Dataset Information

Hepatic miR-192-3p reactivation alleviates steatosis by targeting glucocorticoid receptor.

ABSTRACT:

Background & aims

The paradox of hepatic insulin resistance describes the inability for liver to respond to bioenergetics hormones in suppressing gluconeogenesis whilst maintaining lipid synthesis. Here, we report the deficiency of miR-192-3p in the livers of mice with diabetes and its role in alleviating hepatic steatosis.

Methods

As conventional pre-microRNA (miRNA) stem-loop overexpression only boosts guiding strand (i.e. miR-192-5p) expression, we adopted an artificial AAV(DJ)-directed, RNA Pol III promoter-driven miRNA hairpin construct for star-strand-specific overexpression in the liver. Liver steatosis and insulin resistance markers were evaluated in primary hepatocytes, mice with diabetes, and mice with excessive carbohydrate consumption.

Results

Functional loss of miR-192-3p in liver exacerbated hepatic micro-vesicular steatosis and insulin resistance in either mice with diabetes or wild-type mice with excessive fructose consumption. Liver-specific overexpression of miR-192-3p effectively halted hepatic steatosis and ameliorated insulin resistance in these mice models. Likewise, hepatocytes overexpressing miR-192-3p exhibited improved lipid accumulation, accompanied with decreases in lipogenesis and lipid-accumulation-related transcripts. Mechanistically, glucocorticoid receptor (GCR, also known as nuclear receptor subfamily 3, group C, member 1 [NR3C1]) was demonstrated to be negatively regulated by miR-192-3p. The effect of miR-192-3p on mitigating micro-vesicular steatosis was ablated by the reactivation of NR3C1.

Conclusions

The star strand miR-192-3p was an undermined glycerolipid regulator involved in controlling fat accumulation and insulin sensitivity in liver through blockade of hepatic GCR signalling; this miRNA may serve as a potential therapeutic option for the common co-mobility of diabetic mellitus and fatty liver disease.

Lay summary

The potential regulatory activity of star strand microRNA (miRNA) species has been substantially underestimated. In this study, we investigate the role and mechanism of an overlooked star strand miRNA (miR-192-3p) in regulating hepatic steatosis and insulin signalling in the livers of mice with diabetes and mice under excessive carbohydrate consumption.

SUBMITTER: Wang Z

PROVIDER: S-EPMC7588854 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Hepatic miR-192-3p reactivation alleviates steatosis by targeting glucocorticoid receptor.

Wang Zhangting Z Miu Kai-Kei KK Zhang Xueyan X Wan Angel Tsz-Yau AT Lu Gang G Cheung Hoi-Hung HH Lee Heung-Man HM Kong Alice Pik-Shan AP Chan Juliana Chung-Ngor JC Chan Wai-Yee WY

JHEP reports : innovation in hepatology 20200906 6

<h4>Background & aims</h4>The paradox of hepatic insulin resistance describes the inability for liver to respond to bioenergetics hormones in suppressing gluconeogenesis whilst maintaining lipid synthesis. Here, we report the deficiency of miR-192-3p in the livers of mice with diabetes and its role in alleviating hepatic steatosis.<h4>Methods</h4>As conventional pre-microRNA (miRNA) stem-loop overexpression only boosts guiding strand (<i>i.e.</i> miR-192-5p) expression, we adopted an artificial ...[more]

