Glomerular Area Research Articles

En Bloc Cadaver Kidney Transplantation From a 9-Month-Old Donor to an Adult Recipient: Maturation of Glomerular Size and Podocyte in the Recipient.

BackgroundFavorable outcomes of en bloc pediatric donor kidney transplantation to adult recipients are attributed primarily to grafting of twice the nephron mass of a single kidney.MethodsThe kidneys of a 9-month-old male infant were transplanted en bloc in a 56-year-old man. Biopsies were performed 1 hour postreperfusion, 6 months and 3.5 years posttransplant.ResultsWarm and cold ischemia times were 21 and 426 minutes, respectively. The recipient was released from hemodialysis 10 days posttransplant and discharged 91 days posttransplant when serum creatinine was 0.9 mg/dL. At 4 years and 9 months posttransplant, serum creatinine was 1.0 mg/dL, and estimated glomerular filtration rate was 58.0 mL/min per 1.73 m2. The grafts increased in size until they reached adult size by 3 months posttransplant. The glomerular area and volume, respectively, increased from 5.9 × 103 μm2 and 0.34 × 106 μm3 at 1 hour postreperfusion to 14.9 × 103 μm2 and 1.27 × 106 μm3 at 3.5 years posttransplant, both of which were less than half of adult size. At 1 hour postreperfusion, podocytes were structurally immature. At 6 months posttransplant, podocyte immaturity was still evident. At 3.5 years posttransplant, podocytes were mature.ConclusionsThese findings suggest that podocytes and glomerular size of pediatric donor kidneys can continue to mature in adult recipients at rates appropriate for donor age when transplanted en bloc. The maturational levels of podocytes and glomeruli may also be a factor involved in favorable outcomes of en bloc pediatric donor kidney transplantation.

Betanin, isolated from fruits of Opuntia elatior Mill attenuates renal fibrosis in diabetic rats through regulating oxidative stress and TGF-β pathway

Ethnopharmacological relevanceThe fruits of Opuntia elatior Mill are being used traditionally in different disease condition like diabetes, obesity, asthma, inflammatory disorders, and anemia. Betanin, a compound isolated from fruits of Opuntia elatior Mill has potent anti-oxidative and anti-inflammatory activity. Recent study from our lab indicated the protective effect of betanin against high glucose induced rat renal epithelial cell fibrosis and matrix accumulation, major features of diabetic nephropathy (DN). However the molecular mechanism of betanin in DN has not yet been fully elucidated. Aim of the studyThe aim of the present study was to further investigate the anti-fibrotic mechanisms of betanin against streptozotocin (STZ) induced DN. Materials and methodsBetanin was isolated from fruits of Opuntia elatior Mill (Cactaceae) and structure was elucidated using spectroscopy (UV, IR, 1H-NMR and mass). STZ was injected intraperitoneally with single dose of 50mg/kg for diabetes induction. In order to develop DN the animals were left in diabetes condition without any treatment during the following 4 weeks. Betanin (25, 50 and 100mg/kg/day) and lisinopril (5mg/kg/day, reference compound) were orally administered for 8 weeks after the induction of DN. Renal function, blood glucose, serum creatinine, blood urea nitrogen (BUN) and antioxidant enzyme activities in the kidney tissue were measured. Kidney tissue samples were used for glomerulosclerosis, tubulointerstitial fibrosis and morphometric studies. The expression of transforming growth factor-beta (TGF-β), type IV collagen, alpha-smooth muscle actin (α-SMA) and E-cadherin in kidney tissue were evaluated using reverse transcription-polymerase chain reaction, and immunohistochemistry. ResultsBetanin was successfully isolated from fruits of Opuntia elatior Mill (Cactaceae) and purified by column chromatography. The results showed that betanin attenuated diabetic kidney injury by significantly inhibiting proteinuria, blood glucose, serum creatinine and BUN levels and restored antioxidant enzyme activities in kidney tissue. Histological studies exhibited that betanin treatment reduced the glomerular surface area, glomerulosclerosis and tubulointerstitial fibrosis. Furthermore, betanin modulated mRNA and protein expression of TGF-β, type IV collagen, α-SMA and E-cadherin in kidney. ConclusionsThe results conclude that betanin can effectively suppress renal fibrosis in DN, and may slow down the progression to end-stage renal disease by regulating TGF-β signal pathway.

