Inhibition of autophagy-attenuated calcium oxalate crystal-induced renal tubular epithelial cell injury in vivo and in vitro

Liu, Yunlong; Li, Derong; He, Ziqi; Liu, Quan; Wu, Jianhua; Guan, Xiangdong; Zhou, Tao; Deng, Yaoliang

doi:10.18632/oncotarget.23383

Cited by 34 publications

(37 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Autophagy is also associated with a variety of pathological conditions, such as neurodegeneration, cardiomyopathy’ and cancer [33]. Liu et al clearly demonstrated that autophagy activated via the reactive oxygen species pathway is deleterious in CaOx crystal-induced HK-2 cell injury [22]. The autophagy level that was observed in the current study was directly observed under confocal microscopy but also indirectly by the expression of LC3, Beclin-1, and Bcl-2 (Fig.…”

Section: Discussionsupporting

confidence: 56%

See 1 more Smart Citation

Ethyl Pyruvate Attenuates CaCl2-Induced Tubular Epithelial Cell Injury by Inhibiting Autophagy and Inflammatory Responses

Zhao¹,

Cheng²,

Li³

et al. 2018

Kidney Blood Press Res

View full text Add to dashboard Cite

Background/Aims: Nephrolithiasis is one of the most prevalent diseases of the urinary system. Approximately 80% of human kidney stones are composed of calcium oxalate (CaOx), and hypercalciuria is one of the most common metabolic disorders. Emerging evidence indicates that autophagy and inflammatory responses are related to the formation of CaOx nephrolithiasis. However, the roles of autophagy and inflammation in patients with hypercalciuria remain unclear. Ethyl pyruvate (EP) displays protective effects in experimental models of many illnesses. In this study, we investigated the protective effects of EP in vitro through its inhibition of autophagy and inflammatory responses after CaCl₂-induced tubular epithelial cell injury. Methods: First, we cultured human tubular epithelial (HK-2) cells in the presence of various concentrations of CaCl₂ (0, 0.1, 0.25, 0.5, 1.0, 1.5, and 2.0 mg/ml) for 12 h and EP (0, 1.0, 2.5, 5.0, and 10.0 mM) for 2 h to select the optimum concentration using the Cell Counting Kit-8 assay and lactate dehydrogenase (LDH) assay. Cells in culture were stimulated with CaCl₂ (1.0 mg/ml, 12 h) with or without EP pretreatment (2.5 mM, 2 h). After the exposure, we detected the expression of inflammation-related proteins using an enzyme-linked immunosorbent assay and Western blot analysis. Finally, the levels of autophagy-related proteins were determined through Western blot analysis, and the number of GFP-LC3 dots and autophagic vacuoles was detected under confocal microscopy. Results: With the use of the Cell Counting Kit-8 assay and the LDH assay, we identified the optimum concentration for CaCl₂ (1.0 mg/ml) treatment and EP pretreatment (2.5 mM). Our research indicated that CaCl₂ can induce autophagy and inflammatory responses in HK-2 cells. Furthermore, treatment with EP prior to CaCl₂ stimulation attenuated HK-2 cell injury by inhibiting autophagy and inflammation. Conclusion: Our results provide evidence that EP attenuates CaCl₂-induced injury of HK-2 cells by downregulating the expression of inflammation and autophagy proteins that may be associated with the inhibition of the high-mobility group box-1 (HMGB1)/toll-like receptor 4 (TLR4)/NF-κB pathway and the competitive interaction with Beclin-1 of HMGB1.

show abstract

Section: Discussionsupporting

confidence: 56%

“…Liu et al also demonstrated that CaOx crystal retention can induce autophagy in vivo and in vitro and aggravate HK-2 cell injury. Chloroquine diphosphate inhibits autophagy, and renal injury and the formation of nephrolithiasis markedly decreases [22]. …”

Section: Discussionmentioning

confidence: 99%

Ethyl Pyruvate Attenuates CaCl2-Induced Tubular Epithelial Cell Injury by Inhibiting Autophagy and Inflammatory Responses

Zhao¹,

Cheng²,

Li³

et al. 2018

Kidney Blood Press Res

View full text Add to dashboard Cite

show abstract

“…However, treatment with autophagy inhibitor chloroquine significantly decreased the levels of the above indicators in stone model. In addition, we have previously demonstrated that autophagy was deleterious in CaOx crystal-induced renal tubular epithelial cell injury [20]. Therefore, we inferred that autophagy activation may be further promoted the formation of CaOx kidney stones.…”

Section: Discussionmentioning

confidence: 77%

Inhibition of Autophagy Attenuated Ethylene Glycol Induced Crystals Deposition and Renal Injury in a Rat Model of Nephrolithiasis

