bluuebunny/crossref_metadata_2025_split huggingface dataset:
N‐acetylcysteine Effects on Renal Ischemic Injury Associated to Chronic High Sodium Intake
Authors: Pereira Rafael Canavel , Romão Carolina , Gracioli Fabiana , Shimizu Maria , Oliveira Ivone , Furukawa Luzia 
Published: November 2023

IntroductionAcute episodes of severe renal ischemia result in acute kidney injury. These episodes are followed by a characteristic recovery and repair response, which renal function appear completely restored within one mouth. However, the chronic effects of such an injury associated to high sodium intake and N‐acetylcysteine (NAC) have not been well investigated.AimTo evaluate the NAC effects associated with high sodium diet intake on rats submitted to bilateral ischemia and reperfusion (IR).Material and MethodsEight weeks old male Wistar rats were fed normal sodium diet (1,3% NaCl, 25,0% protein – NSD) or high sodium diet (8% NaCl, 25,0% protein ‐ HSD). Also, at same time, both diet groups received or not NAC (600 mg/L) in the drink water. The animals were submitted or not (SHAM) to bilateral IR for 45 minutes at 10 weeks old. At 22 weeks old, the animals had blood pressure, water intake, urine volume, right kidney and heart masses, plasma renin activity (PRA); serum uric acid, total antioxidant status and thiobarbituric acid reactive species (TBARS) and renal interstitial fibrosis measured.ConclusionThe chronic NAC treatment associated to high sodium intake in renal injury, prevented the increase of blood pressure, kidney and heart masses. Also, it decreased lipid peroxidation and renal interstitial fibrosis, showing that it has beneficial effects in rats fed with high sodium diet.Support or Funding InformationCAPES and FAPESPResults
Variables from male Wistar rats fed normal (NSD) or high (HSD) sodium diet submitted or not (SHAM) to ischemia and reperfusion (IR) and received or not N‐acetylcysteine (NAC) at 22 weeks old.

SHAM
IR
IR+NAC

Variable
NSDn=8
HSDn=7
NSDn=9
HSDn=7
NSDn=10
HSDn=10

Blood‐pressure (mmHg)
147,5±11,2
179,0±10,0*
144,0±12,3
177,0±11,1#
157,8±11,0
170,8±9,8

Water‐intake (mL/24h)
30,0±18,5
87,1±19,8*
30,3±9,0
93,0±45,0#
31,1±16,7
118,6±46,0&

Urine volume (mL/24h)
13,5±3,7
68,9±17,75*
25,2±13,3*
96,0±30,6#
16,0±7,7
71,5±42,3&

Right kidney mass (g/femur)
0,5±0,05
0,6±0,1
0,5±0,2
0,8±0,
0,5±0,2
0,7±0,2

Heart mass (g/femur)
0,4±0,04
0,4±0,1
0,4±0,03
0,69±0,2
0.4±0,04
0,4±0,1

PRA (ngANG/mL/hr)
14,4±7,7
0,09±0,01*
5,6±2,9
0,06±0,1#
9,4±2,0
0,9±1,1&

Uric acid (mg/dL)
1,4±0,2
1,0±0,1#
1,2±0,3
1,8±0,5$
1,5±0,2
2,25±1,4

Total antioxidant status (mmol/L)
1,3±0,1
1,3±0,1
1,3±0,1
1,4±0,03
1,2±0,1
1,3±0,1

TBARS (mmol/L)
2,7±0,65
3,2±0,4
3,7±0,8
3,2±0,7
0,9±0,5*#
0,6±0,4&$

Renal interstitial fibroses (%)
0,6±0,2
0,5±0,2
1,8±0,7*
2,3±1,8#
1,5±0,6
1,5±0,5

Data are expressed as mean ± SD.
p<0.05 vs. Sham‐NSD;
p<0,05 vs. IR‐NSD;
p<0,05 vs. IR+NAC ‐ NSD;
p<0,05 vs. Sham‐HSD.
Selective cannabinoid 2 receptor stimulation reduces tubular epithelial cell injury following renal ischemia‐reperfusion injury
Authors: Pressly Jeffrey , Mustafa Suni , Moore Bob M , Park Frank 
Published: November 2023

