Search

Metformin is widely used for the treatment of type 2 diabetes, and increasing numbers of studies have shown that metformin also ameliorates tumor progression, inflammatory disease, and fibrosis. However, the ability of metformin to improve non-diabetic glomerular disease and chronic kidney disease (CKD) has not been explored. To investigate the effect of metformin on non-diabetic glomerular disease, we used a mouse model of Alport syndrome (Col4a5 G5X) which were treated with metformin or losartan, used as a control treatment. We also investigated the effect of metformin on adriamycin-induced glomerulosclerosis model. Pathological and biochemical analysis showed that metformin or losartan suppressed proteinuria, renal inflammation, fibrosis, and glomerular injury and extended the lifespan in Alport syndrome mice. Transcriptome analysis showed that metformin and losartan influenced molecular pathways-related to metabolism and inflammation. Metformin altered multiple genes including metabolic genes not affected by losartan. Metformin also suppressed proteinuria and glomerular injury in the adriamycin-induced glomerulosclerosis mouse model. Our results showed that metformin ameliorates the glomerular sclerosis and CKD phenotype in non-diabetic chronic glomerular diseases. Metformin may have therapeutic potential for not only diabetic nephropathy but also non-diabetic glomerular disease including Alport syndrome.

Nephrotic syndrome (NS) is a renal disorder that is characterized by massive proteinuria, hypoalbuminemia and edema. One of the main causes of NS is focal segmental glomerulosclerosis (FSGS), which has extremely poor prognosis. Although steroids and immunosuppressants are the first line of treatment, some FSGS cases are refractory, prompting the need to find new therapeutic strategies. We have previously demonstrated that an optimized combination treatment of mild electrical stimulation (MES) and heat shock (HS) has several biological benefits including the amelioration of the pathologies of the genetic renal disorder Alport syndrome. Here, we investigated the effect of MES + HS on adriamycin (ADR)-induced NS mouse model. MES + HS suppressed proteinuria and glomerulosclerosis induced by ADR. The expressions of pro-inflammatory cytokines and pro-fibrotic genes were also significantly downregulated by MES + HS. MES + HS decreased the expression level of cleaved caspase-3 and the number of TUNEL-positive cells, indicating that MES + HS exerted anti-apoptotic effect. Moreover, MES + HS activated the Akt signaling and induced the phosphorylation and inhibition of the apoptotic molecule BAD. In in vitro experiment, the Akt inhibitor abolished the MES + HS-induced Akt-BAD signaling and anti-apoptotic effect in ADR-treated cells. Collectively, our study suggested that MES + HS modulates ADR-induced pathologies and has renoprotective effect against ADR-induced NS via regulation of Akt-BAD axis.

Background Alport syndrome (AS) is a hereditary, progressive nephritis caused by mutation of type IV collagen. Previous studies have shown that activation of signal transducer and activator of transcription 3 (STAT3) exacerbates other renal diseases, but whether STAT3 activation exacerbates AS pathology is still unknown. Here we aim to investigate the involvement of STAT3 in the progression of AS. Method Phosphorylated STAT3 expression was assessed by immunoblotting analysis of kidneys and glomeruli of an AS mouse model (Col4a5 G5X mutant). To determine the effect of blocking STAT3 signaling, we treated AS mice with the STAT3 inhibitor stattic (10 mg/kg i.p., three times per week for 10 weeks; n = 10). We assessed the renal function [proteinuria, blood urea nitrogen (BUN), serum creatinine] and analyzed the glomerular injury score, fibrosis and inflammatory cell invasion by histological staining. Moreover, we analyzed the gene expression of nephritis-associated molecules. Results Phosphorylated STAT3 was upregulated in AS kidneys and glomeruli. Treatment with stattic ameliorated the progressive renal dysfunction, such as increased levels of proteinuria, BUN and serum creatinine. Stattic also significantly suppressed the gene expression levels of renal injury markers (Lcn2, Kim-1), pro-inflammatory cytokines (Il-6, KC), pro-fibrotic genes (Tgf-β, Col1a1, α-Sma) and Mmp9. Stattic treatment decreased the renal fibrosis congruently with the decrease of transforming growth factor beta (TGF-β) protein and increase of antifibrosis-associated markers p-Smad1, 5 and 8, which are negative regulators of TGF-β signaling. Conclusion STAT3 inhibition significantly ameliorated the renal dysfunction in AS mice. Our finding identifies STAT3 as an important regulator in AS progression and provides a promising therapeutic target for AS.

Summary Alport syndrome is a hereditary glomerular disease caused by mutation in type IV collagen α3–α5 chains (α3–α5(IV)), which disrupts trimerization, leading to glomerular basement membrane degeneration. Correcting the trimerization of α3/α4/α5 chain is a feasible therapeutic approach, but is hindered by lack of information on the regulation of intracellular α(IV) chain and the absence of high-throughput screening (HTS) platforms to assess α345(IV) trimer formation. Here, we developed sets of split NanoLuc-fusion α345(IV) proteins to monitor α345(IV) trimerization of wild-type and clinically associated mutant α5(IV). The α345(IV) trimer assay, which satisfied the acceptance criteria for HTS, enabled the characterization of intracellular- and secretion-dependent defects of mutant α5(IV). Small interfering RNA-based and chemical screening targeting the ER identified several chemical chaperones that have potential to promote α345(IV) trimer formation. This split luciferase-based trimer formation assay is a functional HTS platform that realizes the feasibility of targeting α345(IV) trimers to treat Alport syndrome.

