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Urate nephropathy, a common complication of hyperuricemia, has garnered increasing attention worldwide. However, the exact pathogenesis of this condition remains unclear. Currently, inflammation is widely accepted as the key factor in urate nephropathy. Therefore, the aim of this study was to elucidate the interaction of lincRNA-p21/AIF-1/CMPK2/NLRP3 via exosomes in urate nephropathy. This study evaluated the effect of lincRNA-p21/AIF-1/CMPK2/NLRP3 using clinical data collected from patients with urate nephropathy and human renal tubular epithelial cells (HK2) cultured with different concentrations of urate. In clinical research section, the level of lincRNA-p21/AIF-1 in exosomes of urine in patients with hyperuricemia or urate nephropathy was found to be increased, particularly in patients with urate nephropathy. In vitro study section, the level of exosomes, inflammation, autophagy, and apoptosis was increased in HK2 cells induced by urate. Additionally, the expression of lincRNA-p21, AIF-1, CMPK2, and NLRP3 was upregulated in exosomes and HK2 cells. Furthermore, manipulating the activity of lincRNA-p21, AIF-1, CMPK2, and NLRP3 through overexpression or interference vectors regulated the level of inflammation, autophagy, and apoptosis in HK2 cells. In conclusion, the pathway of lincRNA-p21/AIF-1/CMPK2/NLRP3 contributed to inflammation, autophagy, and apoptosis of human renal tubular epithelial cell induced by urate via exosomes. Additionally, the specific exosomes in urine might serve as novel biomarkers for urate nephropathy.
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Apoptosis , Autofagia , Células Epiteliales , Exosomas , Proteína con Dominio Pirina 3 de la Familia NLR , ARN Largo no Codificante , Ácido Úrico , Humanos , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Ácido Úrico/metabolismo , Exosomas/metabolismo , Células Epiteliales/metabolismo , Células Epiteliales/patología , ARN Largo no Codificante/metabolismo , ARN Largo no Codificante/genética , Transducción de Señal , Inflamación/metabolismo , Inflamación/patología , Túbulos Renales/metabolismo , Túbulos Renales/patología , Línea Celular , Masculino , Factor Inductor de la Apoptosis/metabolismo , Femenino , Persona de Mediana Edad , Hiperuricemia/metabolismo , Hiperuricemia/orina , Proteínas de Unión al Calcio , Proteínas de MicrofilamentosRESUMEN
The food crisis has increased demand for agricultural resources due to various factors such as extreme weather, energy crises, and conflicts. A solar greenhouse enables counter-seasonal winter cultivation due to its thermal insulation, thus alleviating the food crisis. The root temperature is of critical importance, although the mechanism of soil thermal environment change remains uncertain. This paper presents a comprehensive study of the soil thermal environment of a solar greenhouse in Jinzhong City, Shanxi Province, employing a variety of analytical techniques, including theoretical, experimental, and numerical simulation, and deep learning modelling. The results of this study demonstrate the following: During the overwintering period, the thermal environment of the solar greenhouse floor was divided into a low-temperature zone, a constant-temperature zone, and a high-temperature zone; the distance between the low-temperature boundary and the southern foot was 2.6 m. The lowest temperature in the low-temperature zone was 11.06 °C and the highest was 19.05 °C. The floor in the low-temperature zone had to be heated; the lowest value of the constant-temperature zone was 18.29 °C, without heating. The minimum distance between the area of high temperature and the southern foot of the solar greenhouse was 8 m and the lowest temperature reading was 19.29 °C. The indoor soil temperature tended to stabilise at a depth of 45 cm, and the lowest temperature reading at a horizontal distance of 1400 mm from the south foot was 19.5 °C. The Fluent and LSTM models fitted well and the models can be used to help control soil temperature during overwintering in extreme climates. The research can provide theoretical and data support for the crop areas and the heating of pipelines in the solar greenhouse.
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Diabetic kidney disease (DKD) is a leading factor in end-stage renal disease. The complexity of its pathogenesis, combined with the limited treatment efficacy, necessitates deeper insights into potential causes. Studies suggest that ferroptosis-driven renal tubular damage contributes to DKD's progression, making its counteraction a potential therapeutic strategy. Quercetin, a flavonoid found in numerous fruits and vegetables, has demonstrated DKD mitigation in mouse models, though its protective mechanism remains ambiguous. In this study, we delved into quercetin's potential anti-ferroptotic properties, employing a DKD rat model and high glucose (HG)-treated renal tubular epithelial cell models. Our findings revealed that HG prompted unusual ferroptosis activation in renal tubular epithelial cells. However, quercetin counteracted this by inhibiting ferroptosis and activating NFE2-related factor 2 (Nrf2) expression in both DKD rats and HG-treated HK-2 cells, indicating its renal protective role. Further experiments, both in vivo and in vitro, validated that quercetin stimulates Nrf2. Thus, our research underscores quercetin's potential in DKD treatment by modulating the ferroptosis process via activating Nrf2 in a distinct DKD rat model, offering a fresh perspective on quercetin's protective mechanisms.
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Diabetes Mellitus Experimental , Nefropatías Diabéticas , Ferroptosis , Ratones , Ratas , Animales , Nefropatías Diabéticas/tratamiento farmacológico , Nefropatías Diabéticas/etiología , Nefropatías Diabéticas/metabolismo , Quercetina/farmacología , Quercetina/uso terapéutico , Estreptozocina , Factor 2 Relacionado con NF-E2/metabolismo , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/metabolismoRESUMEN
Self-compassion, as a personal psychological resource, has been proved to play an important role in coping with suffering. Based on self-determination theory, the present study attempts to establish that self-compassion can promote trust, and the sense of interpersonal responsibility mediates this relationship. Study 1 used cross-sectional data in a community sample of 322 adults to reveal that self-compassion was positively related to trust, and the mediating effects of the sense of interpersonal responsibility were significant. Study 2 used the latent cross-lagged panel model among 1304 college students at three-time points set at six-month intervals to replicate the results and proved the longitudinal mediating effects across groups. Finally, a casual chain design was used to test the mediation effect in Studies 3 and 4. The results indicated that self-compassion induced by writing task resulted in a sense of responsibility in Study 3 (N = 145), and the manipulated sense of responsibility promoted both trust behaviors and beliefs in Study 4 (N = 125). Through four studies, this study highlights a novel but unexpected viewpoint that treating oneself in a self-compassionate way can not only help individuals cope with various challenges but also motivate them to obtain interpersonal benefits. These findings can help motivate community workers and mental health researchers to increase social capital by focusing on self-compassion and interpersonal responsibility.
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Background: To explore the predictive value of the Programme on Research for Integrating Services for the Maintenance of Autonomy 7 (PRISMA-7), quick Sequential Organ Failure Assessment (qSOFA) score, Emergency Severity Index (ESI), and Clinical Frailty Scale (CFS) on the 28-day mortality risk in emergency elderly patients. Methods: A multicenter prospective observational study was conducted to select elderly patients (≥65 years old) admitted to the emergency department of three Grade-A hospitals in different regions of China from January 2020 to March 2022. Primary data were collected at the time of admission. All patients were followed up for 28 days. The primary outcome was 28-day mortality. The predictive value of four scoring systems for 28-day mortality in elderly emergency patients was assessed by receiver operating characteristic (ROC) and logistic regression analysis. Results: A total of 687 elderly emergency patients were enrolled, of whom 66 (9.61%) died within 28 days. Age, ICU admission rate, PRISMA-7, qSOFA, and CFS were significantly higher in the death group than in the survival group (P < 0.05), and ESI was lower than in the survival group (P < 0.001). The AUC for CFS was the largest of the four scoring systems at 0.80. According to the Youden index, the optimal cutoff values for PRISMA-7, qSOFA, ESI, and CFS were >3.5, >0.5, <2.5, and >4.5, respectively. Logistic regression revealed that qSOFA and CFS were the primary risk factors for increased 28-day mortality in elderly emergency patients (P < 0.001). The combined predictor L (L=X1+0.50X2, X1 and X2 are qSOFA and CFS values, respectively) had an AUC of 0.86 and a cutoff value >2.75. Conclusion: PRISMA-7, qSOFA, ESI, CFS, and the combined qSOFA+CFS predictor were all effective predictors of 28-day mortality risk in elderly emergency patients, with the combined qSOFA+CFS predictor having the best predictive power.
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OBJECTIVE: The study's purpose is to explore the link of serum albumin on renal progression in patients with chronic kidney disease (CKD). METHODS: This study was a secondary analysis of a prospective cohort study in which a total of 954 participants were non-selectively and consecutively collected from the research of CKD-ROUTE in Japan between November 2010 and December 2011. We evaluated the association between baseline ALB and renal prognosis (initiation of dialysis or 50% decline in eGFR from baseline) and renal function decline (annual eGFR decline) using the Cox proportional-hazards and linear regression models, respectively. We performed a number of sensitivity analyses to ensure the validity of the results. In addition, we performed subgroup analyses. RESULTS: The included patients had a mean age of (66.86 ± 13.41) years, and 522 (69.23%) were male. The mean baseline ALB and eGFR were (3.89 ± 0.59) g/dL and (33.43 ± 17.97) ml/min/1.73 m2. The annual decline in eGFR was 2.65 mL/min/1.73 m2/year. 218 (28.9%) individuals experienced renal prognosis during a median follow-up period of 36.0 months. The baseline ALB was inversely linked with renal prognosis (HR = 0.61, 95%CI: 0.45, 0.81) and renal function decline (ß = -1.41, 95%CI: -2.11, -0.72) after controlling for covariates. The renal prognosis and ALB had a non-linear connection, with ALB's inflection point occurring at 4.3 g/dL. Effect sizes (HR) were 0.42 (0.32, 0.56) and 6.11 (0.98, 38.22) on the left and right sides of the inflection point, respectively. There was also a non-linear relationship between ALB and renal function decline, and the inflection point of ALB was 4.1 g/dL. The effect sizes(ß) on the left and right sides of the inflection point were -2.79(-3.62, -1.96) and 0.02 (-1.97, 1.84), respectively. CONCLUSION: This study shows a negative and non-linear association between ALB and renal function decline as well as renal prognosis in Japanese CKD patients. When ALB is lower than 4.1 g/dL, ALB decline was closely related to poor renal prognosis and renal function decline. From a therapeutic point of view, reducing the decline in ALB makes sense for delaying CKD progression.
