Oil red O Staining for Lipid Content in Caenorhabditis elegans

An 8-point visual index was developed for Oil Red O staining of neutral lipids in infective juveniles (IJs) of Steinernema carpocapsae (All), S. riobravis (Biosys 355), S. feltiae (UK76) and S. glaseri (NC). The visual index was found to be a reliable and rapid method for determining the relative neutral lipid content of individual IJs and was validated quantitatively by gas chromatography. The relationship between neutral lipid utilization and infectivity of IJs stored in distilled water at 25°C was also investigated and the first quantitative results on neutral lipid utilization in entomopathogenic nematodes are reported. Neutral lipid contents of freshly harvested IJs of S. carpocapsae, S. riobravis, S. feltiae and S. glaseri were 31, 31, 24 and 26% dry wt, respectively. Steinernema carpocapsae showed a sigmoidal pattern for neutral lipid utilization while S. riobravis used neutral lipids at an almost constant rate. Survivorship of these two species ranged between 120 and 135 days, whereas S. feltiae and S. glaseri lived >450 days and had a slower rate of lipid utilization during a 260 day storage period. Oil Red O staining showed that individual IJs in each population utilized lipids at different rates, even though they had the same initial lipid index. The infectivity of S. riobravis, S. feltiae and S. glaseri declined with lipid utilization. In contrast, S. carpocapsae maintained a high level of infectivity even at relatively low lipid levels. Therefore, neutral lipid content was found to be a suitable indicator of infectivity for S. riobravis, S. feltiae and S. glaseri but not for S. carpocapsae.

Download Full-text

Therapeutic effect of Prdx1 on NAFLD mice may be related to the activation of Nrf-2/HO-1 pathway

Current Pharmaceutical Design ◽

10.2174/1381612826666201026151758 ◽

2020 ◽

Vol 26 ◽

Author(s):

Ru-Xue Bai ◽

Ying-Ying Xu ◽

Yan-Ming Chen ◽

Geng Qin ◽

Hui-Fen Wang ◽

...

Keyword(s):

Positive Staining ◽

Wild Type ◽

Mice Model ◽

Alcoholic Fatty Liver ◽

Liver Histopathology ◽

Final Weight ◽

Oil Red O Staining ◽

Alcoholic Fatty Liver Disease ◽

Oil Red O ◽

Liver Tissues

Objective: To investigate the effect of peroxiredoxin1 (Prdx1) on the methionine-choline deficient (MCD)- induced mice model of non-alcoholic fatty liver disease (NAFLD). Methods: Wild type (WT), transgenic Prdx1 over-expressing (TG) and Prdx1 knockout (KO) mice were fed with MCD diet to construct NAFLD model. General parameters was determined followed by detection with HE staining, oil red O staining, Immunofluorescence, Immunohistochemistry, qRT-PCR and Western blotting. The activities of MDA, GPX and SOD were also quantified. Results: Compared with WT + MCD group, mice in KO + MCD group showed the decresed final weight, food intake and the levels of glucose, insulin, total cholesterol and triglyceride, accompanying with the increased FFA, ALT and AST, as well as the aggravated liver histopathology, which was alleviated in TG + MCD group. Also, mice from KO + MCD group had increased F4/80 and CD68 positive staining with the upregulation of pro-inflammatory and fibrogenic factors in liver tissues than those from WT + MCD group, as well as the enhanced MDA and the reduced GPX and SOD, while TG + MCD group demonstrated improvements than the WT + MCD group. Nrf-2/HO-1 pathway in liver tissues from NFALD mice was inhibited, and Prdx1-/- can further reduce the expression of Nrf-2 and HO-1, while Prdx1 overexpression increased Nrf-2 and HO-1 expression. Conclusion: Prdx1 improved oxidative stress, inflammation and fibrosis in liver of NAFLD mice, which may be associate with the activation of Nrf-2/HO-1 pathway.

Download Full-text

Tanshinone IIA Promotes Macrophage Cholesterol Efflux and Attenuates Atherosclerosis of apoE-/- Mice by Omentin-1/ABCA1 Pathway

Current Pharmaceutical Biotechnology ◽

10.2174/1389201020666190404125213 ◽

2019 ◽

Vol 20 (5) ◽

pp. 422-432 ◽

Cited By ~ 3

Author(s):

Yu-lin Tan ◽

Han-xiao Ou ◽

Min Zhang ◽

Duo Gong ◽

Zhen-wang Zhao ◽

...

Keyword(s):

Atherosclerotic Plaque ◽

Lipid Content ◽

Plasma Levels ◽

Cholesterol Efflux ◽

Density Lipoprotein ◽

Tanshinone Iia ◽

Lipoprotein Cholesterol ◽

Mrna Levels ◽

Cellular Lipid ◽

Oil Red O

Background: Tanshinone IIA (Tan IIA) and Omentin-1 have a protective role in the cardiovascular system. However, if and how Tan IIA and Omentin-1 regulate cholesterol metabolism in macrophages has not been fully elucidated. Objective: To investigate the possible mechanisms of Tan IIA and Omentin-1 on preventing macrophage cholesterol accumulation and atherosclerosis development. Methods: The effect of Tan IIA on the protein and mRNA levels of Omentin-1 and ATP-binding cassette transporter A1 (ABCA1) in macrophages was examined by Western blot and qRT-PCR assay, respectively. Cholesterol efflux was assessed by liquid scintillation counting (LSC). Cellular lipid droplet was measured by Oil Red O staining, and intracellular lipid content was detected by high performance liquid chromatography (HPLC). In addition, the serum lipid profile of apoE−/− mice was measured by enzymatic method. The size of atherosclerotic lesion areas and content of lipids and collagen in the aortic of apoE−/− mice were examined by Sudan IV, Oil-red O, and Masson staining, respectively. Results: Tan IIA up-regulated expression of Omentin-1 and ABCA1 in THP-1 macrophages, promoting ABCA1-mediated cholesterol efflux and consequently decreasing cellular lipid content. Consistently, Tan IIA increased reverse cholesterol transport in apoE−/− mice. Plasma levels of high-density lipoprotein cholesterol (HDL-C), ABCA1 expression and atherosclerotic plaque collagen content were increased while plasma levels of low-density lipoprotein cholesterol (LDL-C) and atherosclerotic plaque sizes were reduced in Tan IIA-treated apoE−/− mice. These beneficial effects were, however, essentially blocked by knockdown of Omentin-1. Conclusion: Our results revealed that Tan IIA promotes cholesterol efflux and ameliorates lipid accumulation in macrophages most likely via the Omentin-1/ABCA1 pathway, reducing the development of aortic atherosclerosis.

Download Full-text

Study on the Mechanism of Shugan Xiaozhi Fang on Cells with Non-alcoholic Fatty Liver Disease

Open Chemistry ◽

10.1515/chem-2019-0145 ◽

2019 ◽

Vol 17 (1) ◽

pp. 1328-1338

Author(s):

Yufeng Xing ◽

Chuantao Zhang ◽

Fenfen Zhai ◽

Tianran Zhou ◽

Xiang Cui ◽

...

Keyword(s):