Blocking lysophosphatidic acid receptor 1 signaling inhibits diabetic nephropathy in db/db mice

Lysophosphatidic acid (LPA) is known to regulate various biological responses by binding to LPA receptors. The serum level of LPA is elevated in diabetes, but the involvement of LPA in the development of diabetes and its complications remains unknown. Therefore, we studied LPA signaling in diabetic nephropathy and the molecular mechanisms involved. The expression of autotaxin, an LPA synthesis enzyme, and LPA receptor 1 was significantly increased in both mesangial cells (SV40 MES13) maintained in high-glucose media and the kidney cortex of diabetic db/db mice. Increased urinary albumin excretion, increased glomerular tuft area and volume, and mesangial matrix expansion were observed in db/db mice and reduced by treatment with ki16425, a LPA receptor 1/3 antagonist. Transforming growth factor (TGF)β expression and Smad-2/3 phosphorylation were upregulated in SV40 MES13 cells by LPA stimulation or in the kidney cortex of db/db mice, and this was blocked by ki16425 treatment. LPA receptor 1 siRNA treatment inhibited LPA-induced TGFβ expression, whereas cells overexpressing LPA receptor 1 showed enhanced LPA-induced TGFβ expression. LPA treatment of SV40 MES13 cells increased phosphorylated glycogen synthase kinase (GSK)3β at Ser9 and induced translocation of sterol regulatory element-binding protein (SREBP)1 into the nucleus. Blocking GSK3β phosphorylation inhibited SREBP1 activation and consequently blocked LPA-induced TGFβ expression in SV40 MES13 cells. Phosphorylated GSK3β and nuclear SREBP1 accumulation were increased in the kidney cortex of db/db mice and ki16425 treatment blocked these pathways. Thus, LPA receptor 1 signaling increased TGFβ expression via GSK3β phosphorylation and SREBP1 activation, contributing to the development of diabetic nephropathy.

Effect of high fructose administration on histopathology of kidney, heart and aorta of rats

Objective: This study was conducted to assess the hazardous effects of high fructose administration on kidney, heart and aorta in rats. Materials and methods: Twenty adult healthy male albino rats weighing about 200-220 gm each were used in this study. The rats were divided into 2 duplicate groups; control group and fructose group. Fructose was administered to rats in fresh drinking water daily for 8 weeks (the whole experimental period). Serum urea, creatinine and sodium concentration were determined by using ready-made kits. Spectrophotometric and colorimetric methods were also used for the detection of other serum components. Histopathological examination of the tissues was done by staining with H&E, PAS and Masson trichrome stains. Results : Nephropathy was achieved in fructose group after one month as indicated by biochemical assay. Pathological observation showed that high fructose administration decreased size of cardio-myocytes, increased cardiac interstitial fibrosis score and aortic wall thickness. In kidneys, high fructose administration decreased glomerular tuft area and corpuscular area, increased percentage in the rats affected with interstitial renal fibrosis score 1 and percentage of rats had glomerular sclerosis score 2. Conclusion : High fructose in diet should be avoided because it can damage kidney, heart and aorta in rats. http://doi.org/10.5455/javar.2017.d193

Maternal Hypercholesterolemia Associated with Nicotine Exposure in Adulthood May Induce Kidney Injury in Male Rats if Hypomagnesemia Occurs

Background/Aims: Maternal hypercholesterolemia is a risk factor to renal injury in rat pups at adulthood, especially if they feed a cholesterol-enriched diet after weaning. However, the renal function of male pups of dams with hypercholesterolemia (PH) that were fed a regular chow from weaning to adulthood needs investigation, particularly those exposed to an adverse risk such as nicotine. Methods: We evaluated the renal function of PH animals and we compared the data with those found in male pups of control dams (PC) at 3- and 6-month-old by inulin clearance. Moreover, we investigated the effect of nicotine treatment for 8 days in both PH and PC animals at 6 months old via metabolic function studies and by renal histological analysis. Results: Inulin clearance and other renal function parameters were similar in PH and PC animals at 3 and 6 months old. Nevertheless, the PH group showed significant differences with regard to histological analysis despite a similar number of glomeruli. The glomerular area of PH animals was significantly smaller than that measured in PC animals, and the fractional interstitial area was significantly larger in PH animals than that measured in PC animals at 3 months old. With regard to nicotine treatment, we observed a trend for a reduction in creatinine clearance in both PC and PH groups, but only PH animals showed hypomagnesemia and the highest fractional interstitial area. Conclusions: The offspring exposed to a high cholesterol milieu during intrauterine and neonatal life may show a silent kidney injury at adulthood that may be aggravated by nicotine exposure if hypomagnesemia occurs.

