lactoferrin and Carbon-Tetrachloride-Poisoning

Polyethylene glycol (PEG) is attached to proteins in order to increase their half-life in circulation and reduce their immunogenicity in vivo. The present study was conducted to examine whether two different sizes of PEGylated bovine lactoferrin (40k-PEG-bLf and 20k-PEG-bLf) would enhance the protective effect of native bLf on liver injury induced by carbon tetrachloride (CCl(4)) in rats. Silymarin, a known hepatoprotective drug was used as a positive control. Compared to native bLf, the treatment of PEGylated bLf more markedly prevented the elevation of serum levels of hepatic enzyme markers and inhibited fatty degeneration and the hepatic necrosis induced by CCl(4). 40k-PEG-bLf showed a more significant suppressive effect on CCl(4)-induced hepatic injury than 20k-PEG-bLf. The treatment with PEGylated bLf elevated serum SOD activity reduced by CCl(4) more significantly than native bLf. 40k-PEG-bLf enhanced serum SOD activity more significantly than 20k-PEG-bLf. Immunohistochemical study showed that the PEGylation of bLf enhanced its intracellular transportation to hepatocytes. The increases in intracellular transportation of the PEGylated bLf in order were: 40k-PEG-bLf>20k-PEG-bLf>native bLf. These findings suggested that the mechanism of the enhancement of hepatoprotective effect by PEGylated bLf was associated with an increase in the intracellular transportation of PEGylated bLf in hepatocytes.

Topics: Animals; Apoptosis; Carbon Tetrachloride Poisoning; Chemical and Drug Induced Liver Injury; Fatty Liver; Hepatocytes; Immunohistochemistry; Lactoferrin; Lipid Peroxidation; Liver; Male; Necrosis; Polyethylene Glycols; Protective Agents; Rats; Rats, Wistar; Silymarin; Superoxide Dismutase

It is known that lactoferrin is one of the functional proteins contained in mammalian milk and that it plays an important role in the immune system. In this study, we prepared multi-lamellar liposomal bovine lactoferrin composed of egg yolk phosphatidylcholine and phytosterol for oral delivery, and examined any resulting anti-inflammatory effects. Oral pretreatment of liposomal lactoferrin exhibited more suppressive effects than did non-liposomal lactoferrin on CCl4-induced hepatic injury in rats as well as on lipopolysaccharide-induced TNF-alpha production from mouse peripheral blood mononuclear leukocytes. Further investigation revealed that the liposomalization did not exert influence on the absorbability of lactoferrin to the venous blood or lymph following an intraduodenal administration in rats. Furthermore, there was no significant difference exhibited between the antigenicity of liposomal and non-liposomal lactoferrin, which was measured using the passive cutaneous anaphylaxis reaction following oral sensitization to them in guinea pigs. These results suggest that liposomal lactoferrin might act more effectively than conventional lactoferrin in the intestinal site, which is regarded as an active site of orally administered lactoferrin, although the biological mechanism is not fully understood yet. Consequently we propose that liposomal lactoferrin could be a novel active constituent useful for preventive and therapeutic treatment of inflammatory diseases.

Topics: Absorption; Administration, Oral; Animals; Anti-Inflammatory Agents, Non-Steroidal; Carbon Tetrachloride Poisoning; Chemical and Drug Induced Liver Injury; Drug Carriers; Guinea Pigs; Intestinal Absorption; Jugular Veins; Lactoferrin; Liposomes; Lymph; Mice; Mice, Inbred BALB C; Passive Cutaneous Anaphylaxis; Phosphatidylcholines; Phytosterols; Rats; Rats, Wistar