Metformin/Leucine

Hypercholesterolemia (HLC) was a key risk factor for cardiovascular disease (CVD) characterized by elevated cholesterol levels, particularly LDL. While traditional Chinese medicine preparations Compound Danshen Pills(CDP) has been clinically used for hypercholesterolemia and coronary heart disease, its specific therapeutic effect on HLC remains understudied, necessitating further investigation into its mechanisms.

The aim of this study was to explore the potential of CDP in treating HLC and elucidate its underlying mechanisms and active components.

A hypercholesterolemic lipemia rat model induced by a high-fat diet was employed. Network pharmacology combined with UHPLC-Q exactive orbitrap HRMS technique was used to predict the active components, targets and mechanisms of CDP for HLC. Histological analysis and serum biochemical assays were used to assess the therapeutic effect of CDP and its main active ingredient Sa B on hypercholesterolemic lipemia rat model. Immunofluorescence assays and western blotting were used to verify the mechanism of CDP and Sa B in the treatment of HLC. Metabolomics approach was used to demonstrate that CDP and Sa B affected the metabolic profile of HLC.

Our findings demonstrated that both CDP and its main active ingredient Sa B significantly ameliorated hypercholesterolemic lipemic lesions, reducing levels of TC, LDL, AST, ALT, and ALP. Histological analysis revealed a decrease in lipid droplet accumulation and collagen fiber deposition in the liver, as well as reduced collagen fiber deposition in the aorta. Network pharmacology predicted potential targets such as PPARα and CYP27A1. Immunofluorescence assays and western blotting confirmed that CDP and Sa B upregulated the expression of Adipor1, PPARα and CYP27A1. Metabolomics analyses further indicated improvements in ABC transporters metabolic pathways, with differential metabolites such as riboflavin, taurine, and choline showed regression in levels after CDP treatment and riboflavin, L-Threonine, Thiamine, L-Leucine, and Adenosine showed improved expression after Sa B treatment.

CDP and Sa B have been shown to alleviate high-fat diet-induced hypercholesterolemia by activating the PPAR pathway and improving hepatic lipid metabolism. Our study demonstrated, for the first time, the complex mechanism of CDP, Sa B in the treatment of hypercholesterolemia at the protein and metabolic levels and provided a new reference that could elucidate the pharmacological effects of traditional Chinese medicine on hypercholesterolemia from multiple perspectives.

It is widely known that heat stress (HS) has negative effects on dairy cows, such as a reduction in milk production and milk protein. However, there has been no research yet on the effects of HS at the bovine mammary epithelial cells (MAC-T) level and the function of L-leucine (LEU) and sodium acetate (ACE) in reducing HS. In this study, we evaluated the negative effects of HS at various temperatures on MAC-T and verified whether LEU and ACE are effective at reducing HS and increasing protein synthesis. An experiment was conducted by dividing MAC-T into three groups: 39 °C, 41 °C, and 43 °C. In the case of LEU and ACE supplementation experiments, the cells were supplemented with 0, 0.45, 0.9, 1.8, and 3.6 mM of LEU and ACE to reach the differentiation medium. It was observed that under HS at 41 °C, HSP70, BAX, and eIF4EBP1 gene expression were increased, whereas Bcl-2, eIF4E, and PRKAA1 gene expression were decreased. When 1.8 mM of LEU was added under HS at 41 °C, it suppressed apoptosis by reducing the gene expression of HSP70 and controlling the gene expression of apoptosis-related genes such as BAX and Bcl-2. Additionally, mTOR, P-mTOR, and β-casein proteins were increased. In the case of 0.9 mM of ACE, it was found to decrease the gene expression of HSP70 and BAX and increase the amount of β-casein protein synthesis. Simultaneous supplementation of LEU and ACE has been shown to reduce HS, inhibit apoptosis, and increase β-casein protein expression. In summary, HS at 41 °C began to have a negative effect on MAC-T, while LEU and ACE reduced HS and inhibited apoptosis, alleviating cell damage and effectively increasing β-casein protein synthesis. The results suggest that LEU and ACE have the potential to reduce HS and promote protein synthesis under HS conditions in MAC-T.