Acute kidney injury (AKI) is an increasingly common clinical problem and has been studied for its association with chronic kidney disease progression. Currently, therapies to treat AKI are not available, so further insight into new therapeutic targets are essential in the treatment of this disease. In this study, a novel cannabinoid receptor type 2 (CB2) agonist, SMM‐295, was designed and synthesized for testing as a potential therapeutic to treat acute kidney injury. Molecular docking of SMM‐295 into a CB2 active state model showed that SMM‐295 interacts well with key amino acids to stabilize the active state. In cell culture, SMM‐295 was shown to exhibit ~100‐fold selectivity for CB2‐ versus CB1‐expressing 293 cells. Moreover, the CLogP values were consistent with a more aqueous solubility for SMM‐295 (CLogP=4.96) compared with a commonly used CB2 agonist, JWH‐133 (CLogP=5.72). Subsequent in vivo testing of the CB2 agonist was performed using a mouse model of AKI. Bilateral ischemia‐reperfusion injury was performed in mice, which is a common model to mimic human AKI, and SMM‐295 (6 mg/kg) was immediately administered IP to the mice upon reperfusion of the kidneys following the ischemic episode and every 24 hours until the end of the experiment. Histological damage assessment after 48 hours following reperfusion demonstrated around a 50% reduction (P<0.05) in tubular damage in the presence of SMM‐295 (n=5) compared to vehicle‐treated mice (n=4). This was consistent with a significant reduction (P<0.05) in both plasma creatinine and NGAL by approximately 60% with SMM‐295 administration (n=5) versus vehicle alone (n=7). SMM‐295 also showed better therapeutic results compared to the commercially available CB2 agonist JWH‐133 which also led to a drop in both serum creatinine levels and tubular injury, but did not reach statistical significance (P>0.05). Conversely, CB2 inverse agonist, SMM‐189, showed significant increases (P<0.05) in both serum creatinine (152%; n=4) at 24 hours and tubular injury (47%; n=3) at 48 hours following IRI. SMM‐295 treated mice (n=5) also showed significant reduction (32%; P<0.05) in PCNA‐positive cells in the outer medulla compared to the vehicle treated group (n=4) 48 hours following IRI. Western blot analysis of kidney lysates in SMM‐295 treated mice (n=5) at 48 hours following IRI demonstrate increased levels of cell survival proteins including total and active forms of Akt as well as Bcl‐2 compared to vehicle treated mice (n=3). These data would suggest that selective CB2 activation can play a beneficial role in protecting tubular epithelial cells following transient injury stimulus application to the kidneys, and may open the door to new small molecule activators of the cannabinoid systems for renal‐directed therapies.Support or Funding InformationThis study was partially funded by NIH RO1 DK090123 (F.P.) and institutional funds (F.P.).
Consumption of Gamma‐oryzanol prevents kidney disease and maintains redox balance in a model of obesity induced by high sugar‐fat diet
Authors: Francisqueti Fabiane , Mangegale Maria Clara , Garcia Jessica , Ferron Artur , Moreto Fernando , Silva Vanessa , Ferreira Ana Lucia , Correa Camila 
Published: November 2023

Aimto investigate the effect of Gamma‐oryzanol (yOz) on renal disease, redox state and agressor factors of animals submitted to high sugar‐fat diet (HSF).Methods32 wistar male rats were divided into 4 groups (n=8 animals/group): control (C), control + yOZ (CyOZ), HSF and HSF + yOZ (HSFyOZ). C and CyOZ received standard diet (60% complex carbohydrate‐dextrin, 4% fat‐soybean oil); HSF and HSFyOz received HSF diet (53% simple carbohydrate‐sucrose and fructose,16% fat‐lard) + 25% of sucrose in drinking water. yOz was added to the feed at a dose of 0.5%. During 30 weeks, body weight was measured weekly and feed intake, daily. After the period, it was analysed plasm glucose (mg/dL), HOMA‐IR and systolic blood pressure‐SBP(mmHg). Obesity was characterized by adiposity index (% of body fat) and renal tissue was used to analyse antioxidant protection (%protection/g protein). Renal disease was analyzed by glomerular filtration rate (GFR‐ml/min)) and proteinuria (protein/creatinin ratio).ResultsHSF induced renal disease and risk factors, such as obesity, increased glucose, HOMA‐IR, SBP and renal oxidative stress. yOz was efficient to reduce adipogenesis, improves renal antioxidant protection and prevent renal disease. Table shows the results.ConclusionGamma‐oryzanol prevented renal disease and oxidative stress. Therefore, the compound can be a strategy to prevent these complications related to obesity.Support or Funding InformationFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP‐2015/10626‐0)
Nutritional and metabolic profile and renal function

