Diacerein mitigates renal ischemia/reperfusion injury via inhibition of renal inflammation, dendritic cells maturation and apoptosis: the role of TLR4/NF-kB/NLRP3/IL-1beta signaling pathway / La diacereína mitiga la lesión por isquemia/reperfusión renal a través de la inhibición de la inflamación renal, la maduración de las células dendríticas y la apoptosis: el papel de la vía de señalización TLR4/NF-kB/NLRP3/IL-1beta

SUMMARY: One of the reasons for acute kidney damage is renal ischemia. Nevertheless, there are limited protective and therapeutic approaches for this problem. Diacerein is an anti-inflammatory drug characterized by numerous biological activities. We aimed to determine the ameliorative impact of diacerein on renal ischemia/reperfusion injury (I/R) condition, exploring the underlying mechanisms. Twenty-four male rats were allotted into four groups (n= 6): sham group; Diacerein (DIA) group; I/R group, in which a non-crushing clamp occluded the left renal pedicle for 45 min, and the right kidney was nephrectomized for 5 min before the reperfusion process; I/R + diacerein group, injected intraperitoneally with 50 mg diacerein/kg i.m 30 minutes prior to I/R operation. Ischemia/ reperfusion was found to affect renal function and induce histopathological alterations. The flow cytometry analysis demonstrated an elevated expression of innate and mature dendritic cells in I/R renal tissues. Moreover, upregulation in the expression of the inflammatory genes (TLR4, Myd88, and NLRP3), and overexpression of the pro-inflammatory cytokines (IL-1β), apoptotic (caspase-3) and pyroptotic (caspase-1) markers were observed in I/R-experienced animals. The aforementioned deteriorations were mitigated by pre-I/R diacerein treatment. Diacerein alleviated I/R-induced inflammation and apoptosis. Thus, it could be a promising protective agent against I/R.

La isquemia renal es una de los motivos del daño renal agudo. Sin embargo, los enfoques protectores y terapéuticos para este problema son limitados. La diacereína es un fármaco antiinflamatorio caracterizado por numerosas actividades biológicas. Nuestro objetivo fue determinar el impacto de mejora de la diacereína en la condición de lesión por isquemia/ reperfusión renal (I/R), explorando los mecanismos subyacentes. Veinticuatro ratas macho se distribuyeron en cuatro grupos (n= 6): grupo simulado; grupo de diacereína (DIA); grupo I/R, en el que una pinza no aplastante ocluyó el pedículo renal izquierdo durante 45 min, y el riñón derecho fue nefrectomizado durante 5 min antes del proceso de reperfusión; Grupo I/R + diacereína, inyectado por vía intraperitoneal con 50 mg de diacereína/kg i.m. 30 min antes de la operación I/R. Se encontró que la isquemia/ reperfusión afecta la función renal e induce alteraciones histopatológicas. El análisis de citometría de flujo demostró una expresión elevada de células dendríticas innatas y maduras en tejidos renales I/R. Además, se observó una regulación positiva en la expresión de los genes inflamatorios (TLR4, Myd88 y NLRP3) y una sobreexpresión de las citoquinas proinflamatorias (IL-1β), marcadores apoptóticos (caspasa-3) y piroptóticos (caspasa-1) en animales con experiencia en I/R. Los deterioros antes mencionados fueron mitigados por el tratamiento previo a la diacereína I/R. La diacereína alivió la inflamación y la apoptosis inducidas por I/R. Por lo tanto, podría ser un agente protector prometedor contra I/R.

Vitamin A could be a Therapeutic Agent in Ischemia/Reperfusion Induced Kidney Injury / La vitamina A podría ser un agente terapéutico en la lesión renal inducida por isquemia/reperfusión

ABSTRACT Introduction: Renal ischemia (I) could develop due to decreased or ceased blood flow to the kidney in some clinical conditions such as shock, sepsis, and kidney transplantation. The re-supply of blood to the kidney is called reperfusion (R). Ischemia and reperfusion periods can cause severe kidney damage. Objectives: When we examined the I/R molecular progression, antioxidant molecules such as vitamin A seem promising treatment agents. This study aimed to investigate the effects of vitamin A on renal I/R injury. Material and Methods: In the study, 40 Sprague-Dawley male rats were divided into five groups (n=8): the control group, only I/R, I/R+1000, I/R+3000, and I/R+9000 IU/kg of Vitamin A groups. Vitamin A was administrated to each group for seven days via oral gavage. Blood and kidney tissue samples were collected at the end of the experiment. We took blood samples for Superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), blood urea nitrogen (BUN), and creatinine (Cr) levels, and determined their values. The tissue samples were stained with hematoxylin/eosin to examine the renal changes histopathologically and stereologically under a light microscope. Results: Histopathological changes caused by I/R were decreased with vitamin A administration in a dose-dependent manner (p<0.05). Vitamin A administration decreased MDA levels and increased SOD and CAT activities (p<0.05). The most effective dose among treatment groups was 9000 IU/kg. There was no significant difference between the controls and all other groups regarding BUN and Cr concentrations. Conclusions: Consequently, administration of vitamin A after renal I/R reduced the histological damage and ameliorated the antioxidant state. These results showed that vitamin A could be a promising agent in treating I/R-induced acute kidney injury.

