Lactoferrin

Inhibition of angiogenesis by bovine lactoferrin

Aim
Angiogenesis is deeply related to the increase of micrometastasis. We have found that bovine lactoferrin (bLF) from milk component inhibit the metastases of lung metastasis inflammation colon cancer Co26Lu to lungs by internal administration (oral), and have examined about the action mechanism. This time, we examine the inhibition of angiogenesis.

Method and result
we have used gallinaceum chorioallantoic membrance to examine the inhibition of angiogenesis by bovine lactoferrin. As the result, bLF inhibited the angiogenesis by concentrate dependence. On the other hand, in vitro assey measure by using vascular endothelial cell showed a tendency of inhibiting swarming action, but did not influence increase, enzyme activity of angiogenesis and lumen genesis. We examine cytokine induction by macrofhage because bLF strengthen immunoreaction. Macrophage utilize peritoneal exudates cell of mouse and after adding bLF indicated cytokine in cell and incubator measured by ELISA. As the result, bLF is acting on macrophage and produce IL-18.

Summary
From the above results, we can indicate that metastoses inhibition of bLF is due to the inhibition of angiogenesis by IL-18 produced from macrophage.

Induction of interieukin-18 in intestinal wpithlial cells following orally administered lactoferrin and its antimetastasis.

Aim
Lactoferrin exist in milk and lacrimal fluid and indicate to active action of phylaxis and immunocompetent cell. We have reported the inhibition of calon cancer and lung metastasis then we have found immnocyte active and production of IL-18 from it form the action mechanism. This time, we have examined relation between fluctuate of membrance immnucyte and various cytokine concentration in membrance (epithelium + tunica propria) and serum.

Method
We have measured (tested) blood and membrance cytokine by ELISA. After bLF, pepsin cracked (bLFH) and transferring were orally administered to mause. Then we have examined entero-organization preparation was immuno-colouring by immunocyte.

Result
Il-18: Membrance concentration indicated the highest point after 3-6 hours (approximately 2.5 times more then control group). Serum concentration was significantly increased by bLF. Strong onset on small intestine cell by immno-colouring and IL-18mRNA was confirmed by RT-PCR and Northern blot. IFNr: Membrance concentration was doubled, T and NK cell was also increased. IL-2 and IL-12 wear reduced. TNFa and IL-1b did not chance much.

Mechanistic analysis of bovine lactoferrin inhibition of MelQx-induced Gst-P positive foci in rat liver

Aim
In order to search for bLF inhibition mechanism against MelQx-induced GST-P positive foci in liver, and used CYP1A2 which is a metabolism activity enzyme of MelQx as the index then compared the amount of CYP1A2 and additional weight of MelQx-DNA which is a metabolic product of MelQx.

Method and Result
DEV(or raw diet) were given orally to rats then between 2 - 8 weeks MelQx alone or combination of MelQx and bLF wear given. In third week, part of liver was resected. The liver which was resected on third week and liver after slaughtered were sampled. The amount of CYP1A2 were examined by Western blotting method. As the result rats taking MelQx were 1.5 - 3.4 times more CYP1A2 protein incidence amount were induced than control group. The combination of bLF and MelQx group inhibit 15 - 30% more than MelQx alone. Similar to MelQx-DNA additional weight were measured by post label method, and the combination of bLF and MelQx reduced it by 15 -50 % compared to MwlQx alone. The correlation between CYP1A2 amount and additional weight reduce amount were confirmed.

Conclusion)
As one of the mechanism of MelQx-induced GST-P positive foci in liver by bLF orally administer, it seems that participate in incidence inhibition of CYP1A2.

Inhibitory effect of lactoferrin-producing Bacteroides uniformis cells on formation of azoxymethane-induced aberrant crypt foci in the rat colon

We have showed by orally administer Bacterium Coli which produce lycopene inhibited azoxyrnethane (AOM)-induced aberrant crypt fori in the rat colon formation. (the 57th Annual Meeting of the Japanese Cancer Association, Biochem Biophys Res Commun 262, 322-327, 1997). This time, Plasmio which coding lactoferrin known as prevention effect for colon cancer was introduced to Bacteroides uniformis which is a dominant bacteria of intestinal flora, then examined inhibition of AOM induced rats colon ACF formation by this bacteria strain. PCR amplification products of N-terminal territory in human lactoferrin gane as probe, northern blot was conducted and confirmed band. The band of 80kDa lactoferrin was confirmed at membrance fraction by using weatern blot with rabbit anti-lactoferrin antibody. The culture solution of lactoferrin-producing B. uniformis was orally administer as drinking water to rats and AOM induce ACF formation was significantly reduced when it's compared to vector plasmid introduced functionality intestinal bacterium. At the present, bacterial strain which suit of application the functionality intestinal bacterium are screening, and investigating possible inhibition of AOM induced ACF formation by newly prepared human ractoferrin -producing B. uniforms stain.