Acetoacetate induces hepatocytes apoptosis by the ROS‐mediated MAPKs pathway in ketotic cows

Du, Xiliang; Shi, Zhen; Peng, Zheng; Zhao, Chenxu; Zhang, Yuming; Wang, Zhe; Li, Xiaobing; Li, Xinwei

doi:10.1002/jcp.25773

Cited by 141 publications

(78 citation statements)

References 39 publications

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“…Western blotting analysis was performed as previously described (Du, Chen, et al, ; Du, Zhen, et al, ; Song et al, ; Sun et al, ). Cells were harvested in ice‐cold phosphate‐buffered saline (PBS) and lysed in lysis buffer (Beyotime) with phenylmenthylsulfonyl fluoride (PMSF) (Beyotime).…”

Section: Methodsmentioning

confidence: 99%

Hepatic miR‐125b inhibits insulin signaling pathway by targeting PIK3CD

Chen

et al. 2018

Journal Cellular Physiology

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Insulin resistance is often characterized as the most critical factor contributing to the development of (T2D) type 2 diabetes. MicroRNAs (miRNAs) are endogenous non-coding short single-stranded RNAs that function as negative regulators in many physiological and pathological processes. The objective of this study was to evaluate the roles of miR-125b in the regulation of insulin sensitivity in hepatocytes. We found that hepatic miR-125b levels were significantly increased in the patients with type 2 diabetes, high fat diet (HFD) mice, ob/ob and db/db mice. In vitro, miR-125b was also significantly up-regulated in tumor necrosis factor-alpha- (TNF-α) and glucosamine-induced insulin resistance conditions. Furthermore, miR-125b overexpression impaired the insulin signaling pathway in HepG2 cells, L02c cells, and primary hepatocytes. Inhibition of miR-125b improved insulin sensitivity, especially in insulin-resistant cells induced by either TNF-α or glucosamine. We demonstrated that miR-125b targeted the 3'-untranslated region (3'-UTR) of phosphoinositide 3-kinase catalytic subunit delta (PIK3CD) mRNA. The hepatic PIK3CD protein levels were markedly decreased in patients with type 2 diabetes, HFD, ob/ob, and db/db mice. Inhibition of PIK3CD markedly attenuated the improvement of insulin sensitivity induced by miR-125b inhibitors. More importantly, overexpressing miR-125b in mice causes insulin resistance and impairs glucose homeostasis. Together, these findings indicate that miR-125b inhibits insulin sensitivity by targeting PIK3CD in hepatocytes, supporting hepatic miR-125b, or PIK3CD are potential therapeutic target of insulin resistance.

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Section: Methodsmentioning

confidence: 99%

Hepatic miR‐125b inhibits insulin signaling pathway by targeting PIK3CD

Chen

et al. 2018

Journal Cellular Physiology

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“…As shown in our previous studies, oxidative stress and hepatic damage occur in dairy cows with ketosis [3, 13]. Importantly, Tharwat et al reported that high levels of Caspase 3-positive and single-stranded DNA-(ssDNA)-positive cells in postpartum cows or cows with liver fatty infiltration in the liver, indicating that hepatocyte apoptosis is induced in dairy cows undergoing parturition or with fatty infiltration [14, 15].…”