Effect of curcumin on kidney histopathological changes, lipid peroxidation and total antioxidant capacity of serum in sodium arsenite-treated mice

Sodium arsenite is an environmental pollutant with the ability to generate free radicals and curcumin acts as a potent antioxidant. This study investigates the effect of curcumin on kidney histopathology, lipid peroxidation and antioxidant capacity of serum in the mice treated with sodium arsenite. Adult male mice were divided into four groups: control, sodium arsenite, curcumin and curcumin+sodium arsenite. The treatments were delivered for 5 weeks. After the treatment period, blood samples were collected and the concentrations of malondialdehyde (MDA) and total antioxidant capacity of serum were determined. Left kidney was dissected, weighed and used for histopathological and histomorphometrical studies. Sodium arsenite-treated mice showed a significant decrease in the diameter of glomerulus and proximal tubule, glomerular area, total antioxidant capacity of serum as well as a significant increase in serum concentration of MDA compared to the control group. However, no significant difference was found in kidney weight, area and diameter of Bowman's capsule as well as the diameter of distal tubule in mice treated with sodium arsenite compared to the control. In curcumin+sodium arsenite group, curcumin significantly reversed the adverse effects of sodium arsenite on the diameter of glomerulus and proximal tubule, glomerular area, total antioxidant capacity of serum and serum concentration of MDA compared to the sodium arsenite group. The application of curcumin alone significantly increased the total antioxidant capacity of serum compared to the control. Curcumin compensated the adverse effects of sodium arsenite on kidney tissue, lipid peroxidation and total antioxidant capacity of serum.

Prognostic value of morphologic and morphometric analyses in IgA nephropathy biopsies

IgA nephropathy (IgAN) exhibits a variable course ranging from a benign condition to progressive renal failure with a high proportion of patients undergoing end-stage renal disease in the long term. It is unclear how to predict the risk of this progression. Mesangial C4d deposition has been found to be associated with a poor prognosis. Our aim was to search histological lesions with possible prognostic value in IgAN biopsies performed at the time of the diagnosis. Clinical and laboratory records of 44 patients undergoing renal biopsy were reviewed. IgAN was diagnosed in 32 patients and minimal change disease (MCD) in 12. C4d deposition, glomerular endothelial cell area, glomerular and interstitial macrophages were evaluated in all biopsies. Clinical and laboratory data were available in all IgAN patients at the time of diagnosis and in 21 patients at the end of follow-up (mean follow-up 48.09 ± 19.69 months). All IgAN and MCD biopsies were C4d negative. The number of glomerular macrophages and the area of glomerular endothelial cells in IgAN were greater than in MCD (P < 0.001 and P < 0.0001, respectively), indicating glomerular inflammation associated with endothelial cell enlargement. Glomerular macrophages positively correlated with diffuse IgA and fibrinogen immunoreactivity (P < 0.02 and P < 0.002, respectively). Interstitial macrophages positively correlated with segmental glomerulosclerosis (P < 0.01) and tubular atrophy (P < 0.001), according to the Oxford classification. They correlated with hypertension (P < 0.03) and renal functional impairment both at the time of biopsy and at the end of follow-up. The receiver operating characteristic analysis curve and a cut-off value of ≥19.5 macrophages per high power field showed 75.0% sensitivity and 88.9% specificity in predicting worse clinical outcome. A Poisson generalized regression fit model showed that hypertension and tubular atrophy were the most significant factors in predicting a high number of interstitial macrophages. The number of interstitial macrophages correlated with hypertension, histological damage, functional impairment at the time of biopsy and worse outcome. A high number of interstitial macrophages in C4d negative biopsies may allow the identification of patients at increased risk of progression to renal failure.

Effects of gallic acid on delta - aminolevulinic dehydratase activity and in the biochemical, histological and oxidative stress parameters in the liver and kidney of diabetic rats.

Diabetes mellitus (DM) is characterised by hyperglycaemia associated with the increase of oxidative stress. Gallic acid has potent antioxidant properties. The aim of this study was to evaluate the effect of gallic acid on the biochemical, histological and oxidative stress parameters in the liver and kidney of diabetic rats. Male rats were divided in groups: control, gallic acid, diabetic and diabetic plus gallic acid. DM was induced in the animals by intraperitoneal injection of streptozotocin (65mg/kg). Gallic acid (30mg/kg) was administered orally for 21days. Our results showed an increase in reactive species levels and lipid peroxidation, and a decrease in activity of the enzymes superoxide dismutase and delta-aminolevulinic acid dehydratase in the liver and kidney of the diabetic animals (P<0.05). Gallic acid treatment showed protective effects in these parameters evaluated, and also prevented a decrease in the activity of catalase and glutathione S-transferase, and vitamin C levels in the liver of diabetic rats. In addition, gallic acid reduced the number of nuclei and increased the area of the core in hepatic tissue, and increased the glomerular area in renal tissue. These results indicate that gallic acid can protect against oxidative stress-induced damage in the diabetic state.

Therapeutic Role of Tangshenkang Granule () in Rat Model with Diabetic Nephropathy.

To evaluate the renal protective effect of Tangshenkang Granule () in a rat model of diabetic nephropathy (DN). Forty male Sprague-Dawley rats were randomly divided into control, DN, Tangshenkang and benazepril groups. DN model was established in the rats of DN, Tangshenkang and benazepril groups. Tangshenkang Granule solution and benazepril hydrochloride solution were intragastrically administered daily to the rats in the Tangshenkang and benazepril groups for 8 weeks, respectively. Urinary albumin and creatinine were detected. The albumin/creatinine (ACR) was calculated in addition to 24 h urinary protein (24-h UPr), serum creatinine (Scr), blood urea nitrogen (BUN), total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), and creatinine clearance rate (Ccr). Right kidneys were harvested for pathological observation using periodic acid-silver methenamine-Masson staining. The average glomerular diameter (DG), average glomerular (AG) and mesangial areas (AM) were measured. The thickness of glomerular basement membrane (TGBM) was detected using transmission electron microscope. Compared with rats in the control group, rats in the DN group showed significantly decreased body weight, increased hypertrophy index, 24-h urinary volume, 24-h UPr, ACR, Scr, BUN, Ccr, blood lipids as well as renal pathological indices including DG, AG, AM, AM/AG and TGBM (P <0.05). Compared with the DN group, the weights of rats in the Tangshenkang and benazepril groups were significantly increased, and the renal hypertrophy indices were significantly decreased (P <0.05). The 24-h urinary volumes, ACR, 24-h UPr, Scr, BUN, Ccr, LDL, DG, AG, AM and TGBM were obviously decreased (P <0.05). Compared with the benazepril group, the Tangshenkang group showed significantly decreased levels of ACR, 24-h UPr, AG and AM (P <0.05). Tangshenkang Granule decreased the urinary protein, attenuated the high glomerular filtration rate and improved lipid metabolism in DN rats, and prevented further injury induced by diabetic nephropathy.

Nephron morphometry in mice and rats using tomographic microscopy

The aim was to quantify the glomerular capillary surface area, the segmental tubular radius, length, and area of single nephrons in mouse and rat kidneys. Multiple 2.5-µm-thick serial Epon sections were obtained from three mouse and three rat kidneys for three-dimensional reconstruction of the nephron tubules. Micrographs were aligned for each kidney, and 359 nephrons were traced and their segments localized. Thirty mouse and thirty rat nephrons were selected for further investigation. The luminal radius of each segment was determined by two methods. The luminal surface area was estimated from the radius and length of each segment. High-resolution micrographs were recorded for five rat glomeruli, and the capillary surface area determined. The capillary volume and surface area were corrected for glomerular shrinkage. A positive correlation was found between glomerular capillary area and proximal tubule area. The thickest part of the nephron, i.e., the proximal tubule, was followed by the thinnest part of the nephron, i.e., the descending thin limb, and the diameters of the seven identified nephron segments share the same rank in the two species. The radius and length measurements from mouse and rat nephrons generally share the same pattern; rat tubular radius-to-mouse tubular radius ratio ≈ 1.47, and rat tubular length-to-mouse tubular length ratio ≈ 2.29, suggesting relatively longer tubules in the rat. The detailed tables of mouse and rat glomerular capillary area and segmental radius, length, and area values may be used to enhance understanding of the associated physiology, including existing steady-state models of the urine-concentrating mechanism.

Role of angiotensin II type 1a receptor in renal injury induced by deoxycorticosterone acetate-salt hypertension.

