Effects Of Salvia Miltiorrhiza On NF-κB And CYP450Expressions In Steatosis Hepatocytes Of Dairy Cows

Fatty liver was one of the common diseases in dairy cows, and occurred in the perinatalperiod of high yield dairy cows, mostly. Fatty liver not only affected milk yield, but alsocould induce a variety of other diseases. It caused huge economic losses to farmers. Inrecent years, the incidence of fatty liver in dairy cows showed an increasing trend, but itscause and mechanism were not clear. Many researchers had done a lot of research onrelated pathogenesis, but the research was rare from the cellular level by means ofmolecular biology techniques. Therefore, the relationship between CYP450, NF-κB andfatty liver, was worthy of further exploration and study. Based on the above measured dataand analysis on the pictures stained by oil red, the optimal condition for hepatocytessteatosis model was selected.The primary hepatocytes were isolated and cultured successfully by two-stepcollagenase perfusion method in this study. The hepatocytes were treated withDL-ethionine to be induced steatosis. Then the contents of ALT, AST, TG, TC weredetected. And the hepatocytes viability was measured by MTT method. The lipid dropletswere observed by oil red staining. After modeling, the steatosis hepatocytes were treatedwith gradient concentration of Salvia miltiorrhiza. The contents of ALT, AST, TG, SOD,MDA, TNF-α in the hepatocytes were detected. And the lipid droplets were observed byoil red staining. The expressions of NF-κB and CYP450were determined by Western blot.The purpose of this study was to explore the relationship between TNF-α, NF-κB, CYP450and fatty liver in dairy cows, reveal the causes and the mechanism and provide referencefor prevention and treatment of clinical disease.The results showed that, the hepatocytes isolated by two-step collagenase perfusionmethod had complete morphology, well adherent and high survival rate about94%.Therefore the hepatocytes could be used for further study. After the hepatocytes inducedby DL-ethionine, according to the changes of TG, TC contents, cell survival rate measuredby MTT method, and analysis on the pictures stained by oil red,2mmol/L DL-ethioninetreated the hepatocytes for24h was selected as the optimal condition for steatosis model,eventually. With the treatment of Salvia miltiorrhiza, the contents of ALT, AST, MDA andTG decreased, the release of TNF-α was inhibited, and the activity of SOD increased. The expression of CYP450in hepatocytes decreased gradually. The NF-κB had no significantexpression for24h treatment, but the expression of NF-κB decreased gradually with theconcentration increasing of Salvia miltiorrhiza for48h treatment.These findings suggest that Salvia miltiorrhiza has some therapeutic effect on dairy cowfatty liver, it maybe relate to promote lipid metabolism, inhibit tumor cell necrosis factor,oppose lipid peroxidation and decrease the expressions of NF-κB and CYP450.