Section: Introductionmentioning

confidence: 99%

Elevated Apoptosis in the Liver of Dairy Cows with Ketosis

Chen

Huang

et al. 2017

Cell Physiol Biochem

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Background/Aims: Dairy cows with ketosis are characterized by oxidative stress and hepatic damage. The aim of this study was to investigate hepatic oxidative stress and the apoptotic status of ketotic cows, as well as the underlying apoptosis pathway. Methods: The blood aspartate aminotransferase (AST), alanine aminotransferase (ALT), glutamate dehydrogenase (GLDH) and gamma-glutamyl transferase (GGT) activities and the haptoglobin (HP), serum amyloid A (SAA) and serum apoptotic cytokeratin 18 neo-epitope M30 (CK18 M30) concentrations were determined by commercially available kits and ELISA kits, respectively. Liver histology, TUNEL and Oil red O staining were performed in liver tissue samples. TG contents were measured using an enzymatic kit; Caspase 3 assays were carried out using the Caspase 3 activity assay kit; oxidation and antioxidant markers were measured using biochemical kits; apoptosis pathway were determined by qRT-PCR and western blot. Results: Ketotic cows displayed hepatic fat accumulation. The hepatic malondialdehyde (MDA) content was significantly increased, but the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were markedly decreased in ketotic cows compared with control cows, indicating that ketotic cows displayed severe oxidative stress. Significantly higher serum levels of the hepatic damage markers AST, ALT, GGT and GLDH were observed in ketotic cows than in control cows. The blood concentration of the apoptotic marker CK18 M30 and the number of TUNEL-positive cells in the liver of ketotic cows were 1.19- and 2.61-fold, respectively, higher than the values observed in control cows. Besides, Caspase 3 activity was significantly increased in the liver of ketosis cows. Importantly, the levels of phosphorylated c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK) were significantly increased but the level of phosphorylated extracellular signal-regulated kinase1/2 (ERK1/2) was markedly decreased, which further promoted tumor protein 53 (p53) expression and inhibited nuclear factor E2-related factor 2 (Nrf2) expression. The apoptosis-related molecules p21, MDM2, Caspase 3, Caspase 9 and Bax were expressed at significantly higher levels in ketotic cows than in healthy cows, whereas the anti-apoptosis molecule Bcl-2 was expressed at significantly lower levels. Conclusions: Based on these results, ketotic cows display severe hepatic oxidative stress. The hepatic MAPK-p53-Nrf2 apoptotic pathway is over induced and partially mediated apoptotic damage in the liver.

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“…Thus, the regulation of the proteasome activity depends on ER‐stress but its induction is controlled by further mechanisms able to modulate ROS signaling in the cells, for example mitochondria and peroxisomes (Sandalio & Romero‐Puertas, ). Some researchers have demonstrated that ROS‐mediated MAPKs pathway induces ER stress and apoptosis (Du, Shi, et al, ), which could be a mechanism regulated by SFN and MG‐132.…”

Section: Discussionmentioning

confidence: 99%

Proteasome inhibition promotes autophagy and protects from endoplasmic reticulum stress in rat alveolar macrophages exposed to hypoxia‐reoxygenation injury

Fan

Huang

Wang

et al. 2018

Journal Cellular Physiology

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Alveolar macrophages play vital roles in acute lung injury, and macrophage response to hypoxia play relevant roles to disease mechanisms. There is growing evidence that cell death pathways play crucial roles in physiological and pathological settings and that the ubiquitin-proteasome system is involved in the regulation of these processes. However, the functional role of proteasome in alveolar macrophages exposed to hypoxia-reoxygenation (H/R) injury is unknown. We aimed to investigate the function of proteasome on alveolar macrophages exposed to H/R and the underlying mechanisms. NR8383 cells were pretreated with proteasome activator sulforaphane (SFN) or inhibitor MG-132 for 1 hr, and then submitted to 2/6 hr, 4/6 hr, and 6/6 hr H/R treatment. Cell viability was assessed with MTT assay. Autophagy was monitored using electron transmission microscope and flow cytometry and western blotting. The endoplasmic reticulum (ER) stress and unfolded protein response (UPR) pathways were equally analyzed by western blotting. Cell apoptosis was detected by immunohistochemistry, caspase3/7 activity, and western blotting. The viability of NR8383 cells exposed to H/R was affected by proteasome activity and proteasome inhibition significantly inhibited cell death. Treatment with MG-132 led to autophagy activation and induced the survival of NR8383 cells exposed to H/R. Pretreatment with SFN significantly decreased cell autophagy and induced cell death. ER stress was activated in H/R-treated NR8383 cells, and SFN further promoted ER stress whereas proteasome inhibition led to contrary results. Proteasome inhibtion hindered cell apoptosis as demonstrated by decreased caspase-3/7 activity, immunolabelling, and western blot results. Proteasome inhibition might be a promising approach for treating H/R injury-related lung diseases.

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Acetoacetate induces hepatocytes apoptosis by the ROS‐mediated MAPKs pathway in ketotic cows

Cited by 141 publications

References 39 publications

Hepatic miR‐125b inhibits insulin signaling pathway by targeting PIK3CD

Hepatic miR‐125b inhibits insulin signaling pathway by targeting PIK3CD

Elevated Apoptosis in the Liver of Dairy Cows with Ketosis

Proteasome inhibition promotes autophagy and protects from endoplasmic reticulum stress in rat alveolar macrophages exposed to hypoxia‐reoxygenation injury

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