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Insuficiencia Renal Crónica , Albúmina Sérica , Humanos , Masculino , Persona de Mediana Edad , Anciano , Anciano de 80 o más Años , Femenino , Albúmina Sérica/análisis , Estudios Prospectivos , Progresión de la Enfermedad , Diálisis Renal , Tasa de Filtración Glomerular , Factores de Riesgo , Riñón , Insuficiencia Renal Crónica/terapia , PronósticoRESUMEN
The microRNA-214 (miR-214) precursor is formed by the DNM3 gene on human chromosome 1q24.3, which is encoded and transcribed in the nucleus and processed into mature miR-214 in the cytoplasm. Association of miR-214 with the interstitial fibrosis of the kidney has been reported in existing research. Renal interstitial fibrosis is considered necessary during the process of various renal injuries in chronic kidney disease (CKD). One of the important mechanisms is the TGF- (transforming growth factor-) ß1-stimulated epithelial interstitial transformation (EMT). The specific mechanisms of miR-214-3p in renal interstitial fibrosis and whether it participates in EMT are worthy of further investigation. In this paper, we first demonstrated modulation of the downstream PI3K/AKT axis by miR-214-3p through targeting phosphatase and tension protein homologues (PTEN), indicating the miRNA's participation in unilateral ureteral obstruction (UUO) nephropathy and TGF-ß1-induced EMT. We overexpressed or silenced miR-214-3p and PTEN for probing into the correlation of miR-214-3p with PTEN and the downstream PI3K/AKT signalling pathways. According to the results of the study, miR-214-3p overexpression silenced PTEN, activated the PI3K/AKT signalling pathway, and exacerbated EMT induced by TGF-ß1, while miR-214-3p knockdown had the opposite effect. In miR-214-3p knockdown mice, the expression of PTEN was increased, the PI3K/AKT signalling pathway was inhibited, and fibrosis was alleviated. In conclusion, miR-214-3p regulates the EMT of renal tubular cells induced by TGF-ß1 by targeting PTEN and regulating the PI3K/AKT signalling pathway. Furthermore, miR-214-3p knockdown can reduce renal interstitial fibrosis through the PTEN/PI3K/AKT pathway.
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Enfermedades Renales , MicroARNs , Humanos , Ratones , Animales , Factor de Crecimiento Transformador beta1/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Línea Celular , Fosfohidrolasa PTEN/genética , Fosfohidrolasa PTEN/metabolismo , Fibrosis , Enfermedades Renales/genética , Enfermedades Renales/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Factores de Crecimiento Transformadores/farmacología , Transición Epitelial-Mesenquimal/genéticaRESUMEN
Increasing evidence suggests that aldosterone (Aldo) plays an essential role in vascular calcification which is a serious threat to cardiovascular disease (CVD) developed from chronic kidney disease (CKD). However, the exact pathogenesis of vascular calcification is still unclear. First, we established CKD-associated vascular calcification mice model and knockout mice model to investigate the causal relationship between allograft inflammatory factor 1 (AIF-1) and vascular calcification. Then, endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) co-culture experiments were performed to further explore the mechanisms of calcification. The results of the Aldo intervention mice model and transgenic mice model showed that Aldo could cause calcification by increasing the AIF-1 level. The results of in vitro co-culture model of ECs and VSMCs showed that AIF-1 silence in ECs may alleviate Aldo-induced calcification of VSMCs. In conclusion, our study indicated that Aldo may induce vascular calcification related to chronic renal failure via the AIF-1 pathway which may provide a potential therapeutic target.