Liu

Xiang

et al. 2018

Kidney Blood Press Res

Self Cite

View full text Add to dashboard Cite

Background/Aims: Nephrolithiasis is a common and frequently occurring disease, its exact pathogenesis is remains unclear. Emerging data suggest that autophagy plays a vital role in the pathophysiological processes of kidney diseases. Therefore, this study was designed to investigate the potential role of autophagy in the formation of calcium oxalate (CaOx) kidney stones in rat model. Methods: Thirty-two rats were randomly divided into four groups (eight rats/group): untreated control group, stone model group, rapamycin-treated group, chloroquine-treated group. Rat models of CaOx nephrolithiasis was administration of 0.75% ethylene glycol (EG) in their drinking water for 4 weeks. Western blot and transmission electron microscope (TEM) were used to detect the expression of autophagy related protein LC3-II, BECN1 and p62 and autophagic vacuoles respectively. Renal function was evaluated by measuring the levels of serum CRE and BUN. Renal tubular injury markers NGAL and Kim-1 was determined by ELISA kits. Von Kossa staining was used to assess crystal deposits and histological tissue injury. TUNEL staining was employed to assess apoptosis of the renal tubular cell. Results: Compare with the controls, the expression of autophagy related protein LC3-II, BECN1 and number of autophagic vacuoles were increased significantly, whereas the p62 protein level was decreased in the stone model group. The levels of apoptosis, serum CRE and BUN, NGAL and Kim-1 in the stone model group were increased compared with the control group and crystals deposition and renal injury were increased significantly. However, the levels of autophagy, kidney injury and crystal deposition were decreased by chloroquine but increased by rapamycin. Conclusion: These findings suggested that rats were administration of ethylene glycol could lead to the formation of CaOx nephrolithiasis and autophagy activation. Inhibiting autophagy could be an effective therapeutic approach for decreasing the formation of nephrolithiasis.

show abstract

“…CaOx‐induced kidney stone formation is a very complex process involved with oxidative stress, inflammation, autophagy, and exocrine body, and so on . Recently, the interaction between the crystal and renal tubular epithelial HK‐2 cells has been considered as an important factor in the process of stone formation, such as the adhesion and endocytosis of crystals to cells.…”

Section: Discussionmentioning

confidence: 99%

Atorvastatin inhibits renal inflammatory response induced by calcium oxalate crystals via inhibiting the activation of TLR4/NF‐κB and NLRP3 inflammasome

et al. 2020

Self Cite

View full text Add to dashboard Cite

This study aimed to investigate the renal protective effect of atorvastatin (ATV) on the kidney inflammation induced by calcium oxalate (CaOx) crystals. A cell model of cell‐crystal interactions and a rat model of CaOx kidney stone were established. The expressions of TLR4, NF‐κB, NLRP3, and cleaved caspase‐1 in cells and rat kidney tissues were detected using Western blot, immunohistochemical, and/or immunofluorescence. The concentrations of malondialdehyde (MDA), superoxide dismutase (SOD), reactive oxygen species (ROS) in cells, and lactic acid dehydrogenase (LDH) in the culture medium were measured. The secreted levels of interleukin (IL)‐1β, IL‐18, IL‐6, and tumor necrosis factor‐α (TNF‐α) were examined by ELISA. The serum levels of creatinine (CRE) and blood urea nitrogen (BUN) were measured. von Kossa staining was used for the evaluation of renal lens deposition. The CaOx model group showed significantly decreased SOD level; increased concentrations of MDA; ROS and LDH; elevated expressions of TLR4, NF‐κB, NLRP3, and cleaved caspase‐1; and the elevated release of IL‐1β, IL‐18, IL‐6, and TNF‐ α as compared to the control group. The treatment with ATV significantly inhibited the formation of CaOx kidney stone by increasing the level of SOD; downregulating MDA, ROS, and LDH; inhibiting the expressions of TLR4, NF‐κB, NLRP3 and cleaved caspase‐1; and blocking the secretion of inflammatory cytokines. In addition, the serum levels of CRE and BUN, and the intrarenal crystal deposition were also significantly decreased in ATV‐treated rats. In summary, oxidative stress, TLR4/NF‐κB, and NLRP3 inflammasome pathways are involved in renal inflammatory responses induced by CaOx crystals. ATV treatment significantly suppressed oxidative stress, inhibited the activation of TLR4/NF‐κB and NLRP3 inflammasome pathways, and decreased the release of inflammatory mediators, thereby ameliorating CaOx crystal‐induced damage and crystal deposition in HK‐2 cells and rat kidney tissues.

show abstract

Inhibition of autophagy-attenuated calcium oxalate crystal-induced renal tubular epithelial cell injury in vivo and in vitro

Cited by 34 publications

References 42 publications

Ethyl Pyruvate Attenuates CaCl<sub>2</sub>-Induced Tubular Epithelial Cell Injury by Inhibiting Autophagy and Inflammatory Responses

Ethyl Pyruvate Attenuates CaCl<sub>2</sub>-Induced Tubular Epithelial Cell Injury by Inhibiting Autophagy and Inflammatory Responses

Inhibition of Autophagy Attenuated Ethylene Glycol Induced Crystals Deposition and Renal Injury in a Rat Model of Nephrolithiasis

Atorvastatin inhibits renal inflammatory response induced by calcium oxalate crystals via inhibiting the activation of TLR4/NF‐κB and NLRP3 inflammasome

Contact Info

Product

Resources

About