A seminal study recently demonstrated that bromide (Br-) has a critical function in the assembly of type IV collagen in basement membrane (BM), and suggested that Br- supplementation has therapeutic potential for BM diseases. Because salts of bromide (KBr and NaBr) have been used as antiepileptic drugs for several decades, repositioning of Br- for BM diseases is probable. However, the effects of Br- on glomerular basement membrane (GBM) disease such as Alport syndrome (AS) and its impact on the kidney are still unknown. In this study, we administered daily for 16 weeks 75 mg/kg or 250 mg/kg (within clinical dosage) NaBr or NaCl (control) via drinking water to 6-week-old AS mice (mouse model of X-linked AS). Treatment with 75 mg/kg NaBr had no effect on AS progression. Surprisingly, compared with 250 mg/kg NaCl, 250 mg/kg NaBr exacerbated the progressive proteinuria and increased the serum creatinine and blood urea nitrogen in AS mice. Histological analysis revealed that glomerular injury, renal inflammation and fibrosis were exacerbated in mice treated with 250 mg/kg NaBr compared with NaCl. The expressions of renal injury markers (Lcn2, Lysozyme), matrix metalloproteinase (Mmp-12), pro-inflammatory cytokines (Il-6, Il-8, Tnf-α, Il-1β) and pro-fibrotic genes (Tgf-β, Col1a1, α-Sma) were also exacerbated by 250 mg/kg NaBr treatment. Notably, the exacerbating effects of Br- were not observed in wild-type mice. These findings suggest that Br- supplementation needs to be carefully evaluated for real positive health benefits and for the absence of adverse side effects especially in GBM diseases such as AS.

This study was conducted to assess whether respiratory frequency can be used as a valid parameter for estimating ventilatory threshold and for examining differences in exercise modes such as a cycle ergometer and a treadmill. 24 men and 12 women performed an incremental exercise test to exhaustion on a cycle ergometer and on a treadmill. Oxygen uptake, carbon dioxide output, pulmonary ventilation, ventilatory frequency, and heart rate were measured continuously every 30 sec. during the test. Three different and independent reviewers detected the ventilatory threshold point and break point of respiratory rate, which were then compared. Analysis indicated that (1) ventilatory threshold was well correlated with break point of respiratory rate for both cycle ( r = .88, p

This study was undertaken to establish an approach for the investigation of age-related changes in indices of body composition during childhood in Japan. It provides current reference values for total body fat mass (TBFM) and lean body mass (LBM) as indices of body composition in an urban population of 3- to 6-year-old Japanese children. Moreover, we assessed the age-specific patterns of body fat distribution [subcutaneous fat mass (SFM) and internal fat mass (IFM)] during childhood. Measurements of body composition by bioelectrical impedance were made in 141 boys and 139 girls, all apparently healthy, aged 3-6 years. Determinations of impedance were made using a four-terminal impedance analyzer (TP-95K; Toyo Physical, Inc., Fukuoka, Japan). LBM was calculated using the equation of Kushner et al. (1992) and Goran et al. (1993). SFM was calculated using a modification of the equation derived by Skerjl et al. (1953). IFM was calculated as the difference between TBFM and SFM. From ages 3 through 6 years, the mean LBM increased with age in boys and girls, and showed significant age differences. Between the ages of 3 and 6, the average increment in LBM was 5.1 kg in boys and 4.4 kg in girls. On average, boys gained 0.5 kg of TBFM each year, whereas girls gained 0.4 kg of TBFM each year. Furthermore, both groups gained 0.3 kg of SFM each year. Percentage body fat decreased in both genders until approximately the age of 5, and increased again slightly at the age of 6. The age-specific pattern of fat accumulation during childhood was characterized by an almost linear increase in SFM in girls, but a transient decrease in IFM in boys. We conclude that further research is required, including longitudinal assessment of body composition variables, in order to unravel the dynamics of body composition change in Japanese children.

The purpose of this study was to evaluate the differential perceived exertion measured using a new set of Visual Analogue Scales (VAS) during pedaling and running. The subjects were eleven healthy males. They performed an incremental maximal test and then three 4-min stages of exercise, for both pedaling and running. During the tests, VO2, V(CO2), V(E), f, and HR were monitored continuously. Bla and perceptual variables including VAS consisting of four scales (VAS 1-VAS 4) and Borg's RPE were measured at the end of each stage. Although the VO2 (%VO2max)) and HR for both pedaling and running were not significantly different, Bla in pedaling was significantly higher than that in running. A significant interaction (mode, stage) was also obtained. The VAS 1 of pedaling was significantly higher than that of running. A significant interaction in VAS 1 (mode, stage) was obtained. The VAS 2 of pedaling was significantly higher than that of running. The subjects indicated that local pain became stronger than central pain in pedaling, but they were almost equal in running. In both pedaling and running, leg pain became stronger than arm pain (VAS 3). VAS 4 showed that during running, breathing difficulty and heart pain were almost equal in perceived intensity. However, during pedaling, breathing difficulty became greater than heart pain. Thus, a new four-part visual analogue scale was found to be useful for monitoring exercise intensity. In addition, the new VAS gave us more information in relation to the differential perceived exertion reflected in the different physiological responses obtained by different exercise modes.

