scholarly journals Extract of Acalypha australis L. inhibits lipid accumulation and ameliorates HFD-induced obesity in mice through regulating adipose differentiation by decreasing PPARγ and CEBP/α expression

2021 ◽  
Author(s):  
Lang You ◽  
Fengxia Li ◽  
Yan Sun ◽  
Liang Luo ◽  
Jian Qin ◽  
...  
Keyword(s):  
Lipid Accumulation ◽  
Water Soluble ◽  
Pathological Examination ◽  
Chow Diet ◽  
Obese Mice ◽  
Food Ingredient ◽  
Gene And Protein Expression ◽  
Adipose Differentiation ◽  
Oil Red O Staining ◽  
Oil Red O

Background: Obesity is a principal risk factor for the development of type 2 diabetes and cardiovascular diseases. Natural plants and/or foods play an important role in the management of obesity. Acalypha australis L. (AAL) is a kind of potherb popular among Asian populations, and it is also consumed as a food ingredient and traditional herbal medicine. Objective: We investigated the effects of water extract from AAL on high-fat-diet (HFD)-induced obese mice and 3T3-L1 adipocytes to develop a new functional food material. Design: Nine-week-old male mice were randomly divided into control (chow diet, n = 6) and HFD (n = 30) group. From 12-weeks onward, mice in the HFD group were further separated into model (saline, 6 mL/ kg), simvastatin (0.11 mg/mL, 6 mL/kg), and AAL treatment (low, middle, and high dosage: 300, 600, and 900 mg/kg) group, with 6 animals per group, while mice in the control group were treated with saline (6 mL/ kg). Food intake, body/fat weight, liver/kidney indexes, and lipid profiles were determined. Tissues were fixed with formalin for pathological examination. Western blotting and PCR were performed to evaluate the protein and mRNA expression in 3T3-L1 adipocytes. Oil Red O staining was used to determine lipid accumulation. Results: AAL administration significantly suppressed body weight gain, and reduced fat pad weight and Lee’s index in obese mice, but had no effect on liver/kidney index. AAL also reduced serum cholesterol, triglyceride, and LDL-C and increased HDL-C levels. Histological analysis revealed that AAL significantly ameliorated lipid accumulation in the liver and subcutaneous adipose tissue. In vitro, Oil Red O staining showed that AAL inhibited adipose differentiation by down-regulating the gene and protein expression of PPARγ and C/EBPα. AAL also reversed HFD-induced intestinal dysbacteriosis. Conclusion: AAL water-soluble extract has a significant anti-adipogenic effect in the HFD-induced obese mice model.

Inhibition of 3T3-L1 adipose differentiation by 2,3,7,8-tetrachlorodibenzo-p-dioxin

1995 ◽  
Vol 108 (1) ◽  
pp. 395-402 ◽  
Author(s):  
M. Phillips ◽  
E. Enan ◽  
P.C. Liu ◽  
F. Matsumura
Keyword(s):  
Northern Blotting ◽  
Ah Receptor ◽  
Reporter Construct ◽  
Fat Cell ◽  
Camp Signaling Pathway ◽  
Adipose Differentiation ◽  
Tetrachlorodibenzo P Dioxin ◽  
Oil Red O Staining ◽  
Oil Red O ◽  
Do So

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-induced toxicity is particularly striking in adipose tissue, where it causes severe wasting. This phenomenon suggests that TCDD could have effects on adipocyte differentiation, now demonstrated using 3T3-L1 cells as a model system. When cells were treated with 10 nM TCDD before differentiation or during the first two days of induction in the presence of dexamethasone (dex) and isobutylmethylxanthine (IBMX), a reduction occurred in the number of fat cell colonies measured 7–10 days later by Oil Red O staining. Northern blotting showed an accompanying reduction in amounts of mRNA encoding several adipocyte markers. In contrast, when TCDD was added after differentiation, it had no effect on the maintenance of the adipose phenotype. Dose-response and structure-activity relationships were consistent with a process mediated by interaction of TCDD with the Ah receptor. The possibility that TCDD acts by inhibiting the signaling pathways activated by dex and IBMX was investigated. TCDD did not interfere with glucocorticoid-inducible transcription as judged by the unimpaired responsiveness of a transfected reporter construct. Treatment of cells with TCDD augmented the increase in protein kinase A (PKA) activity elicited by either IBMX or forskolin; therefore, if TCDD disrupts the cAMP signaling pathway, it must do so at a step after activation of PKA.