C
CyOz
HSF
HSFyOz

food intake (g/day)
20 ± 3
19 ± 3
8 ± 1#
8 ± 2$

caloric intake (kcal/day)
71 ± 10
68 ± 11
71 ± 6
61 ± 22

final body weight (g)
493 ± 51
497 ± 54
583 ± 76#
494 ± 75&

adiposity index (%)
5 ± 1
4 ± 1
9 ± 2#
6 ± 2&,$

plasm glucose (mg/dl)
71 ± 20
78 ± 21
106 ± 18#
89 ± 7

HOMA‐IR
6 ± 3
7 ± 4
22 ± 22#
26 ± 17$

SBP (mmHg)
121 ± 6
127 ± 9
140 ± 12#
145 ± 13$

GFR (ml/min)
2.5 ± 1
3 ± 0*
1.6± 1#
2.3 ± 1&,$

protein/creatinin ratio
0.0022 ± 0.005
0.0027 ± 0.002
0.0031 ± 0.005#
0.0025 ± 0.002&

renal antioxidant protection (%/g protein)
51 ± 7
52 ± 5
41 ± 6#
54 ± 18&

compares C to CyOZ
compares C to HSF
compares HSF to HSFyOZ
compares CyOz to HSFyOz
Pretreatment with estrogen protects HK‐2 cells from cisplatin‐induced cytotoxicity
Authors: Chen Yu‐Tong , Fu Yaw‐Syan , Shih Hou‐Yu , Huang Bin , Lu Tzongshi , Wu Chia‐Chun 
Published: November 2023

Introduction and hypothesisEstrogen is a major sex hormone in females, and exerts multiple physiological functions. Cisplatin is a platinum‐containing anti‐cancer drug that is widely used as a chemotherapeutic agent. Nephrotoxicity of cisplatin is a dose‐limiting side effect and that can cause significant morbidity. HK‐2 cell line is an immortalized proximal tubule epithelial cell line from normal male adult human kidneys. In this study, we investigated the protective roles of estrogen on HK‐2 cells using a cisplatin‐induced nephrotoxicity model.MethodsHK‐2 Cells were treated at time 0 with: 1) 17β‐estradiol (0~20 nM); 2) estrogen receptor agonists; 3) estrogen receptor antagonists for 12 hours respectively. Then, the all groups of cells were treated with cisplatin (0–15 mg/ml) for 24 hours. The cell survival rate, proliferation, morphology were analyzed by ELISA and histology studies and, cell apoptosis/necrosis were analyzed by Annexin V/PI staining using flow cytometry. Estrogen receptors and apoptosis related proteins expressions were further confirmed by Western blotting.ResultsOur data indicates that HK‐2 cells express all 3 types of estrogen receptors. HK‐2 cells pretreated with 10 nM 17β‐estradiol have higher cell survival rate (87%) in comparison with untreated cells (60%). And, both Annexin V positive apoptotic cells and caspase 3 expressions are significant lower in 17β‐estradiol pretreated HK‐2 cells. All 3 types of estrogen receptors exert the protective effects in cisplatin‐induced cytotoxicity, and the G protein‐coupled estrogen receptor (GPER) is the most effective estrogen receptor. In addition, GPER antagonist also significantly accentuates cytotoxicity of cisplatin treated HK‐2 cells.ConclusionsOur data suggested that estrogen pretreatment could protect HK‐2 cells from cisplatin induced cytotoxicity, and this protective effect occurs mainly through the GPER pathway. It may offer a novel therapeutic strategy for cisplatin‐induced nephrotoxicity. Further investigation into the molecular mechanisms and clinical applications are needed.Support or Funding InformationKMU‐TP104E34
Docosahexaenoic acid attenuates renal injury and inflammation in LPS‐induced acute kidney injury via increased resolvin D2
Authors: Katary Mohamed , Elsherbini Nehal M. , Ibrahim Ahmed S. , Al‐Shabrawey Mohamed , Elmarakby Ahmed A. 
Published: November 2023