The aim of this study was to investigate the in vivo role of angiotensin II type 1a (AT1a) receptor in renal damage as a result of hypertension by using transgenic mice with AT1a receptor gene disruption. Transgenic mice that express human liver-type fatty acid binding protein (L-FABP) with or without disruption of the AT1a receptor gene (L-FABP+/− AT1a−/−, and L-FABP+/− AT1a+/+, respectively) were used with urinary L-FABP as an indicator of tubulointerstitial damage. Those female mice were administered subcutaneously deoxycorticosterone acetate (DOCA)–salt tablets plus drinking water that contained 1% saline for 28 d after uninephrectomy. In L-FABP+/− AT1a+/+ mice that received DOCA-salt treatment, hypertension was induced and slight expansion of glomerular area, glomerular sclerosis, and tubulointerstitial damage were observed. In L-FABP+/− AT1a−/− mice that received DOCA-salt treatment, hypertension was similarly induced and the degree of glomerular damage was significantly more severe than in L-FABP+/− AT1a+/+-DOCA mice. Urinary L-FABP levels were significantly higher in L-FABP+/− AT1a−/−-DOCA mice compared with those in L-FABP+/− AT1a+/+-DOCA mice. Hydralazine treatment significantly attenuated renal damage that was found in L-FABP+/− AT1a−/−-DOCA mice along with a reduction in blood pressure. In summary, activation of the AT1a receptor may contribute to maintenance of the glomerular structure against hypertensive renal damage.—Hisamichi, M., Kamijo-Ikemori, A., Sugaya, T., Ichikawa, D., Natsuki, T., Hoshino, S., Kimura, K., Shibagaki, Y. Role of angiotensin II type 1a receptor in renal injury induced by deoxycorticosterone acetate–salt hypertension.

Insulin Prevents Hyperfiltration and Proteinuria but Not Glomerular Hypertrophy and Increases Mesangial Matrix Expansion in Diabetic Rats

Objective: The aim of this work was to study the effect of 7 days of strict glycemic control with insulin on glomerular function and structure in streptozotocin (STZ)-diabetic rats. Materials and Methods: Three groups of adult male Fischer rats were studied: controls (n = 15), diabetics (n = 15), and insulin-treated diabetics (n = 15). Diabetes was induced by treating the rats with STZ (55 mg/kg i.p.). One week after the induction of diabetes, blood glucose, protein excretion rate (PER), glomerular filtration rate (GFR), and renal plasma flow (RPF) were estimated in each group. Furthermore, morphometric analysis was performed to estimate the tuft volume and changes in mesangial matrix area. The results are expressed as the mean ± SEM. Results: STZ diabetes caused significant increases in GFR (0.89 ± 0.1 to 1.21 ± 0.1 mL/min/100 g; p < 0.01) and RPF (1.78 ± 0.37 to 3.32 ± 0.6 mL/min/100 g; p < 0.05). Furthermore, the diabetic rats had higher glomerular volumes but mesangial matrix areas similar to controls. Insulin treatment prevented the increases in blood glucose (4.5 ± 0.2 m<smlcap>M</smlcap>), PER (66.1 ± 7.8 mg/day), GFR (0.6 ± 0.07 mL/min/100 g), and RPF (1.72 ± 0.36 mL/min/100 g), but did not prevent glomerular hypertrophy (21.7% increase), but induced mesangial matrix expansion (25% increase). Conclusions: Insulin prevented the diabetes-induced hyperfiltration and proteinuria, but did not prevent glomerular growth, and induced mesangial expansion. Hyperglycemic episodes could be partly responsible for persistent glomerular growth and accelerated mesangial growth.

PS 08-52 BLOOD PRESSURE RELATED TO GLOMERULOMEGALY AND TUBULOINTERSTITIAL FIBROSIS IN CHRONIC GLOMERULONEPHRITIS

Objective: Several factors contribute to the development of hypertension in patients with chronic glomerulonephritis (GN). This study was conducted to find the relationships between baseline blood pressure (BP) and clinico-pathological findings in patients with chronic GN. Design and Method: Clinico-pathological findings were analyzed in a total of 233 patients with chronic GN from The Kyung-Hee Cohort of GN. The glomerular surface area (GSA) was determined using imaging analysis software. The serum and urine angiotensinogen (AGT) concentrations were measured using human ELISA kits. Results: Mean serum creatinine concentration was 1.19 (0.5∼6.0) mg/dL. Systolic BP was > = 130 mmHg in 124 patients (53%) and > = 140 mmHg in 77 (33%). Systolic BP was positively correlated with serum uric acid concentrations and renal tissue findings including mean GSA and the degree of tubulointerstitial fibrosis and deposits of complement C3 and IgM and negatively with follow-up glomerular filtration rate and the slope of change in 1/serum creatinine for 2 years, while it has no significant relationships with serum and urine AGT and 24 hour urinary sodium excretion. Patients with systolic BP > = 130 mmHg as compared with those < 130 mmHg are older age and showed severer degree of clinico-pathological findings; baseline and follow-up serum creatinine, the amount of proteinuria, mean GSA, tubulointerstitial fibrosis and deposits of IgM and C3. Conclusions: This study showed that systolic BP was related to renal progression and pathological findings, glomerulomegaly and tubulointerstitial fibrosis, in patients with chronic GN.

