MOLECULAR AND CELLULAR ENDOCRINOLOGY
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2020
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IF 3.871
Ziwen Wei,Xiaoli Qin,Xiaojie Kang,Haixia Zhou,Shaodan Wang,Dong Wei
Background MiR-142-3p has been shown to be suppressed in obese patients , while the underlying regulatory mechanism is unclear. Methods Body shape indexes as well as peripheral blood for biochemical parameter analysis were obtained from obese and healthy subjects. When 3T3-L1 cells were induced to differentiate, miR-142-3p expression was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The effects of miR-142-3p on triglyceride (TG) and adipogenic differentiation-related genes during the adipogenic differentiation of 3T3-L1 cells were detected by transfection, Oil Red O staining, and Western blot . The targeting relationship between miR-142-3p and Krueppel-like transcription factor 9 (KLF9) was verified by TargetScan and dual-luciferase experiment. The specific regulatory effects of miR-142-3p on cell adipogenic differentiation and autophagy were analyzed by rescue experiments. In vivo experiments further validated the results of in vitro experiments through obese mouse models. Results Obesity-marked biochemical indicators increased whereas high density lipoprotein and miR-142-3p decreased in obese patients. The content of miR-142-3p gradually decreased with cell lipid differentiation. Overexpression of miR-142-3p reduced TG deposition in cells by down-regulating lipid formation and fatty acid synthesis genes and up-regulating fatty acid oxidation genes. KLF9 targeting miR-142-3p was suppressed by miR-142-3p. KLF9 overexpression partially reversed the inhibitory effect of miR-142-3p mimic on adipogenic differentiation and the expressions of autophagy related-genes in 3T3-L1 cells. MiR-142-3p overexpression also inhibited fat cell differentiation and autophagy in obese mice . Conclusion Overexpressed miR-142-3p inhibited adipogenic differentiation and autophagy through targeting KLF9.
研究领域:
分子生物学
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细胞生物学
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内分泌学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2018
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IF 5.667
Leiliang He,Lihua Ding,Songcheng Yu,Fei Yu,Yongmei Tian,Xinsheng Xie,Lingbo Qu,Lie Liu,Yongjun Wu
The establishment of a highly sensitive and efficient platform for detecting circulating miRNA in serum is important for disease diagnosis and prognosis. To this end, a self-assembled horseradish peroxidase (HRP)-based polymer (poly-HRP) dual signal amplification strategy, via skillful combination of functionalized magnetic nanoparticles as the capture nanoprobes and the poly-HRP as signal amplification probes, has been developed for highl-sensitive and accurate detection of circulating miR-142-3p in human serum. Just by mixing building blocks together at one time, the poly-HRP used for dual signal amplification can be engineered by only one-step assembly. As the poly-HRP can be directly prepared ahead, avoiding chain amplification reaction in the detection process, the assay can be rapidly accomplished in about one hour. The measured limit-of-detection (LOD) is down to 100 fM. Standard miR-142-3p spiked in human serum is measured with ideal recovery rates ranging from 97.46% to 115.90%. More importantly, this method was validated to be applicable to direct detection of miR-142-3p in serums, and the results suggested that the miR-142-3p levels of serum samples obtained from T-cell acute lymphoblastic leukemia children (n = 25) are significantly higher than those from healthy control groups (n = 25) ( P < 0.01). To conclude, the ingenious combination of functionalized magnetic nanoparticles and the poly-HRP presented in this work brings forth a rapid, convenient, yet simple, sensitive, and specific biosensing method that can potentially be applied for clinical diagnosis.
研究领域:
生物化学
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分子生物学
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临床诊断
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产品关联
DOI
高影响因子
Journal of Stomatology Oral and Maxillofacial Surgery
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2022
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IF 2.48
Yi Sun,Jianlu Shi,Xiaoan Luo,Xuehong Xu
Objective Human periodontal ligament stem cells (hPDLSCs) refer to one kind of somatic stem cells that are capable of differentiating into multiple cell kinds and undergoing robust clonal self-renewal. This work was unearthed to elucidate the possible molecular mechanism of miR-142–3p in mediating osteogenic differentiation of hPDLSCs by targeting SGK1 . Methods The hPDLSCs were isolated, cultured, and identified. hPDLSCs were identified by immunofluorescence staining and multiple differentiation ability detection. Cell proliferation ability was assessed by CCK-8 assay. hPDLSCs were induced using osteogenic differentiation medium. ALP activity was detected by alkaline phosphatase (ALP) staining and ALP activity assay, and mineralized nodule formation was determined by alizarin red staining. The expression levels of osteogenic differentiation marker proteins ALP, RUNX2, and OCN were measured by RT-qPCR. miR-142–3p candidate targets were obtained through bioinformatics analysis. The relationship between miR-142–3p and SKG1 was verified. Results miR-142–3p in hPDLSCs after osteogenic induction was down-regulated. Elevated miR-142–3p restricted hPDLSCs proliferation, and diminished ALP activity and mineralized nodule formation, as well as the expression of ALP, RUNX2, and OCN, while miR-142–3p inhibition led to inverse results. miR-142-3p inhibited SKG1 expression. SKG1 overexpression promoted hPDLSC proliferation and osteogenic differentiation, and reversed the inhibitory function of miR-142–3p on hPDLSCs. Conclusion This study highlights that miR-142–3p represses osteogenic differentiation of hPDLSCs by reducing SGK1 expression.
