Effects and Mechanism of Salvianolic Acid B on the Injury of Human Renal Tubular Epithelial Cells Induced by Iopromide

With the increasing application of medical imaging contrast materials, contrast-induced nephropathy (CIN) has become the third major cause of iatrogenic renal insufficiency. CIN is defined as an absolute increase in serum creatinine levels of at least 0.50 mg/dl or an increase >25% of serum creatinine from baseline after exposure to contrast. In this study, the protective effects of salvianolic acid B (Sal B) were detected in human renal tubular epithelial cells (HK-2) exposed to iopromide. The results showed that different concentrations of Sal B counteract the loss of cell viability induced by iopromide, and reduce cell apoptosis, the reactive oxygen species (ROS) levels, and the levels of endoplasmic reticulum stress (ERS)–related and apoptosis-related proteins such as p-IRE-1α, p-eIF-2α/eIF-2α, p-JNK, CHOP, Bax/Bcl-2, and cleaved caspase-3. In addition, Sal B at a concentration of 100 μmol/L inhibited ERS and reduced cell damage to a similar extent as the ERS inhibitor 4-PBA. Importantly, treatment with Sal B could abolish the injury induced by ERS agonist tunicamycin, increasing cell viability and the mitochondrial membrane potential, as well as significantly reducing ROS levels and the expression of Bax/Bcl-2, cleaved-caspase-3, GRP78, p-eIF2α, p-JNK, and CHOP. These results suggested that the protective effect of Sal B against HK-2 cell injury induced by iopromide may be related to the inhibition of ERS.

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Mcl-1 is downregulated in cisplatin-induced apoptosis, and proteasome inhibitors restore Mcl-1 and promote survival in renal tubular epithelial cells

AJP Renal Physiology ◽

10.1152/ajprenal.00505.2006 ◽

2007 ◽

Vol 292 (6) ◽

pp. F1710-F1717 ◽

Cited By ~ 28

Author(s):

Cheng Yang ◽

Varsha Kaushal ◽

Sudhir V. Shah ◽

Gur P. Kaushal

Keyword(s):

Epithelial Cells ◽

Cytochrome C ◽

Caspase 3 ◽

Proteasome Inhibitors ◽

Mrna Levels ◽

Dependent Manner ◽

Tubular Epithelial Cells ◽

Renal Tubular Epithelial Cells ◽

Renal Tubular ◽

Induced Apoptosis

Mcl-1 is an antiapoptotic member of the Bcl-2 family that plays an important role in cell survival. We demonstrate that proteasome-dependent regulation of Mcl-1 plays a critical role in renal tubular epithelial cell injury from cisplatin. Protein levels of Mcl-1 rapidly declined in a time-dependent manner following cisplatin treatment of LLC-PK1cells. However, mRNA levels of Mcl-1 were not altered following cisplatin treatment. Expression of other antiapoptotic members of the Bcl-2 family such as Bcl-2 and BclxL was not affected by cisplatin treatment. Cisplatin-induced loss of Mcl-1 occurs at the same time as the mitochondrial release of cytochrome c, activation of caspase-3, and initiation of apoptosis. Treatment of cells with cycloheximide, a protein synthesis inhibitor, revealed rapid turnover of Mcl-1. In addition, treatment with cycloheximide in the presence or absence of cisplatin demonstrated that cisplatin-induced loss of Mcl-1 results from posttranslational degradation rather than transcriptional inhibition. Overexpression of Mcl-1 protected cells from cisplatin-induced caspase-3 activation and apoptosis. Preincubating cells with the proteasome inhibitor MG-132 or lactacystin not only restored cisplatin-induced loss of Mcl-1 but also resulted in an accumulation of Mcl-1 that exceeded basal levels; however, Bcl-2 and BclxL levels did not change in response to MG-132 or lactacystin. The proteasome inhibitors effectively blocked cisplatin-induced mitochondrial release of cytochrome c, caspase-3 activation, and apoptosis. These studies suggest that proteasome regulation of Mcl-1 is crucial in the cisplatin-induced apoptosis via the mitochondrial apoptotic pathway and that Mcl-1 is an important therapeutic target in cisplatin injury to renal tubular epithelial cells.

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Astragalosides IV Improves Palmitic Acid-induced Renal Tubular Epithelial Cells Injury Due to Inhibition of NLRP3 Inflammasome

10.21203/rs.3.rs-122884/v1 ◽

2020 ◽

Author(s):

Yanhua Zhang ◽

Yan Li ◽

Yong Su ◽

Yunfeng Zhu ◽

Yuli Han ◽

...

