| ObjectiveMenopause is the period of women who have the transition of reproductive age to the old age. In this period, ovarian function declines stage by stage and the estrogen level decreases significantly. These cause the change of the coordination fuction of the hypothalamic-pituitary-gonadal axis. Part of those women will suffer from the related syndromes, which is called menopausal syndrome. Our Objective is to investigate the influence of the ovariectomy and the emotional stimulation on female rats. We discuss the intervention role of Kidney Yin Invigorating Pill either.MethodsForty eight two months old famale SD rats weighing 180-220g were divided into six groups by randomized block design and the weight as the single factor: sham group, ovariectomy group, ovariectomy+fear group, zuoguiwan group, alprazolam group, premarin group. Each group had eight rats. Ovariectomy groups ovariectomy+fear group,zuoguiwan group,alprazolam group and premarin group implemented bilateral ovarian enucleation. Sham group was opened the abdomen in the same operation method without removing ovaries. From the nineth day after the operation, ovariectomy+fear group,zuoguiwan group,alprazolam,group and premarin group were frightened by a cat one hour daily as the emotional stimulation, altogether 21 days. Each group also began accepting intragastric administration once a day for 21 days From the nineth day after the operation, sham group, ovariectomy group, ovariectomy+fear group, were treated by distilled water by 9.45ml per kg. Zuoguiwan group was treated by Kidney Yin Invigorating Pill Suspension (including crude drug 1g per ml) by 9.45ml per kg. Alprazolam grou was treated by alprazolam dstilled water (concentration 0.0076mg per ml) by 9.45ml per kg. Premarin group was treated by premarin distilled water (concentration 5.952 u g per ml) by 9.45ml per kg. The elevated T maze test and open field test were carried out two hours after the last administration. Twenty four hours later, each rat was executed.The serum sex hormone was quantitated by CLIA; 5-HT, NE,5-HIAA were quantitated by HPLC-ECD; estrogen receptor was quantitated by IHCA.Result1 In open field test, the crossing score of ovariectomy group was significantly lower than the sham group(P<0.05), but there was no significant difference between ovariectomy group and ovariectomy+fear group(P>0.05). The rearing score of ovariectomy group was significantly lower than the sham group(P<0.05) and the rearing score of ovariectomy+fear group was significantly lower than the ovariectomy group(P<0.05). Compared with the ovariectomy+fear group, the crossing scores of zuoguiwan group, alprazolam group, premarin group had no significant difference(P>0.05), but the rearing scores were significantly higher (P<0.05). The rearing score of zuoguiwan group was significantly higher than the premarin group(P<0.05). In the elevated T maze test, the one-way escape time had no significant difference between ovariectomy group and the sham group(P>0.05). The one-way escape time of ovariectomy+fear group was significantly shorter than the ovariectomy group(P<0.05). Compared with the ovariectomy+fear group, the one-way escape time of zuoguiwan group, premarin group were significantly longer(P<0.05), but there was no significant difference between zuoguiwan group and sham group(P>0.05). The one-way escape time of zuoguiwan group was significantly longer than the premarin group(P<0.05).2 The content of serum E2 of the ovariectomy group had been descended significantly comparing with the sham group (P<0.05), but there was no significant difference between ovariectomy group and ovariectomy+fear group(P >0.05). Compared with the ovariectomy+fear group, the content of serum E2 of zuoguiwan group and premarin group had been risen significantly(P<0.05). Compared with the sham group, the content of serum E2 of zuoguiwan group and premarin group had no significant difference (P>0.05)and there was no significant difference between zuoguiwan group and premarin group(P>0.05).3 The contents of monoamine neurotransmitters in hippocampus:the contents of 5-HT, NE of the ovariectomy group were significantly higher than the sham group (P<0.05), but the content of 5-HIAA had no significant difference (P >0.05). The contents of 5-HT, NE of the ovariectomy+fear group was significantly higher than the sham group (P<0.05), but the content of 5-HIAA had no significant difference (P>0.05). The contents of 5-HT, NE of the ovariectomy+fear group were significantly lower than the ovariectomy group (P<0.05), and 5-HT/NE value was significantly risen(P<0.05). The contents of 5-HT, NE of the zuoguiwan group were significantly lower than the ovariectomy+fear group group (P<0.05), but the content of 5-HIAA had no significant difference (P>0.05). The contents of 5-HT, NE of the zuoguiwan group and the sham group had no significant difference (P>0.05). The content of 5-HT of the zuoguiwan group and the premarin group had no significant difference (P>0.05), but the content of NE of the zuoguiwan group was significantly lower than the premarin group (P<0.05)。