The polyunsaturated omega‐3 fatty acid, docosahexaenoic acid (DHA), has a promising role in preventing acute kidney injury (AKI); however; DHA reno‐protective mechanism remains to be elucidated. Our aim is to explore how DHA attenuates lipopolysaccharide (LPS)‐induced AKI. Four groups of wild type C57BL/6 (WT) mice were used in the current study (n=4); control, LPS injected (4 mg/kg i.p), LPS treated with the 12/15‐lipoxygenase (12/15‐LO) inhibitor baicalein (20 mg/kg i.p for 6 days) and LPS treated with DHA (50 mg/kg i.p for 6 days). LPS injection in WT mice showed a significant elevation in markers of renal injury compared to WT control mice (albuminuria was 112±4 vs. 8±1 mg/day and podocalyxin was 3.5±0.6 vs. 0.6±0.1 mg/day in LPS injected mice vs. WT control, P< 0.05). The elevation in renal injury markers were also associated with increases in oxidative stress and inflammation as evident by increased urinary thiobarbituric acid reactive substance (TBARs) and monocyte chemoattractant protein‐1 (MCP‐1), respectively, in LPS injected mice. Treatment of LPS injected mice with either DHA or baicalein significantly reduced markers of renal injury and inflammation. Although DHA or baicalein treatment significantly reduced the elevation in renal intercellular adhesion molecule 1 (ICAM‐1) and tumor necrosis factor‐α (TNF‐α) mRNA expression levels in LPS injected mice, DHA was superior over baicalein in reducing Interleukin‐6 (IL‐6) and IL‐1β and in increasing the anti‐inflammatory IL‐10 in LPS injected mice. DHA reduced renal tubular necrosis and vaculated cells and lowered renal expression of CD45, a marker of leucocyte adhesion, in LPS injected mice. DHA also restored the decrease in plasma resolvin D2 in LPS injected mice. Finally, injection of resolvin D2 decreased markers of renal injury, oxidative stress and inflammation and reduced renal tubular degeneration in LPS‐induced AKI. In conclusion, our data suggest that DHA is a superior treatment over 12/15‐LO inhibitor in preventing LPS‐induced AKI, at least in part, via increased resolvin D2.Support or Funding InformationAugusta University intramural Funding
Mitochondrial Function in Long‐term Consequences of Renal Ischemia‐Reperfusion Injury
Authors: Hepokoski Mark , Bullen Alexander , Li Ying , Hall Elanore , Pham Hai , Singh Prabhleen 
Published: November 2023