RESUMEN Introducción: La isquemia renal (I) puede desarrollarse debido a la disminución o interrupción del flujo sanguíneo al riñón en algunas condiciones clínicas como shock, sepsis y trasplante renal. El reabastecimiento de sangre al riñón se denomina reperfusión (R). Tanto la isquemia como los períodos de reperfusión pueden causar graves daños renales. Objetivos: Cuando examinamos la progresión molecular I/R, las moléculas antioxidantes como la vitamina A parecen agentes de tratamiento prometedores. El objetivo de este estudio fue investigar los efectos de la vitamina A sobre la lesión renal I/R. Material y Métodos: En el estudio, 40 ratas macho Sprague-Dawley se dividieron en 5 grupos (n=8) como: control, solo I/R, I/R+1000, I/R+3000 e I/R+9000 UI/kg de la Vitamina A. La vitamina A se administró a cada grupo durante 7 días por vía oral forzada. Al final del experimento se recolectaron muestras de sangre y tejido del riñón. A partir de muestras de sangre se determinaron los niveles de superóxido dismutasa (SOD), malondialdehído (MDA), catalasa (CAT), nitrógeno ureico en sangre (BUN) y creatinina (Cr). Las muestras de tejido se tiñeron con hematoxilina/eosina y los cambios en la histología renal se examinaron histopatológicamente y estereológicamente al microscopio de luz. Resultados: Los cambios histopatológicos causados por I/R disminuyeron con la administración de la vitamina A de manera dependiente de la dosis (p<0,05). La administración de la vitamina A disminuyó los niveles de MDA, aumentó las actividades de SOD y CAT (p<0,05). La dosis más eficaz entre los grupos del tratamiento fue de 9000 UI/kg. No hubo una diferencia significativa entre el grupo control y todos los demás grupos con respecto a las concentraciones de BUN y Cr. Conclusiones: Consiguientemente, la administración de la vitamina A, después de I/R renal, redujo el daño histológico y mejoró el estado antioxidante. Estos resultados mostraron que la vitamina A puede ser un agente promisorio en el tratamiento de la lesión renal aguda (LRA) inducida por I/R.

Protective effects of diosmetin on renal ischemia-reperfusion injury in mice / 现代泌尿外科杂志

【Objective】 To investigate whether diosmetin has protective effects on renal ischemia-reperfusion (I/R) injury in mice and explore the potential mechanism. 【Methods】 A total of 45 BALB/c male mice were randomly divided into the sham group, I/R group and diosmetin treatment group. The protective effects of diosmetin on the renal function of mice was evaluated by detecting blood biochemical indexes and renal tissue HE staining injury score. The expression of p-P65, a key molecule of NF-κB family, was detected with immunohistochemistry and western blotting, and the downstream inflammatory factors of NF-κB signaling pathway were detected with quantitative polymerace chain reaction (qPCR). 【Results】 Compared with the sham group, the I/R group had significantly increased levels of serum creatinine and urea nitrogen, significantly damaged morphological construction, and significantly increased expressions of p-P65 and inflammatory cytokines, while the diosmetin treatment group had decreased levels of serological indexes, improved structure of kidney tissue and reduced expressions of p-P65 and inflammatory cytokines. 【Conclusion】 Diosmetin can protect kidney from I/R-induced acute kidney injury by inhibiting NF-κB pathway in mice.

Role of TXNIP/NLRP3 pathway in renal interstitial fibrosis after renal ischemia-reperfusion injury / 中华急诊医学杂志

Objective:To explore the role of thioredoxin interaction protein (TXNIP)/NOD-like receptor protein 3 (NLRP3) pathway in renal interstitial fibrosis induced by renal ischemia-reperfusion injury (IRI) in mice.Methods:Adult male C57BL/6J mice aged 6 to 8 weeks and TXNIP knockout mice with the same genetic background were selected. The wild type mice were divided into the sham operation (Sham) group and renal IRI group. The TXNIP knockout mice were divided into the sham+TXNIP KO group and IRI+TXNIP KO group, with 12 mice in each group. The model of renal ischemia-reperfusion injury was established by clamping bilateral renal pedicles for 45 min and then restoring perfusion. The sham operation model was only dissociated bilateral renal arteries without other treatment. Blood creatinine, urea nitrogen, kidney injury molecule-1 (Kim-1) and neutrophil gelatinase-associated lipocalin (NGAL), blood transforming growth factor-β (TGF-β) and interleukin 6 (IL-6) were measured on the 1st, 7th and 28th days after reperfusion. The renal cortex was taken on the 1st and 28th days for Masson staining, in which the renal tubule-interstitial injury score was obtained. TGF-β and IL-6 mRNA expression were detected by qPCR, TXNIP, NLRP3, Pro-IL-1β, IL-1β and α-SMA protein expression were detected by Western blot, and MDA and SOD levels were detected by ELISA. Homogeneity test of variance was performed before the statistics of normal distribution measurement data, one-way ANOVA was used for the comparison between multiple groups, and LSD- t test was used for the comparison between the two groups. Results:On the 1st, 7th and 28th days after IRI, compared with the sham group, the Scr, BUN, Kim-1, NGAL, TGF-β and IL-6 were increased continuously in the IRI group ( P<0.05). On the 28th day after IRI, large areas of collagen fibers and inflammatory cell infiltration were observed in the renal interstitium of the IRI group. In the IRI group, the scores of renal tubular injury and renal interstitial fibrosis on the 28th day were significantly higher than those on the 1st day (all P<0.05). On the 1st, 7th and 28th days after IRI, compared with the IRI group, the levels of Scr, BUN, Kim-1, NGAL, TGF-β and IL-6 were significantly decreased in the IRI+TXNIP KO group (all P<0.05). On the 1st and 28th days after IRI, compared to the IRI group, the areas of collagen fibers and inflammatory cell infiltration in the renal interstitium of the IRI+TXNIP KO group were decreased. The renal tubule injury score [Day 1, (192.2 ± 62.4) vs. (103.2 ± 49.1); Day 28, (154.3 ± 93.6) vs. (64.3 ± 24.8), both P<0.05] and interstitial fibrosis score [Day 1, (7.3 ± 3.2) vs. (4.8 ± 1.7); Day 28, (12.8 ± 3.9) vs. (2.3 ± 0.8), both P<0.05] were all decreased. The expression of TGF-β, IL-6 mRNA, TXNIP, NLRP3, Pro-IL-1 β, IL-1 β and α-SMA protein in renal cortex were significantly decreased (both P<0.05). In renal cortex, MDA level was decreased and SOD level was increased (all P<0.05). Conclusions:TXNIP/NLRP3 pathway is involved in the development of renal interstitial inflammation and fibrosis after renal ischemia and reperfusion. Knockout or inhibition of TXNIP can inhibit the progression of acute renal injury to chronic renal disease.