PMID: 33134908

Similar Datasets

Project description:Glucocorticoid receptor (GR) plays crucial roles in various processes, including stress response, inflammatory response and toxin response, making it a therapeutic target for numerous diseases. microRNA (miRNA) can target and negatively regulate the expression of GR, thus interfering with its normal function. Aflatoxin B1 (AFB1) seriously affects poultry health and productivity, exhibiting hepatotoxicity and cytotoxicity. However, the expression of GR and GR-targeting miRNAs in poultry after AFB1 poisoning has not been studied. In this study, four-day-old broiler chicks were randomly divided into control group (CON) and AFB1 group (AFB1). After a 3-day pre-feed, the AFB1 group was given 0.25 mg/kg of AFB1 for 18 days. Hematoxylin and eosin (HE) staining revealed AFB1-induced hepatocyte damage accompanied by inflammatory cell infiltration. Liver oxidative stress enzymes superoxide dismutase (SOD) and catalase (CAT) decreased significantly. RT-qPCR showed decreased mRNA expression of Phase I and II metabolic detoxification enzymes. Western blot analysis indicated reduced GR protein levels, while miRNA PCR revealed upregulation of GR-targeting miR-124a-3p, miR-142-3p, miR-18b-5p, and miR-183. After 24 h of 40 μM AFB1 treatment in LMH cells, Edu and flow cytometry confirmed inhibition of cell proliferation and promotion of apoptosis. Additionally, AFB1 induced oxidative stress and DNA damage. RT-qPCR showed reduced expression of certain Phase I/II metabolic detoxification enzymes and GR. Western blot confirmed a significant decrease in GR protein. miRNA PCR revealed upregulation of miR124a-3p. We found that transfection of LMH cells with miR124a-3p inhibitor alleviated the changes in GR and metabolic detoxification enzyme gene expression induced by AFB1. Additionally, AFB1-induced reductions in cell viability, increased apoptosis, inhibited proliferation, oxidative stress, and DNA damage were also alleviated. Overall, our findings suggest that inhibition of miR124a-3p, which targets GR, can ameliorate AFB1-induced hepatotoxicity. This study highlights GR and its miRNA as potential therapeutic targets for AFB1-induced liver disease, providing new insights into therapeutic strategies to mitigate the harmful effects of AFB1 exposure in poultry.

Project description:Background: Kawasaki disease (KD) is a self-limiting illness with acute systematic vascular inflammation. It causes pathological changes in mostly medium and small-sized arteries, especially the arteria coronaria, which adds the risk of developing coronary heart disease in adults. Materials and methods: We detected the miR-223-3p expression in 30 KD patients combined with 12 normal controls using miRNA microarrays and RT-PCR. A KD mouse model was constructed using Candida albicans water insoluble substance (CAWS). We also checked the miR-223-3p's expression using qRT-PCR. The Luciferase reporting system was implemented to validate the correlation between miR-223-3p and Interleukin-6 receptor subunit beta (IL-6ST). TNF-α was used to stimulate human coronary artery endothelial cells (HCAECs), and miR-223-3p activator or inhibitor and KD serum were used to treat HCAECs. A Western blotting automatic quantitative analysis protein imprinting system was used to test the expression of signal transducer and the activator of transcription 3 (STAT3), phosphorylated-signal transducer and the activator of transcription 3 (pSTAT3) and NF-κB p65. Results: Clinical trials found that miR-223-3p expressions were markedly different (more than 2-fold) between the acute KD group and the control group. E-selectin and intercellular cell adhesion molecule-1 (ICAM-1) levels were also significantly higher (about 2-fold) in KD especially with coronary artery lesions. MiR-223-3p could alleviate vascular endothelial damage in KD mice, and IL-6 (Interleukin-6), E-selectin and ICAM-1 were simultaneously negative. The values of IL-6, E-selectin, and ICAM-1 mRNA expressions decreased, while the value of IL-6ST was increased between the agonist treated mice and KD mice. The RT-qPCR consequences displayed that miR-223-3p explored the highest expression on the third day in both the KD mice as well as the agonist group. MiR-223-3p can directly combine with IL-6ST 3' untranslatable regions (UTR) and held back the IL-6's expression. Overexpression of miR-223 down regulated IL6ST expression and decreased the expression of p-STAT3 and NF-κB p65, while the miR-223 inhibitor could reverse the above process. Conclusion: MiR-223-3p is an important regulatory factor of vascular endothelial damage in KD and could possibly become a potential target of KD treatment in the future.

Dataset Information

Hepatic miR-192-3p reactivation alleviates steatosis by targeting glucocorticoid receptor.

Background & aims

Methods

Results

Conclusions

Lay summary

Publications

Hepatic miR-192-3p reactivation alleviates steatosis by targeting glucocorticoid receptor.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets