Protective Effects of Supercritical CO2-Extracted Mulberry Leaf Extract against Non-Alcoholic Fatty Liver Disease in Mice

Abstract

Purpose

Alcohol or other known liver toxins are not the cause of non-alcoholic fatty liver disease (NAFLD), which is defined by the buildup of fat in the liver cells. Numerous studies have indicated that compounds found in mulberry leaves exhibit hepatoprotective effects. This study aims to optimize the extraction process of mulberry leaves and demonstrate the hepatoprotective properties of the extract. The Further objective is to explore the potential biological advantages of mulberry leaf extracts in the treatment of NAFLD.

Methods

In this research, we optimized the extraction process of mulberry leaves using response surface methodology. The advantages of supercritical carbon dioxide extraction were investigated through LC-MS analysis. Following the establishment of a NAFLD mouse model, the mice were treated with mulberry leaf extract(108, 162, 217 mg/kg/d) for 12 weeks. Post-treatment analyses included histopathological examination, lipid profile assessment and immunofluorescence staining to confirm the antioxidant capacity and therapeutic efficacy of the mulberry leaf extract against NAFLD.

Results

We identified the optimal extraction conditions for both supercritical carbon dioxide extraction and ethanol reflux methods in obtaining mulberry leaf extract. Ultimately, the extraction rates of the two methods reached 13.76% and 12.60% respectively. Research shows that the mulberry leaf extracts obtained by the two methods contain 54 active components, and the extract obtained by the supercritical carbon dioxide extraction method has a higher content of active components. Furthermore, studies have shown that mulberry leaf extracts can improve cellular morphology, decrease the quantity of lipid droplets and the degree of hepatic steatosis, improve mice’s immunity, and decrease weight gain and liver weight in mice. They can also lower serum levels of TC, TG, ALT, and AST in diseased mice. In addition, we found that the treatment effect of medium and high concentrations of mulberry leaf extracts is better.

Conclusion

Our investigation enhanced the process of extracting mulberry leaves. We determined that the extract is safe to use, has potent antioxidant qualities, and shields the liver. Additionally, compared to the ethanol reflux approach, the supercritical CO2 extraction method extracted greater types and quantities of chemicals from the leaves.