Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: SNS activated FXR to modulate LDs transport. A – C Protein expression levels of GPAT4 and LDAH in liver tissues. D IHC staining demonstrates GPAT4 expression levels and localization in liver tissues. E , F ORO staining demonstrated successful model establishment, and SNS and OCA reduced LDs deposition in cells. G – L Western blot analysis was performed to confirm that SNS and OCA activated FXR and inhibited GPAT4 expression in vitro. Scale bars, 50 μm, 100 μm and 200 μm. Data are presented as mean ± SD (n = 5). * p < 0.05. ** p < 0.01

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot, In Vitro

Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: Knockdown of GPAT4 reduced LDs deposition, while knockdown of FXR increased GPAT4 expression. A , B The knockdown efficiency of GPAT4 was detected by Western blot assay. C The knockdown efficiency of GPAT4 was verified by PCR experiments, and 1505 was selected as the effective GPAT4 knockdown for subsequent experiments. D , E ORO staining was used to analyze the effect of GPAT4 knockdown on LDs deposition in cells. F , G , H Western blot and PCR experiments were used to screen and verify the knockdown efficiency of FXR, and 786 was selected for subsequent experiments. I , J , K After knocking down FXR, western blot and PCR experiments were performed to detect the expression level of GPAT4. L , M ORO staining was used to analyze the effect of FXR knockdown on LDs deposition in cells. Scale bar, 50 μm. Data are expressed as mean ± SD (n = 3). * p < 0.05. ** p < 0.01

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Knockdown, Expressing, Western Blot, Staining

Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: Database predictions of FXR binding to GPAT4, validated by DL. A , B The UCSC database was used to predict correlation scores and their corresponding p-values. C Predicted binding sequence of FXR. D The JASPAR database analyzed the binding correlation and potential binding sites between FXR and GPAT4. E Schematic diagram of constructing the plasmid for overexpressing FXR. F Schematic diagram of FXR binding sites in the three predicted GPAT4 promoter regions. G , H Western blot was used to detect the expression level of FXR in cells transfected with the plasmid overexpressing FXR. I DL was used to detect the binding of transcription factor FXR to the GPAT4 promoter region. Data are presented as mean ± SD (n = 3). * p < 0.05. ** p < 0.01

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Binding Assay, Sequencing, Plasmid Preparation, Western Blot, Expressing, Transfection

Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: The active components of SNS could bind to FXR and GPAT4. A – G Visualization of the binding of Glycyrrhizin, Hesperidin, Liquiritin, Neohesperidin, Isorhamnetin, Peoniflorin and Nobiletinto to FXR. H – N Visualization of the binding of Glycyrrhizin, Hesperidin, Liquiritin, Neohesperidin, Isorhamnetin, Peoniflorin and Nobiletinto to GPAT4. O – Q The RMSD, Rg and RMSF values of the FXR-Liquiritin complex over time. R – T The RMSD, Rg and RMSF values of the GPAT4-Neohesperidin complex over time

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Binding Assay

Journal: Chinese Medicine

Article Title: Si-Ni-San improves the deposition of lipid droplets in MAFLD through modulating the FXR-GPAT4 axis

doi: 10.1186/s13020-025-01309-5

Figure Lengend Snippet: As illustrated in the figure, SNS activates hepatic FXR, inhibits the LDs transport protein GPAT4, and modulates the expression of proteins involved in lipolysis and lipophagy, including P62, Beclin1, LC3Ⅰ/Ⅱ, HSL, ATGL, and MAGL. Activation of hepatic FXR inhibits GPAT4 expression. Conversely, when FXR is inhibited, LDs deposition in liver cells worsens, leading to feedback upregulation of GPAT4 expression. The FXR-GPAT4 axis was validated using a DL

Article Snippet: The paraffin sections of liver were performed dewaxing followed by antigen retrieval, then add FXR primary antibody (bs-12867R, Bioss, 1:200) and GPAT4 primary antibody (bs-1924R, Bioss, 1:200), and incubate overnight at 4 °C.