Naringin and trimetazidine improve baroreflex sensitivity and nucleus tractus solitarius electrical activity in renal ischemia-reperfusion injury / Naringin and trimetazidine improve baroreflex sensitivity and nucleus tractus solitarius electrical activity in renal ischemia-reperfusion injury

Resumo Fundamento: O núcleo do trato solitário (NTS) é uma área do cérebro que desempenha um papel fundamental na regulação renal e cardiovascular através dos impulsos dos barorreceptores. Objetivos: O objetivo deste estudo foi avaliar o efeito da Naringina (NAR) e trimetazidina (TMZ), isoladamente e combinadas, na atividade elétrica do NTS e na sensibilidade barorreflexa (SBR) na lesão de isquemia e reperfusão (I/R) renal. Métodos: Foram utilizados quarenta ratos machos Sprague-Dawley (200-250 g), alocados em 5 grupos com 8 ratos cada. Grupos: 1) Sham; 2) I/R; 3) TMZ 5 mg/kg; 4) NAR 100 mg/kg; e 5) TMZ5 + NAR100. A veia femoral esquerda foi canulada para infundir a solução salina ou droga e avaliar a SBR. A I/R foi induzida por oclusão dos pedículos renais por 45 min, seguida de reperfusão de 4 horas. O eletroencefalograma local do NTS foi registrado antes, durante a isquemia e durante a reperfusão. A fenilefrina foi injetada por via intravenosa para avaliar a SBR ao final do tempo de reperfusão. Os dados foram analisados por ANOVA de duas vias com medidas repetidas seguida pelo teste post hoc de Tukey. Um valor de p<0,05 foi considerado como significativo. Resultados: As ondas elétricas do NTS não se alteraram durante o tempo de isquemia, mas diminuíram significativamente durante todos os tempos de reperfusão. A atividade elétrica do NTS e a SBR foram reduzidas drasticamente em ratos com lesão I/R; no entanto, a administração de NAR e TMZ, isoladamente e combinadas, melhorou significativamente essas alterações em ratos com lesão I/R. Conclusões: Os resultados mostraram que a lesão de I/R leva à redução da atividade elétrica da SBR e do NTS, e pode haver uma ligação entre a I/R e a diminuição da SBR. Além disso, a NAR e a TMZ são agentes promissores para tratar complicações de I/R.

Abstract Background: Nucleus tractus solitarius (NTS) is a brain area that plays a key role in kidney and cardiovascular regulation via baroreceptors impulses. Objectives: The aim of this study was to evaluate the effect of naringin (NAR) and trimetazidine (TMZ) alone and their combination on NTS electrical activity and baroreceptor sensitivity (BRS) in renal ischemia- reperfusion (I/R) injury. Methods: Forty male Sprague-Dawley rats (200- 250 g) were allocated into 5 groups with 8 in each. 1) Sham; 2) I/R; 3) TMZ 5 mg/kg; 4) NAR 100 mg/kg; and 5) TMZ5+ NAR100. The left femoral vein was cannulated to infuse saline solution or drug and the BRS was evaluated. I/R was induced by occlusion of renal pedicles for 45 min, followed by 4 hours of reperfusion. The NTS local electroencephalogram (EEG) was recorded before, during ischemia and throughout the reperfusion. Phenylephrine was injected intravenously to evaluate BRS at the end of reperfusion time. The data were analyzed by two-way repeated measurement ANOVA followed by Tukey's post hoc test. A p-value <0.05 was considered significant. Results: NTS electrical waves did not change during ischemia time, while they significantly decreased during the entire reperfusion time. NTS electrical activity and BRS dramatically reduced in rats with I/R injury; however, administration of NAR, TMZ alone or their combination significantly improved these changes in rats with I/R injury. Conclusions: The results showed that I/R injury leads to reduced BRS and NTS electrical activity and there may be an association between I/R and decreased BRS. In addition, NAR and TMZ are promising agents to treat I/R complications.

Prophylactic effect of Potentilla fulgens on renal ischemia-reperfusion injury in rats / Efecto profiláctico de Potentilla fulgens sobre la lesión por reperfusión por isquemia renal en rata

SUMMARY: We aimed to investigate the possible protective effects of Potentilla fulgens on kidney tissue with ischemia- reperfusion using immunohistochemical methods. Wistar rats were grouped as sham, ischemia, ischemia-reperfusion (I/R) and I/R treated with Potentilla fulgens. Renal vessels of the left rat kidney were clamped for 60 minutes for ischemia, IR group had 6 h of reperfusion. 400 mg/kg Potentilla fulgens were given intraperitoneally 5 days before ischemia+reperfusion procedure. Biochemical analysis (MDA, GSH and MPO) of samples were performed. Kidney tissues were fixed with 10 % neutral formalin and routine paraffin tissue follow-up protocol was applied, stained with routine Hematoxylin and Eosin. ADAMTS-5 and Caspase-3 immunostaining was applied for immunohistochemistry and examined under a light microscope. In the ischemia group, inflammation and congestion in the vessels and increased ADAMTS-5 expression in glomerular cells and tubule cells were observed. In reperfusion, an increase in degenerative glomerular cells, tubule cells and intertubular connective tissue and inflammatory cells ADAMTS-5 expression was observed. In the P. fulgens group, degeneration and inflammation decreased and positive ADAMTS-5 expression was observed. In the ischemia and ischemia reperfusion group, increased apoptotic appearance and Caspase-3 positive expression in glomerular and tubular cells, and negative expression in most cells in the P. fulgens group. Potentilla fulgens are thought to stop apoptotic cell development at a certain stage, which affects the cytokine mechanism and plays an important role in the reduction of inflammatory cells and angiogenic regulation.