PS 10-03 EFFECTS OF LCZ696 ON BLOOD PRESSURE AND RENAL INJURY IN TYPE 2 DIABETIC RATS WITH OVERT PROTEINURIA

Objective: Clinical studies have indicated that LCZ696, an angiotensin receptor-neprilysin inhibitor, exerts beneficial effects in patients with heart failure. Here, we aimed to examine the effect of LCZ696 on blood pressure and renal injury in type 2 diabetic Otsuka-Long-Evans-Tokushima-Fatty (OLETF) rats with overt proteinuria. Design and Method: In OLETF rats, vehicle (n = 10), valsartan (30 mg/kg/day, n = 10), LCZ696 (68 mg/kg/day, n = 9) or valsartan plus hydralazine (3 mg/kg/day, n = 10) was treated from 56 to 80 weeks of age. Results: At baseline (56-week-old), diabetic OLETF rats showed hypertension, overt proteinuria, glomerular injury (glomerular PAS positive area) and tubulointerstitial fibrosis (Sirius Red-positive area). At 80-week-old, vehicle-treated OLETF rats showed developed hypertension and proteinuria, severe glomerular injury and tubulointerstitial fibrosis, and increases in plasma BUN and creatinine levels. Treatment with LCZ696 or valsartan plus hydralazine similarly caused greater blood pressure reduction than valsartan. On the other hand, LCZ696 caused greater reductions in proteinuria, glomerular injury and tubulointerstitial fibrosis than valsartan or valsartan plus hydralazine. LCZ696 also prevented the increases in plasma BUN and creatinine levels. Conclusions: These data suggest that LCZ696 elicits a reno-protective effect in type 2 diabetes with hypertension and nephrotic-range proteinuria.

Effect of the oral iron chelator deferiprone in diabetic nephropathy rats.

The aim of the present study was to investigate the effects of the iron chelator deferiprone in diabetic nephropathy (DN) rats and the mechanisms involved. Thirty-two male Wistar rats (180-220 g, 6 weeks old) were randomly divided into a control group, a DN group and two DN groups treated with either 50 or 100 mg/kg per day deferiprone. The DN group was established by feeding of a high-carbohydrate-fat diet and injection of 35 mg/kg streptozotocin into the vena caudalis. The duration of deferiprone treatment was 20 weeks. Histopathological changes were detected by hematoxylin-eosin and Masson staining, as well as transmission electron microscopy. Levels of nuclear factor (NF)-κB, monocyte chemotactic protein (MCP)-1, matrix metalloproteinase (MMP)-9, tissue-specific inhibitor of metalloproteinase (TIMP)-1, cyclo-oxygenase (COX)-2, and nitrotyrosine were determined in kidney tissues using reverse transcription-polymerase chain reaction (RT-PCR), western blotting, and immunohistochemistry. Histopathological observations showed that deferiprone treatment alleviated inflammation infiltrates and collagenous fibrosis in DN rats. Results from RT-PCR and western blotting indicated that deferiprone inhibited the expression of NF-κB, MCP-1, COX-2, and nitrotyrosine, which were overexpressed in DN rats. Immunohistochemistry showed that the mechanism of deferiprone action may involve regulation of MMP-9 and TIMP-1. Decreased MMP-9 expression and increased TIMP-1 expression in DN rats were significantly promoted and inhibited by deferiprone, respectively. Iron chelation by oral deferiprone has a renoprotective effect in DN rats by relieving oxidative stress, inflammation, and fibrosis, which is related to the cytokines NF-κB, MCP-1, MMP-9, TIMP-1, COX-2, and nitrotyrosine.

Blood capillary rarefaction and lymphatic capillary neoangiogenesis are key contributors to renal allograft fibrosis in an ACE inhibition rat model.