Techniques: Expressing, Activation Assay

Journal: Investigative Ophthalmology & Visual Science

Article Title: Alteration in Lysophospholipids and Converting Enzymes in Glaucomatous Optic Nerves

doi: 10.1167/iovs.61.6.60

Figure Lengend Snippet: Evaluation of mRNA expression levels of LPL conversion enzymes in glaucomatous and control ON samples. The GEO dataset GSE45570 was used for these analyses. The dataset provided mRNA amounts for each protein in ( n = 6) control and ( n = 6) POAG ONHs. The RNA amount for genes of interest was obtained for each sample and averaged across control and glaucomatous samples, respectively. A significant overexpression of LPIN2, PLPP3, and LCAT, and a significant downregulation of GPAT4 protein corresponding mRNA was found in glaucomatous samples. We performed an unpaired parametric Student's t -test with Gaussian distribution between control and glaucoma (** P < 0.01; * P < 0.02).

Article Snippet: The primary antibodies against 1-acyl-sn-glycerol-3-phosphate acyltransferase gamma or AGPAT3 (Proteintech; rabbit polyclonal), Autotaxin or ATX/ENPP2 (Abcam, Cambridge, MA, USA; mouse polyclonal), Glycerophosphodiester phosphodiesterase 1 or GDE1 (Proteintech; rabbit polyclonal), Glyceraldehyde-3-phosphate dehydrogenase or GAPDH (Abcam; mouse monoclonal), Glycerol-3-phosphate acyltransferase 4 or GPAT4 (Proteintech; rabbit polyclonal), Phosphatidylcholine sterol acyltransferase or LCAT (Proteintech, Rosemont, IL; rabbit polyclonal), Lysophosphatidylcholine acyltransferase 1 or LPCAT1 (Proteintech; mouse monoclonal), Phosphatidate phosphatase LPIN2 or LPIN2 (Abcam; rabbit polyclonal), Calcium-dependent phospholipase A2 or PLA2G5 (Thermo Fisher Scientific; mouse monoclonal), and Phospholipid phosphatase 3 or PLPP3 (Thermo Fisher Scientific; mouse monoclonal) were used for protein detection.

Techniques: Expressing, Control, Over Expression

Journal: Investigative Ophthalmology & Visual Science

Article Title: Alteration in Lysophospholipids and Converting Enzymes in Glaucomatous Optic Nerves

doi: 10.1167/iovs.61.6.60

Figure Lengend Snippet: A schematic diagram depicting our findings of the aberrations present in the metabolism of LPA, LPE, and LPC. A depiction of how the pathways of the respective LPLs are related, and the enzymes of interest that help catalyze each metabolic reaction. The enzymes depicted in red are upregulated in glaucoma, whereas the enzymes depicted in blue are downregulated. PLA2G5 and LCAT are extracellular, whereas GPAT4, LPIN2, and PLPP3 are cytosolic.

Article Snippet: The primary antibodies against 1-acyl-sn-glycerol-3-phosphate acyltransferase gamma or AGPAT3 (Proteintech; rabbit polyclonal), Autotaxin or ATX/ENPP2 (Abcam, Cambridge, MA, USA; mouse polyclonal), Glycerophosphodiester phosphodiesterase 1 or GDE1 (Proteintech; rabbit polyclonal), Glyceraldehyde-3-phosphate dehydrogenase or GAPDH (Abcam; mouse monoclonal), Glycerol-3-phosphate acyltransferase 4 or GPAT4 (Proteintech; rabbit polyclonal), Phosphatidylcholine sterol acyltransferase or LCAT (Proteintech, Rosemont, IL; rabbit polyclonal), Lysophosphatidylcholine acyltransferase 1 or LPCAT1 (Proteintech; mouse monoclonal), Phosphatidate phosphatase LPIN2 or LPIN2 (Abcam; rabbit polyclonal), Calcium-dependent phospholipase A2 or PLA2G5 (Thermo Fisher Scientific; mouse monoclonal), and Phospholipid phosphatase 3 or PLPP3 (Thermo Fisher Scientific; mouse monoclonal) were used for protein detection.

Techniques: