Apigenin suppresses epithelial-mesenchymal transition in high glucose-induced retinal pigment epithelial cell by inhibiting CBP/p300-mediated histone acetylation.

Epithelial mesenchymal transition (EMT) is a critical process implicated in the pathogenesis of retinal fibrosis and the exacerbation of diabetic retinopathy (DR) within retinal pigment epithelium (RPE) cells. Apigenin (AP), a potential dietary supplement for managing diabetes and its associated complications, has demonstrated inhibitory effects on EMT in various diseases. However, the specific impact and underlying mechanisms of AP on EMT in RPE cells remain poorly understood. In this study, we have successfully validated the inhibitory effects of AP on high glucose-induced EMT in ARPE-19 cells and diabetic db/db mice. Notably, our findings have identified CBP/p300 as a potential therapeutic target for EMT in RPE cells and have further substantiated that AP effectively downregulates the expression of EMT-related genes by attenuating the activity of CBP/p300, consequently reducing histone acetylation alterations within the promoter region of these genes. Taken together, our results provide novel evidence supporting the inhibitory effect of AP on EMT in RPE cells, and highlight the potential of specifically targeting CBP/p300 as a strategy for inhibiting retinal fibrosis in the context of DR.

Paeoniflorin promotes PPARγ expression to suppress HSCs activation by inhibiting EZH2-mediated histone H3K27 trimethylation.

BACKGROUND: The alleviating effect of paeoniflorin (Pae) on liver fibrosis has been established; however, the molecular mechanism and specific target(s) underlying this effect remain elusive. PURPOSE: This study was to investigate the molecular mechanism underlying the regulatory effect of Pae on hepatic stellate cells (HSCs) activation in liver fibrosis, with a specific focus on the role of Pae in modulating histone methylation modifications. METHODS: The therapeutic effect of Pae was evaluated by establishing in vivo and in vitro models of carbon tetrachloride (CCl4)-induced mice and transforming growth factor ß1 (TGF-ß1)-induced LX-2 cells, respectively. Molecular docking, surface plasmon resonance (SPR), chromatin immunoprecipitation-quantitative real time PCR (ChIP-qPCR) and other molecular biological methods were used to clarify the molecular mechanism of Pae regulating HSCs activation. RESULTS: Our study found that Pae inhibited HSCs activation and histone trimethylation modification in liver of CCl4-induced mice and LX-2 cells. We demonstrated that the inhibitory effect of Pae on the activation of HSCs was dependent on peroxisome proliferator-activated receptor γ (PPARγ) expression and enhancer of zeste homolog 2 (EZH2). Mechanistically, Pae directly binded to EZH2 to effectively suppress its enzymatic activity. This attenuation leaded to the suppression of histone H3K27 trimethylation in the PPARγ promoter region, which induced upregulation of PPARγ expression. CONCLUSION: This investigative not only sheds new light on the precise targets that underlie the remission of hepatic fibrogenesis induced by Pae but also emphasizes the critical significance of EZH2-mediated H3K27 trimethylation in driving the pathogenesis of liver fibrosis.

Si-Ni-San inhibits hepatic Fasn expression and lipid accumulation in MAFLD mice through AMPK/p300/SREBP-1c axis.

BACKGROUND: Soothing the liver and regulating qi is one of the core ideas of traditional Chinese medicine (TCM) in the treatment of fatty liver. Si-Ni-San (SNS) is a well-known herbal formula in TCM for liver soothing and qi regulation in fatty liver treatment. However, its efficacy lacks modern scientific evidence. PURPOSE: This study was aimed to investigate the impact of SNS on metabolic associated fatty liver disease (MAFLD) in mice and explore the underlying molecular mechanisms, particularly its effects on lipid metabolism in hepatocytes. METHODS: The therapeutic effect of SNS was evaluated using in vivo and in vitro models of high-fat/high-cholesterol (HFHC) diet-induced mice and palmitic acid (PA)-induced hepatocytes, respectively. Molecular biological techniques such as RNA-sequencing (RNA-seq), co-immunoprecipitation (co-IP), and western blotting were employed to elucidate the molecular mechanism of SNS in regulating lipid metabolism in hepatocytes. RESULTS: Our findings revealed that SNS effectively reduced lipid accumulation in the livers of HFHC diet-induced mice and PA-induced hepatocytes. RNA-seq analysis demonstrated that SNS significantly down-regulated the expression of fatty acid synthase (Fasn) in the livers of HFHC-fed mice. Mechanistically, SNS inhibited Fasn expression and lipid accumulation by activating adenosine monophosphate (AMP)-activated protein kinase (AMPK). Activation of AMPK suppressed the activity of the transcriptional coactivator p300 and modulated the protein stability of sterol regulatory element-binding protein-1c (SREBP-1c). Importantly, p300 was required for the inhibition of Fasn expression and lipid accumulation by SNS. Furthermore, SNS activated AMPK by decreasing adenosine triphosphate (ATP) production in hepatocytes. CONCLUSION: This study provided novel evidence on the regulatory mechanisms underlying the effects of SNS on Fasn expression. Our findings demonstrate, for the first time, that SNS exerts suppressive effects on Fasn expression through modulation of the AMPK/p300/SREBP-1c axis. Consequently, this regulatory pathway mitigates excessive lipid accumulation and ameliorates MAFLD in mice.

Essential oil extracted from Quzhou Aurantii Fructus prevents acute liver failure through inhibiting lipopolysaccharide-mediated inflammatory response.

Quzhou Aurantii Fructus (QAF) has a long history as a folk medicine and food for the treatment of liver diseases. While our earlier study provided evidence of hepatoprotective properties contained within the flavonoids and limonins constituents in QAF, the potential preventative effects afforded by essential oil components present within QAF remains enigmatic. In this study, we prepared Quzhou Aurantii Fructus essential oil (QAFEO) and confirmed its anti-inflammatory effects on liver inflammation through experimentation on lipopolysaccharide and D-galactosamine (LPS/D-GalN) induced acute liver failure (ALF) mouse models. Using RNA-sequence (RNA-seq) analysis, we found that QAFEO prevented ALF by systematically blunting the pathways involved in response to LPS and toll-like receptor signaling pathways. QAFEO effectively suppressed the phosphorylation of tank-binding kinase 1 (TBK1), TGF-beta activated kinase 1 (TAK1), interferon regulatory factor 3 (IRF3), and the activation of mitogen activated kinase-like protein (MAPK) and nuclear factor-kappa B (NF-κB) pathways in vivo and in vitro. Importantly, QAFEO substantially reduced myeloid differentiation primary response gene 88 (MyD88)- toll-like receptor 4 (TLR4) interaction levels. Moreover, 8 compounds from QAFEO could directly bind to REAL, TAK1, MyD88, TBK1, and IRF3. Taken together, the results of our study support the notion that QAFEO exerts a hepatoprotective effect through inhibiting LPS-mediated inflammatory response.

Saikosaponin A triggers cell ferroptosis in hepatocellular carcinoma by inducing endoplasmic reticulum stress-stimulated ATF3 expression.

Ferroptosis is a type of nonapoptotic necrotic cell death characterized by iron-dependent lipid peroxidation. Saikosaponin A (SsA), a natural bioactive triterpenoid saponin extracted from Radix Bupleuri, has shown potent antitumor activity against various tumors. However, the underlying mechanism of the antitumor activity of SsA remains unclear. Here, we discovered that SsA induced HCC cell ferroptosis in vitro and in vivo. Using RNA-sequence analysis, we found that SsA mainly affected the glutathione metabolic pathway and inhibited the expression of cystine transporter solute carrier family 7 member 11 (SLC7A11). Indeed, SsA increased intracellular malondialdehyde (MDA) and iron accumulation, while it decreased the levels of reduced glutathione (GSH) in HCC. Deferoxamine (DFO), ferrostatin-1 (Fer-1) and GSH could rescue SsA-induced cell death, whereas Z-VAD-FMK was found ineffective in inhibiting SsA-induced cell death in HCC. Importantly, our result indicated that SsA induced the expression of activation transcription factor 3 (ATF3). SsA-induced cell ferroptosis and suppression of SLC7A11 are dependent on ATF3 in HCC. Moreover, we revealed that SsA induced ATF3 upregulation via activation of endoplasmic reticulum (ER) stress. Taken together, our findings support that ATF3-dependent cell ferroptosis mediated the antitumor effects of SsA, opening the possibility to explore SsA as a ferroptosis inducer in HCC.

Analysis of retinal arteriolar and venular parameters in primary open angle glaucoma.

AIM: To measure the retinal vessels of primary open angle glaucoma (POAG) patients on spectral domain optical coherence tomography (SD-OCT) with a full-width at half-maximum (FWHM) algorithm to better explore their structural changes in the pathogenesis of POAG. METHODS: In this retrospective case-control study, the right eyes of 32 patients with POAG and 30 healthy individuals were routinely selected. Images of the supratemporal and infratemporal retinal vessels in the B zones were obtained by SD-OCT, and the edges of the vessels were identified by the FWHM method. The internal and external diameters, wall thickness (WT), wall cross-sectional area (WCSA) and wall-to-lumen ratio (WLR) of the blood vessels were studied. RESULTS: Compared with the healthy control group, the POAG group showed a significantly reduced retinal arteriolar outer diameter (RAOD), retinal arteriolar lumen diameter (RALD) and WSCA in the supratemporal (124.22±12.42 vs 138.32±10.73 µm, 96.09±11.09 vs 108.53±9.89 µm, and 4762.02±913.51 vs 5785.75±1148.28 µm2, respectively, all P<0.05) and infratemporal regions (125.01±15.55 vs 141.57±10.77 µm, 96.27±13.29 vs 110.83±10.99 µm, and 4925.56±1302.88 vs 6087.78±1061.55 µm2, all P<0.05). The arteriolar WT and WLR were not significantly different between the POAG and control groups, nor were the retinal venular outer diameter (RVOD), retinal venular lumen diameter (RVLD) or venular WT in the supratemporal or infratemporal region. There was a positive correlation between the arteriolar parameters and visual function. CONCLUSION: In POAG, narrowing of the supratemporal and infratemporal arterioles and a significant reduction in the WSCA is observed, while the arteriolar WT and WLR do not change. Among the venular parameters, the external diameter, internal diameter, WT, WLR, and WSCA of the venules are not affected.

Author Correction: A full-width half-maximum method to assess retinal vascular structural changes in patients with ischemic heart disease and microvascular angina.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

A full-width half-maximum method to assess retinal vascular structural changes in patients with ischemic heart disease and microvascular anginga.

Chest pain patients without obstructive ischemic heart disease (IHD) have increased attention in the clinical practice as carrying higher cardiovascular (CV) risk and impaired life quality. Retinal vasculature is a novel but reliable risk factor of atherosclerosis and systemic vascular diseases. However, the association of retinal blood vessels and unobstructed IHD, as known as microvascular anginga (MA) is poorly understood. This study compared retinal vascular structures of obstructive IHD and MA using spectral domain optical coherence tomography (SD-OCT) and full-width half-maximum (FWHM) methods to provide new risk predictive evidence of MA. Fundus vessels of 120 IHD patients, including 91epicardial IHD and 29 MA patients, and 66 control subjects were evaluated. Significant differences in the retinal arterial lumen diameter (RALD), retinal arterial outer diameter (RAOD), and arteriovenous ratio (AVR) have been found (P < 0.05). The severity of IHD was negatively correlated with diameters of RAOD, RALD and AVR (P < 0.05). In conclusion, there were significant differences in the retinal vascular structure between IHD patients and patients with MA. Thus, assessment of retinal vascular structure is suggested to evaluate CV risk of IHD patients, despite having no obstructive IHD.

Retinal Arteriolar Morphometry Based on Full Width at Half Maximum Analysis of Spectral-Domain Optical Coherence Tomography Images.

OBJECTIVES: In this study, we develop a microdensitometry method using full width at half maximum (FWHM) analysis of the retinal vascular structure in a spectral-domain optical coherence tomography (SD-OCT) image and present the application of this method in the morphometry of arteriolar changes during hypertension. METHODS: Two raters using manual and FWHM methods measured retinal vessel outer and lumen diameters in SD-OCT images. Inter-rater reproducibility was measured using coefficients of variation (CV), intraclass correlation coefficient and a Bland-Altman plot. OCT images from forty-three eyes of 43 hypertensive patients and 40 eyes of 40 controls were analyzed using an FWHM approach; wall thickness, wall cross-sectional area (WCSA) and wall to lumen ratio (WLR) were subsequently calculated. RESULTS: Mean difference in inter-rater agreement ranged from -2.713 to 2.658 µm when using a manual method, and ranged from -0.008 to 0.131 µm when using a FWHM approach. The inter-rater CVs were significantly less for the FWHM approach versus the manual method (P < 0.05). Compared with controls, the wall thickness, WCSA and WLR of retinal arterioles were increased in the hypertensive patients, particular in diabetic hypertensive patients. CONCLUSIONS: The microdensitometry method using a FWHM algorithm markedly improved inter-rater reproducibility of arteriolar morphometric analysis, and SD-OCT may represent a promising noninvasive method for in vivo arteriolar morphometry.

Update on retinal vessel structure measurement with spectral-domain optical coherence tomography.

