Clinical characteristics of hepatocellular carcinoma patients and controls (mean ± SD).

1<p>Significantly different from the controls at <i>P</i><0.001. ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; AFP: Alpha-fetoprotein.</p

Serum ochratoxin A levels in healthy subjects and in hepatocellular carcinoma patients.

<p>A: HPLC chromatograms for OTA in a representative control; B: HPLC chromatograms for OTA in a representative HCC serum; C: Dispersion diagram for OTA concentration in human sera from healthy and HCC groups; D: Statistical analysis of OTA levels in HCC and normal groups. HPLC: High-performance liquid chromatography; OTA: Ochratoxin A; HCC: Hepatocellular carcinoma.</p

Receiver operating characteristic curve of optimal cut-off value dichotomizing patients with high or low ochratoxin A exposure.

<p>OTA: Ochratoxin A.</p

Rate of ochratoxin A-positive samples and risk of hepatocellular carcinoma in patients presenting ochratoxin A levels above or below the cut-off of 0.207 (calculated by the receiver operating characteristic curves) compared with controls.

1<p>OR relative to an increase of 1 standard deviation among controls.</p>2<p><i>P</i> = 0.0006.</p>3<p><i>P</i><0.001 Compared to the control group.</p><p>OR: Odd ratio.</p><p>CI: Confidence interval; OTA: Ochratoxin A.</p

Potential effect of curcumin on iron accumulation within liver and spleen tissues.

<p><b>A)</b> Structural characteristic of curcumin. <b>B, C)</b> Fold change in the level of iron (μmole/g tissue) in liver and spleen relative to that of normal control, which was assigned a value of 1. <b>D, E)</b> Histological examination of liver and spleen for iron overloaded rats with or without curcumin treatment using Prussian blue staining (blue stain).</p

Curcumin boosts activities of endogenous enzymatic antioxidants that were depleted with chronic iron overload.

<p><b>A, B)</b> Fold change in catalase (CAT) activity (U/g tissue) in liver and spleen relative to that of normal control, which was assigned a value of 1; <b>C, D)</b> Fold change in Superoxide dismutase (SOD) activity (U/g tissue) in liver and spleen relative to that of normal control which was assigned a value of 1. Activities of CAT and SOD in these tissues were significantly decreased in iron overloaded rats than controls. Curcumin treatment significantly boosted activities of both CAT and SOD in iron overloaded rats even to higher levels than those of control rats. Data shown are the mean±SD (n = 5).</p

Validation of the analytical method.

<p>A: HPLC chromatograms of OTA-free serum; B: OTA standard solution 20 ng/mL; C: 20 ng/mL OTA-spiked serum, OTA retention time was 4.2±0.3 min; D: Dose–response curve for OTA spiked in serum. Plotted experimental points are the average of three determinations. HPLC: High-performance liquid chromatography; OTA: Ochratoxin A.</p

Curcumin’s modulation of copper content and antioxidant protein (ceruloplasmin) activity in liver and spleen of iron overloaded rats (A, B).

<p>Levels of copper in both liver and spleen homogenates of iron overloaded rats treated with or without curcumin; (<b>C, D)</b> ceruloplasmin activity (U/g tissue) in liver and spleen relative to normal control, which was assigned a value of 1. Compared to the corresponding values of the control rat group, the administration of curcumin in iron overloaded rats significantly enhanced the decrease in copper levels and ceruloplasmin activity in both liver and spleen homogenates. Data shown are the mean±SD (n = 5).</p

Effect of iron overloading on oxidative stress and NO levels in liver and spleen and potential protective effect of curcumin.

<p><b>A, B)</b> Fold change in the level of malonyldialdehyde (MDA) as a marker of oxidative stress in liver and spleen (nmole/ g wet tissue), respectively, relative to that of normal control, which was assigned a value of 1. Levels of MDA in liver and spleen were significantly increased in iron overloaded rats relative to normal controls and curcumin treatment significantly decreased MDA levels in these tissues during iron overload. <b>C, D)</b> Fold change in the level of NO (nmole/ g wet tissue) in liver and spleen tissues, respectively, relative to that of normal control. Levels of NO in these tissues were significantly increased in iron overloaded rats than in control rats and was significantly reduced by curcumin treatment in iron overloaded rats. Data shown are the mean ± SD (n = 5).</p

3‑Acetyl-11-keto-β-boswellic Acid-Based Hybrids Alleviate Acetaminophen-Induced Hepatotoxicity in HepG2 by the Regulation of Inflammatory and Oxidative Stress Pathways: An Integrated Approach

In an effort to develop new compounds for managing drug-induced liver injury, we prepared 23 novel hybrids based on 3-acetyl-11-keto-β-boswellic acid (AKBA) using various biocompatible linkers. A bioguided approach was employed to identify the most promising hybrid. Eight compounds exhibited superior anti-inflammatory activity compared to the parent compound. Two of these hybrids (5b and 18) were able to reduce gene expression of TNF-α in LPS-induced inflammation in RAW 264.7 cells, similar to dexamethasone. Subsequently, the hepatoprotective potential of these hybrids was evaluated against acetaminophen (APAP) toxicity in HepG2 cells at doses of 1 and 10 μM. Both hybrids effectively restored cytokine levels, which had been elevated by APAP, to normal levels. Furthermore, they normalized depleted superoxide dismutase and reduced glutathione levels while significantly reducing malondialdehyde (MDA) levels. Network pharmacology analysis suggested that AKBA-based hybrids exert their action by regulating PI3K and EGFR pathways, activating anti-inflammatory mechanisms, and initiating tissue repair and regeneration. Molecular docking studies provided insights into the interaction of the hybrids with PI3K. Additionally, the hybrids demonstrated good stability at different pH levels, following first-order kinetics, with relatively long half-lives, suggesting potential for absorption into circulation without significant degradation