Journal: Lipids in Health and Disease

Article Title: Dysregulated lipid metabolism is associated with kidney allograft fibrosis

doi: 10.1186/s12944-024-02021-3

Figure Lengend Snippet: Primer sequences used for RT-qPCR in rat

Article Snippet: The cells in some experiments were pretreated with the Carnitine palmitoyltransferase 1 A (CPT1A) activator C75 (10 μM, HY-12,364, MedChemExpress, USA) or the CPT1A antagonist Etomoxir sodium salt (80 μM, HY-50,202 A, MedChemExpress, USA) for 2 h [ ].

Techniques:

Journal: Lipids in Health and Disease

Article Title: Dysregulated lipid metabolism is associated with kidney allograft fibrosis

doi: 10.1186/s12944-024-02021-3

Figure Lengend Snippet: Immunohistochemical analysis of EMT and CPT1A expression in a rat model of kidney transplantation. (a) The gene expression level of α-SMA in the indicated kidney tissues was analyzed by RT-qPCR. (b) The gene expression level of vimentin in the indicated kidney tissues was analyzed by RT-qPCR. (c) The gene expression level of CPT1A in the indicated kidney tissues was analyzed by RT-qPCR. (d) Representative IHC image of α-SMA. (e) The percentages of α-SMA-positive cells were calculated. (f) Representative IHC image of E-cadherin. (g) The percentages of E-cadherin-positive cells were calculated. (h) Representative IHC image of CPT1A. (i) The intensity of CPT1A expression was calculated. * P < 0.05, ** P < 0.01, **** P < 0.0001, unpaired t test

Article Snippet: The cells in some experiments were pretreated with the Carnitine palmitoyltransferase 1 A (CPT1A) activator C75 (10 μM, HY-12,364, MedChemExpress, USA) or the CPT1A antagonist Etomoxir sodium salt (80 μM, HY-50,202 A, MedChemExpress, USA) for 2 h [ ].

Techniques: Immunohistochemical staining, Expressing, Transplantation Assay, Gene Expression, Quantitative RT-PCR

Journal: Lipids in Health and Disease

Article Title: Dysregulated lipid metabolism is associated with kidney allograft fibrosis

doi: 10.1186/s12944-024-02021-3

Figure Lengend Snippet: Pathological staining and Immunohistochemical analysis of EMT and CPT1A expression in human kidney allograft fibrosis. (a) Representative HE staining. (b) Representative Trichrome-Masson staining. (c) Representative IHC image of α-SMA. (d) Representative IHC image of E-cadherin. (e) Representative IHC image of CPT1A. (f) The percentages of α-SMA-positive cells were calculated. (g) The percentages of E-cadherin-positive cells were calculated. (h) The intensity of CPT1A expression was calculated. ** P < 0.01, unpaired t test

Article Snippet: The cells in some experiments were pretreated with the Carnitine palmitoyltransferase 1 A (CPT1A) activator C75 (10 μM, HY-12,364, MedChemExpress, USA) or the CPT1A antagonist Etomoxir sodium salt (80 μM, HY-50,202 A, MedChemExpress, USA) for 2 h [ ].

Techniques: Staining, Immunohistochemical staining, Expressing

Journal: Lipids in Health and Disease

Article Title: Dysregulated lipid metabolism is associated with kidney allograft fibrosis

doi: 10.1186/s12944-024-02021-3

Figure Lengend Snippet: In vitro model of EMT induced by TGF-β1. (a) A scratch healing assay showed that C75 could inhibit HK-2 cell migration induced by TGF-β1 at 24 h. (b) Comparison of migration indices among the four intervention groups. The migration index is defined as maximum migration distance at 24 h; *** P < 0.001, **** P < 0.0001; one-way ANOVA. (c) Changes in HK-2 cell morphology at 48 h in four intervention groups. (d) Volcano plot showing the DEGs between untreated group and TGF-β1 group. (e) GO analysis of the DEGs shown in Fig. 7D

Article Snippet: The cells in some experiments were pretreated with the Carnitine palmitoyltransferase 1 A (CPT1A) activator C75 (10 μM, HY-12,364, MedChemExpress, USA) or the CPT1A antagonist Etomoxir sodium salt (80 μM, HY-50,202 A, MedChemExpress, USA) for 2 h [ ].