BackgroundRenal tissue hypoxia is a common link between AKI and CKD. The kidney has high mitochondrial oxidative metabolism, and is exquisitely sensitive to hypoxic injury. Mitochondria play a central role in cell injury and their dysfunction has been implicated in early AKI, but limited information is available regarding the long‐term consequences of AKI.MethodsWe performed 10min and 15min of bilateral renal artery clamping (IR) in mice and then serial GFR measurement by FITC inulin kinetics at days 1, 3, 7, 14 days, 4 weeks and 14 weeks after ischemia and protein expression of mitochondrial oxidative phosphorylation complexes (Ox phos), fission, fusion and AMPK at the same time points.ResultsSerial GFRsIn the 10min group, GFR was significantly lower at days 1 and 3 but improved after that to levels similar in shams. In the 15min group, GFR was dramatically reduced with partial recovery over 4 and 14 weeks.Ox Phos expressionSignificant reduction in expression for both 10 and 15 min IR at 1–2 weeks for most complexes, with recovery in expression at 4 weeks for 10min, but persistent reduction for 15min IR. Mitochondrial Proteins at 4weeks, 15min IR showed significant reduction in Mfn2 and increase in Fis1 expression. Total AMPK expression was increased at day1, but no increase in pAMPK was seen in the 15min IR kidney. In 10min IR, pAMPK was increased at 24 hrs and persisted at 4 weeks.ConclusionsOur findings show duration of IR corresponding with severity of injury and impaired recovery in GFR, and mitochondrial dysfunction with decreased Ox phos proteins and altered dynamics with impaired AMPK activation in IR kidneys, corresponding to severity of injury, but in some instances despite functional recovery. These may be important in transition of AKI to CKD. Additional studies to compare these two models when manipulated with maneuvers to worsen or improve recovery from IR are ongoing.Support or Funding InformationNIDDK O'Brien Center for AKI Research P30DK079337.Veterans Affairs (VA) Merit Award BX002175 (P. Singh)NIDDK R03 DK101841 (P.Singh)NIDDK R01 DK107852 (P.Singh)
Automated Glomerular Localization in Bright‐field, Trichrome‐stained Rat Kidneys
Authors: Bukowy John D , Evans Louise C , Dayton Alex , Cowley Allen W 
Published: November 2023

Histological examination of fixed renal tissue is widely used to assess kidney morphology and the progression of renal disease. Common renal metrics include glomerular number, cortical area/volume, protein cast area, and glomerular damage. Evaluation of these metrics is a time consuming process requiring a trained observer and manual recording/counting. The purpose of this study is to implement an automated pipeline to localize glomeruli in trichrome‐stained, bright‐field, whole renal slices resulting in minimization of human‐hours required to process experimental data groups, elimination of human observer variability, and expansion of glomerular‐specific renal metrics.The dataset for this study consisted of a population of histologically prepared rat kidney slices with varying degrees of protein casts and glomerular damage. All kidney slices were prepared from formalin fixed and paraffin embedded kidneys cut in 4 μm slices and mounted to glass slides. Slices were then stained with trichrome and imaged in bright‐field on a Nikon Ni‐E automated microscope with a resolution of 0.68 μm/pixel. Feature based thresholding and selection was performed over images of whole renal slices to nominate glomerular candidates which were subsequently cropped into individual images. From 14 separate kidney slices, initial filtering produced over 6,500 images (observations) containing >1,400 glomeruli. From this dataset, manual separation of glomerular positive images was performed creating a training dataset for supervised learning methods to classify glomerular candidates. The trained classifier was tested on a naïve dataset of 619 observations summarized by:

Sensitivity ~ 98% | Ratio of the number of images with glomeruli correctly identified by algorithm to number identified by human observer.
Specificity ~ 94% | Ratio of the number of images without glomeruli correctly identified by algorithm to number detected by human observer.
Precision ~ 79% | Ratio of the number of images with glomeruli correctly identified by algorithm to total number of images with glomeruli identified by the algorithm.
False Discovery Rate ~ 21% | Ratio of the number of images without glomeruli incorrectly identified by algorithm to number of glomeruli detected by algorithm.
Successful localization of glomeruli provides a framework for future workflows targeted to reduce processing time including semi‐automated scoring of identified glomeruli. Given the widespread usage of trichrome staining in renal morphology/pathology, this technique has the potential to greatly reduce the time and effort spent on renal dataset processing.Support or Funding InformationAmerican Heart Association | 16PRE29700006 ‐‐ National Institutes of Health | HL‐116264
Diminished TRPV4 activity contributes to compromised [Ca2+]i homeostasis in human ADPKD cells
Authors: Tomilin Viktor N , Zaika Oleh , Mamenko Mykola , Reif Gail , Wallace Darren , Pochynyuk Oleh 
Published: November 2023