研究领域:
干细胞生物学
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分子生物学
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口腔生物学
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产品关联
DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
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2026
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IF 3.5
Li Yuejin,Ouyang Yiming,Zhu Yu,Hou Dezhi,Li Xing,Guo Xiaodong,Sheng Ping,Wu Hailong,Liao Yingzhong,Gong Kunmei,Li Linhai
Natural killer (NK) cells play a crucial role in combating gastric cancer (GC). However, they undergo ferroptosis in the tumor microenvironment of GC, which weakens their antitumor effects. Exosomal circRNA influences GC progression. Previous studies have reported the cancer-promoting role of circPDSS1 in GC, but whether it is involved in NK cell ferroptosis remains unclear. This study aimed to investigate the role of GC cell-derived exosomal circPDSS1 in NK cell ferroptosis. A noncontact coculture system of GC cells (HGC-27, AGS, and MKN-45 cells) and human NK-92 cells was established, and humanized mouse and cell-derived xenograft (CDX) tumor models were constructed for joint investigation. Cell death and viability were assessed via flow cytometry, LDH release assays and CCK-8 assays; circPDSS1 stability was determined by agarose gel electrophoresis and actinomycin D treatment; ferroptosis was determined by measuring lipid ROS, MDA, and Fe 2+ levels; and key molecular interactions were determined by performing dual-luciferase reporter assays and RIP and RNA pull-down experiments. CD56 and CD16 expression was reduced in clinical tumor tissues. In vitro experiments revealed that the proportion of 7-AAD + NK-92 cells cocultured with GC cells was significantly increased, accompanied by increased cell mortality and decreased IFN-γ and TNF-α secretion. Further studies revealed that circPDSS1 expression was abnormally elevated in GC tissues and cells and was particularly enriched in MKN-45 cell-derived exosomes (Exos). Knockdown of circPDSS1 alleviated MKN-45-Exo-induced cell damage; reduced lipid ROS, MDA, and Fe 2+ levels in NK-92 cells; and delayed tumor progression. Furthermore, a ferroptosis inhibitor suppressed circPDSS1 overexpression-induced cell death. Mechanistically, circPDSS1 sponged miR-142-3p to upregulate ACSL4 expression, promoting NK cell ferroptosis. Exosomal circPDSS1 derived from GC cells contributes to NK cell ferroptosis by sponging miR-142-3p to increase ACSL4 expression and exacerbate GC.
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DOI
PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
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2023
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IF 5.4
Chuiyang Zheng,Huan Nie,Maoxing Pan,Wen Fan,Dajin Pi,Zheng Liang,Dongdong Liu,Fengzhen Wang,Qinhe Yang,Yupei Zhang
Ethnopharmacological relevance Chaihu Shugan powder (CSP) plays an important role in the prevention and treatment of nonalcoholic fatty liver disease (NAFLD) through a variety of biological mechanisms. However, whether the mechanism involves microRNA (miRNA) regulation remains unknown. Aim of the study To investigate the effects of CSP on the miRNA expression profile of rats with NAFLD induced by high-fat diet (HFD), and to explore the mechanism of CSP in the treatment of NAFLD. Methods NAFLD rat models were established by an 8-week HFD. The therapeutic effects of CSP on NAFLD were evaluated by physiological, biochemical and pathological analysis and hepatic surface microcirculation perfusion test. MicroRNA sequencing was used to study the effect of CSP on the miRNA expression profile of NAFLD rats, and the target genes of differentially expressed (DE) miRNAs were predicted for further function enrichment analysis. Next, targets of CSP and NAFLD were collected by a network pharmacological approach, and Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analysis were performed for the common target genes of CSP, NAFLD and DE miRNAs, and the expression levels of key genes and proteins were verified by quantitative Real-time PCR and Western blot . Finally, a network among formula-herb-compound-miRNA-target-biological processes-disease was established to explained the complex regulation mechanism of CSP on NAFLD. Results The results showed that CSP significantly improved liver lipid accumulation, serum lipid and transaminase levels and liver surface microcirculation disturbance in HFD-induced NAFLD rats. The intervention of CSP reversed the high expression of 15 miRNAs in liver tissues induced by HFD, including miR-34a-5p, miR-146a-5p, miR-20b-5p and miR-142-3p. The results of pathway and functional enrichment analysis showed that, CSP might play an anti-NAFLD role via regulating DE miRNAs related to fatty acid metabolic process. Combined with the network pharmacological analysis, it was found that the DE miRNAs might affected the fatty acid biosynthesis pathway in the treatment of NAFLD by CSP. Molecular biology experiments have conformed the decreased the gene and protein levels of acetyl-CoA carboxylase alpha (ACACA), fatty acid synthase (FASN) and other fatty acid biosynthesis related enzymes on NAFLD rats after intervention of CSP. Conclusions CSP can significantly reduce hepatic lipid accumulation of NAFLD rat model induced by HFD, and its mechanism may be through the action of 15 miRNAs such as miR-34a-5p, miR-146a-5p, miR-20b-5p and miR-142-3p. Reduce the gene and protein expression levels of ACACA, FASN and other fatty acid biosynthesis related enzymes, thus reducing fatty acid biosynthesis. Based on an epigenetic perspective, this study explains the key anti-NAFLD mechanism of CSP via combination of microRNA sequencing and network pharmacological analysis, providing a new reference for the modernization of traditional Chinese medicine .
研究领域:
药理学
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分子生物学
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中医药
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表观遗传学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）