Keyword(s):

Epithelial Cells ◽

Cell Viability ◽

Palmitic Acid ◽

Cell Counting ◽

Renal Tubules ◽

Tubular Epithelial Cells ◽

Renal Tubular Epithelial Cells ◽

Caspase 1 ◽

Renal Tubular

Abstract BackgroundDiabetic nephropathy (DN) is a serious complication of type 2 diabetes mellitus (T2DM). Hyperlipidemia plays a key role in occurrence and development of DN. The main saturated fatty acids of palmitic acid (PA) is closely related to glomeruli and tubules injury in T2DM. Astragaloside IV (AS-IV) has comprehensive pharmacological effects, such as anti-inflammation, anti-oxidation and anti-apoptosis. However, whether AS-IV can attenuate PA-induced renal tubular epithelial cells damages and its underlying mechanisms remain unclear.MethodsIn vitro, the HK-2 cells were stimulated with PA (200 μM) for 6, 9, 12, and 24 h respectively, then treated with AS-IV (10, 20 or 40 μM) and Apo (100 μM) for 24 h. In vivo, we investigated the effects of AS-IV in high-fat diets (HFD) and low-dose streptozotocin (STZ)-induced type 2 DN rats. Cell viability was detected by Cell Counting Kit-8 (CCK-8) assay kit. The lipid deposition was detected by ORO Staining assay kit. Annexin V-FITC/PI staining was used to detect the cell apoptosis. Immunofluorescence staining and Immunohistochemistry were used to measure the expressions of NLRP3 and NOX4. The ELISA kits were conducted to assess the levels of IL-1β, IL-18, GSH and SOD. Western blotting was used to assess the expressions of NLRP3, caspase-1, ASC, 1L-1β, NOX4, p22phox and p47phox proteins.ResultsOur results indicated that PA exposure not only significantly decreased cell viability, GSH and SOD expressions, and increased lipids deposition and apoptosis, but also increased the level of ROS and the expressions of IL-1β and IL-18. Moreover, PA exposure significantly increased the expressions of NADPH oxidase 4 (NOX4), NLRP3, ASC and caspase-1. However, all these abnormalities were significantly ameliorated by AS-IV and apocynin (Apo) treatment. In addition, our results indicated that AS-IV (80 mg/kg) could attenuate the renal tubular pathological injury and lipids deposition, and decrease the expressions of NLRP3, ASC, caspase-1, NOX4, p22phox and p47phox in renal tubules. ConclusionsThe study suggested that AS-IV could improve PA-induced HK-2 cells injury and renal tubular damage in T2DM rats, and the mechanism may be related to inhibition of NOX4-mediated NLRP3 inflammasome activation.

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Effect of Forkhead Box O1 in Renal Tubular Epithelial Cells on Endotoxin-Induced Acute Kidney Injury

AJP Renal Physiology ◽

10.1152/ajprenal.00289.2020 ◽

2020 ◽

Author(s):

Mengxi Zhang ◽

Wei Dong ◽

Zhilian Li ◽

Zhenmeng Xiao ◽

Zhiyong Xie ◽

...

Keyword(s):

Acute Kidney Injury ◽

Epithelial Cells ◽

Cell Viability ◽

Kidney Diseases ◽

Kidney Injury ◽

Mitochondrial Damage ◽

Tubular Epithelial Cells ◽

Renal Tubular Epithelial Cells ◽

Forkhead Box ◽

Renal Tubular

Mitochondrial damage in renal tubular epithelial cells (RTECs) is a hallmark of endotoxin-induced acute kidney injury (AKI). Forkhead box O1 (FOXO1) is responsible for regulating mitochondrial function and is involved in several kidney diseases. Herein, we investigated the effect of FOXO1 on endotoxin-induced AKI and the related mechanism. In vivo, FOXO1 downregulation in mouse RTECs and mitochondrial damage were found in endotoxin-induced AKI. Overexpression of FOXO1 by kidney focal adeno-associated virus (AAV) delivery improved renal function and reduced mitochondrial damage. PGC1-α, a master regulator of mitochondrial biogenesis and function, was reduced in endotoxin-induced AKI, but the reduction was reversed by FOXO1 overexpression. In vitro, exposure to LPS led to a decline in HK-2 cell viability, mitochondrial fragmentation, and mitochondrial superoxide accumulation, as well as downregulation of FOXO1, PGC1-α and mitochondrial complex I/V. Moreover, overexpression of FOXO1 in HK-2 cells increased HK-2 cell viability and PGC1-α expression, and it alleviated the mitochondrial injury and superoxide accumulation induced by LPS. Meanwhile, inhibition of FOXO1 in HK-2 cells by siRNA treatment decreased PGC1-α expression and HK-2 cell viability. Chromatin immunoprecipitation assays and PCR analysis confirmed that FOXO1 bound to the PGC1-α promoter in HK-2 cells. In conclusion, downregulation of FOXO1 in RTECs mediated endotoxin-induced AKI and mitochondrial damage. Overexpression of FOXO1 could improve renal injury and mitochondrial dysfunction, and this effect occurred at least in part as a result of PGC1-α signaling. FOXO1 might be a potential target for the prevention and treatment of endotoxin-induced AKI.