RESUMEN: El objetivo de este estudio fue investigar los posibles efectos protectores de Potentilla fulgens en el tejido renal con isquemia-reperfusión utilizando métodos inmunohistoquímicos. Se agruparon ratas Wistar como simulación, isquemia, isquemia-reperfusión (I / R) e I / R tratadas con Potentilla fulgens. Los vasos renales del riñón iz- quierdo de las ratas se fijaron durante 60 min por isquemia, el grupo de IR tuvo 6 h de reperfusión. Se administraron 400 mg / kg de Potentilla fulgens por vía intraperitoneal 5 días antes del procedimiento de isquemia + reperfusión. Se realizaron análisis bioquímicos (MDA, GSH y MPO) de muestras. Los tejidos renales se fijaron con formalina neutra al 10 % y se aplicó el protocolo de seguimiento de tejido de parafina de rutina y teñido con hematoxilina y eosina. Se aplicó inmunotinción de ADAMTS-5 y Caspasa-3 para inmunohistoquímica y se examinó con un microscopio óptico. En el grupo de isquemia, se observó inflamación y congestión en los vasos y el aumento de la expresión de ADAMTS-5 en células glomerulares y células tubulares. En la reperfusión, se observó un aumento en la expresión de ADAMTS-5 de células glomerulares degenerativas, células tubulares y tejido conjuntivo intertubular y células inflamatorias. En el grupo de Potentilla fulgens, la degeneración y la inflamación disminuyeron y se observó expresión positiva de ADAMTS-5. En el grupo de isquemia y reperfusión de isquemia, aumentó la apariencia apoptótica y expresión positiva de Caspasa-3 en células glomerulares y tubulares, y expresión negativa en la mayoría de las células del grupo de Potentilla fulgens. Se cree que Potentilla fulgens detiene el desarrollo de las células apoptóticas en una determinada etapa, lo que afecta el mecanismo de las citocinas y juega un papel importante en la reducción de las células inflamatorias y la regulación angiogénica.

The progress on the mechanism of NLRP3 inflammasome in renal ischemia-reperfusion injury / 中华泌尿外科杂志

Renal ischemia-reperfusion injury often occurs during operations that need to block the blood supply of the kidneys. It is an important pathophysiological process that affects the recovery of kidney function and causes acute renal failure. Activated NLRP3 inflammasome can mediate the maturation and release of a variety of pro-inflammatory factors, and participate in the inflammatory response in renal ischemia-reperfusion injury, thereby regulating body inflammation and related cell functions. This article summarized how NLRP3 inflammasome mediated the occurrence of inflammation in renal ischemia reperfusion injury, and further provied a new basis for the prevention and treatment of renal ischemia-reperfusion injury.

Study on therapeutic effect and its related mechanism of anemoside B4 on ischemia reperfusion injury induced by renal artery and vein ligation in rats / 中国中药杂志

The aim of this paper was to investigate the effect and mechanism of anemoside B4 on renal ischemia reperfusion injury in rats. A total of 50 rats were randomly divided into the model group(NS) and anemoside B4 low-dose(1.25 mg·kg~(-1)), medium-dose(2.5 mg·kg~(-1)) and high-dose(5 mg·kg~(-1)) groups after the right kidney was removed and the left kidney was ligated to make the ischemia reperfusion model. Another 10 rats were selected as sham operation group only for normal control group(NS, received normal saline). Automatic biochemical analyzer was used to measure serum blood urea nitrogen(BUN), creatinine(Cre), cerebrospinal fluid(CSF) and urinemicroalbumin(mALB) levels after 5 days of tail vein injection treament. Total urine protein and total urinary albu-min were calculated and kidney samples were collected. Histopathological changes of renal tissues were observed by PAS staining. Western blot analysis was performed to detect the protein expressions of TLR4 and NF-κB in renal inflammatory factors related to NLRP3 pathway and TLR4/NF-κB pathway. The results showed that the levels of BUN, Cre, urinary total protein and urinary total albumin in the model group were significantly increased(P<0.01), with severe renal tubule injury was serious, manifested by obvious expansion of renal tubules, more serious tubular proteins, and some tubular epithelial cells were exfoliated. At the same time, the expression of inflammatory factors related to NLRP3 pathway and TLR4/NF-κB pathway increased significantly(P<0.01 or P<0.05). The levels of BUN, Cre were reduced in different doses of anemoside B4(P<0.05). The levels of total urinary protein and total urinary albumin were decreased in the low and high dose groups of anemoside B4.The level of total urinary albumin in the high-dose group of anemoside B4 was significantly reduced(P<0.05).Renal tubular injury was alleviated, tubular epithelial cell exfoliation was reduced, and the expression of related inflammatory factors was reduced in different degrees(P<0.01 or P<0.05). This study showed that anemoside B4 could alleviate renal ischemia-reperfusion injury in rats. And its mechanism may be related to the inhibition of inflammatory factors related to response mediated by NLRP3 pathway and TLR4/NF-κB pathway by anemoside B4.

Effects of phosphcreatine preconditioning on lung injury induced by renal ischemia-reperfusion in rats / 临床麻醉学杂志

Objective To investigate the effects of phosphcreatine preconditioning on lung injury induced by renal ischemia-reperfusion (IR) in rats.Methods Forty-five SPF male Sprague-Dawley rats, aged 8-10 weeks, weighing 180-220 g, were randomly divided into 3 groups using a random number table:sham operation group (group S), renal IR group (group IR), and phosphcreatine preconditioning group (group PCr), 15 cases in each group.The rats in group S recieved dissoci ation of renal pedicles and right nephrectomy, on top of which renal IR model was prepared in group IR and group PCr.phosphcreatine 150 mg/kg was injected in group PCr for 30 minutes before ischemia, where as rats in group S and group I/R recieved the normal saline at the same time.The blood samples were obtained from left ventricle at 6 hours after reperfusion, the arterial blood gas analysis was performed in order to determined the oxygen partial pressure (PaO2).Serum levels of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were also determined.Fluo 3-AM staining and flow cytometry were used to measure the concentration of alveolar macrophage calcium ions.The lung tissue was obtained with HE staining for determination of microscope examination of pathologic changes, and weight/dry (W/D) ratio were also determined.The lung tissue cell apoptotic rate was measured by Annexin V/PI apoptosis detection reagent staining and flow cytometry.Fluo 3-AM staining and flow cytometry were used to measure the concentration of alveolar macrophage calcium ions.Results Compared with group S, the histopathological demages, W/D ratio, lung tissue cell apoptotic rate, the serum levels of MDA and the concentration of alveolar macrophage calcium ions were signifcant increased (P<0.05), whereas the PaO2 and the activity of SOD were signifcantly decreased in group IR and group PCr (P<0.05).Compared with group IR, the histopathological demages, W/D ratio, lung tissue cell apoptotic rate, the serum levels of MDA and the concentration of alveolar macrophage calcium ions were signifcant decreased (P<0.05), whereas the PaO2 and the activity of SOD were signifcantly increased in group PCr (P<0.05).Conclusion Phosphcreatine preconditioning can attenuate lung injury induced by renal I/R, the mechanism is related to inhabit oxidative stress, and reduce cell apopotosis and calcium overload.