scholarly journals LncRNA-H19 promotes hepatic lipogenesis by directly regulating miR-130a/PPARγ axis in non-alcoholic fatty liver disease

2019 ◽  
Vol 39 (7) ◽  
Author(s):  
Jun Liu ◽  
Tao Tang ◽  
Guo-Dong Wang ◽  
Bo Liu
Keyword(s):  
Liver Disease ◽  
Fatty Liver ◽  
Lipid Accumulation ◽  
Fatty Liver Disease ◽  
Hepatic Lipogenesis ◽  
Alcoholic Fatty Liver ◽  
Oil Red O Staining ◽  
Alcoholic Fatty Liver Disease ◽  
Oil Red O ◽  
Liver Tissues

Abstract Background: As one of the most common liver disorders worldwide, non-alcoholic fatty liver disease (NAFLD) begins with the abnormal accumulation of triglyceride (TG) in the liver. Long non-coding RNA-H19 was reported to modulate hepatic metabolic homeostasis in NAFLD. However, its molecular mechanism of NAFLD was not fully clear. Methods: In vitro and in vivo models of NAFLD were established by free fatty acid (FFA) treatment of hepatocytes and high-fat feeding mice, respectively. Hematoxylin and Eosin (H&E) and Oil-Red O staining detected liver tissue morphology and lipid accumulation. Immunohistochemistry (IHC) staining examined peroxisome proliferator-activated receptor γ (PPARγ) level in liver tissues. ELISA assay assessed TG secretion. Luciferase assay and RNA pull down were used to validate regulatory mechanism among H19, miR-130a and PPARγ. The gene expression in hepatocytes and liver tissues was detected by quantitative real-time PCR (qRT-PCR) and Western blotting. Results: H19 and PPARγ were up-regulated, while miR-130a was down-regulated in NAFLD mouse and cellular model. H&E and Oil-Red O staining indicated an increased lipid accumulation. Knockdown of H19 inhibited steatosis and TG secretion in FFA-induced hepatocytes. H19 could bind to miR-130a, and miR-130a could directly inhibit PPARγ expression. Meanwhile, miR-130a inhibited lipid accumulation by down-regulating NAFLD-related genes PPARγ, SREBP1, SCD1, ACC1 and FASN. Overexpression of miR-130a and PPARγ antagonist GW9662 inhibited lipogenesis and TG secretion, and PPARγ agonist GW1929 reversed this change induced by miR-130a up-regulation. Conclusion: Knockdown of H19 alleviated hepatic lipogenesis via directly regulating miR-130a/PPARγ axis, which is a novel mechanistic role of H19 in the regulation of NAFLD.

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 < 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 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.

Influences of Unmodified and Carboxylated Carbon Nanotubes on Lipid Profiles in THP-1 Macrophages: A Lipidomics Study

2021 ◽  
pp. 109158182110566
Author(s):  
Lanjie Pei ◽  
Wenxiang Yang ◽  
Yi Cao
Keyword(s):  
Er Stress ◽  
Lipid Profile ◽  
Lipid Accumulation ◽  
Lipid Profiles ◽  
Lipid Classes ◽  
Foam Cell Formation ◽  
Stress Genes ◽  
Profile Changes ◽  
Oil Red O Staining ◽  
Oil Red O