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Insuficiencia Renal Crónica , Calcificación Vascular , Aldosterona/metabolismo , Animales , Técnicas de Cocultivo , Modelos Animales de Enfermedad , Células Endoteliales/metabolismo , Ratones , Ratones Noqueados , Músculo Liso Vascular/metabolismo , Insuficiencia Renal Crónica/complicaciones , Insuficiencia Renal Crónica/metabolismo , Calcificación Vascular/inducido químicamenteRESUMEN
INTRODUCTION: Classic hemodialysis schedules present inadequate middle-molecular-weight toxin clearance due to limitations of membrane-based separation processes. Accumulation of uremic retention solutes may result in specific symptoms (e.g., pruritus) and may affect clinical outcome and patient's quality of life. Hemoperfusion (HP) is a blood purification modality based on adsorption that can overcome such limitations, and thus, it may be interesting to test the efficacy of at least one session per week of HP combined with hemodialysis. This is a randomized, open-label trial, controlled, multicenter clinical study to investigate the effect of long-term HP combined with hemodialysis on middle-molecular-weight toxins and uremic pruritus in maintenance hemodialysis (MHD) patients. METHODS: 438 MHD patients from 37 HD centers in China with end-stage kidney disease (63.9% males, mean age 51 years) suffering from chronic intractable pruritus were enrolled in the study. Eligible patients were randomized into four groups: low-flux hemodialysis (LFHD), high-flux hemodialysis (HFHD), HP + LFHD, and HP + HFHD at a 1:1:1:1 ratio. Beta-2 microglobulin (ß2M) and parathyroid hormone (PTH) were measured at baseline, 3-6, and 12 months. At the same time points, the pruritus score was evaluated. The primary outcome was the reduction of ß2M and PTH, while the secondary outcome was the reduction of the pruritus score. RESULTS: In the two groups HP + LFHD and HP + HFHD, there was a significant decrease of ß2M and PTH levels after 12 months compared to the control groups. No significant differences were noted between HP + LFHD and HP + HFHD. Pruritus score reduction was 63% in the HP + LFHD group and 51% in the HP + HFHD group, respectively. CONCLUSION: The long-term HP + HD can reduce ß2M and PTH levels and improve pruritus in MHD patients independently on the use of high- or low-flux dialyzers, showing that the results are linked to the effect of adsorption.
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Inflammation and glomerular endothelial dysfunction promote diabetic kidney disease (DKD) progression, but the mechanisms are not fully understood. Allograft inflammatory factor-1 (AIF-1) is a protein that regulates inflammatory reactions and immune responses. This study aimed to explore the mechanism of AIF-1 in a DKD animal model and mouse renal glomerular endothelial cells (MRGECs). We injected AIF-1-shRNA into the tail vein to knockdown AIF-1 in db/db mice. Metabolic index, renal pathological changes and inflammatory factors were measured in each group. Lentiviral transfection was used to overexpress AIF-1 in MRGECs. Inflammatory factors, oxidative stress and nuclear factor-κB (NF-κB) pathway-related proteins were examined. AIF-1 expression was upregulated in glomerular endothelial cells in renal tissues of db/db mice. Knockdown of AIF-1 reversed kidney injury and renal inflammation in db/db mice. In a 30 mM high-glucose environment, overexpression of AIF-1 in MRGECs activated the NF-κB pathway and induced inflammation and oxidative stress. Moreover, this damage could be attenuated by the addition of an NF-κB inhibitor (BAY 11-7082). In conclusion, AIF-1 facilitates glomerular endothelial cell inflammation and oxidative stress in DKD via the NF-κB signaling pathway. Our results provide evidence for the molecular mechanism of DKD and may offer a potential target for DKD treatment.
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Proteínas de Unión al Calcio/metabolismo , Nefropatías Diabéticas , Inflamación/metabolismo , Proteínas de Microfilamentos/metabolismo , Aloinjertos , Animales , Diabetes Mellitus/metabolismo , Nefropatías Diabéticas/metabolismo , Nefropatías Diabéticas/patología , Células Endoteliales/metabolismo , Inflamación/patología , Riñón/metabolismo , Ratones , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Estrés OxidativoRESUMEN
Vascular calcification is a major complication of maintenance hemodialysis patients. Studies have confirmed that calcification mainly occurs in the vascular smooth muscle cells (VSMC) of the vascular media. However, the exact pathogenesis of VSMC calcification is still unknown. This study shows that the crosstalk between calcium and aldosterone via the allograft inflammatory factor 1 (AIF-1) pathway contributes to calcium homeostasis and VSMC calcification, which is a novel mechanism of vascular calcification in uremia. In vivo results showed that the level of aldosterone and inflammatory factors increased in calcified arteries, whereas no significant changes were observed in peripheral blood. However, the expression of inflammatory factors markedly increased in the peripheral blood of uremic rats without aortic calcification and gradually returned to normal levels with aggravation of aortic calcification. In vitro results showed that there was an interaction between calcium ions and aldosterone in macrophages or VSMC. Calcium induced aldosterone synthesis, and in turn, aldosterone also triggered intracellular calcium content upregulation in macrophages or VSMC. Furthermore, activated macrophages induced inflammation, apoptosis, and calcification of VSMC. Activated VSMC also imparted a similar effect on untreated VSMC. Finally, AIF-1 enhanced aldosterone- or calcium-induced VSMC calcification, and NF-κB inhibitors inhibited the effect of AIF-1 on VSMC. These in vivo and in vitro results suggest that the crosstalk between calcium ions and aldosterone plays an important role in VSMC calcification in uremia via the AIF-1/NF-κB pathway. Local calcified VSMC induced the same pathological process in surrounding VSMC, thereby contributing to calcium homeostasis and accelerating vascular calcification.
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Aldosterona/metabolismo , Apoptosis , Proteínas de Unión al Calcio/metabolismo , Calcio/metabolismo , Proteínas de Microfilamentos/metabolismo , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/metabolismo , FN-kappa B/metabolismo , Transducción de Señal , Uremia/metabolismo , Calcificación Vascular/metabolismo , Animales , Proteínas de Unión al Calcio/genética , Línea Celular , Femenino , Humanos , Inflamación/genética , Inflamación/metabolismo , Inflamación/patología , Masculino , Proteínas de Microfilamentos/genética , Músculo Liso Vascular/patología , Miocitos del Músculo Liso/patología , FN-kappa B/genética , Ratas , Ratas Sprague-Dawley , Uremia/genética , Uremia/patología , Calcificación Vascular/genética , Calcificación Vascular/patologíaRESUMEN
OBJECTIVE: To explore the potential mechanism of KMUP-1 in the vascular calcification of chronic renal failure (CRF) through mediating NO/cGMP/PKG pathway, and provide novel insights into the CRF treatment. METHODS: CRF rats were treated by KMUP-1 with/without L-NNA (a NOS inhibitor) and then performed by ELISA, alizarin red staining, Von Kossa staining, Masson's trichrome, Sirius red staining and CD3 immunohistochemical staining. Simultaneously, vascular smooth muscle cells (VSMCs) were collected from rats to confirm the effect of KMUP-1 on vascular calcification in vitro via NO/cGMP/PKG pathway. Besides, protein and mRNA expressions were determined via Western blotting and qRT-PCR, respectively. RESULTS: CRF rats were elevated in 24-h urine protein, blood urea nitrogen (BUN), serum creatinine, Cys-C levels and inflammatory cytokines. Besides, CRF rats also showed increased calcium content and ALP level with up-regulated mRNA of osteogenic differentiation-related markers. Furthermore, the up-regulated expressions of eNOS and PKG, as well as down-regulated levels of NOx and cGMP were also found in CRF rats. However, renal failure and vascular calcification of CRF were improved significantly by KMUP-1 treatment via activation of NO/cGMP/PKG pathway. Moreover, KMUP-1 treatment attenuated calcified VSMCs, accompanied by the decreases in the calcified nodules, level of calcium and activity of ALP. In addition, either L-NNA treatment for CRF rats or the calcified VSMCs could antagonize the improving effect of KMUP-1. CONCLUSION: KMUP-1 can improve the renal function and vascular calcification in CRF rats at least in part by activating NO/cGMP/PKG pathway.
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Fallo Renal Crónico/tratamiento farmacológico , Miocitos del Músculo Liso/efectos de los fármacos , Piperidinas/farmacología , Calcificación Vascular/tratamiento farmacológico , Xantinas/farmacología , Animales , Calcio/metabolismo , GMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Modelos Animales de Enfermedad , Fallo Renal Crónico/fisiopatología , Masculino , Miocitos del Músculo Liso/patología , Óxido Nítrico/metabolismo , Osteogénesis/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Calcificación Vascular/patologíaRESUMEN
Diabetic nephropathy (DN) represents one of the most devastating complications for patients with diabetes. The anti-diabetic activities of Magnoflorine (MF) were reported, with underlying mechanism unknown. Lysine-specific demethylase 3A (KDM3A) was identified in the renal injuries. In the current study, we investigated the functional role of MF in DN progression with the involvement of KDM3A. We reported that in the animal model of DN induced by streptozotocin (STZ) injection, MF attenuated inflammatory response and fibrosis in the kidneys. In cultured mesangial cells, MF similarly ameliorated abnormal proliferation and lowered the expression of inflammation- and fibrosis-related factors stimulated by high glucose (HG) treatment. Upon MF treatment, there was a decline in KDM3A-positive cells in renal tissues of rats, accompanying an augment in KDM3A ubiquitination. KDM3A upregulation in vitro by a proteasome inhibitor MG132 comparably dampened the inhibitory role of MF in inflammatory response and fibrosis. Further analyses revealed that MF increased transforming growth factor ß-induced factor 1 (TGIF1) transcriptional activity by promoting ubiquitination and degradation of KDM3A, thus inhibiting the activation of TGF-ß1/Smad2/3 signaling pathway. TGIF1 silencing weakened the repressive role of MF in mesangial cells as well. In conclusion, MF contributes to TGIF1 transcription via an epigenetic mechanism.
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Angiotensin II (Ang II) is an important profibrotic factor, and the tumor-promoting microRNA miR-21 was recently linked to fibrotic disorders. We aimed to investigate whether and how miR-21 mediates Ang II-induced renal fibrosis. In renal tubular epithelial cells, Ang II upregulated miR-21 and fibrosis-related indicators but decreased PPARα expression. miR-21 overexpression promoted PPARα downregulation, activated the TGF-ß1/Smad3 pathway and induced fibrogenesis, while miR-21 suppression exerted opposite effects. In Ang II-treated cells, reduced PPARα expression, TGF-ß1/Smad3 pathway activation and fibrogenesis were all exacerbated by miR-21 upregulation but alleviated by miR-21 inhibition. The dual-luciferase assay confirmed PPARα as the target of miR-21. PPARα silencing alone could overactivate the TGF-ß1/Smad3 pathway in the presence or absence of Ang II. Importantly, the regulatory effects of miR-21 knockdown and the angiotensin type 1 receptor blocker losartan alone or in combination on the PPARα/TGF-ß1/Smad3 pathway in Ang II-treated cells were almost the same. More crucially, PPARα restoration abolished the profibrotic effect of miR-21 overexpression. In addition, inhibiting miR-21 in Ang II-treated mice effectively ameliorated the abnormally activated PPARα/TGF-ß1/Smad3 pathway, albuminuria, and renal fibrosis without lowering blood pressure. These results demonstrated that miR-21 extensively mediates Ang II-induced kidney fibrosis via amplifying the TGF-ß1/Smad3 pathway by targeting PPARα.
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Angiotensina II/efectos adversos , Expresión Génica , Enfermedades Renales/genética , Enfermedades Renales/patología , Riñón/patología , MicroARNs/genética , MicroARNs/metabolismo , PPAR alfa/metabolismo , Transducción de Señal/fisiología , Proteína smad3/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Células Cultivadas , Fibrosis , Riñón Fusionado , Humanos , Enfermedades Renales/terapia , Terapia Molecular DirigidaRESUMEN
INTRODUCTION: Starting dialysis early or late both result in a low quality of life and a poor prognosis in patients undergoing haemodialysis. However, there remains no consensus on the optimal timing of dialysis initiation, mainly because of a lack of suitable methods to assess variations in dialysis initiation time. We have established a novel equation named DIFE (Dialysis Initiation based on Fuzzy-mathematics Equation) through a retrospective, multicentre clinical cohort study in China to determine the most suitable timing of dialysis initiation. The predictors of the DIFE include nine biochemical markers and clinical variables that together influence dialysis initiation. To externally validate the clinical accuracy of DIFE, we designed the assessment of DIFE (ADIFE) study as a prospective, open-label, multicentre, randomised controlled trial to assess the clinical outcomes among patients who initiate dialysis in an optimal start dialysis group and a late-start dialysis group, based on DIFE. METHODS AND ANALYSIS: A total of 388 enrolled patients with end-stage renal disease will be randomised 1:1 to the optimal start dialysis group, with a DIFE value between 30 and 35, or the late-start dialysis group, with a DIFE value less than 30, using the Randomization and Trial Supply Management system. Participants will be assessed for changes in signs and symptoms, dialysis mode and parameters, biochemical and inflammatory markers, Subjective Global Assessment, Kidney Disease Quality of Life Short Form, Cognitive Assessment, medical costs, adverse events and concomitant medication at baseline, predialysis visiting stage and postdialysis visiting stage, every 12-24 weeks. The following data will be recorded on standardised online electronic case report forms. The primary endpoint is 3-year all-cause mortality. The secondary endpoints include non-fatal cerebrocardiovascular events, annual hospitalisation rate, quality of life, medical costs and haemodialysis related complications. ETHICS AND DISSEMINATION: Ethical approval was obtained from the Ethics Committee of the First Affiliated Hospital of Dalian Medical University China (registration no: YJ-KY-2017-119) and the ethics committees of all participating centres. The final results of the ADIFE trial will be presented to the study sponsor, clinical researchers and the patient and public involvement reference group. Findings will be disseminated through peer-reviewed journals, Clinical Practice Guidelines and at scientific meetings. TRIAL REGISTRATION NUMBER: ClinicalTrial.gov. Registry (NCT03385902); pre-results.
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Fallo Renal Crónico/terapia , Ensayos Clínicos Controlados Aleatorios como Asunto , Diálisis Renal/normas , Tiempo de Tratamiento/normas , Adulto , Algoritmos , Estudios de Cohortes , Femenino , Lógica Difusa , Humanos , Masculino , Estudios Prospectivos , Proyectos de InvestigaciónRESUMEN
A number of studies have shown that aldosterone serves an important role in promoting renal interstitial fibrosis, although the specific mechanism remains to be elucidated. A previous study revealed that the fibrotic effect of aldosterone was associated with the expression of allograft inflammatory factor 1 (AIF1) in RAW264.7 macrophage cells, in a time and concentrationdependent manner. However, the exact mechanism through which aldosterone promotes renal interstitial fibrosis remains unknown. In the present study, the effects of aldosterone on renal inflammatory cell infiltration, collagen deposition and the expression levels of AIF1, phosphatidylinositol 3kinase (PI3K), AKT serine/threonine kinase (AKT), mammalian target of rapamycin (mTOR), the oxidative stress factor NADPH oxidase 2 (NOX2) and nuclear transcription factor erythroidrelated factor 2 (Nrf2) were assessed in normal rats, rats treated with aldosterone, rats treated with aldosterone and spironolactone and those treated with spironolactone only (used as the control). The effect of aldosterone on these factors was also investigated in the renal interstitium of unilateral ureteral obstruction (UUO) rats. Additionally, the AIF1 gene was overexpressed and knocked down in macrophage RAW264.7 cells, and the effects of aldosterone on PI3K, AKT, mTOR, NOX2 and Nrf2 were subsequently investigated. The results showed that aldosterone promoted inflammatory cell infiltration, collagen deposition and the expression of AIF1, PI3K, AKT, mTOR and NOX2, but inhibited the expression of Nrf2. In the UUO rats, aldosterone also promoted renal interstitial inflammatory cell infiltration, collagen deposition and the expression of AIF1, NOX2, PI3K, AKT and mTOR, whereas the expression of Nrf2 was downregulated by aldosterone compared with that in the UUOonly group; the influence of aldosterone was counteracted by spironolactone in the normal and UUO rats. In vitro, aldosterone upregulated the expression levels of AKT, mTOR, NOX2 and Nrf2 in RAW264.7 cells compared with those in untreated cells. Suppressing the expression of AIF1 inhibited the effects of aldosterone, whereas the overexpression of AIF1 enhanced these effects in RAW264.7 cells. These findings indicated that aldosterone promoted renal interstitial fibrosis by upregulating the expression of AIF1 and that the specific mechanism may involve AKT/mTOR and oxidative stress signaling.
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Aldosterona/metabolismo , Proteínas de Unión al Calcio/metabolismo , Proteínas de Microfilamentos/metabolismo , Nefritis Intersticial/etiología , Nefritis Intersticial/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismo , Animales , Colágeno/metabolismo , Biología Computacional , Fibrosis , Humanos , Masculino , Ratones , Nefritis Intersticial/patología , Estrés Oxidativo , Células RAW 264.7 , RatasRESUMEN
In order to develop an equation that integrates multiple clinical factors including signs and symptoms associated with uraemia to assess the initiation of dialysis, we conducted a retrospective cohort study including 25 haemodialysis centres in Mainland China. Patients with ESRD (n = 1281) who commenced haemodialysis from 2008 to 2011 were enrolled in the development cohort, whereas 504 patients who began haemodialysis between 2012 and 2013 were enrolled in the validation cohort comprised. An artificial neural network model was used to select variables, and a fuzzy neural network model was then constructed using factors affecting haemodialysis initiation as input variables and 3-year survival as the output variable. A logistic model was set up using the same variables. The equation's performance was compared with that of the logistic model and conventional eGFR-based assessment. The area under the bootstrap-corrected receiver-operating characteristic curve of the equation was 0.70, and that of two conventional eGFR-based assessments were 0.57 and 0.54. In conclusion, the new equation based on Fuzzy mathematics, covering laboratory and clinical variables, is more suitable for assessing the timing of dialysis initiation in a Chinese ESRD population than eGFR, and may be a helpful tool to quantitatively evaluate the initiation of haemodialysis.
Asunto(s)
Fallo Renal Crónico/patología , Redes Neurales de la Computación , Adulto , Anciano , Área Bajo la Curva , Femenino , Tasa de Filtración Glomerular , Humanos , Fallo Renal Crónico/mortalidad , Fallo Renal Crónico/terapia , Modelos Logísticos , Masculino , Persona de Mediana Edad , Curva ROC , Diálisis Renal , Estudios Retrospectivos , Factores de TiempoRESUMEN
Peritoneal dialysis (PD) is often used to treat patients with end stage renal disease, and its long-term complications include excessive inflammation and oxidative stress. Allograft inflammatory factor 1 (AIF-1), as a cytoplasmic protein, is originally identified from infiltrating macrophages, and it was associated with inflammation in the cells other than macrophages, such as endothelial cells and vascular smooth muscle cells. To clarify the molecular mechanisms of AIF-1-modulated pathological changes in the peritoneum during PD, we first detected the AIF-1 expression in peritoneal tissues from PD mice. Results revealed that the pro-fibrotic stimulation caused AIF-1 upregulation and triggered inflammation in peritoneal tissues, and that AIF-1 co-expressed with pan-cytokeratin (a marker of peritoneal mesothelial cells). We next treated primary mouse peritoneal mesothelial cells (pan-cytokeratin and intercellular adhesion molecule 1 positive cells) with 50 or 100 ng/mL recombinant AIF-1, and evaluated the direct effects of AIF-1 on these cells in vitro. We found that exogenous AIF-1 treatment induced inflammation and oxidative stress in mesothelial cells. Apart from the augmented IL-6 and TNF-α secretion, the level of ROS was upregulated and the activity of anti-oxidative SOD was reduced in cells exposed to AIF-1. Moreover, AIF-1 simulation triggered the activation of NF-κB pathway-enhanced the conversion of IκB to phosphorylated IκB and promoted the translocation of NF-κB p65 from cytoplasm into nucleus. Additionally, AIF-1-evoked inflammation in peritoneal mesothelial cells was attenuated by the addition of NF-κB inhibitor (BAY 11-7082). In brief, this study provides us novel information to understand the molecular regulation mechanisms of AIF-1 in peritoneal fibrosis.