Zinc Supplementation and Ischemia Pre-conditioning in Renal Ischemia/ Reperfusion Injury

@#Backgrounds: Renal ischemia/reperfusion (RIR) is a major cause of kidney dysfunction in clinic. The main objective of this study was to investigate the effect of pre-conditioning ischemia (IPC) and zinc (Zn) supplementation on renal RIR injury. Methods: A total of 63 unilateral nephrectomised male and female Wistar rats were divided into five groups. Group 1 (ShOPR): Rats as sham-operated group were subjected to surgical procedure without RIR. Group 2 (Isch): Rats underwent RIR (left kidney ischemia for 30 min followed by 48 h reperfusion). Group 3 (Zn+Isch): Rats were treated as group 2 but they received Zn sulphate (30 mg/kg) 1 h before induction of RIR. Group 4 (IPC+Isch): Rats were treated as group 2 but they underwent 1 min of ischemia followed by 3 min reperfusion as IPC, which was repeated for three times before induction of RIR. Group 5 (Zn+IPC+Isch): Rats were subjected to receive both Zn sulphate and IPC before induction of RIR. Urine samples were collected in the last 6 h of reperfusion, and finally biochemical and histological measurements were performed. Results: The serum level of creatinine (Cr), normalised kidney weight (KW) and kidney tissue damage score (KTDS) increased by RIR alone significantly (P < 0.05). These parameters were attenuated statistically by Zn supplementation (P < 0.05). However, IPC alone or cotreatment of Zn and IPC did not improve the biochemical and histological markers altered by RIR injury. Conclusion: Zn supplementation had a protective role against RIR while such protective effect was not observed by IPC alone or by co-treatment of Zn and IPC.

Effect of protein kinase C β inhibitor on renal ischemia-reperfusion injury and expression level of macrophage subtypes in rat models / 器官移植

Objective To investigate the effect of protein kinase C (PKC) β inhibitor on the renal ischemia-reperfusion injury (RIRI) in rat models and detect the expression level of macrophage subtypes. Methods Eighteen healthy SD male rats were randomly divided into the Sham operation group (Sham group, n=6), RIRI group (n=6), PKCβ inhibitor +RIRI group (Inhibitor+RIRI group, n=6). Serum and left renal tissue samples were collected at postoperative 24 h. The contents of serum creatinine (Scr) and blood urea nitrogen (BUN) were detected by automatic biochemical analyzer. The infiltration of inflammatory cells and pathological injury in the renal tissues were observed by hematoxylin-eosin (HE) staining. The expression levels of CD68, inducible nitric oxide synthase (iNOS) and CD206 proteins in the renal tissues of rats in each group were assessed by immunohistochemistry and Western blot. Results The contents of serum Scr and BUN in the RIRI group were significantly higher than those in the Sham group (both P < 0.05). The contents of serum Scr and BUN in the Inhibitor+RIRI group were considerably lower than those in the RIRI group (both P < 0.05). No obvious renal injury was noted in the Sham group, whereas renal inflammatory cell infiltration and renal tubular structural damage were observed in the RIRI group. The renal inflammatory cell infiltration and renal tubular structural damage in the Inhibitor+RIRI group was slighter than that in the RIRI group. The protein expression levels of CD68, iNOS and CD206 in the renal tissue of rats in the RIRI group were significantly higher than those in the Sham group (all P < 0.05). The protein expression levels of CD68 and iNOS in the Inhibitor+RIRI group were remarkably lower than those in the RIRI group (all P < 0.05). The expression level of CD206 protein in the Inhibitor+RIRI group was significantly higher than that in the RIRI group (P < 0.05). Conclusions PKC β inhibitor can alleviate the RIRI in rat models to certain extent, which may be correlated with the role of PKC β inhibitor in mitigating inflammatory cell infiltration in ischemic renal tissues and promoting the expression of alternatively activated macrophage

Study on acute kidney injury model induced by renal ischemia-reperfusion in rats / 中国中药杂志

In this study,in-depth systematic evaluation of rat of acute kidney injury(AKI) caused by renal arteriovenous ligation was conducted to better master and apply this model for drug research. Male SD rats of 2-3 months old were employed in this study.The left kidney was removed,and the right kidney received ligation for 40 min and reperfusion for 24 h. Serum creatinine(Crea),urea nitrogen(BUN) and the renal tissue sections were assayed as the basic indicators to evaluate their renal function. The mRNA expression of inflammatory necrosis factors and apoptotic factors was used to evaluate the mechanism of molecular pathophysiological changes. The results showed that the serum Crea and BUN caused by ligation of both renal arteries and veins were significantly higher than those of rats with renal artery ligation. After renal arteriovenous ligation for 40 min and reperfusion for 24 h in rats,the serum Crea of the rats varied from less than 100 μmol·L-1 to more than 430 μmol·L-1. Among them,5 rats showed less than 100 μmol·L-1 serum Crea,20 rats with 100-200 μmol·L-1 serum Crea and 12 rats with more than 430 μmol·L-1. Rats with serum Crea between 300-430 μmol·L-1 accounted for 66.3%(122/184) of the total number of the experiment rats. After 72 h reperfusion,serum Crea in the group of Crea 370-430 μmol·L-1 continued to increase,while the serum Crea in the group of Crea 200-300 μmol·L-1 and the group of Crea 300-370 μmol·L-1 recovered quickly. No matter serum Crea was elevated or decreased,the renal tubules showed pathological changes such as vacuolar degeneration or even necrosis. The mRNA expression levels of Toll-like receptor(TLR4),tumor necrosis factor(TNF-α) and interleukin(IL-6) in renal tissueswere significantly up-regulated,and the effect was most obvious in the group of serum Crea 370-430 μmol·L-1. The study indicated that the model for AKI caused by renal arteriovenous ligation and reperfusion is easy to operate,and the serum Crea and BUN have the characteristics of continuous increase,beneficial to the observation of drug effects. This acute kidney injury is mainly related to the pathophysiological response of inflammatory necrosis.