Since the possible roles of surface modifications in determining multi-walled carbon nanotube (MWCNT)–promoted endoplasmic reticulum (ER) stress-mediated lipid-laden macrophage foam cell formation are still in debate, we compared unmodified and carboxylated MWCNT-induced cytotoxicity, lipid profile changes, and expression of ER stress genes in THP-1 macrophages. Particularly, we focused on lipid profile changes by using lipidomics approaches. We found that unmodified and carboxylated MWCNTs significantly decreased cellular viability and appeared to damage the cellular membrane to a similar extent. Likewise, the results from Oil Red O staining showed that both types of MWCNTs slightly but significantly induced lipid accumulation. In keeping with Oil Red O staining results, lipidomics data showed that both types of MWCNTs up-regulated most of the lipid classes. Interestingly, almost all lipid classes were relatively higher in carboxylated MWCNT-exposed THP-1 macrophages compared with unmodified MWCNT-exposed cells, indicating that carboxylated MWCNTs more effectively changed lipid profiles. But in contrast to our expectation, none of the MWCNTs significantly induced the expression of ER stress genes. Even, compared with carboxylated MWCNTs, unmodified MWCNTs induced higher expression of lipid genes, including macrophage scavenger receptor 1 and fatty acid synthase. Combined, our results suggested that even though carboxylation did not significantly affect MWCNT-induced lipid accumulation, carboxylated MWCNTs were more potent to alter lipid profiles in THP-1 macrophages, indicating the need to use omics techniques to understand the exact nanotoxicological effects of MWCNTs. However, the differential effects of unmodified and carboxylated MWCNTs on lipid profiles might not be related with the induction of ER stress.

scholarly journals GLP-1/GLP-1R Signaling in Regulation of Adipocyte Differentiation and Lipogenesis

2017 ◽  
Vol 42 (3) ◽  
pp. 1165-1176 ◽  
Author(s):  
Jicui Chen ◽  
Huichen Zhao ◽  
Xiaoli Ma ◽  
Yuchao Zhang ◽  
Sumei Lu ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Lipid Accumulation ◽  
Fatty Acid Synthase ◽  
Body Weight Gain ◽  
Mapk Signaling ◽  
Marker Genes ◽  
Down Regulation ◽  
Adipose Tissues ◽  
Oil Red O Staining ◽  
Oil Red O

Background/Aims: The aim of this study was to determine the direct role of liraglutide (LG) in adipogenesis and lipid metabolism. Methods: Lipid accumulation was evaluated by oil red O staining, quantitative real-time PCR (qPCR) was performed to determine glucagon-like peptide 1 receptor (GLP-1R), fatty acid synthase (FASN) and adipose triglyceride lipase (ATGL) expression in 3T3-L1 preadipocytes, differentiated adipocytes and in adipose tissues from mice. The effects of LG on 3T3-L1 adipogenesis and lipid metabolism were analyzed with qPCR, Western Blotting, oil red O staining, immunohistochemistry (IHC) and immunofluorescence (IF). All measurements were performed at least three times. Results: LG increased the expression of differentiation marker genes and lipid accumulation during preadipocyte differentiation. In differentiated adipocytes, LG decreased FASN expression, and simultaneously led to CREB phosphorylation and ERK1/2 activation which were abolished by a GLP-1R antagonist, exendin (9-39). LG induced-FASN down-regulation was partially reversed by PKA and ERK1/2 inhibitors. Consistent with above in vitro findings, LG treatment significantly reduced FASN expression in visceral adipose tissues of ob/ob mice, and reduced body weight gain. Conclusion: LG promotes preadipocytes differentiation, and inhibits FASN expression in adipocytes. LG induced down-regulation of FASN is at least partially mediated by PKA and MAPK signaling pathways.

scholarly journals Dietary Oxysterol, 7-Ketocholesterol Accelerates Hepatic Lipid Accumulation and Macrophage Infiltration in Obese Mice

2021 ◽  
Vol 11 ◽  
Author(s):  
Jiuyang Chang ◽  
Masahiro Koseki ◽  
Ayami Saga ◽  
Kotaro Kanno ◽  
Tomoaki Higo ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Hepatic Steatosis ◽  
Lipid Accumulation ◽  
Lipid Extraction ◽  
Acid Oxidation ◽  
Chow Diet ◽  
Obese Mice ◽  
Mice Model ◽  
Alcoholic Fatty Liver ◽  
Hepatic Lipid Accumulation

Non-alcoholic fatty liver disease is strongly associated with obese and type 2 diabetes. It has been reported that an oxidized cholesterol, 7-ketocholesterol (7KC), might cause inflammatory response in macrophages and plasma 7KC concentration were higher in patients with cardiovascular diseases or diabetes. Therefore, we have decided to test whether small amount of 7KC in diet might induce hepatic steatosis and inflammation in two types of obese models. We found that addition of 0.01% 7KC either in chow diet (CD, regular chow diet with 1% cholesterol) or western type diet (WD, high fat diet with 1% cholesterol) accelerated hepatic neutral lipid accumulation by Oil Red O staining. Importantly, by lipid extraction analysis, it has been recognized that triglyceride rather than cholesterol species was significantly accumulated in CD+7KC compared to CD as well as in WD+7KC compared to WD. Immunostaining revealed that macrophages infiltration was increased in CD+7KC compared to CD, and also in WD+7KC compared to WD. These phenotypes were accompanied by inducing inflammatory response and downregulating fatty acid oxidation. Furthermore, RNA sequence analysis demonstrated that 7KC reduced expression of genes which related to autophagy process. Levels of LC3-II protein were decreased in WD+7KC compared to WD. Similarly, we have confirmed the effect of 7KC on acceleration of steatohepatitis in db/db mice model. Collectively, our study has demonstrated that small amount of dietary 7KC contributed to accelerate hepatic steatosis and inflammation in obese mice models.

scholarly journals Telithromycin Treatment of Chronic Chlamydia pneumoniae Infection in C57BL/6J mice

2004 ◽  
Vol 48 (10) ◽  
pp. 3655-3661 ◽  
Author(s):  
Liisa Törmäkangas ◽  
Hannu Alakärppä ◽  
Denise Bem David ◽  
Maija Leinonen ◽  
Pekka Saikku
Keyword(s):  
Lung Tissue ◽  
Lipid Accumulation ◽  
Chlamydia Pneumoniae ◽  
Cholesterol Diet ◽  
Aortic Sinus ◽  
Geometric Means ◽  
Once Daily ◽  
Inflammatory Reactions ◽  
Oil Red O Staining ◽  
Oil Red O

ABSTRACT Chronic Chlamydia pneumoniae infections have been associated with atherosclerosis, but clear knowledge about how these infections should be treated is lacking. We studied the effect of a new ketolide antibiotic, telithromycin, on chronic C. pneumoniae lung infection. Female C57BL/6J mice on a 0.2% cholesterol diet were inoculated intranasally with C. pneumoniae either two or three times every fourth week. Telithromycin was given to the mice subcutaneously at 75 mg/kg of body weight once daily for 5 or 10 days, starting at 3 days after the last inoculation. Samples were taken at 4 and 12 weeks after the last inoculation. The presence of C. pneumoniae DNA in lung tissue was demonstrated by PCR and the detection of lipid accumulation in the aortic sinus by Oil-Red-O staining. C. pneumoniae DNA positivity and inflammatory reactions in the lung tissue of the mice inoculated twice were significantly affected by treatment after both inoculations or only after the second inoculation at 12 weeks. Intimal lipid accumulation in the aortic sinus was also slightly but significantly less abundant in the mice treated after both inoculations compared to the levels in those treated only after the second inoculation for 10 days (geometric means, 823 and 4,324 μm2, respectively; P = 0.033). No differences between the infected, untreated controls and the group inoculated three times and treated for 5 days were seen. We conclude that telithromycin is effective in preventing the development of chronic C. pneumoniae infection and intimal lipid accumulation in C56BL/6J mice when the treatment is given after each inoculation.

Study on the anti-inflammatory and lipid-lowering activities of novel chimeric peptide

2020 ◽  
Vol 10 (3) ◽  
pp. 389-395
Author(s):  
Cong Liu ◽  
Meijun Tan ◽  
Kaixiang Cao ◽  
Jingwei Yan ◽  
Kuikui Hu ◽  
...  
Keyword(s):  
Propionibacterium Acnes ◽  
Water Soluble ◽  
Lipid Lowering ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dilution Method ◽  
Tetrazolium Salt ◽  
Hacat Cells ◽  
Anti Inflammatory ◽  
Oil Red O Staining ◽  
Oil Red O

Acne was treated by combining the N-terminal of HPA3NT3 (where Lys in position 13 was replaced by Asn) and the C-terminal antibacterial peptide HAG of GLP-1 (32–36) amide pentapeptide after amino replacement. The minimal inhibitory concentration (MIC) was measured using the broth dilution method to evaluate HAG’s antibacterial activity. HAG’s cytotoxicity was determined using water-soluble tetrazolium salt (WST-1). Interleukin-8 (IL-8) expression in human immortalized keratinocytes (HaCaT) cells stimulated by Propionibacterium acnes and treated with HAG was measured by Enzyme-linked immunosorbent assay (ELISA). IL-8 and toll-like receptor 2 (TLR2) expression was detected using real-time reverse transcriptase polymerase chain reaction (qRTPCR) to analyze HAG’s anti-inflammatory effect in vitro. The total RNA was extracted using SiO2 nanoparticles. Oil red O staining was used to detect the intracellular lipid drop of Sebaceous Gland Cell Line (SZ95), and ELISA was used to detect triglyceride levels. Hematoxylin-eosin (HE) staining was used to evaluate ear edema induced by Propionibacterium acne in mice. The MIC of HAG in Propionibacterium acnes was 6.3 μg/mL, and 50 μg/mL of HAG showed cytotoxicity. HAG significantly reduced TLR2 and IL-8 expression in HaCaT cells. Oil red O staining showed that the lipid distribution of HAG-treated SZ95 and triglyceride secretion decreased. HAG reduced ear swelling induced by Propionibacterium acnes. HAG has anti-bacterial and anti-inflammatory effects, excellent hypolipidemic function, and low cytotoxicity. Further development of HAG could be a promising and effective reagent for acne therapy.

scholarly journals Agastache rugosa Extract and Its Bioactive Compound Tilianin Suppress Adipogenesis and Lipogenesis on 3T3-L1 Cells

2021 ◽  
Vol 11 (16) ◽  
pp. 7679
Author(s):  
Jae Min Hwang ◽  
Mun-Hoe Lee ◽  
Jin-Hee Lee ◽  
Jong Hun Lee
Keyword(s):  
Lipid Accumulation ◽  
Antioxidant Properties ◽  
Bioactive Compound ◽  
Positive Control ◽  
Mrna Levels ◽  
Agastache Rugosa ◽  
Related Factors ◽  
Freeze Dried ◽  
Oil Red O Staining ◽  
Oil Red O

Agastache rugosa, or Korean mint, is an herb used as a spice, food additive and traditional medicinal ingredient. It has desirable effects, such as its antibacterial, antifungal and antioxidant properties. A. rugosa contains many phenolic compounds studied for their various health benefits, with the primary components being tilianin. A. rugosa extract (ARE), which was extracted with ethanol and freeze-dried, contained 21.14 ± 0.15 mg/g of tilianin with a total polyphenol content of 38.11 ± 0.88 mg/g. Next, the antiadipogenic effect of A. rugosa and tilianin was clarified using 3T3-L1 cells, which differentiate into adipocytes and develop lipid droplets. 3T3-L1 cells were treated with ARE or tilianin and lipid accumulation (%) was calculated through oil red O staining. Tilianin elicited dose-dependent decrease in lipid accumulation (% of positive control) (30 μM 92.10 ± 1.19%; 50 μM 69.25 ± 1.78%; 70 μM 54.86 ± 1.76%; non-differentiation 18.10 ± 0.32%), assessed by oil-red-O staining, whereas ARE treatments caused consistent diminution in lipid accumulation regardless of dose (100 μM 86.90 ± 4.97%; 200 μM 87.25 ± 4.34%; 400 μM 88.54 ± 2.27%; non-differentiation 17.96 ± 1.30%), indicating that both compounds have anti-obesity effects on adipocytes. Treatment with ARE lowered the mRNA (PPARγ; C/EBPα; FABP4; SREBP1; ACC; FAS) and protein (PPARγ; C/EBPα; SREBP1) levels of adipogenesis and lipogenesis-related factors. Tilianin showed a greater effect on the mRNA levels compared with ARE. Thus, tilianin and ARE may have anti-adipogenic and anti-lipogenic effects on 3T3-L1 cells and be possible candidates of obesity-related supplements.

Sign in / Sign up

Export Citation Format

Share Document