Abstract
The present study aimed to investigate the effects of bovine placental preparation under in vitro and in vivo conditions. Cell Proliferation Kit I (MTT) was used for in vitro study of placental preparation effect to proliferate lymphocytes. Lymphocytes were isolated from spleen of Balb C mice, 100 μl cell was added to each well of 96 well culture plate, followed by addition of 10 μl placental preparation and mitogen (concanavalin-A) separately and in combination and cell culture only as control was used. Results were obtained by measuring and comparing the absorbance reading of the wells of the samples against the standards with ELISA microplate reader. Effect of placental preparation to proliferate lymphocytes in vivo condition was investigated in mice, which were divided into 4 groups and 4 subgroups. The results were estimtaed with spleen weight, spleen index and splenocyte counts. Results of in vitro study demonstrated that stimulation index increased by 1.19 or 19% for cell division in wells to which no mitogen was added, but the preparation was added as compared to control wells, cell division index increased with 1.38 or 38% for cell division in wells to which mitogen was added as compared to control wells and stimulation index was higher by 1.68 or 68% for cell division in wells with cells to which both mitogen and preparation were added than control wells. For in vivo experiments, spleen index and splenocyte count for animals of positive control subgroup-1 treated once by 0.2 ml sheep red blood cells were greater by 1.38 and 1.5 times respectively than relevant negative control animals, whereas spleen index and splenocyte count for animals of experimental subgroup -1 were greater by 3.09 and 2.2 times respectively. For animals of positive control subgroup -2, both spleen index and splenocyte count decreased by 1.35 and 1.3 times respectively than negative control animals, whereas they dropped by 1.1 and 1.17 times respectively in mice of experimental subgroup -2. Spleen index and splenocyte count in mice treated with the preprartion only increased by 1.2 times or 20% as compared to negative control animals. From above results, it is shown that bovine placental preparation is able to exert immunomodulatory effect regardless of antigen under both in vitro and in vivo conditions.
Highlights
Bioactive peptides play an important role in metabolic regulation and modulation, including antioxidant, antihypertensive, antimicrobial and immunomodulatory activities [9].Immunomodulatory peptides can enhance immune cell functions such as lymphocyte proliferation, natural killer (NK) cell activity, antibody synthesis, and cytokine regulation [13]
Preparation of splenocytes: Spleens of Balb C mice were taken under sterile condition, lymphocytes were isolated and suspended, and cell suspension was dispensed in 100 μl aliquots into a well of 96 well cell culture plate and placental preparation in 10 μl aliquots into a well both separately and in combination
Lymphocyte proliferation effect of placental preparation was investigated under the condition the same in above studies and the preparation added to splenocyte culture increased splenocyte division by 19% as compared to variant where no preparation was added
Summary
Immunomodulatory peptides can enhance immune cell functions such as lymphocyte proliferation, natural killer (NK) cell activity, antibody synthesis, and cytokine regulation [13]. According to studies performed by a number of authors, such immunomodulatory peptides can be obtained from animal and plant sources. According to studies of a number of scientists, preparations from placenta of both human and animals, including cattle, horse, sheep, goat and pig, which were obtained by various methods, exert immunomodulatory effects [3]. Chinese researchers isolated and purified several fractions of immunomodulatory peptide from water soluble extract of cow placenta and characterized their effects under in vitro conditions by using lymphocyte proliferation MTT assay [3]. The present study aimed to prepare aqueous extract of bovine placenta and investigate its effect on animal body immune system
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