Chronic allograft fibrosis is the major cause of graft loss in kidney transplantation. Progression can only be reduced by inhibition of the renin-angiotensin system (RAS). We tested the hypothesis that the protection provided by angiotensin-converting enzyme (ACE) inhibition also decreases capillary rarefaction, lymphangiogenesis, and podocyte injury in allograft fibrosis. Fisher kidneys were transplanted into bilaterally nephrectomized Lewis rats treated with enalapril (60 mg/kg per day) (ACE inhibitor, ACEi) or vehicle. Proteinuria, blood urea nitrogen, and plasma creatinine were regularly assessed, and grafts were harvested for morphological and immunohistological analysis at various times up to 32 wk. In the vehicle group, many new lymphatic capillaries and severe and diffuse mononuclear infiltration of allografts were observed already 1 wk after transplantation. Lymphangiogenesis increased until week 4, by which time inflammatory infiltration became focal. Lymphatic capillaries were often located at sites of inflammation. Progressive interstitial fibrosis, glomerulosclerosis, capillary rarefaction, and proteinuria appeared later, at weeks 4-12 The number of lymphatic capillary cross sections strongly correlated with the interstitial fibrosis score. Podoplanin immunostaining, a marker of healthy podocytes, disappeared from inflamed or sclerotic glomerular areas. ACEi protected from lymphangiogenesis and associated inflammation, preserved glomerular podoplanin protein expression, and reduced glomerulosclerosis, proteinuria, tubulointerstitial fibrosis, and blood capillary rarefaction at 32 wk. In conclusion, ACEi considerably decreased and/or delayed both glomerulosclerosis and tubulointerstitial injury. Prevention of glomerular podoplanin loss and proteinuria could be attributed to the known intraglomerular pressure-lowering effects of ACEi. Reduction of lymphangiogenesis could contribute to amelioration of tubulointerstitial fibrosis and inflammatory infiltration after ACEi.

Sleep restriction during pregnancy and its effects on blood pressure and renal function among female offspring.

The influence of sleep restriction (SR) during pregnancy on blood pressure and renal function among female adult offspring was investigated. Pregnant Wistar rats were distributed into control and SR groups. The SR was performed between the 14th and 20th days of pregnancy (multiple platforms method for 20 h/day). At 2 months of age, half of the offspring from both groups were subjected to an ovariectomy (ovx), and the other half underwent sham surgery. The groups were as follows: control sham (Csham), control ovx (Covx), SR sham (SR sham), and SR ovx (SR ovx). Renal function markers and systolic blood pressure (BPi, indirect method) were evaluated at 4, 6, and 8 months. Subsequently, the rats were euthanized, kidneys were removed, and processed for morphological analyses of glomerular area (GA), number of glomeruli per mm3 (NG), and kidney mass (KM). Increased BPi was observed in the Covx, SR sham, and SR ovx groups compared to Csham at all ages. Increased plasma creatinine concentration and decreased creatinine clearance were observed in the SR sham and SR ovx groups compared to the Csham and Covx groups. The SR ovx group showed higher BPi and reduced creatinine clearance compared to all other groups. The SR ovx group showed reduced values of GA and KM, as well as increased NG, macrophage infiltration, collagen deposit, and ACE1 expression at the renal cortex. Therefore, SR during pregnancy might be an additional risk factor for developing renal dysfunction and increasing BP in female adult offspring. The absence of female hormones exacerbates the changes caused by SR.

Aging increases the susceptibility of hepatic inflammation, liver fibrosis and aging in response to high-fat diet in mice.

