| Soybean isoflavones,own abundant biological activities,like preventing cancer,oxidation,osteoporosis,and enhancing the immune enhancement and thin hormone,and it is as a kind of feed additives,catchs the more and more attention of people on livestock and poultry production,especially it has been widely applied in the pig production practice,however,soybean isoflavones are potentially toxic,especially the possible harm to the reproductive system should not be ignored,while there are few researches about the effects of soybean isoflavones on the tissues structure and morphology of reproductive system,as well as the relationship of endocrine and immune function,thus this paper is carried out to study the effect of soybean isoflavones to the reproductive performance of female bama pigs and its mechanism from the following two aspects.One is the experiment in vivo to study the effects of soy isoflavones on uterus structure and morphology and serum immune parameters.Single factor design was used,and select 4female bama pigs per litter form 12 nest 35 day-old piglets whose gestational ages are the same and parity are similar,randomly assigned to 4 treatments with 12 repetitions per treatment and a piglet for one repetition.Feed the pigs basal diet supplemented with soybean isoflavones,0,500,1250 mg/kg and 0.66 mg/kg estradiol valerate(similarity to 0.5 mg/kg),one by one,noted as the Control,SI-500,SI-1250 and EV-0.66,one by one.Animal feeding trials was carried out in the crop and animal experiment building of the institute of subtropical agriculture and and ecology of the chinese academy of sciences.Every half month blood from vena cava anterior were collected when pigs of groups were on an empty stomach in the morning,for the measurement of immune items in serum.During the experiment,detect puberty by boars,if sows appear still reflection,meaning sows get their puberty,as the first day of the puberty,respectively.In the 4th day of the puberty,6 pigs were chosen for each group to slaughter in the morning on an empty stomach,collect samples of uterus for tissue section,hemotoxylin and eosin staining(HE staining)and morphological analysis.The results were following: 1)Compared to the basal diet,theeffects of SI to the thickness of endometrium and uterine muscle layer,as well as the number of glands unit area,are insignificant(P>0.05),while 500 mg/kg SI significantly shortened the height of epithelial cells of endometrial free surface(P<0.05);2)In contrast to the blank group,SI disturbed the shape of uterus and the arrangement of epithelial cells from cavity and glands of uterus endometrium,caused vascular hemorrhage,and harm to the development of gland.3)Made contrast to the control treatment,500 mg/kg SI significantly or quite significantly elevated the concentration of immune globulin G(Ig G)in serum from50-d-old or 95-d-old pigs(P>0.05 or P<0.05);50-d-old pigs,500 mg/kg SI extremely significantly or 1250 mg/kg SI significantly reduced the concentration of tumor necrosis factor(TNF)(P<0.01 or P<0.05),but 500 mg/kg SI extremely significantly reduced the content of interleukin-6(IL-6)and interleukin-2(IL-2)of 95-d-old pigs(P<0.01).The results of the investigation in vivo show that the effect of soybean isoflavones on the structure and morphology of uterine and immune indexes in serum of female bama pigs,exists time and dose effect,taking security into consideration,suggest adding soy isoflavones to the basic diet of female bama pigs about 50 days old with dosage less than 500 mg/kg.The other one is the experiment in vitro to study the effect of different level of soybean isoflavones(daidzin and genistein)on fetal trophoblast cells vitality,extra-cellular hormone secretion,cells cycle,and the mRNA expression of genes closely related to the embryo growth from 35-day old pregnant sows.The results were following: 1)In contrast to the control experiment,the effects of genistein to cell vitality for 24 h were obvious than daidzin,0.01 uM genistein extremely significantly improve the cells vitality while 10 uM and 100 uM genistein extremely significantly inhibited it(P < 0.01);2)The effects of soybean isoflavones to the secretion of extra-cellular hormone of trophoblast cells for 24 h were insignificant(P > 0.05),while 0.1 uM soybean isoflavones to trophoblast cells for 48 h extremely significantly inhibited secretion of extra-cellular hormone thyroxine-4(T4)(P<0.01);10 uM genistein to trophoblast cells for 24 h extremely significantly extended the cell cycle(P<0.01);3)10 uM daidzin and genistein significantly or very significantly risen the transcription of Gn RHR(P<0.05 or P<0.01),while 100 uM daidzin very significantly increased the transcription of gene Gn RHR(P < 0.01),10 uM daidzin extremely significantly raised the expression of CYP11A1 mRNA(P<0.01),but 100 uM daidzin in great degree lowed it(P<0.05),extremely significantly lowed the expression of 3β-HSDmRNA(P<0.01),and the high concentration 10 uM genistein extremely significantly risen the expression of CYP11A1 mRNA(P<0.01).4)0.1 uM daidzin significantly reduced the expression of TGFβ3 mRNA(P<0.05),10 uM daidzin significantly lowed the expression of TGFβ1 mRNA(P<0.05),and 100 uM daidzin extremely significantly lowed the expression of mRNA of TGFβ1and IFN(P < 0.01),the higher concentration 10 uM genistein significantly risen the expression of TGFβ3 mRNA(P<0.05);5)1 uM daidzin significantly reduced the expression of mRNA of VEGF and MMP9(P<0.05),but 100 uM daidzin very significantly reduced the expression of mRNA of MMP9(P<0.01);6)10 uM and 100 uM daidzin extremely significantly lowed the expression of mRNA of IL6(P<0.01);at the meantime,the low concentration of 0.01 uM and 0.1 uM genistein greatly risen the transcription of IL6(P < 0.05);the high concentration 10 uM genistein extremely significantly lowed the expression of mRNA of IL6 and IL1α(P<0.01).The results of the investigation of cells culture in vitro also show that the effect of soybean isoflavones on the trophoblast cells exists dose and biphasic effect,furthermore genistein is more obvious than daidzin,and the threshold content of daidzin,which is benefit for the fetal development to trophoblast cells for 24 h,is among 10-100 uM,while for genistein is very likely among 1-10 uM.An analysis was carried out on the basis of the relationship between the endocrine and immune secretion extra-cellular hormone is less,hormone acts in the intracellular is more,so better increase the body’s sensitivity to the inflammatory cytokines,indicates that there exists certain positive correlation between endocrine and immune function.In a word,the content of this paper is very important,and the results can be important reference basis for the safety dosage of soybean isoflavones as feed additives in the pig production,as well as the mechanism of interactions between endocrine and immune. |
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