Techniques: In Vitro, Migration, Comparison

Journal: Lipids in Health and Disease

Article Title: Dysregulated lipid metabolism is associated with kidney allograft fibrosis

doi: 10.1186/s12944-024-02021-3

Figure Lengend Snippet: Immunofluorescence analysis of EMT and CPT1A expression in the in vitro model. (a) Immunofluorescence analysis of fibronectin expression. (b) The relative fluorescence intensity of fibronectin in Fig. 8A was determined by ImageJ. (c) Immunofluorescence analysis of α-SMA expression. (d) The relative fluorescence intensity of α-SMA in Fig. 8C was determined by ImageJ. (e) Immunofluorescence assay of CPT1A expression. (F) The relative fluorescence intensity of α-SMA in Fig. 8E was determined by ImageJ. ** P < 0.01, *** P < 0.001, **** P < 0.0001, one-way ANOVA

Article Snippet: The cells in some experiments were pretreated with the Carnitine palmitoyltransferase 1 A (CPT1A) activator C75 (10 μM, HY-12,364, MedChemExpress, USA) or the CPT1A antagonist Etomoxir sodium salt (80 μM, HY-50,202 A, MedChemExpress, USA) for 2 h [ ].

Techniques: Immunofluorescence, Expressing, In Vitro, Fluorescence

Journal: Foods

Article Title: Sea Buckthorn Pericarp Flavonoids Improve Diet-Induced Hyperlipidemia via Coordinated Modulation of Hepatic Lipid Metabolism and Gut Microbiota

doi: 10.3390/foods15061049

Figure Lengend Snippet: TFSP modulates the expression of ACC, FAS, CPT-1α, PPARα and ATGL proteins in the livers of HFD mice. n = 6 per group. Data are presented as mean ± SD. Different letters above bars indicate statistically significant differences ( p < 0.05) by one-way ANOVA followed by LSD post hoc test.

Article Snippet: Peroxisome proliferator-activated receptor alpha (PPARα) primary antibody (Cat. No. A00600-2), carnitine palmitoyltransferase-1 alpha (CPT-1α) primary antibody (Cat. No. A00917-3), acetyl-CoA carboxylase (ACC) primary antibody (Cat. No. M01802-2), fatty acid synthase (FAS) primary antibody (Cat. No. BA0484), adipose triglyceride lipase (ATGL) primary antibody (Cat. No. A01800-1), and GAPDH primary antibody (Cat. No. BM1623) were obtained from BOSTER Biological Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing

Journal: Biomolecules & Therapeutics

Article Title: Dietary Potassium Synergistically Enhances Anti-Obesity Efficacy of Garcinia Cambogia Complex in High-Fat Diet-Induced Obese Mice

doi: 10.4062/biomolther.2025.069

Figure Lengend Snippet: The synergistic effect of potassium supplement on lipid accumulation in liver and adipose tissues. (A) Hematoxylin and eosin (H&E) staining of liver, epididymal white-adipose tissue, and inter-scapular brown adipose tissue. Oil staining of liver sections from the same groups and the quantification of hepatic steatosis. (B) Quantification of lipid droplet area in liver, WAT, and BAT. Values are expressed as mean ± SD (n=5 per group). (C) Western blot analysis of lipid metabolism-related proteins including FAS, CPT1, UCP-1, and PGC1α in liver, WAT, and BAT. β-actin was used as a loading control. The significance of the difference between the four groups was analyzed by one-way ANOVA analysis of variance. * p <0.05 and *** p <0.001 versus HFD group.

Article Snippet: Antibodies used included sterol regulatory element-binding protein 1 (SREBP1, 1:50, Invitrogen, Waltham, MA, USA, cat. No. PA1-337), phosphorylated acetyl-CoA carboxylase1 (p-ACC1, 1:30, Cell Signaling, cat. No. 3661), FAS (1:50, Santa Cruz Biotechnology, cat. No. sc20140), phosphorylated AMP-activated protein kinase (p-AMPK, 1:30, cat. No. 2575), and CPT1 (1:50, Novus Biologicals, cat. No. NB100-53791).