Liver Disease ◽

Fatty Liver Disease ◽

Dose Group ◽

Control Group ◽

High Dose ◽

Model Group ◽

Alcoholic Fatty Liver ◽

Oil Red O Staining ◽

Alcoholic Fatty Liver Disease ◽

Oil Red O

AbstractCells with non-alcoholic fatty liver disease (NAFLD) were studied to determine the mechanism of liver deficiency via the AdipoR2-PPARa pathway. NAFLD cells were randomly divided into a normal control group, blank control group, model group, low dose group, medium dose group, and high dose group. The NAFLD models were established by incubating the cells with linoleic acid (LA) and palmitic acid (PA) (2:1) for 24 h. The test groups were incubated with different doses of Shugan Xiaozhi Fang extract. The pathological changes in cells that accumulated lipids were detected by Oil Red O staining. Malondialdehyde (MDA) and triglyceride (TG) levels were measured. The apoptosis of cells was evaluated by flow cytometry. The levels of AdipoR2, PPARa, CD36, acyl-CoA mRNA, and protein were confirmed by RT- PCR and Western blot. The results of the Oil Red O staining demonstrated that the NAFLD cell model was successfully established. Compared with the model group, the levels of TG and MDA in the groups that received low, medium, and high doses of Shugan Xiaozhi were significantly lower (P<0.01), and a dose effect was evident. In addition, the expression of AdipoR2, PPARa, CD36, acyl-CoA protein, and mRNA in the Shugan Xiaozhi-treated groups was upregulated. Furthermore, the levels of AdipoR2, PPAR, CD36, acyl-CoA protein, and mRNA in all drug treatment groups that were extracted from L-O2 normal human hepatocytes were significantly upregulated (P<0.01). Moreover, the factor pattern of HepG2 human liver carcinoma cells was similar to that of L-O2. The levels of AdipoR, CD36, acyl-CoA, and AdipoR mRNA in the HepG2 low group were increased (P<0.05). AdipoR, PPAR, CD36, and acyl-CoA protein levels and AdipoR mRNA expression were significantly increased in the intermediate dose group and high dose group (P<0.01). Shugan Xiaozhi Fang attenuates hepatic lipid deposition in NAFLD induced by incubating with LA and PA for 24 h, which is associated with the activation of the AdipoR2-PPARα pathway.

Download Full-text

Type III Collagen is Required for Adipogenesis and Actin Stress Fibre Formation in 3T3-L1 Preadipocytes

Biomolecules ◽

10.3390/biom11020156 ◽

2021 ◽

Vol 11 (2) ◽

pp. 156

Author(s):

Mohammad Al Hasan ◽

Patricia E. Martin ◽

Xinhua Shu ◽

Steven Patterson ◽

Chris Bartholomew

Keyword(s):

Quantitative Polymerase Chain Reaction ◽

Type Iii Collagen ◽

Type Iii ◽

Actin Stress Fibre ◽

Matrix Gene ◽

Gene Transcripts ◽

Polymerase Chain ◽

Oil Red O Staining ◽

Oil Red O

GPR56 is required for the adipogenesis of preadipocytes, and the role of one of its ligands, type III collagen (ColIII), was investigated here. ColIII expression was examined by reverse transcription quantitative polymerase chain reaction, immunoblotting and immunostaining, and its function investigated by knockdown and genome editing in 3T3-L1 cells. Adipogenesis was assessed by oil red O staining of neutral cell lipids and production of established marker and regulator proteins. siRNA-mediated knockdown significantly reduced Col3a1 transcripts, ColIII protein and lipid accumulation in 3T3-L1 differentiating cells. Col3a1−/− 3T3-L1 genome-edited cell lines abolished adipogenesis, demonstrated by a dramatic reduction in adipogenic moderators: Pparγ2 (88%) and C/ebpα (96%) as well as markers aP2 (93%) and oil red O staining (80%). Col3a1−/− 3T3-L1 cells displayed reduced cell adhesion, sustained active β-catenin and deregulation of fibronectin (Fn) and collagen (Col4a1, Col6a1) extracellular matrix gene transcripts. Col3a1−/− 3T3-L1 cells also had dramatically reduced actin stress fibres. We conclude that ColIII is required for 3T3-L1 preadipocyte adipogenesis as well as the formation of actin stress fibres. The phenotype of Col3a1−/− 3T3-L1 cells is very similar to that of Gpr56−/− 3T3-L1 cells, suggesting a functional relationship between ColIII and Gpr56 in preadipocytes.