We aimed to investigate whether aging increases the susceptibility of hepatic and renal inflammation or fibrosis in response to high-fat diet (HFD) and explore the underlying genetic alterations. Middle (10months old) and old (20months old) aged, male C57BL/6N mice were fed either a low-fat diet (4% fat) or HFD (60% fat) for 4months. Young (3months old) aged mice were included as control group. HFD-induced liver and kidney injuries were analyzed by serum and urine assay, histologic staining, immunohistochemistry, and reverse-transcription real-time quantitative polymerase chain reaction. Total RNA sequencing with next-generation technology was done with RNA extracted from liver tissues. With HFD feeding, aged was associated with higher serum alanine aminotransferase levels, marked infiltration of hepatic macrophages, and increased expression of inflammatory cytokines (MCP1, TNF-α, IL-1β, IL-6, IL-12, IL-17A). Importantly, aged mice showed more advanced hepatic fibrosis and increased expression of fibrogenic markers (Col-I-α1, αSMA, TGF-β1, TGF-β2, TGFβRII, PDGF, PDGFRβII, TIMP1) in response to HFD. Aged mice fed on HFD also showed increased oxidative stress and TLR4 expression. In the total RNA seq and gene ontology analysis of liver, old-aged HFD group showed significant up-regulation of genes linked to innate immune response, immune response, defense response, inflammatory response compared to middle-aged HFD group. Meanwhile, aging and HFD feeding showed significant increase in glomerular size and mesangial area, higher urine albumin/creatinine ratio, and advanced renal inflammation or fibrosis. However, the difference of HFD-induced renal injury between old-aged group and middle-aged group was not significant. The susceptibility of hepatic fibrosis as well as hepatic inflammation in response to HFD was significantly increased with aging. In addition, aging was associated with glomerular alterations and increased renal inflammation or fibrosis, while the differential effect of aging on HFD-induced renal injury was not remarkable as shown in the liver.

The efficacy of Aesculus hippocastanum seeds on diabetic nephropathy in a streptozotocin-induced diabetic rat model.

Cytokines, such as transforming growth factor (TGF)-ß1, and increased oxidative stress are considered to be responsible for the development of diabetic nephropathy. We hypothesized that Aesculus hippocastanum (AH) seeds may have preventive effects on oxidative stress and TGF-β-related diabetic nephropathy in streptozotocin (STZ)-induced diabetic nephropathy in rats. Twenty-one male Sprague-Dawley albino rats were divided into three groups (n=7). Except for the control group, they all had diabetic nephropathy induced by an intraperitoneal injection of STZ. While the diabetes group did not receive any medication, the diabetes+AH group was given the medication for 4 weeks. After the experiment, analyses were performed to evaluate the glomerular area, severity of sclerosis, and fibronectin immunoexpression, as well as levels of malondialdehyde (MDA), TGF-β, blood urea nitrogen (BUN), blood glucose, creatinine, and proteinuria. It was found that glomerular area, severity of sclerosis, fibronectin immunoexpression, and levels of MDA, TGF-β, BUN, creatinine, and proteinuria were decreased in the diabetes+AH group. It is known that diabetic nephropathy is induced, to a large extent, by hyperglycemia. In the present study, AH extract ameliorated diabetic nephropathy without decrease in blood glucose levels. In the study, AH seeds showed beneficial effects on the functional properties of the kidney and microscopic improvements in diabetic nephropathy.

Morphometry Predicts Early GFR Change in Primary Proteinuric Glomerulopathies: A Longitudinal Cohort Study Using Generalized Estimating Equations.

ObjectiveMost predictive models of kidney disease progression have not incorporated structural data. If structural variables have been used in models, they have generally been only semi-quantitative.MethodsWe examined the predictive utility of quantitative structural parameters measured on the digital images of baseline kidney biopsies from the NEPTUNE study of primary proteinuric glomerulopathies. These variables were included in longitudinal statistical models predicting the change in estimated glomerular filtration rate (eGFR) over up to 55 months of follow-up.ResultsThe participants were fifty-six pediatric and adult subjects from the NEPTUNE longitudinal cohort study who had measurements made on their digital biopsy images; 25% were African-American, 70% were male and 39% were children; 25 had focal segmental glomerular sclerosis, 19 had minimal change disease, and 12 had membranous nephropathy. We considered four different sets of candidate predictors, each including four quantitative structural variables (for example, mean glomerular tuft area, cortical density of patent glomeruli and two of the principal components from the correlation matrix of six fractional cortical areas–interstitium, atrophic tubule, intact tubule, blood vessel, sclerotic glomerulus, and patent glomerulus) along with 13 potentially confounding demographic and clinical variables (such as race, age, diagnosis, and baseline eGFR, quantitative proteinuria and BMI). We used longitudinal linear models based on these 17 variables to predict the change in eGFR over up to 55 months. All 4 models had a leave-one-out cross-validated R2 of about 62%.ConclusionsSeveral combinations of quantitative structural variables were significantly and strongly associated with changes in eGFR. The structural variables were generally stronger than any of the confounding variables, other than baseline eGFR. Our findings suggest that quantitative assessment of diagnostic renal biopsies may play a role in estimating the baseline risk of succeeding loss of renal function in future clinical studies, and possibly in clinical practice.