Techniques: Staining, Western Blot, Control

Journal: Biomolecules & Therapeutics

Article Title: Dietary Potassium Synergistically Enhances Anti-Obesity Efficacy of Garcinia Cambogia Complex in High-Fat Diet-Induced Obese Mice

doi: 10.4062/biomolther.2025.069

Figure Lengend Snippet: Inhibition of lipogenesis by potassium supplementation through the IGF1R/PI3K/AKT/GSK3β signaling in combination with the GC complex. (A) The immunohistochemical staining of FAS, SREBP1, CPT1 and p-ACC1 in liver sections from each group. (B) Quantification of immunohistochemical (IHC) scoring for positive staining of the four proteins. The expression of lipogenesis-related proteins was significantly decreased by potassium and GC complex compared to the HFD group. Values are expressed as mean ± SD (n=5 per group). (C) Western blot analysis of IGF1R, PI3K, p-AKT, p-GSK3β in liver tissue of each group. β-actin was used as a loading control. The significance of the difference between the four groups was analyzed by one-way ANOVA analysis of variance. * p <0.05 and ** p <0.01 versus HFD group.

Article Snippet: Antibodies used included sterol regulatory element-binding protein 1 (SREBP1, 1:50, Invitrogen, Waltham, MA, USA, cat. No. PA1-337), phosphorylated acetyl-CoA carboxylase1 (p-ACC1, 1:30, Cell Signaling, cat. No. 3661), FAS (1:50, Santa Cruz Biotechnology, cat. No. sc20140), phosphorylated AMP-activated protein kinase (p-AMPK, 1:30, cat. No. 2575), and CPT1 (1:50, Novus Biologicals, cat. No. NB100-53791).

Techniques: Inhibition, Immunohistochemical staining, Staining, Expressing, Western Blot, Control

Journal: Nature communications

Article Title: PD-1 alters T-cell metabolic reprogramming by inhibiting glycolysis and promoting lipolysis and fatty acid oxidation.

doi: 10.1038/ncomms7692

Figure Lengend Snippet: Figure 3 | PD-1 induces upregulation of CPT1A and fatty acid b-oxidation. (a) CPT1A mRNA was analysed by real-time quantitative PCR and relative expression of mRNA of each time point and culture condition over the levels expressed in unstimulated cells (defined as 1) is shown (*Po0.05, TCC þ PD1 versus TCC, Student’s t-test; ~Po0.05, TCC þ PD1 versus UT and TCC versus UT, analysis of variance (ANOVA); n ¼ 3 experiments). (b) CD4 þ primary human T cells were cultured under the indicated conditions. Cell lysates were prepared at the indicated time points and expression of CPT1A and b-actin were assessed by SDS–PAGE and immunoblot. Results are representative of three experiments. (c) Fatty acid b-oxidation rate after culture under the indicated conditions for various time intervals was examined. Values of TCC and TCC þ PD1 cells were compared with unstimulated (UT) cells (~Po0.05, ANOVA; n ¼ 3) and values of TCC þ PD1 were compared with TCC cells (*Po0.05, Student’s t-test; n ¼ 3).

Article Snippet: The anti-FASN (#3189) was from Cell Signaling Technologies, Danvers, MA, and the anti-CPT1A (#TA303144) antibody was from OriGene Technologies Inc., Rockville, MD.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Cell Culture, SDS Page, Western Blot

Journal: Nature communications

Article Title: PD-1 alters T-cell metabolic reprogramming by inhibiting glycolysis and promoting lipolysis and fatty acid oxidation.

doi: 10.1038/ncomms7692

Figure Lengend Snippet: Figure 6 | Inhibition of PI3K/Akt and MEK/Erk pathways is involved in the metabolic reprogramming of activated Tcells. (a) CD4 þ primary human T cells were cultured with tosyl-activated magnetic beads conjugated with aCD3/aCD28/IgG (TCC) in the presence of either LY294002 (LY, 10 mM), UO126 (UO, 10 mM) or their combination. Cell lysates were prepared at the indicated time points and expression of CPT1A and b-actin was assessed by SDS–PAGE and immunoblot. Results are representative of three experiments. (b–d) In parallel experiments, rate of fatty acid b-oxidation was examined. Values of TCC þ inhibitor cultures were compared with TCC (*Po0.05, Student’s t-test; n ¼ 3) and values of TCC and TCC þ inhibitor cultures were compared with unstimulated (UT) (~Po0.05, analysis of variance (ANOVA); n ¼ 3). (e) At 72 h of culture under the indicated conditions, spare respiratory capacity (SRC) was determined. Values of TCC and TCC þ inhibitor cells were compared with unstimulated (UT) cells (~Po0.05, ANOVA; n ¼ 3) and values in TCC þ inhibitor cells were compared with TCC (*Po0.05, Student’s t-test; n ¼ 3). (f) T cells were cultured with tosyl-activated magnetic beads conjugated with aCD3/aCD28/PD-L1-Ig or with tosylatcivated magnetic beads conjugated with aCD3/aCD28/IgG in the presence of either LY294002 (LY, 10 mM), UO126 (UO, 10 mM) or their combination. Cell lysates were prepared at the indicated time points and expression of ATGL and b-actin was assessed by SDS–PAGE and immunoblot.

