Fish liver oil and propolis as protective natural products against the effect of the anti-epileptic drug valproate on immunological markers of bone formation in rats.

Epilepsy is a major public health problem affecting nearly 50 million people world wide. Treatment with anti-epileptic drugs (AEDs) is generally chronic if not life long and may be associated with significant metabolic effects including decreased bone mass and increased fractures. The aim of this work was to investigate the protective role of fish liver oil and propolis against the effect of the drug valproate that is widely used for treatment of epilepsy. Group of 40 rats was divided into four groups each contain 10 rats. The first group (group I) is healthy normal rats, as control. Epilepsy was conducted in the rest of the rats. The epileptic rats were divided into three subgroups: group II was epileptic group, supplemented orally with valproate. The third group was epileptic group which supplemented orally with valproate in concomitant with fish liver oil, the last group; group IV was epileptic group which supplemented orally with valproate in concomitant with propolis. In the present study oral administration of valproate to the epileptic rats by a dose of 400mg/kg/daily for six months (group II) resulted in a significant increase of bone alkaline phosphatase, osteocalcin and N-telepeptide of type 1 collagen (NTX) relative to the control group. There were increase of receptor activator of NF kappa B ligand (RANKL), tumor necrosis factor - alpha (TNF-alpha) and decrease of osteoprotegrin (OPG) compared to normal control. Administration of fish liver oil orally in a dose of 0.4mg/kg daily in concomitant with valproate 400mg/kg daily for six months (group III), result in reduction of N-telepeptide of type 1 collagen (NTX) in comparison to group II and with no significant increase than the control (group I). There were high significant increase of bone alkaline phosphatase and osteocalcin compared to control group I. There was high significant increase of bone alkaline phosphatase than group II and increase in osteocalcin, and decrease in N-telepeptide of type 1 collagen (NTX) compared to group II. A significant increase in osteoprotegrin (OPG) in comparison to group II and to control (group I) with a decrease in RANKL compared to group II and with no significant increase than normal control (group I). The TNF-alpha showed a significant decrease compared to group II with no significant increase than normal control. Administration of propolis orally in a dose of 50mg/kg daily in combination with valproate 400mg/kg/daily for six months (group IV) cause increase in bone alkaline phosphatase with no statistical difference between osteocalcin and N-telepeptide of type 1 collagen (NTX) and normal control (group I). There were increase in bone alkaline phosphatase than group II but less than group III. The increase in osteocalcin in-group III (fish oil group) was significantly higher than in-group IV and there was no statistical difference between it and group II. Where the N-telepeptide of type 1 collagen (NTX) the bone resorption marker was significantly higher than Group III and significantly lower than group II. There was a decrease of RANKL in comparison to group II with no significant difference than group III and a significant increase than control group. There was an increase in osteoprotegrin (OPG) in comparison to control (group I), group II and from group III. There was decrease in TNF-alpha than group III, group I and group II. In conclusion, in epileptic rats treated with valproate (which cause osteoporosis) fish liver oil and propolis increase the bone formation markers and decrease the bone resorption one's. They increase the OPG and decrease TNF-alpha, and RANKL which inhibit the osteoclastogenesis. We recommend the use of Fish Oil, or propolis as a prophylactic treatment for epileptic patients using valproate against the side effect of valproate on bone.