Polycystic kidney disease (PKD) is a devastating clinically relevant pathology causing graduate decline in kidney function due to development and growth of cysts filled with fluid. No effective pharmacological treatments exist for PKD patients. At the cellular level, defective flow‐mediated [Ca2+]i responses and disrupted [Ca2+]i homeostasis have been repeatedly associated with the development of PKD. Our previous work in rodents demonstrated that the activity of the Ca2+‐permeable TRPV4 channel is imperative for flow‐mediated [Ca2+]i responses in the distal renal tubule. Moreover, TRPV4 function is dramatically decreased in isolated cyst monolayers and systemic stimulation of TRPV4 interferes with PKD progression in PCK 453 rats, an autosomal recessive PKD model. To probe translational implication of this mechanism, we used primary cultured normal human kidney (NHK) and autosomal dominant PKD (ADPKD) cells isolated from patients and determined TRPV4 function and its role in [Ca2+]i signaling. As expected, ADPKD cells failed to respond to elevated flow and had significantly lower basal [Ca2+]i levels of 43±1 nM, when compared to NHK cells (91±1 nM). Application of a selective TRPV4 activator, GSK1016790A (30 nM) elicited more than two times higher [Ca2+]i response in NHK than in ADPKD cells. GSK1016790A‐mediated responses were precluded upon pretreatment with a specific TRPV4 inhibitor, HC‐067047 (4 μM) or a Ca2+‐free media, suggesting a direct TRPV4‐dependent Ca2+ influx. While total TRPV4 levels were comparable in NHK and ADPKD cells, we detected a marked decrease in TRPV4 glycosylation. Consistently, treatment with tunicomycin for 24 hours to prevent glycosylation significantly inhibited TRPV4 activity pointing to a mechanism of reduced TRPV4 function in ADPKD cells. Finally, acute application of TRPV4 antagonist, HC‐067047 significantly reduced basal [Ca2+]i levels in NHK cells but had no measurable effect in ADPKD cells. Overall, we demonstrate that similarly to studies in rodents, TRPV4 function and glycosylation are greatly diminished in primary cultured human ADPKD cells and this is at least partially responsible for their distorted [Ca2+]i signaling. We speculate that TRPV4 stimulation might be beneficial in restoring [Ca2+]i homeostasis in cyst cells thereby counteracting ADPKD progression in humans.Support or Funding InformationThis research was supported by NIH‐NIDDK DK095029 (to O. Pochynyuk), AHA‐15SDG25550150 (to M. Mamenko)
Chitooligosaccharide inhibits cyst formation and enlargement through calcium restoration and AMPK activation
Authors: Tuangpholkrung Phattharaphon , Muanprasat Chatchai , Chatsudthipong Varanuj 
Published: November 2023

Autosomal dominant polycystic kidney disease (ADPKD) is an inherited disease. It is developed from mutation in renal Pkd1or/and Pkd2 gene leading to decreased intracellular calcium and consequently cysts formation. These renal cysts replace normal parenchyma cells disrupting its function that could lead to end‐stage renal failure. Currently, no effective therapy is available for PKD treatment. Recently, activation of AMP‐activated protein kinase (AMPK) by Metformin was found to inhibit cell proliferation and also reduce fluid secretion via CFTR in Madin‐Darby Canine Kidney (MDCK) cell. Interestingly, our group has shown that chitooligosaccharide or COS, the derivative of chitosan, could activate AMPK via the mechanism involved an elevation of intracellular calcium in colon cancer cell. Therefore, we hypothesized that COS‐activated AMPK may inhibit cyst formation and growth in MDCK cyst model. The results of toxicity test indicated that COS at the concentrations of 50, 100, 200 and 500 μg/ml produced no toxic effect to cells. In contrast, it was found that the treatments with COS at concentrations of 100, 200 and 500 μg/ml markedly decreased both cyst formation and cyst growth. Minimum effective dose of COS (100 μg/ml) was selected to study for its underlying mechanisms. Western blot analysis showed that COS at concentration of 100 μg/ml could activate AMPK and its downstream protein, acetyl Co‐A carboxylase (ACC). COS also induced an elevation in intracellular calcium. These data indicated that COS was able to inhibit cyst formation and retard cyst enlargement, in part, by the restoration of intracellular calcium and activation of AMPK.Support or Funding InformationThis work was supported by Development and Promotion of Science and Technology Talents Project (DPST), Institute for the Promotion of Teaching Science and Technology (to PT), and Faculty of Science, Mahidol University, Thailand.