Dexmedetomidine hydrochloride up-regulates expression of hypoxia inducible factor-1α to alleviate renal ischemiareperfusion injury in diabetic rats / 南方医科大学学报

OBJECTIVE@#To verify whether dexmedetomidine hydrochloride (Dex) alleviates renal ischemia-reperfusion (IR) injury in diabetic rats by increasing the expression of hypoxia inducible factor-1 (HIF-1).@*METHODS@#A rat model of type 2 diabetes mellitus was established by high-fat diet and streptozotocin injection. The rats were subjected to daily intragastric administration of 0.05 mg/kg digoxin for 7 consecutive days and intraperitoneal injection of Dex 2 h before renal IR injury induced by ligation of the bilateral renal arteries for 60 min followed by reperfusion for 120 min. After reperfusion, blood samples were taken for detection of serum creatinine (Scr) and urea nitrogen (BUN) levels. Western blotting was used to detect the expression of HIF-1, cleaved caspase-3, Bcl-2, and Bax in the renal tissues; the expression of the HIF-1, p-eNOS, and eNOS were detected using ELISA. The percentage of apoptotic glomerular cells was assessed using TUNEL assay.@*RESULTS@#The levels of Scr, BUN, HIF-1, p-eNOS, and eNOS and the percentage of apoptotic cells in both normal and diabetic rats increased significantly after renal IR injury ( < 0.05). The expressions of Scr, BUN, p-eNOS, and eNOS decreased while HIF-1 expression increased significantly in Dex-treated rats with renal IR injury ( < 0.05). Compared with the non-diabetic rats, the diabetic rats showed more obvious increase in the expressions of Scr, BUN, p-eNOS, and eNOS following renal IR injury. In the diabetic rats with renal IR injury, Dex treatment prior to the injury significantly lowered the expressions of Scr, BUN, p-eNOS, eNOS, cleaved caspase-3, and Bax, decreased the percentage of apoptotic cells, and increased the levels of HIF-1a and Bcl-2 ( < 0.05). Digoxin treatment significantly antagonized the effects of Dex in the diabetic rats with renal IR injury by increasing the expressions of cleaved caspase-3 and Bax, promoting glomerular cell apoptosis, and decreasing renal expressions of HIF-1 and Bcl-2 ( < 0.05).@*CONCLUSIONS@#Dex alleviates renal IR injury in diabetic rats probably by inhibiting renal expression of HIF-1 and glomerular cell apoptosis.

Study on the Effects of Prophylactic Administration of Ramulus mori Polysaccharides on Inflammatory Responses of Renal Ischemia Reperfusion Injury Model Mice and Its Mechanism / 中国药房

OBJECTIVE： To study on the effects of prophylactic administration of Ramulus mori polysaccharides （RMP） on inflammatory response of renal ischemia reperfusion injury （RIRI） model mice and to explore its possible mechanism. METHODS： Totally 60 C57BL/6 mice were randomly divided to sham operation group， model group， atorvastatin group （positive control， 15 mg/kg）， RMP low-dose， medium-dose and high-dose groups （300， 600， 1 200 mg/kg）. Except for sham operation group， RIRI model was induced in other 5 groups. 24 h before surgery， they were given relevant medicine intragastrically， once a day， for consecutive one week. 24 h after reperfusion， the mice were sacrificed. The serum levels of Scr and BUN were detected. The morphological changes of renal tissue were observed under optical microscope. The serum levels of IL-1β， IL-6， IL-10 and TNF-α were determined by ELISA. Western blot assay was used to determine the protein expressions of Toll-like receptor 4 （TLR4）， p38 mitogen-activation protein kinase （p38MAPK） and p-p38MAPK. RESULTS： Compared with sham operation group， the serum levels of Scr and BUN were significantly elevated in model group  （P＜0.01）. RIRI led to typical inflammatory response of renal tissue， widespread renal tubular epithelial cell degeneration and necrosis， and inflammatory cells infiltration. Serum levels of IL-1β， IL-6， IL-10 and TNF-α were increased significantly （P＜0.01）. The protein expressions of TLR4， p38MAPK and p-p38MAPK were increased significantly in renal cortex （P＜0.01）. Compared with model group， serum levels of Scr and BUN were decreased significantly in administration groups （P＜0.05 or P＜0.01）. The pathological damage of renal tissue was improved in varying degrees， especially in the RMP medium-dose and high-dose groups. Serum levels of IL-1β and IL-6 were decreased significantly in administration groups （P＜0.05 or P＜0.01）. Serum levels of IL-10 were further increased in atorvastatin group and RMP high-dose group （P＜0.01）， and serum level of TNF-α was decreased significantly in atorvastatin group and RMP medium-dose and high-dose groups （P＜0.05 or P＜0.01）. The protein expressions of TLR4 and p-p38MAPK in renal cortex were decreased significantly in administration groups （P＜0.05 or P＜0.01）. CONCLUSIONS： RMP prophylactic administration can improve RIRI of mice， the mechanism of which may be associated with relieving the inflammatory response through inhibition of TLR4/p38MAPK signaling pathway.

