Curcumin Modulates NOX Gene Expression and ROS Production via P-Smad3C in TGF-ß-Activated Hepatic Stellate Cells.

Background: Liver fibrosis, associated with hepatic stellate cells (HSCs), occurs when a healthy liver sustains damage, thereby impairing its function. NADPH oxidases (NOXs), specifically isoforms 1, 2, and 4, play a role in reactive oxygen species (ROS) production during hepatic injuries, resulting in fibrosis. Curcumin has shown strong potential in mitigating liver fibrosis. Our research aimed to investigate the effects of curcumin on lowering NOX and ROS levels. This compound was also studied for its effects on NOXs, ROS concentrations through the inhibition of Smad3 phosphorylation in transforming growth factor beta (TGF-ß)-activated human HSCs. Methods: MTT assay investigated the cytotoxic effects of curcumin on HSCs. The cells were activated by exposure to TGF-ß (2 ng/mL) for 24 hours. After activating, the cells were treated with curcumin at 25-150 µM concentrations. After administering curcumin to the cells, we employed RT-PCR and Western blot techniques to evaluate the related gene and protein expression levels. This evaluation was primarily focused on the mRNA expression levels of NOX1, NOX2, NOX4 and phosphorylated Smad3C. Results: The mRNA expression level of aforesaid NOXs as well as α-smooth muscle actin (α-SMA), collagen1-α, and ROS levels were significantly reduced following 100 µM curcumin treatment. Furthermore, curcumin significantly decreased the p-Smad3C protein level in TGF-ß-activated cells, with fold changes of 3 and 2 observed at 75 and 100 µM, respectively. Conclusion: Curcumin decreased the levels of ROS and NOX, as well as the expression of α-SMA and collagen1-α. The primary mechanism for this reduction could be linked to the level of p-Smad3C. Hence, curcumin could serve as an effective therapeutic agent for liver fibrosis.

Examining dyslipidaemia, metabolic syndrome and liver enzyme levels in patients with prediabetes and type 2 diabetes in population from Hoveyzeh cohort study: A case-control study in Iran.

INTRODUCTION: Type 2 diabetes mellitus (T2DM) is among the world's top 10 leading causes of death. Additionally, prediabetes is a major risk factor for diabetes. Identifying diabetes co-occurring disorders can aid in reducing adverse effects and facilitating early detection. In this study, we evaluated dyslipidaemia, metabolic syndrome (MetS), and liver enzyme levels in pre-diabetic and T2DM patients in the Persian cohort compared to a control group. MATERIALS AND METHODS: In this cross-sectional study, 2259 pre-diabetes, 1664 T2DM and 5840 controls (35-70 years) who were selected from the Hoveyzeh cohort centre were examined. Body mass index, blood pressure, fasting blood glucose (FBG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), triglyceride (TG) and liver enzymes: γ-glutamyltransferase (GGT), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined using the standard protocols. MetS subjects were also identified based on the National Cholesterol Education Program guidelines. RESULTS: Prediabetes and T2MD were closely correlated with the lipid profile, MetS, and liver enzymes (ALT, GGT, ALT/AST). MetS increases the risk of T2DM by 12.45 [95% CI: 10.88-14.24] fold, while an increase in ALT/AST ratio increases the risk of T2DM by 3.68 [95% CI: 3.159-4.154] fold. ROC curve analysis also revealed the diagnostic roles of GGT, ALT, AST and the ALT/AST ratio among pre-diabetics, diabetics and the control group. The GGT level corresponds to the highest AUCs (0.685) with the highest sensitivity (70.25%). CONCLUSIONS: Our results indicated a significant increase in liver enzymes, lipid profile and MetS status in both pre-diabetic and T2MD subjects, with the differences being more pronounced in diabetic individuals. Consequently, on the one hand, these variables may be considered predictive risk factors for diabetes, and on the other hand, they may be used as diagnostic factors. In order to confirm the clinical applications of these variables, additional research is required.

Exosomes of Whartons' jelly mesenchymal stem cell reduce the NOX genes in TGF-ß-induced hepatic fibrosis.

Objectives: Activated cells which are called star-shaped cells, are some of the key factors in the development of liver fibrosis. Activation of NADPH oxidase (NOX) is associated with increased HSCs activity and progression of hepatic fibrosis. In this study, the effects of human exosomes derived from WJ-MSCs on NOX1, NOX2, and NOX4 gene expression in TGF-ß-induced hepatic fibrosis were investigated. Materials and Methods: LX2 cell line was treated with 2 ng/ml TGF-ß for 24 hr, in order to induce liver fibrosis after starvation. In the next step, the cells were treated with several concentrations of the exosomes derived from WJ-MSCs (10, 20, 30, 40, and 50 µg/ml). Finally, Smad3C phosphorylated protein expression level and NOX1, NOX2, and NOX4 gene expression levels were measured. Results: The results demonstrated that the level of NOX1, NOX2, and NOX4 mRNA expressions decreased significantly during 24 hrs at concentrations of 40 and 50 µg/ml of WJ-MSCs exosomes in TGF-ß-induced-HSCs. The p-Smad3C proteins were significantly decreased (fold change: 1.83, P-value<0.05) after exposure to WJ-MSC-derived exosomes. Conclusion: Treatment with exosomes prevents further activation of HSCs by inhibiting the level of Smad3C phosphorylation. The experimental data of our study suggested that in liver fibrosis, the protection of HSCs activation against TGF-ß by inhibiting the NOX pathway via human exosomes of WJ-MSCs is extremely important. It needs further research as a treatment method.