p<0.05) compare to K(+) and X2. In addition, X1 showed a decreased level of Interleukin-6 expression (p<0.05) than K(+), X2, and X3 groups but higher than K(-). Also, it showed the highest total SCFA, acetate, and propionate concentration (p<0.05). However, a moderately negative correlation was discovered between the pair of total SCFA and Interleukin-6 expression, acetate and Interleukin-6 expression, SCFA and body weight, propionate and body weight, butyrate and body weight. On the contrary, a strong positive correlation was observed between the pair of Interleukin-6 expression levels and body weight. Conclusion: This study shows that fortified dadih with vitamin D3 from fermented foods improve the expression level of Interleukin-6 and increase the production of SCFA. Also, they improve intestinal homeostasis because of the increased SCFA production.


2016 ◽  
Vol 66 (3) ◽  
pp. 294-303 ◽  
Author(s):  
Emília Hijová ◽  
Jozef Kuzma ◽  
Ladislav Strojný ◽  
Alojz Bomba ◽  
Izabela Bertková ◽  
...  

Abstract The effect of the probiotic strain Lactobacillus plantarum LS/07 on intestinal bacterial enzyme activities – β-glucuronidase (β-GLUCUR), β-galactosidase (β-GAL), and β-glucosidase (β-GLU) in the prevention of cancer, atherosclerosis and dysbiosis was investigated. Male Sprague-Dawley rats were randomly divided into 12 experimental groups: C (control group), AT (atherosclerotic group), CC (carcinogenic group), and then each group in combination with antibiotics and probiotics individually and each group in double combination on antibiotic and probiotic. In the control group the β-glucuronidase activity did not change throughout the experiment. High fat diet in the atherosclerotic group significantly increased the activity of β-glucuronidase (p<0.001) and β-glucosidase (p<0.01). Azoxymethane application in the carcinogenic group significantly increased β-glucuronidase (p<0.01), but reduced β-glucosidase (p<0.01). Daily application of probiotics individually and in double combination with antibiotics increased the activity of β-galactosidase, and β-glucosidase, and positively decreased the level of β-glucuronidase. In the control antibiotic group β-glucuronidase was significantly increased (p<0.05), and β-glucosidase decreased (p<0.01) which can be caused by a change of microflora in favor of coliform bacteria. These finding indicate the positive effects of probiotic Lactobacillus plantarum LS/07 which allows its use in disease prevention in human and veterinary medicine.


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