Article Snippet: The anti-FASN (#3189) was from Cell Signaling Technologies, Danvers, MA, and the anti-CPT1A (#TA303144) antibody was from OriGene Technologies Inc., Rockville, MD.

Techniques: Inhibition, Cell Culture, Magnetic Beads, Expressing, SDS Page, Western Blot

Journal: Diabetes

Article Title: Hepatic CPT1A Facilitates Liver-Adipose Cross-Talk via Induction of FGF21 in Mice.

doi: 10.2337/db21-0363

Figure Lengend Snippet: Figure 1—Diet-induced obesity and insulin resistance are improved upon hepatocyte-restricted deletion of Cpt1a. Liver-specific Cpt1a KO (LKO) mice and WT littermates were fed with HFD for 8 weeks. A: Food intake. B: Body weight. C: Representative images of WT and LKO mice (n 5 7). D: Body composition (n 5 5). GTT (E) and area under the curve (AUC) (F) of the mice after 8 weeks of HFD (n 5 7). G: Fasting insulin level (n 5 7). ITT (H) and AUC (n 5 7) (I). J–L: Western blotting of insulin stimulated phospho-S473 (p-S473) and total (t)-AKT in epididymal WAT (eWAT), skeletal muscle, and liver. M: Densitometry quantification of p-Akt/t-Akt in tissues (n 5 4). Data represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The membrane was then blotted with antibodies against CPT1A (NBP1–59608; Novus Biologicals); UCP1 (ab10983; Abcam); and GAPDH (cat. no. 5174T), AKT (cat. no. 4685), p-AKT (cat. no. 4060), eIF2a (cat. no. 5324T0), p-eIF2a (cat. no. 5324T), ATF6 (cat. no. 65880S) and CHOP (cat. no. 5554T) (all from Cell Signaling Technology).

Techniques: Western Blot

Journal: Diabetes

Article Title: Hepatic CPT1A Facilitates Liver-Adipose Cross-Talk via Induction of FGF21 in Mice.

doi: 10.2337/db21-0363

Figure Lengend Snippet: Figure 2—CPT1A deficiency in liver leads to more severe hepatosteatosis but protects mice against ER stress and liver injury. LKO and WT mice were fed with HFD for 8 weeks. A: Liver weight (n 5 5). B: Gross picture of liver. H-E (C) and Oil Red O (D) staining of liver sec- tions. E: TG content in WT and LKO mice (n 5 5). Serum AST (F) and ALT (G) of the mice (n 5 6). H: Real-time PCR of inflammatory markers in liver (n 5 5). Western blotting (I) and corresponding quantification (J). I: ER stress–related proteins in liver. K: Real-time PCR of ER stress–related genes in liver (n 5 5). Data represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The membrane was then blotted with antibodies against CPT1A (NBP1–59608; Novus Biologicals); UCP1 (ab10983; Abcam); and GAPDH (cat. no. 5174T), AKT (cat. no. 4685), p-AKT (cat. no. 4060), eIF2a (cat. no. 5324T0), p-eIF2a (cat. no. 5324T), ATF6 (cat. no. 65880S) and CHOP (cat. no. 5554T) (all from Cell Signaling Technology).

Techniques: Staining, Real-time Polymerase Chain Reaction, Western Blot

Journal: Diabetes

Article Title: Hepatic CPT1A Facilitates Liver-Adipose Cross-Talk via Induction of FGF21 in Mice.

doi: 10.2337/db21-0363

Figure Lengend Snippet: Figure 3—Adipose tissue browning and energy expenditure are increased upon hepatic Cpt1a deletion in mice. LKO and WT mice were fed with HFD for 8 weeks. Adipose tissue weight (n 5 5) (A) and gross images of inguinal (iWAT) and epididymal (eWAT) white adipose tis- sue (B). H-E staining of adipose tissues (C) and quantification of adipocyte area (D). Whole-body oxygen consumption (E and F) and heat production (G and H) of the mice (n 5 5). Real-time PCR for brown markers (I) and Western blotting of UCP1 in brown adipose tissue (J) (n 5 5). Real-time PCR for brown markers (K) and immunohistochemical staining for UCP1 in iWAT (L) (n 5 5). Data represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The membrane was then blotted with antibodies against CPT1A (NBP1–59608; Novus Biologicals); UCP1 (ab10983; Abcam); and GAPDH (cat. no. 5174T), AKT (cat. no. 4685), p-AKT (cat. no. 4060), eIF2a (cat. no. 5324T0), p-eIF2a (cat. no. 5324T), ATF6 (cat. no. 65880S) and CHOP (cat. no. 5554T) (all from Cell Signaling Technology).

Techniques: Staining, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemical staining

Journal: Diabetes

Article Title: Hepatic CPT1A Facilitates Liver-Adipose Cross-Talk via Induction of FGF21 in Mice.

doi: 10.2337/db21-0363

Figure Lengend Snippet: Figure 4—PPARa-FGF21 axis is stimulated in CPT1-deficient liver. A–C: RNA-seq analysis of LKO vs. WT liver from HFD-treated mice (n 5 5). A: Kyoto Encyclopedia of Genes and Genomes pathway analysis of DEGs. B: Heat map of PPARa downstream genes in liver from control and LKO mice. C: Volcano plot of genes mapped in RNA-seq. mRNA level of Fgf21 (D) and Western blotting for FGF21 (E) in HFD- treated WT and LKO mice liver (n 5 5). F: Quantification of E. G: Plasma FGF21 concentration (n 5 8). H: Liver explants from WT and LKO mice on HFD were cultured ex vivo and the concentration of FGF21 in conditioned medium was measured by ELISA (n 5 5). I: Primary hepatocytes were isolated from obese WT and LKO mice and treated with vehicle or GW6471, and Fgf21 mRNA was examined 12 h later. Data represent mean ± SEM. **P < 0.01; ***P < 0.001.

Article Snippet: The membrane was then blotted with antibodies against CPT1A (NBP1–59608; Novus Biologicals); UCP1 (ab10983; Abcam); and GAPDH (cat. no. 5174T), AKT (cat. no. 4685), p-AKT (cat. no. 4060), eIF2a (cat. no. 5324T0), p-eIF2a (cat. no. 5324T), ATF6 (cat. no. 65880S) and CHOP (cat. no. 5554T) (all from Cell Signaling Technology).

Techniques: RNA Sequencing, Control, Western Blot, Clinical Proteomics, Concentration Assay, Cell Culture, Ex Vivo, Enzyme-linked Immunosorbent Assay, Isolation

Journal: Diabetes

Article Title: Hepatic CPT1A Facilitates Liver-Adipose Cross-Talk via Induction of FGF21 in Mice.

doi: 10.2337/db21-0363

Figure Lengend Snippet: Figure 7—Schematic showing how CPT1A deficiency results in hepatosteatosis and improved metabolic phenotype. CPT1 defi- ciency causes lipid accumulation in hepatocytes and subsequent activation of PPARa. In response, hepatic Fgf21 is transactivated, leading to increased FGF21, which then acts autonomously to antagonize hepatic ER stress and cellular damage evoked by HFD. FGF21 also serves as a mediator to engage liver–adipose interor- gan communication, leading to adipose tissue browning and improved insulin sensitivity.

Article Snippet: The membrane was then blotted with antibodies against CPT1A (NBP1–59608; Novus Biologicals); UCP1 (ab10983; Abcam); and GAPDH (cat. no. 5174T), AKT (cat. no. 4685), p-AKT (cat. no. 4060), eIF2a (cat. no. 5324T0), p-eIF2a (cat. no. 5324T), ATF6 (cat. no. 65880S) and CHOP (cat. no. 5554T) (all from Cell Signaling Technology).

Techniques: Activation Assay