The purpose of this study was to examine the physiological responses and RPE during water walking using the Flowmill, which has a treadmill at the base of a water flume, in order to obtain basic data for prescribing water walking for people of middle and advanced age. Twenty healthy female volunteers with an age of 59.1 +/- 5.2 years took part in this study. They belonged to the same swimming club and regularly swam and exercised in water. Walking in water took place in the Flowmill. Subjects completed four consecutive bouts of 4 min duration at progressively increasing speeds (20, 30, 40 and 50 m/min) with 1 min rest between each bout. In addition, water velocity was adjusted to the walking speed of each bout. Subjects were instructed to swing both arms in order to maintain their balance during walking in water. The water depth was to the level of the xiphoid process and the water temperature was 30.31 +/- 0.08 degrees C. Both heart rate (HR) and oxygen uptake (VO2) increased exponentially as walking speed increased. HR was 125 +/- 15 bpm, and VO2 was 18.10 +/- 2.72 ml/kg.min-1 during walking in water at 50 m/min, which was the highest speed. The exercise intensity at this speed was equivalent to 5.2 +/- 0.8 Mets. The relationship between HR and VO2 during walking in water showed a highly significant linear relationship in each subject. There was also a highly significant linear relationship in the mean HR and VO2 of all subjects. Blood lactate concentration (LA) measured at rest and immediately after each bout was 1.1 +/- 0.4 mmol/l at rest, 1.0 +/- 0.2 mmol/l at 20 m/min, 1.0 +/- 0.3 mmol/l at 30 m/min, 1.1 +/- 0.2 mmol/l at 40 m/min, and 2.4 +/- 0.7 mmol/l at 50 m/min. LA at 50 m/min was significantly higher than at rest and at the other speeds. The relationship between HR and RPE during walking in water showed a highly significant linear relationship. The relationship between walking speed and energy expenditure calculated from VO2 and the respiratory exchange ratio (R) showed a high significant exponential relationship. These results suggested that HR and RPE can be effective indices for exercise prescription during Flowmill walking as with land walking.

The primary objective of this study was to determine gender differences in total body fat mass (TBFM) and body fat distribution (subcutaneous fat mass, SFM; and internal fat mass, IFM) in a cross-sectional sample of 280 children. Measurements of the body composition of 141 boys and 139 girls, all apparently healthy and aged 3-6 years were made using bioelectrical impedance. Determinations of impedance were made using a four-terminal impedance analyzer (TP-95K; Toyo Physical, Fukuoka, Japan). Lean body mass (LBM) was calculated using a previously published equation [Goran MI, Kaskoun MC, Carpenter WH, Poehlman ET, Ravussin E, Fontvieikke A-M (1993) Estimating body composition of young children by using bioelectrical resistance. J Appl Physiol 75: 1776-1780]. SFM was calculated using a modification of the equation derived by Skerjl [Skerjl B, Brozek J, Hunt EE (1953) Subcutaneous fat and age changes in body build and body form in women. Am J Phys Anthrop 11: 577-580] and Davies [Davies PSW, Jones PRM, Norgan NG (1986) The distribution of subcutaneous and internal fat in man. Ann Hum Biol 13: 189-192]. The main modifications of the equation in the present study were the introduction of: (1) mean thickness of adipose tissue over body surface/2, and (2) skin mass. IFM was calculated as the difference between TBFM and SFM. The body mass index (BMI; kg/m2) was calculated from the formula: body mass/height2. For each gender, the subjects in the lowest and highest 25th percentiles were designated as "low body mass" and "high body mass", respectively. In the present study, no gender differences in absolute TBFM, SFM and IFM were observed in either of these groups. In contrast, gender differences in relative TBFM (%Fat) and SFM (SFM/mass) were evident in girls. However, the four subgroups were similar in terms of relative IFM (IFM/mass). The TBFM was independently related to SFM, IFM and %Fat in both genders after adjustment for BMI; however, there was no significant association of SFM with IFM after adjustment for BMI in any group. Even after adjustment for BMI, IFM was independently related to %Fat in both genders, although SFM was not independently related to %Fat in any group except low-body-mass boys. This study shows that relative TBFM and SFM are higher in high-body-mass groups and tend to be higher in girls than in boys, and that the higher %Fat in high-body-mass girls than in high-body-mass boys appears to be associated with internal adipose tissue deposits. External adipose tissue mass does not appear to be related to the higher %Fat levels in high-body-mass girls. In addition, subcutaneous fat mass appears to be higher in low-body-mass girls than in low-body-mass boys, although this observation needs confirmation using more valid measures of subcutaneous fat such as computerized tomography and magnetic resonance imaging.