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Insulin Inhibits Caspase-3 Activity in Human Renal Tubular Epithelial Cells via the PI3-Kinase/Akt Pathway

Cellular Physiology and Biochemistry ◽

10.1159/000129386 ◽

2008 ◽

Vol 21 (4) ◽

pp. 279-286 ◽

Cited By ~ 8

Author(s):

Markus Meier ◽

Martin Nitschke ◽

Caroline Hocke ◽

Jan Kramer ◽

Wolfram Jabs ◽

...

Keyword(s):

Epithelial Cells ◽

Caspase 3 ◽

Pi3 Kinase ◽

Akt Pathway ◽

Tubular Epithelial Cells ◽

Renal Tubular Epithelial Cells ◽

Renal Tubular

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Caspase-11 promotes cisplatin-induced renal tubular apoptosis through a caspase-3-dependent pathway

AJP Renal Physiology ◽

10.1152/ajprenal.00091.2017 ◽

2018 ◽

Vol 314 (2) ◽

pp. F269-F279 ◽

Cited By ~ 8

Author(s):

Naijun Miao ◽

Bao Wang ◽

Dan Xu ◽

Yanzhe Wang ◽

Xinxin Gan ◽

...

Keyword(s):

Epithelial Cells ◽

Cell Apoptosis ◽

Caspase 3 ◽

Tubular Epithelial Cells ◽

Tubular Injury ◽

Renal Tubular Epithelial Cells ◽

Small Interference Rna ◽

Fluorescence Confocal Microscopy ◽

Renal Tubular ◽

Interference Rna

Renal tubular injury is the hallmark of cisplatin-induced nephrotoxicity. Caspase-11, a member of the caspase family, plays an important role in inflammation and cell death. However, its role in cisplatin-induced renal tubular injury remains unclear. In cisplatin-treated mice, caspase-11 expression was significantly elevated and the expression of caspase-11 was mainly located in renal tubule. Inhibition of caspase-11 by small-interference RNA or its inhibitor wedelolactone attenuated cisplatin-induced renal dysfunction and tubular injury. In cultured primary renal tubular epithelial cells, cisplatin significantly promoted the expression and activation of caspase-11. Inhibition of caspase-11 by small-interference RNA reduced cisplatin-induced cell apoptosis. Overexpression of caspase-11 promoted cell apoptosis by activating the caspase-3-related cell apoptosis. Furthermore, coimmunoprecipitation results showed there was a direct interaction between caspase-11 and caspase-3, and the interaction was enhanced by cisplatin. The fluorescence confocal microscopy results showed that caspase-11 and caspase-3 were colocalized in the cytoplasm of renal tubular epithelial cells. These results demonstrate that caspase-11 plays an important role in cisplatin-induced renal tubular injury. Caspase-11 promotes renal epithelial cell apoptosis by activating the caspase-3-dependent apoptotic pathway. Caspase-11 might be a potential target for therapeutic treatment against cisplatin-induced nephrotoxicity.

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Caspase 3/GSDME-dependent pyroptosis contributes to chemotherapy drug-induced nephrotoxicity

Cell Death and Disease ◽

10.1038/s41419-021-03458-5 ◽

2021 ◽

Vol 12 (2) ◽

Author(s):

Xiujin Shen ◽

Haibing Wang ◽

Chunhua Weng ◽

Hong Jiang ◽

Jianghua Chen

Keyword(s):