Download Full-text

Genetic characteristics of Bursaphelenchus xylophilus third-stage dispersal juveniles

Scientific Reports ◽

10.1038/s41598-021-82343-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Qiaoli Chen ◽

Ruizhi Zhang ◽

Danlei Li ◽

Feng Wang

Keyword(s):

Fat Metabolism ◽

Bursaphelenchus Xylophilus ◽

Pinewood Nematode ◽

Genetic Characteristics ◽

Highest Weight ◽

Homologous Genes ◽

Third Stage ◽

Highly Correlated ◽

Oil Red O Staining ◽

Oil Red O

AbstractThe third-stage dispersal juvenile (DJ3) of pinewood nematode (PWN) is highly associated with low-temperature survival and spread of the nematode. Oil-Red-O staining showed that its lipid content was significantly higher compared with other PWN stages. Weighted gene coexpression network analysis identified that genes in the pink module were highly related to DJ3 induced in the laboratory (DJ3-lab). These genes were arranged according to their gene significance (GS) to DJ3-lab. Of the top 30 genes with the highest GS, seven were found to be highly homologous to the cysteine protease family cathepsin 1 (CATH1). The top 30 genes with the highest weight value to each of the seven genes in the pink module were selected, and finally 35 genes were obtained. Between these seven CATH1 homologous genes and their 35 highly related genes, 15 were related to fat metabolism or autophagy. These autophagy-related genes were also found to be highly correlated with other genes in the pink module, suggesting that autophagy might be involved in the mechanism of longevity in DJ3 and the formation of DJ3 by regulating genes related to fat metabolism.

Download Full-text

Pathogenic Microenvironment from Diabetic–Obese Visceral and Subcutaneous Adipocytes Activating Differentiation of Human Healthy Preadipocytes Increases Intracellular Fat, Effect of the Apocarotenoid Crocetin

Nutrients ◽

10.3390/nu13031032 ◽

2021 ◽

Vol 13 (3) ◽

pp. 1032

Author(s):

Lesgui Alviz ◽

David Tebar-García ◽

Raquel Lopez-Rosa ◽

Eva M. Galan-Moya ◽

Natalia Moratalla-López ◽

...

Keyword(s):

Adipose Tissue ◽

Subcutaneous Adipose Tissue ◽

Bioactive Components ◽

Hypertrophic Growth ◽

Visceral Adipose ◽

Subcutaneous Adipose ◽

Oil Red O Staining ◽

Oil Red O ◽

Excessive Accumulation

In diabetes mellitus type 2 (DM2), developed obesity is referred to as diabesity. Implementation of a healthy diet, such as the Mediterranean, prevents diabesity. Saffron is frequently used in this diet because of its bioactive components, such as crocetin (CCT), exhibit healthful properties. It is well known that obesity, defined as an excessive accumulation of fat, leads to cardiometabolic pathology through adiposopathy or hypertrophic growth of adipose tissue (AT).This is related to an impaired adipogenic process or death of adipocytes by obesogenic signals. We aimed to evaluate the effect of the pathogenic microenvironment and CCT, activating differentiation of healthy preadipocytes (PA). For this, we used human cryopreserved PA from visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) depots obtained from healthy and obese-DM2 donors. We studied the effect of a metabolically detrimental (diabesogenic) environment, generated by obese-DM2 adipocytes from VAT (VdDM) or SAT (SdDM), on the viability and accumulation of intracellular fat of adipocytes differentiated from healthy PA, in the presence or absence of CCT (1 or 10 μM). Intracellular fat was quantified by Oil Red O staining. Cytotoxicity was measured using the MTT assay. Our results showed that diabesogenic conditions induce cytotoxicity and provide a proadipogenic environment only for visceral PA. CCT at 10 μM acted as an antiadipogenic and cytoprotective compound.

Download Full-text

Evaluation of Hepatic Steatosis in Dogs With Congenital Portosystemic Shunts Using Oil Red O Staining

Veterinary Pathology ◽

10.1177/0300985813481609 ◽

2013 ◽

Vol 50 (6) ◽

pp. 1109-1115 ◽

Cited By ~ 11

Author(s):

G. B. Hunt ◽

J. A. Luff ◽

L. Daniel ◽

R. Van den Bergh

Keyword(s):

Hepatic Steatosis ◽

Portosystemic Shunts ◽

Oil Red O Staining ◽

Oil Red O

Download Full-text

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

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2022.2960 ◽

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.

Download Full-text