Intramammary inoculation of Panax ginseng extract in cows at drying off enhances early mammary involution

This study was designed to evaluate the effects of a single intramammary infusion of Panax ginseng extract on cell proliferation and death mechanism in bovine mammary gland during early involution. Eight mammary quarters from six non-pregnant cows in late lactation were infused with 10 ml of ginseng solution (3 mg/ml), six quarters were treated with 10 ml of placebo (vehicle alone) and six quarters were maintained as uninoculated controls. Milking was interrupted after infusion. Animals included in the three groups were slaughtered 7 d after inoculation and samples for histological analysis were taken. Morphometric analysis showed a significant increase in percentages of mammary tissue area occupied by stroma in ginseng-treated quarters compared with controls. A significant increase of immunostained area for bax protein and active caspase-3 was observed in ginseng-treated quarters compared with controls, whereas no differences were observed for bcl-2 immunostaining. Expression of bax mRNA was significantly higher in ginseng-treated quarters than in controls. The bax/bcl-2 ratio indicated a significant predominance of bax over bcl-2 mRNA expression in ginseng-treated quarters compared with controls. The rise of epithelial and stromal cell apoptosis in situ by TUNEL was more marked in quarters treated with ginseng than in controls. Ginseng inoculation had no effect on the number of epithelial and stromal proliferating cells labelled with Ki-67 antibody. Ratio of apoptotic to proliferating cells was higher in quarters treated with ginseng compared with controls, indicating a net loss of cells in parenchymal components. Also, the intramammary inoculation of ginseng extract at drying off increased the rate of mammary cell apoptosis without inhibiting cell proliferation. Taken together, these changes are indicative of mammary regression enhancement during early involution.