This study was conducted to demonstrate a new scan method for retinal vessel structure measurement in a specific region of fundus (zone B) using spectral-domain optical coherence tomography (SD-OCT), and to assess its reliability. One temporal superior retinal vessel pair passing through a concentric ring (zone B), which was defined between half and one disc distance from the optic disc border, was chosen for the measurement using a volume scan in SD-OCT. On the SD-OCT image, retinal arteriolar outer diameter (RAOD), retinal arteriolar lumen diameter (RALD), retinal venular outer diameter (RVOD) and retinal venular lumen diameter (RVLD) were measured. Retinal vessel diameters on color fundus photographs were also analyzed. Fifty-five healthy individuals were recruited to evaluate intraobserver and interobserver reproducibility between the two examiners. The intraobserver intraclass correlation coefficient (ICC) ranged from 0.972 to 0.981, and the interobserver ICC ranged from 0.968 to 0.980. In the Bland-Altman plot, the 95% limits of interobserver agreement for the RAOD, RALD, RVOD and RVLD were -5.60 to 4.84µm, -5.78 to 5.05µm, -7.52 to 5.62µm and -7.10 to 5.63µm, respectively. The retinal arteriolar and venular lumen diameters on the SD-OCT image were close to the mean arteriolar and venular diameters obtained from the color fundus photographs. Volume scan method produced better images of retinal vessels showing the fine structures of the vessel wall, and provided reliable retinal vessel structure measurement in zone B with good repeatability and reproducibility.

[Spectral domain optical coherence tomography to assess the association between the inner caliber of large retinal vessel and the primary hypertension].

OBJECTIVE: To assess the inner caliber of large retinal vessel quantitatively using spectral domain optical coherence tomography (SD-OCT) and to reveal the association between changes in the inner caliber of large retinal vessel and the primary hypertension. METHODS: A retrospective case-control study was carried out including 215 cases (with primary hypertension) and 210 controls (without primary hypertension) admitted to our hospital since 2009 and all the cases and controls were grouped according to age. SD-OCT was performed to assess the inner caliber of large retinal vessel quantitatively including retinal artery inner caliber (RAIC), retinal vein inner caliber (RVIC) and retinal arterio-venous inner caliber ratio (RAVICR). The differences in the inner caliber of large retinal vessel between the cases and the controls in each age groups were analyzed by t test. In all cases, multiple comparisons were performed according to their blood pressure level by SNK test of one way ANOVA. The RAVICR was also correlated with the following relevant determinants via multiple stepwise regression analysis: age, diastolic and systolic blood pressure. RESULTS: In each age group of cases (< 40, 40 to 49, 50 to 59, 60 to 69, ≥ 70 years), the values of RAIC were (93.0 ± 6.3), (86.2 ± 6.1), (84.5 ± 5.1), (84.0 ± 5.5), and (81.7 ± 5.4) µm respectively, and the values of RVIC were (129.4 ± 5.8), (130.7 ± 6.5), (129.6 ± 5.4), (132.2 ± 6.4), and (131.6 ± 5.1) µm respectively, and the values of RAVICR were (0.720 ± 0.07), (0.661 ± 0.06), (0.653 ± 0.04), (0.637 ± 0.06), and (0.621 ± 0.05) µm respectively. Compared with controls, RAIC (t = -4.813, -10.893, -15.689, -8.811, and -10.151 respectively; P < 0.05) and RAVICR (t = -3.276, -8.654, -13.470, -7.801, and -9.210 respectively; P < 0.05) were significantly decreased in each age group of cases. Multiple comparisons were made among each systolic and diastolic pressure groups in all cases. In systolic groups, difference of RAIC or RAVICR were significant (SNK test)between 140 to 149 mm Hg (1 mm Hg = 0.133 kPa) and 170 to 179 mm Hg group (q = 9.46, 10.61; P < 0.05), 140 to 149 mm Hg and ≥ 180 mm Hg group (q = 11.03, 13.98; P < 0.05), 150 to 159 mm Hg and 170 to 179 mm Hg group (q = 8.13, 8.82; P < 0.05), 150 to 159 mm Hg group and ≥ 180 mm Hg group (q = 9.01, 9.97; P < 0.05). In diastolic groups, difference of RAIC or RAVICR were significant (SNK test) between 90 to 99 mm Hg and 100 to 109 mm Hg group (q = 6.79, 5.95;P < 0.05), 90 to 99 mm Hg and ≥ 110 mm Hg group (q = 9.72, 10.21; P < 0.05), 100 to 109 mm Hg and ≥ 110 mm Hg group (q = 5.93, 6.07; P < 0.05). RAVICR was associated with the diastolic and systolic blood pressure revealed by the multiple stepwise regression analysis (ANOVA: F = 11.231; Standardized regression coefficient: ß = -0.024, -0.019, respectively; P < 0.05). CONCLUSIONS: Quantitative assessment for the inner caliber of large retinal vessel can be done by SD-OCT. The value of RAI and RAVICR were correlated with diastolic and systolic blood pressure in primary hypertension.