Epithelial Cells ◽

Caspase 3 ◽

Tubular Epithelial Cell ◽

Tubular Epithelial Cells ◽

Chemotherapeutic Drug ◽

Renal Tubular Epithelial Cells ◽

Renal Tubular Epithelial Cell ◽

Drug Induced ◽

Jnk Signaling ◽

Renal Tubular

AbstractChemotherapy drug-induced nephrotoxicity limits clinical applications for treating cancers. Pyroptosis, a newly discovered programmed cell death, was recently reported to be associated with kidney diseases. However, the role of pyroptosis in chemotherapeutic drug-induced nephrotoxicity has not been fully clarified. Herein, we demonstrate that the chemotherapeutic drug cisplatin or doxorubicin, induces the cleavage of gasdermin E (GSDME) in cultured human renal tubular epithelial cells, in a time- and concentration-dependent manner. Morphologically, cisplatin- or doxorubicin-treated renal tubular epithelial cells exhibit large bubbles emerging from the cell membrane. Furthermore, activation of caspase 3, not caspase 9, is associated with GSDME cleavage in cisplatin- or doxorubicin-treated renal tubular epithelial cells. Meanwhile, silencing GSDME alleviates cisplatin- or doxorubicin-induced HK-2 cell pyroptosis by increasing cell viability and decreasing LDH release. In addition, treatment with Ac-DMLD-CMK, a polypeptide targeting mouse caspase 3-Gsdme signaling, inhibits caspase 3 and Gsdme activation, alleviates the deterioration of kidney function, attenuates renal tubular epithelial cell injury, and reduces inflammatory cytokine secretion in vivo. Specifically, GSDME cleavage depends on ERK and JNK signaling. NAC, a reactive oxygen species (ROS) inhibitor, reduces GSDME cleavage through JNK signaling in human renal tubular epithelial cells. Thus, we speculate that renal tubular epithelial cell pyroptosis induced by chemotherapy drugs is mediated by ROS-JNK-caspase 3-GSDME signaling, implying that therapies targeting GSDME may prove efficacious in overcoming chemotherapeutic drug-induced nephrotoxicity.

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Syndecan-1 (CD138) immunohistochemical expression patterns in lupus nephritis; reflections on different clinicopathological parameters

Journal of Nephropathology ◽

10.34172/jnp.2021.06 ◽

2020 ◽

Vol 10 (1) ◽

pp. e06-e06

Author(s):

Marwa M Shakweer ◽

Lobna S Shash

Keyword(s):

Epithelial Cells ◽

Lupus Nephritis ◽

Serum Creatinine ◽

Urinary Albumin ◽

Clinicopathological Parameters ◽

Tubular Epithelial Cells ◽

Renal Tubular Epithelial Cells ◽

Immunohistochemical Expression ◽

Biopsy Specimens ◽

Renal Tubular

Introduction: Syndecan 1 (SCD1) is a lectin expressed at the surface of renal tubular epithelial cells and plasma cells. In epithelial cells, cell surface syndecan1 is cleaved by inflammation-induced proteases (eg, ADAMTS, MMP), since loss of cell surface syndecan1 is associated with higher susceptibility to cell damage. Objectives: To explore a potential additional value of SCD1 immunohistochemical expression in lupus nephritis specimens of different ISN/RPS classes and NIH activity and chronicity indices. Patients and Methods: This retrospective study included 50 renal biopsy specimens diagnosed as lupus nephritis at the pathology laboratory, and electron microscopy (EM) laboratory of Ain-Shams University specialized hospitals. Data were collected from records as personal data, medical history and laboratory results including serum creatinine and proteinuria. Immunohistochemical expression of syndecan-1 was evaluated in renal tubular epithelial cells (TECs) followed by correlation with different clinicopathological parameters. Results: Fifty renal biopsy specimens with lupus nephritis including 14 cases of class II, 4 cases of class III, 20 cases of class IV and 12 cases of class V were re-evaluated. The mean serum creatinine was 1.57 ± 0.67 mg/dL. Nine cases (18%) were negative for proteinuria, while 41 cases (82%) were presented with proteinuria with a mean of 1.5 ± 0.9 g/24 h. There was no statistically significant difference in the percentage of SCD-1 expression with different lupus classes. Serum creatinine and albumin showed a statistically significantly different correlation with semiquantitative score of SCD-1 expression. The highest value of creatinine detected with score 1 of SCD-1 expression (P=0.038) and the highest value of urinary albumin was recorded with score 1 of SCD-1 expression. Accordingly, the lowest mean of urinary albumin recorded in SCD-1 score 3 (P<0.001). There was a weekly negative association between loss of SCD-1 expression and increased NIH activity and chronicity indices. Conclusion: Loss of syndecan immunohistochemical expression in renal TECs in lupus nephritis is highly associated with proteinuria and elevated serum creatinine and can be used as a predictive marker for disease severity and progression.

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