Effect Of Alfalfa Flavonoids On Reproductivity And Gene Expression From The Hypothalamus-pituitarv-ovarian Axis At Difference Physiological Stages

The impact of different doses of alfalfa flavonoids on reproductive performanceand gene expression in hypothalamus-pituitary-ovarian axis of female rats at mature,immature, pregnant stages were studied. The results of three experiments indicated asfollows:The results of the first experiment showed that the feed intake and daily body gainof rats were influenced by alfalfa flavonoids, but there was no significant difference.There was a decreasing trend in the daily body gain among three groups of differentdoses of alfalfa flavonoids, compared with control group. We analyzed data of wetweight ratio of uterine to ovarian. Low doses of alfalfa flavonoids significantlyincreased E2of level and uterine wet weight/coefficient (P <0.05) and significantlyreduced ovarian wet weight/coefficient (P <0.05) in mature female rats. There was aincrease trend in the litter size and newborn litter weight Litter size of middle andHigh dose group increase0.4%and2.2%than control group (P>0.05). Newborn litterweight of middle and High dose group increased5.14%and0.1%than control group(P>0.05). But litter size and Newborn litter weight of diethylstilbestrol was evidentlyreduced (P<0.05). We analyzed data of gene expression in hypothalamus-pituitary-ovarian axis, the expression of GnRH in hypothalamus was significant reduced byalfalfa flavonoids and diethylstilbestrol (P<0.05). The expression of FSHR in ovarianwas reduced than control group, but there were no significant difference. Theexpression of LHR was significant increased6times by low doses group alfalfaflavonoid（sP<0.01）. Moreover, the expression of cAMP had no significant difference,compared with control group (P>0.05). The expression of CYP19A-1was significantincreased8times by low doses group alfalfa flavonoids (P<0.05), and the expressionof other group also high compared with control group (P>0.05). The expression ofERα were increased by alfalfa flavonoids (P>0.05), and it significantly increased14times by diethylstilbestrol (P<0.05).In the second experiment, we analyzed feed intake of immature female rats, theresult was in accord with mature rats. There was no evidence difference in the dailybody gain, but it was obviously reduced by diethylstilbestrol (P<0.05). Uterine andovarian wet weight/coefficient and E2had positive correlation with dose of alfalfaflavonoids in immature rats, but there were no significant difference (P>0.05). Weanalyzed data of gene expression in hypothalamus-pituitary-ovarian axis, theexpression of GnRH was significantly reduced by middle and high dose of alfalfa flavonoids and diethylstilbestrol (P<0.05). The expression of FSH in pituitaryincreased by alfalfa flavonoids (P>0.05). The expression of LH in low dose evidentlyincreased than in high dose (P<0.05). The expression of FSHR was significantlyincreased by high dose alfalfa flavonoids (P<0.01), There was increase than controlgroup in the expression of LHR (P>0.05). The expression of CYP19A-1wassignificantly increased11times by low dose group (P<0.01), and was significantlyincreased8times by diethylstilbestrol (P<0.05). The expression of ERα had nosignificant difference in all group (P>0.05).In the third experiment, the middle and high dose alfalfa flavonoids evidentlyincreased E2of level (P<0.05), the low dose alfalfa flavonoids significant increasedE2of level (P<0.01). The level of Progesterone showed no difference between allgroups (P>0.05). The number of live fetus evidently increased in three does of alfalfaflavonoids (P<0.05), the weight of live fetus was significant increased by high dosealfalfa flavonids (P<0.01). There were no evident difference in the number of luteum,but the trend was fall. Moreover, Diethylstilbestrol evidently reduced the number ofluteum than control. Litter size of three does of alfalfa flavonoids was highersignificantly than control group (P<0.05), and the newborn litter weight were highsignificant increased than control group (P<0.01). We analyzed data of geneexpression in hypothalamus-pituitary-ovarian axis, the expression of GnRH inhypothalamus and FSH in pituitary were significantly reduced by alfalfa flavonoidsand diethylstilbestrol (P<0.01), the expression of FSHR in ovarian were evidentlyreduced by the high dose alfalfa flavonoids, but the expression of LHR were evidentlyincreased, and the expression other gene showed no significant difference comparedwith control group.