Asunto(s)
Proteínas de Unión al Calcio/fisiología , Proteínas de Microfilamentos/fisiología , Fibrosis Peritoneal/metabolismo , Peritoneo/metabolismo , Aloinjertos/inmunología , Animales , Proteínas de Unión al Calcio/metabolismo , Células Epiteliales/metabolismo , Fibrosis/metabolismo , Proteínas I-kappa B/metabolismo , Inflamación/metabolismo , Queratinas/análisis , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas de Microfilamentos/metabolismo , FN-kappa B/metabolismo , Estrés Oxidativo/fisiología , Diálisis Peritoneal/efectos adversos , Fibrosis Peritoneal/fisiopatología , Peritoneo/fisiología , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Factor de Transcripción ReIA/metabolismoRESUMEN
Macrophages have been identified as a key cell type in the pathogenesis of renal interstitial fibrosis (RIF). However, the mechanism through which macrophages drive fibrosis remains unclear. The current study focuses on the effects and possible underlying mechanism of allograft inflammatory factor1 (AIF1), an inflammationresponsive scaffold protein expressed and secreted by macrophages, in promoting fibroblasts to a profibrotic phenotype. In vivo experiments indicated that AIF1, CD68 and αsmooth muscle actin (αSMA) were upregulated in kidney tissues of mice subjected to unilateral ureteric obstruction, while their expressions were inhibited by an aldosterone receptor antagonist, spironolactone. Double immunofluorescence staining revealed that AIF1 expression colocalized with CD68positive macrophages in the renal interstitium, indicating that AIF1 expression in macrophages was increased in the RIF animal model. Furthermore, to identify the role of AIF1 in promoting fibrosis, its expression and secretion by the RAW264.7 macrophage cell line were detected in vitro. The expression levels of αSMA, phosphorylated p38 (pp38) and fibronectin (FN) in fibroblasts were examined subsequent to coculture with macrophages. The increase in AIF1 expression and secretion was confirmed in RAW264.7 cells in response to aldosterone. After 72 h of coculture between fibroblasts and macrophages stimulated with aldosterone, the αSMA expression was induced in fibroblasts, with significantly increased expression levels of FN and pp38 observed. In addition, AIF1 expression was reduced by stable transfection of RAW264.7 cells with AIF1 small interfering RNA, resulting in significantly reduced expression levels of αSMA, pp38 and FN in fibroblasts cocultured with macrophages as compared with normal macrophages. These findings indicate that the expression of AIF1 in macrophages is critical for the activation of renal fibroblasts to a profibrotic phenotype. AIF1 expression was upregulated in macrophages, and may be a novel mechanism linking macrophages to the promotion of RIF via the p38 signaling pathway.
Asunto(s)
Aldosterona/metabolismo , Proteínas de Unión al Calcio/genética , Fibroblastos/patología , Enfermedades Renales/patología , Riñón/patología , Macrófagos/patología , Proteínas de Microfilamentos/genética , Regulación hacia Arriba , Animales , Proteínas de Unión al Calcio/análisis , Línea Celular , Fibroblastos/metabolismo , Fibrosis , Riñón/metabolismo , Enfermedades Renales/genética , Enfermedades Renales/metabolismo , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas de Microfilamentos/análisis , Células RAW 264.7RESUMEN
TGF-ß1-induced excessive deposition of ECM and EMT process of tubular epithelial cells play critical roles in the development and progression of fibrosis in diabetic nephropathy (DN). Orai1 has been demonstrated to be involved in TGF-ß1-induced EMT via TGF-ß/Smad3 pathway. We are aimed to explore the effects of miR-93 on TGF-ß1-induced EMT process in HK2 cells. In this study, our data showed that miR-93 was dramatically decreased in renal tissues of patients with DN and TGF-ß1-stimulated HK2 cells. Moreover, the decreased level of miR-93 was closely associated with the increased expression of Orai1. Overexpression of miR-93 decreased Orai1 expression, and then suppressed TGF-ß1-mediated EMT and fibrogenesis. Next, we predicted that the Orai1 was a potential target gene of miR-93, and demonstrated that miR-93 could directly target Orai1. SiRNA targeting Orai1 was sufficient to suppress TGF-ß1-induced EMT and fibrogenesis in HK2 cells. Furthermore, Overexpression of Orai1 partially reversed the protective effect of miR-93 overexpression on TGF-ß1-mediated EMT and fibrogenesis in HK2 cells. Taken together, Orai1 and miR-93 significantly impact on the progression of TGF-ß1-mediated EMT and fibrogenesis in HK2 cells, and they may represent novel targets for the prevention strategies of fibrosis in the context of DN.