Protective effects of D-005, a lipid extract from Acrocomia crispa fruits, against ischemia/reperfusion-induced acute kidney injury in rats

BACKGROUND: Acute kidney injury (AKI) induced by renal ischemia/reperfusion (IR) is associated with enhanced production of reactive oxygen species in renal tissues. D-005, a lipid extract obtained from Acrocomia crispa fruit, has previously shown antioxidant effects. The aim of this work was to evaluate the effects of D-005 on renal IR-induced AKI in rats.METHODS: Rats were randomized into seven groups including a negative control group (vehicle) without AKI and six groups with renal IR-induced AKI as follows: a positive control (vehicle); D-005 treatment at 25, 100, 200, or 400 mg/kg; and dexamethasone at 3 mg/kg. All treatments were orally administered as single doses 1 hour before AKI induction. Biomarkers (serum creatinine, urea, and uric acid concentrations), oxidative variables, and histopathological AKI changes were evaluated in blood and kidney tissues.RESULTS: All D-005 doses protected against IR-induced AKI in rats by significantly decreasing biomarkers and histopathological AKI changes as assessed by reduced serum concentrations of creatinine, urea, and uric acid. In addition, all D-005 doses decreased tubular damage, as shown by fewer detached cells and casts in the tubular lumen. D-005 reversed oxidation disturbance markers by decreasing malondialdehyde and sulfhydryl group concentrations in plasma and in kidney homogenates and by increasing kidney catalase activity. Dexamethasone, the reference substance, protected against IR-induced AKI in rats by reducing biochemical and histological variables of renal damage in a similar manner.CONCLUSION: Administration of single oral doses of D-005 markedly and significantly protected against renal IRinduced AKI, possibly due to its known antioxidant effects.

Sulforaphane attenuates renal ischemia reperfusion injury in mice by Nrf-2 against inflammation / 西安交通大学学报(医学版)

Objective: To explore the effect and mechanism of sulforaphane on renal ischemia reperfusion injury (IRI) in mice to provide experimental evidence for effective intervention in renal IRI. Methods: Totally 24 male C57 mice were randomly divided into four groups: control group (Ctrl), IRI group, sulforaphane intervention IRI group (IRI + SFN), and sulforaphane alone group (SFN). SFN was injected into the mice at the dosage of 500 μg/kg 24 h before surgery. The renal IRI model was established by using the bilateral renal pedicles of mice with non-invasive vascular clamping for 45 min. After 24 h of renal blood perfusion, we sacrificed the mice and collected the whole blood and kidney tissue samples for pathological and biological examinations. The levels of TNF-α, IL-1β and IL-6 in murine serum were tested by ELISA. The expression levels of Nrf-2, keap-1, p-iκBα, IκBα and NFκB p-p65 in murine kidney were detected by Western blot. Results: Sulforaphane could improve renal function in mice with ischemia reperfusion and reduce apoptosis of renal tubular epithelial cells. Sulforaphane could also decrease the levels of TNF-α, IL-1β and IL-6 in murine serum and increase the expression of Nrf-2 in murine kidney tissue after ischemia reperfusion. Moreover, sulforaphane decreased the expression of p-iκBα in total protein and NFκB p-p65 in nucleus protein of renal tissue. Conclusion: Sulforaphane could inhibit phosphorylation, thus inhibiting the phosphorylation of NFκB p65 to translocation into the nucleus. Further it prevented the expressions of downstream inflammatory factors such as TNF-α, IL-1β and IL-6. Finally sulforaphane attenuated renal IRI in mice by Nrf-2 against inflammation.

Remifentanil alleviates renal ischemia/reperfusion injury by activating PI3K/Akt/eNOS pathway / 第二军医大学学报

Objective: To explore the effect and mechanism of intraoperative remifentanil (RF) on renal ischemia/reperfusion (I/R) injury. Methods: Fifty male C57/BL mice were randomly divided into 5 groups: sham group, I/R group, I/R+LY294002 (a phosphatidylinositol 3-kinase [PI3K] inhibitor) group, I/RRF group and I/R + RF+LY294002 group, with 10 mice in each group. Venous blood or renal tissue samples were collected from the mice of each group 6 h after I/R operation. The levels of blood urea nitrogen (BUN) and serum creatinine (SCr) were detected using automatic biochemical analyzer. The expression levels of PI3K/protein kinase B (Akt)/endothelial nitric oxide synthase (eNOS) pathway-related proteins in renal tissues of mice were detected using Western blotting. The aggregation of inflammatory cells was observed by H-E staining. The expression levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6 and IL-10 in renal tissues of mice were detected by enzyme-linked immunosorbent assay. The mRNA expression levels of anti-apoptotic factor Bcl2 and apoptotic factor Caspase-3 in renal tissues were determined by real-time quantitative PCR. Results: Compared with the sham group, the BUN and SCr levels in venous blood were increased in the I/R group, the PI3K expression, phosphorylated-Akt (p-Akt)/Akt ratio and phosphorylated-eNOS (p-eNOS)/eNOS ratio in renal tissues were decreased, the release levels of inflammatory factors (TNF-α, IL-1β, IL-6 and IL-10) were increased, Bcl2 mRNA expression was decreased, and Caspase-3 mRNA expression was increased; and the differences were significant (all P < 0.05). The mice of the I/R group had increased inflammatory cell recruitment in renal tissues. After RF treatment, the mice of the I/R + RF group had decreased levels of BUN and SCr in venous blood, increased PI3K expression, p-Akt/Akt ratio and p-eNOS/eNOS ratio in renal tissues, decreased release levels of inflammatory factors (TNF-α, IL-1β, IL-6 and IL-10), increased Bcl2 mRNA expression, and decreased Caspase-3 mRNA expression; and the differences were significant compared with the mice of the I/R group (all P < 0.05). The inflammatory cell recruitment was decreased in the I/R RF group. Moreover, compared with the mice of the I/RRF group, the mice of the I/RRF LY294002 group had increased levels of BUN and SCr in venous blood, decreased p-eNOS/eNOS ratio in renal tissues, increased IL-1β and IL-6 release, and increased Caspase-3 mRNA expression; and the differences were significant (all P<0.05). The inflammatory cell recruitment was increased in the I/R + RF + LY294002 group. Conclusion: RF exerts protective effect on kidney with I/R injury by alleviating renal inflammation and cell apoptosis through activating PI3K/Akt/eNOS pathway.

