scholarly journals miR-208b Regulates Rabbit Preadipocyte Proliferation and Differentiation

Genes ◽  
2021 ◽  
Vol 12 (6) ◽  
pp. 890
Author(s):  
Jiahao Shao ◽  
Ting Pan ◽  
Jie Wang ◽  
Tao Tang ◽  
Yanhong Li ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Target Gene ◽  
Cell Counting ◽  
Signal Pathways ◽  
Preadipocyte Differentiation ◽  
Proliferation And Differentiation ◽  
Oil Red O Staining ◽  
Oil Red O ◽  
Differentiation Model ◽  
Gene Mrna

microRNAs (miRNAs) play an important role in gene regulation in animals by pairing with target gene mRNA. Many miRNAs are differentially expressed in the adipose tissue, often with conserved expression. In our study, we found that miR-208b expression was observed differently in the preadipocyte differentiation model. When miR-208b was overexpressed in the preadipocyte differentiation model, the overexpressed group displayed higher expression of PPARγ and FABP4—the markers of preadipocyte differentiation. Oil Red O staining revealed that the count of lipid droplets was increased in the overexpressed group. When the expression of miR-208b was inhibited, the above indicators showed an opposite trend. Moreover, results from both 5-ethynyl-2’-deoxyuridine (EDU) and cell counting kit (CCK) analysis showed that miR-208b promoted the proliferation of preadipocyte. Expression of gene CSNK2A2, a direct miR-208b target, was downregulated in the overexpressed group, providing a possible link to multiple signal pathways. Overall, our data indicate that miR-208b play a positive regulatory effect on the proliferation and differentiation of rabbit preadipocyte.

scholarly journals UHRF1 Promotes Proliferation of Human Adipose-Derived Stem Cells and Suppresses Adipogenesis via Inhibiting Peroxisome Proliferator-Activated Receptor γ

2019 ◽  
Vol 2019 ◽  
pp. 1-9
Author(s):  
Ke Chen ◽  
Zi Guo ◽  
Yufang Luo ◽  
Jingjing Yuan ◽  
Zhaohui Mo
Keyword(s):  
Stem Cells ◽  
Ring Finger ◽  
Cell Counting ◽  
Peroxisome Proliferator ◽  
Adipose Derived Stem Cells ◽  
Peroxisome Proliferator Activated Receptor ◽  
Proliferation And Differentiation ◽  
Oil Red O Staining ◽  
Oil Red O

Once the adipose tissue is enlarged for the purpose of saving excess energy intake, obesity may be observed. Ubiquitin-like with PHD and RING Finger domains 1 (UHRF1) is helpful in repairing damaged DNA as it increases the resistance of cancer cells against cytocidal drugs. Peroxisome proliferator-activated receptor γ (PPARγ), an important nucleus transcription factor participating in adipogenesis, has been extensively reported. To date, no study has indicated whether UHRF1 can regulate proliferation and differentiation of human adipose-derived stem cells (hADSCs). Hence, this study aimed to utilize overexpression or downregulation of UHRF1 to explore the possible mechanism of proliferation and differentiation of hADSCs. We here used lentivirus, containing UHRF1 (LV-UHRF1) and siRNA-UHRF1 to transfect hADSCs, on which Cell Counting Kit-8 (CCK-8), cell growth curve, colony formation assay, and EdU proliferation assay were applied to evaluate proliferation of hADSCs, cells cycle was investigated by flow cytometry, and adipogenesis was detected by Oil Red O staining and Western blotting. Our results showed that UHRF1 can promote proliferation of hADSCs after overexpression of UHRF1, while proliferation of hADSCs was reduced through downregulation of UHRF1, and UHRF1 can control proliferation of hADSCs through transition from G1-phase to S-phase; besides, we found that UHRF1 negatively regulates adipogenesis of hADSCs via PPARγ. In summary, the results may provide a new insight regarding the role of UHRF1 on regulating proliferation and differentiation of hADSCs.

Isolation and in vitro culture of intramuscular pre-adipocytes from Luxi adult Yellow cattle

2007 ◽  
Vol 4 (3) ◽  
pp. 229-232
Author(s):  
Wan Rong ◽  
Ding Jian ◽  
Zhou Zhen-Ming ◽  
Ren Li-Ping ◽  
Meng Qing-Xiang
Keyword(s):  
Lipid Droplets ◽  
Morphological Changes ◽  
Local Breed ◽  
Chinese Adult ◽  
Yellow Cattle ◽  
Proliferation And Differentiation ◽  
Cell Strains ◽  
Oil Red O Staining ◽  
Oil Red O

AbstractThree Luxi adult Yellow steers were used to isolate and culture intramuscular pre-adipocytes in vitro as well as to examine factors influencing their proliferation and differentiation. The intramuscular pre-adipocytes were taken from adipose tissues within muscles between the sixth and seventh rib and cultured after digestion with collagenase I. The results showed that the separated cell populations were highly homogeneous, proliferative and doubled within 62 h. When the confluent pre-adipocytes were treated with 10 μg/ml insulin and 0.25 μmol/l dexamethasone, small lipid droplets appeared on day 2 and the number of lipid droplets rapidly increased around the nuclei on day 6. Their dynamic morphological changes, growth curve, Oil Red O staining, and reaction to insulin and dexamethasone all verified their pre-adipocyte identity. Under controlled conditions, the intramuscular pre-adipocytes resumed proliferating and differentiating in vitro. Interestingly, the proportion of cultured diploid pre-adipocytes reached more than 90% after six repeated cultures. This study confirms the existence of functionally active pre-adipocytes within the muscles of Chinese adult local breed cattle. These cell strains are a potentially useful model for understanding further the mechanism of intramuscular adipose deposition in tissues, in order to improve beef quality based on Chinese local breed beef cattle.

Stard 3 (Steroidogenic Acute Regulatory-Related Lipid Transfer Domain-Containing Protein 3) Play Important Role in Preadipocyte Differentiation

2022 ◽  
Vol 12 (4) ◽  
pp. 827-833
Author(s):  
Zhonge Chen ◽  
Yanhua Tang ◽  
Wenyong Jiang ◽  
Xiaoqian Zhou
Keyword(s):  
Fluorescence Intensity ◽  
Lipid Transfer ◽  
Gene Expressions ◽  
Preadipocyte Differentiation ◽  
Protein Expressions ◽  
Positive Rate ◽  
Steroidogenic Acute Regulatory ◽  
Oil Red O Staining ◽  
Oil Red O

Aim: To evaluate Stard 3’s effects and relative mechanisms in preadipocyto differentiation by vitro study. Materials and Methods: The 3T3-L1 cell were divided into 5 groups as NC, si-Stard 3, ROS agonist, ROS inhibitor and si-Stard 3+ROS agonist groups. The cell of different groups were evaluated by Oil red O staining and Triglyceride. Evaluating ROS production by DHE and NBT assay. Using RT-qPCR and WB methods to evaluate gene and protein expressions. Results: Compared with NC group, Triglyceride, DHE fluorescence intensity and NBT positive rate were significantly down-regulation in si-Stard 3 and ROS inhibitor groups (P < 0.001, respectively), and were significantly up-regulation in ROS agonist group (P < 0.001, respectively); However, with si-Stard 3 transfection and ROS agonist treatment, compared with si-Stard 3 group, Triglyceride, DHE fluorescence intensity and NBT positive rate were significantly increased in si-Stard 3+ROS agonist group (P < 0.001, respectively). With RT-qPCR and WB assay, Compared with NC group, Stard 3 gene and protein expressions of si-Stard 3 and si-Stard 3+ROS agonist group were significantly depressed (P < 0.001, respectively), AMPK, PPARγ, CEBPα and FABP4 gene expressions were significantly differences in si-Stard 3, ROS agonist and ROS inhibitor groups (P < 0.001, respectively) and p-AMPK, PPARγ, CEBPα and FABP4 protein expressions were significantly differences in si-Stard 3, ROS agonist and ROS inhibitor groups (P < 0.001, respectively), with si-Stard 3 transfection and ROS agonist the relative gene and protein expressions were significantly resumed compared with si-Stard 3 group (P < 0.001, respectively). Conclusion: Stard 3 knockdown had effects to suppress 3T3-L1 cells transformation into adipocytes in vitro study.

scholarly journals ANTIHYPERGLYCEMIC ACTIVITY OF TINOCRISPOSIDE BY STIMULATING 3T3-L1 ADIPOCYTE CELL DIFFERENTIATION

2018 ◽  
Vol 11 (11) ◽  
pp. 494
Author(s):  
Adek Zamrud Adnan ◽  
Muhammad Taher ◽  
Annisa Fauzana ◽  
Tika Afriani ◽  
Dewi Imelda Roesma ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Lipid Droplets ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
Antihyperglycemic Activity ◽  
Significant Difference ◽  
Adipocyte Cell ◽  
Oil Red O Staining ◽  
Oil Red O

Objective: The aim of this study is to investigate the antihyperglycemic activity of tinocrisposide by stimulating 3T3-L1 adipocyte cell differentiation. Tinocrisposide is a furanoditerpene glycoside that was isolated from Tinospora crispa (Menispermaceae).Methods: Adipocyte cell differentiation activity of tinocrisposide in interval concentrations of 50, 25, 12.5, and 6.25 μg/ml has been investigated on 3T3-L1 cell line using insulin of 1 μg/ml as a positive and Dulbecco’s modified Eagle media (DMEM) as a negative control group. The effect of tinocrisposide was quantified with oil red O staining method by measuring an absorbance of lipid solution in isopropanol at a wavelength (λ) of 520 nm.Results: Tinocrisposide in the concentrations of 50, 25, 12.5, and 6.25 μg/ml insulin of 1 μg/ml and DMEM groups showed absorbance value of 0.7669, 0.7253, 0.6563, 0.6481, 0.954, and 0.2653, respectively. It was found that there was a significant difference statistically in lipid droplets accumulation among all groups (p<0.05) and tinocrisposide at a concentration of 50 μg/ml showed the highest lipid droplets accumulation in 3T3-L1 adipocyte cells.Conclusion: From the study, it could be concluded that tinocrisposide was able to stimulate the differentiation of adipocyte cell and had antihyperglycemic activity.

scholarly journals Korean Traditional Fermented Soybean Paste (Doenjang) Regulate Renin-Angiotensin System (RAS) in 3T3-L1 Adipocytes

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 791-791
Author(s):  
Hayoung Woo ◽  
Jung Eun Park ◽  
Youn-Soo Cha
Keyword(s):  
Blood Pressure ◽  
Lipid Accumulation ◽  
Lipid Droplets ◽  
Renin Angiotensin System ◽  
Angiotensin System ◽  
Renin Angiotensin ◽  
Fermented Soybean ◽  
Soybean Paste ◽  
Oil Red O Staining ◽  
Oil Red O

Abstract Objectives Doenjang, the Korean traditional fermented soybean paste, contains much salt. There is a concern that cardiovascular disease may occur due to such high salinity. Nevertheless, previous studies have demonstrated functional properties of doenjang anti-obesity and anti-cancer effects. Furthermore, in our recent studies, we showed that the anti-hypertensive effect of doenjang through renin-angiotensin system (RAS) regulation. Doenjang regulated the RAS to improve lipid metabolism in adipose tissue, which had a positive effect on blood pressure control. Therefore, we expected to find the exact mechanism of action or target point of doenjang in adipocyte using 3T3-L1 cells. Methods In this study, 3T3-L1 cells were treated with doenjang and RAS blockers, Losartan (10−4 M), and Captopril (10−4 M), were treated as positive control which suppresses AT1R and ACE, respectively. Non-cytotoxic concentrations of samples were selected as per MTT assay and added with induction media, harvested after 4 days for RNA extraction. Lipid droplets were detected by Oil Red O staining. Results Doenjang downregulated mRNA levels of peroxisome proliferator-activated receptor-γ (Pparg), RAS related genes such as angiotensinogen (Agt), Renin (Ren), and aldosterone-releasing factors (P &lt; 0.05). Especially, angiotensin convert enzyme (Ace) and angiotensin II receptor 2 (Agtr2) levels were decreased by doenjang treatment. Doenjang reduced the lipid accumulation, which was confirmed from the Oil Red O staining of lipid droplets. As a result, it is revealed that doenjang not only inhibits lipid accumulation in adipocytes but also may inhibit ACE in 3T3-L1 adipocytes through a mechanism similar to the effect of Captopril. Conclusions These data are consistent with our animal study. It have been shown to regulate blood pressure through lipid improvement and ACE inhibition despite high salt content in doenjang. Funding Sources This work was supported by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korea government (MEST) (No. 2018R1A2B6006477).

scholarly journals Upregulated microRNA-106a Promotes Porcine Preadipocyte Proliferation and Differentiation by Targeting Different Genes

Genes ◽  
2019 ◽  
Vol 10 (10) ◽  
pp. 805 ◽  
Author(s):  
Kuilong Huang ◽  
Xin’e Shi ◽  
Jie Wang ◽  
Ying Yao ◽  
Ying Peng ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Lipid Droplets ◽  
Luciferase Reporter ◽  
Cell Cycles ◽  
Lipogenic Genes ◽  
Proliferation And Differentiation ◽  
Luciferase Reporter Vector ◽  
Membrane Bound ◽  
Porcine Adipocytes ◽  
Oil Red O

Adipose tissue is one of the main organs for the energy storage and supply of organisms. Adipose deposition and metabolism are controlled by a cascade of transcription factors and epigenetic regulatory mechanisms. Previous studies have also shown that miR-106a plays a considerable role in the development of organisms. The regulatory mechanism of miR-106a on porcine preadipocytes is still not clear. In this study, preadipocytes were isolated from the neck subcutaneous deposits of 3–5-day old Chinese native Guanzhong black pigs using 5-ethynyl-20-deoxyuridine (EdU) staining and a CCK-8 assay to detect the number of proliferous cells and real-time qPCR (RT-qPCR) and western blot analysis to detect gene expression, as well as Oil Red O and BODIPY staining dye lipid droplets and flow cytometry (FCM) to detect cell cycles. We also used the double luciferase method to detect the relative luciferase activities. Upregulated miR-106a increased the number of proliferous cells and enhanced the expression of cell proliferation-related genes in porcine adipocytes. The double luciferase reporter vector confirmed that p21 was a target gene of miR-106a in the cell proliferation phase. miR-106a upregulation increased the number of lipid droplets and the expression of lipogenic genes and directly targeted BMP and activin membrane-bound inhibitor (BAMBI) in the process of differentiation. Our results indicated that miR-106a promotes porcine preadipocyte proliferation and differentiation by targeting p21 and BAMBI.

Secreted-Osteopontin Contributes to Brown Adipogenesis In Vitro via a CD44-Dependent Pathway

2019 ◽  
Vol 51 (11) ◽  
pp. 741-748
Author(s):  
Mengxi Wang ◽  
Yaoyao Guo ◽  
Yumeng Zhou ◽  
Wanwan Yuan ◽  
Huixia Li ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Biological Activities ◽  
Tumor Formation ◽  
Brown Adipocyte ◽  
Brown Adipocytes ◽  
Protein Levels ◽  
Infected Cells ◽  
Oil Red O Staining ◽  
Oil Red O

AbstractOsteopontin (OPN), a secreted glycoprotein, is involved in various pathophysiological processes including immune response, inflammation, tumor formation, and metabolism. OPN exists in 2 forms, secreted-OPN (sOPN) and intracellular-OPN (iOPN). While they might have different biological activities, it remains largely unknown whether sOPN and iOPN induce the differentiation of brown adipocytes. To test this possibility, 3T3-L1 cells were induced by DMI induction with or without recombinant human OPN (rhOPN, 10, 50, 100, 200 μM), respectively. Meanwhile, another batch of 3T3-L1 cells were infected with Ad-GFP-ap2-OPN and followed by DMI differentiation. Subsequently, the infected cells were treated with either anti-CD44 antibody or immunoglobulin G (Ig G). Accumulation of lipid droplets was visualized by Oil red O staining and protein levels were assayed by western blotting analysis. The results showed that sOPN and not rhOPN, notably increased the accumulation of lipid droplets and the expression of brown adipocyte-related genes. Moreover, neutralization of CD44 partially abrogated the effects induced by sOPN. These data demonstrate that sOPN and not rhOPN has the capacity to induce the differentiation of white preadipocytes into brown adipocytes through a CD44-dependent mechanism. The findings might provide a potential target for sOPN to combat obesity.

scholarly journals Mechanism of lncRNA H19 in the Regulation of Lipid Accumulation and Inflammatory Response in Macrophages

2020 ◽  
Author(s):  
Xue-Mei Wang ◽  
Xiao-Ming Gao ◽  
Fen Liu ◽  
Ying Cao ◽  
Jie-Ying Wang ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Lipid Accumulation ◽  
Target Gene ◽  
Molecular Mechanisms ◽  
Cholesterol Concentration ◽  
Abca1 Expression ◽  
Intracellular Lipid Accumulation ◽  
Oil Red O Staining ◽  
Oil Red O

Abstract Background and aims: Lipid accumulation of macrophages caused by oxidative stress is the key reason for the early pathological changes of atherosclerosis. LncRNA H19 repression downregulated NF-κB activation, upregulated ABCA1 expression, intracellular lipid accumulation increased, but the role of lncRNA H19 in atherogenesis and the molecular mechanisms have not been defined. We aimed to explore if and how lncRNA H19 affects lipid accumulation of macrophages by regulating lipid metabolism and inflammatory response.Methods and results: THP-1 macrophages were cultured with ox-LDL to form foam cells. THP-1-derived macrophages were incubated with H19 siRNA or not. Oil Red O staining was used for the determination lipid accumulation in macrophages. Enzymatic methods were performed to analyze cholesterol concentration. Both western blot and qRT-PCR were applied to detect target gene expression. ELISA was used to examine the levels of oxidative and inflammatory mediators. We found that lncRNA H19 repression reduced lipid accumulation by elevating efficiency of RCT and via upregulation of ABCA1 and PPARα expression in THP-1 derived macrophages. Further, lncRNA H19 repression upregulated PGC-1α and downregulated NF-κB signaling pathway.Conclusion: These results suggest that lncRNA H19 repression inhibits atherosclerosis by promoting RCT process and reducing inflammatory response via PGC-1α and NF-κB pathways, respectively.

scholarly journals Embryonic cardiospecific knockout of α-E-catenin gene leads to alteration of energy metabolism in adult heart

2017 ◽  
Vol 23 (2) ◽  
pp. 65-69
Author(s):  
V. Balatskyy ◽  
L. Macewicz ◽  
O. Piven
Keyword(s):  
Lipid Metabolism ◽  
Energy Metabolism ◽  
Transgenic Mice ◽  
Lipid Droplets ◽  
Metabolic Disorders ◽  
Conditional Knockout ◽  
Heart Hypertrophy ◽  
Oil Red O Staining ◽  
Oil Red O

Previously we have shown that the α-E-catenin knockout in the embryonic heart leads to hypertrophy in adult and activation of canonical Wntsignaling. Heart hypertrophy is also accompanied by metabolic disorders, but role of the α-E-catenin in these processes is not known. Aim of our work is to study the effect of α-E-catenin deletion on the lipid metabolism in the heart. Methods. In our experiment we have used α-Е-catenin conditional knockout and αMHC-Cre transgenic mice. We have utilized histological (Oil Red O staining) and molecular biological (Western blot) methods. Results. α-Е-catenin deletion leads to accumulation of lipid droplets in myocardium, and to violation of expression and phosphorylation of key regulators of lipid metabolism (Ampk, Pparα, Acc, Hsl). Conclusions. Ous results suggest that α-Е-catenin deletion leads to inhibition of lipid metabolism in the heart.

scholarly journals MiR-146a-5p Targeting SMAD4 and TRAF6 Inhibits Adipogenensis Through TGF-β and NF-κB Signal Pathways in Porcine Intramuscular Preadipocytes

2020 ◽  
Author(s):  
Que Zhang ◽  
Rui Cai ◽  
Guorong Tang ◽  
Wanrong Zhang ◽  
Weijun Pang
Keyword(s):  
Cell Proliferation ◽  
Adipogenic Differentiation ◽  
Signal Pathways ◽  
Pork Quality ◽  
Sequencing Analysis ◽  
Tnf Receptor ◽  
Preadipocyte Differentiation ◽  
Novel Mirna ◽  
Proliferation And Differentiation ◽  
Imf Content

Abstract Background: Intramuscular fat (IMF) content is a vital parameter to assess pork quality. Increasing evidences have shown that microRNAs (miRNAs) play an important role in regulating porcine IMF deposition. Here, a novel miRNA implicated in porcine IMF adipogenesis was found, and its effect and regulatory mechanism was further explored on the proliferation and differentiation of intramuscular preadipocytes.Results: By miRNAs sequencing analysis in porcine adipose tissues, we found that miR-146a-5p was a potential regulator of porcine IMF adipogenesis. Further study showed that miR-146a-5p mimics inhibited the proliferation of porcine intramuscular preadipocytes, but its inhibitor promoted cell proliferation. Interestingly, miR-146a-5p mimics also repressed preadipocyte differentiation, whereas its inhibitor promoted adipogenic differentiation. Mechanistically, miR-146a-5p suppressed cell proliferation by directly targeting SMAD family member 4 (SMAD4) to attenuate the TGF-β signal. Moreover, miR-146a-5p inhibited the differentiation of intramuscular preadipocytes by targeting TNF receptor associated factor 6 (TRAF6) to weaken the NF-κB signaling of the TRAF6 downstream pathway.Conclusions: MiR-146a-5p targeting SMAD4 and TRAF6 inhibited porcine intramuscular adipogenesis through attenuating TGF-β and NF-κB signals, respectively. These findings provided a novel miRNA biomarker for regulating intramuscular adipogenesis to promote pork quality.

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