Study on Protective Mechanism of Panax Notoginseng Saponins on Rats with Renal Ischemia Reperfusion Injury Based on Klotho / 中国中医药信息杂志

Objective To investigate the effects of Panax Notoginseng saponins (PNS) on protein expression of Klotho in rats with renal ischemia reperfusion injury; To discuss its protective mechanism for model rats. Methods Experimental rats were randomly divided into sham-operation group, model group, positive medicine group, PNS high-, medium- and low-dosage groups. Each administration group was given relevant medicine for gavage, once a day. Renal ischemia reperfusion injury model was established. Rats were sacrificed by taking blood from abdominal aorta after 4 hours of modeling. Serum levels of blood urea nitrogen (BUN), creatinine (SCr), malondialdehyde (MDA) content in kidney tissue, superoxide dismutase (SOD) activity and glutathione peroxidase (GSH-Px) activity were measured. HE staining was used to observe the morphological changes of renal tissue. The protein expressions of Klotho and NF-κB p65 were measured by immunohistochemical method. Results Compared with the sham-operation group, the levels of BUN and SCr in the model group increased significantly (P<0.05); protein expression of Klotho in renal tissue decreased and the protein expression of NF-κB p65 increased (P<0.05). Compared with the model group, the expression of Klotho increased but protein expression of NF-κB p65 decreased in each administration group (P<0.05); Compared with the positive medicine group, the expression of Klotho in PNS high-dosage group increased but protein expression of NF-κB p65 decreased (P<0.05). The protein expression of NF-κB p65 was negatively related to protein expression of Klotho (r=-0.895, P<0.05). Conclusion PNS can inhibit oxidative stress and anti-inflammatory effects through upregulating protein expression of Klotho, and reduce the protein expression of NF-κB p65, and thus exerts renal protective effects.

Protective Effect of Fingolimod on Renal Ischemia Reperfusion Injury Model Mice and Its Mechanism Study / 中国药房

OBJECTIVE:To study the protective effect of fingolimod on renal ischemia reperfusion injury (RIRI) model mice and its mechanism.METHODS:A total of 60 mice were randomly divided into sham operation group,model group,fingolimod group (1 mg/kg) and fingolimod+wortmannin group [fingolimod 1 mg/kg+phosphatidylinositol 3-kinase (PI3K) specific blocker wortmarmin 1.4 mg/kg],with 15 mice in each group.Except for sham operation group,RIRI model was induced in other 3 groups,and those model mice were given relevant medicine via caudal vein at once 24 h before surgery.Serum of mice were collected in each group after 24 h perfusion.Serum levels of Scr and BUN were measured by automatic biochemical analyzer.The pathological changes of renal tissue were observed under light microscope.The protein expression of intercellular cell adhesion molecule-1 (ICAM-1),monocyte chemoattractant protein-1 (MCP-1) and phosphorylated protein kinase B (p-Akt) in renal tissue were measured by Western blot assay.RESULTS:Compared with sham operation group,the serum levels of Scr and BUN in model group were increased significantly (P＜0.01).Pathological changes were found in the kidney,and RIRI led to widespread renal tubular epithelial cell injury,apoptosis and inflammatory cells infiltration.The protein expression of ICAM-1 and MCP-1 in renal tissue were increased significantly (P＜0.01),the protein expression of p-Akt was increased slightly (P＞0.05).Compared with model group,other indexes of fingolimod group were improved significantly (P＜0.01) except that the protein expression of p-Akt in renal tissue was increased significantly (P＜0.01).Compared with fingolimod group,above indexes of fingolimod+wortmannin group were reversed (P＜0.05 or P＜0.01).CONCLUSIONS:Fingolimod can obviously ameliorate renal injury induced by RIRI in mice,the mechanism of which may be associated with the activation of PI3K/Akt signaling pathway.

Effect of 17β-estradiol on renal tubular cells apoptosis and inflammation induced by renal ischemia /reperfusion injury in rats / 临床与实验病理学杂志

Purpose To investigate the effects and mechanism of 17β-estradiol on the apoptosis and inflammation of renal tubular cells in rats with renal ischemia/reperfusion injury. Methods All the female Sprague-Dawley rats were ovariectomized and randomly divided into four groups: Control group, Sham group, I/R group and estrogen plus I/R (E2 + I/R) group (n = 8). Right kidney of the rat was excised and artery of the left kidney was blockaded for 45 min.24 h after the reperfusion, we collected the blood and nephridial tissue of each group. An automatic biochemical analyzer was used to measure the expression level of BUN and Cr in blood. Hematoxylin-eosin (HE) staining was used to observe the pathological changes and the degree of inflammatory reaction of the ischemia/reperfusion injury kidney. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method was used to detect the apoptosis of renal tubular cells. The expression levels of Cleaved-Caspase-3 protein were measured by Western blot, while the numbers of CD4+ T lymphocyte infiltration in each group were tested by immunofluorescence (IF). Results Compared with the Sham group, expression level of BUN, Cr and Cleaved-Caspase-3 in I/R group significantly increased (P＜0.05) as well as the number of apoptotic cells (P＜0.05). In the meantime, inflammatory reaction significantly aggravated (P＜0.05) and the number of CD4 + T lymphocytes increased remarkably (P＜0.05). However, expression level of BUN, Cr and Gleaved-Caspase-3 in E2 + I/R group decreased significantly (P＜0.05) and the pathological damage in the kidney was alleviated (P＜0.05) compared with I/R group, furthermore, the number of apoptotic cells decreased (P＜0.05) compared with I/R group. The inflammatory reaction significantly blunted (P＜0.05) and the infiltration of CD4 + T lymphocytes decreased remarkably (P＜0.05) compared with I/R group. Conclusion Estrogen can inhibit the expression of Cleaved-Caspase-3 in renal tissue during ischemia/reperfusion injury and reduce the apoptosis of renal tubular cells. It can also reduce the infiltration of CD4 + T lymphocytes, thus playing a protective role on renal ischemia/reperfusion injury.