5-HT/NE value between the zuoguiwan group and the sham group had no significant difference (P> 0.05).The contents of monoamine neurotransmitters in the frontal cortex:the contents of 5-HT, NE of the ovariectomy+fear group were significantly higher than the sham group (P<0.05), and the content of 5-HIAA was significantly lower (P<0.05).5-HT/Ne of the ovariectomy+fear group was significantly higher than the sham group(P<0.05). The contents of 5-HT of the zuoguiwan group were significantly lower than the ovariectomy+fear group group(P< 0.05), but the content of 5-HIAA had been risen significantly (P<0.05). The content of 5-HT had no significant difference between the zuoguiwan group and the sham group (P>0.05), but the content of 5-HIAA had been risen significantly (P<0.05). The content of NE had no significant difference between the zuoguiwan group and the sham group (P>0.05), and 5-HT/NE value had no significant difference (P>0.05).4 The expression of Erβin hippocampus:The IOD value of Erβof the ovariectomy group was significantly higher than the sham group (P<0.05). The IOD value of Erβof the ovariectomy+fear group was significantly higher than the ovariectomy group (P<0.05). The IOD value of Erβof the ovariectomy+fear group was significantly higher than the sham group (P<0.05). The IOD value of Erβof the zuoguiwan group was significantly lower than the ovariectomy+fear group (P<0.05), but there was no significant difference between the zuoguiwan group and the sham group (P>0. 05). The IOD value of Erβof the premarin group was significantly lower than the ovariectomy+fear group (P<0. 05), but there was no significant difference between the premarin group and the sham group had no significant difference (P>0.05).Conclusion1 In open field test, the crossing and rearing score of ovariectomized rats decended, indicating that the investigation was inhibited, and the emotional stimulation could inhibit it further. In the elevated T maze test, ovariectomy and emotional stimulation could shorten the one-way escape time, indicating that the panic level was increased. With the intervention of Kidney Yin Invigorating Pill, the rearing score and the one-way escape time were increased, indicating that the Kidney Yin Invigorating Pill could enhance the investigating activity ability and decrease the panic level.2 The serum E2 of ovariectomized rats significantly decended, emotional stimulation had no significant effect on serum E2 of ovariectomized rats. Kidney Yin Invigorating Pill can significantly rose the serum E2 of ovariectomy+fear group to the level of sham group.3 The study revealed that ovariectomy and emotional stimulation could significantly increase the monoamine neural transmitter 5-HT, NE and the 5-HT/NE value in hippocampus, while the content of 5-HIAA had no significant change. The increase of the content of 5-HT may ascribed to the synthetic enhancement by ovariectomy and emotional stimulation. With the intervention of Kidney Yin Invigorating Pill,the content of 5-HT, NE in hippocampus decreased to an approximate level of the sham group and return to balance in proportion. The decrease of the content of 5-HT may ascribed to inhibiting one or more links in the synthetic route.Ovariectomy and emotional stimulation could significantly increase the content of 5-HT,NE and the 5-HT/NE value, but significantly decrease the content of 5-HIAA. The degradation of 5-HT may slow down or accompanied with synthetic increase due to ovariectomy and emotional stimulation. With the intervention of Kidney Yin Invigorating Pill, the content of 5-H,NE in hippocampus decreased to an approximate level of the sham group and return to balance in proportion. The decrease of the content of 5-HT may ascribed to acceleratng degradation of 5-HT.4 The expression of ERβrose in ovariectomized rats, the mechanism may be compensatory rise owing to lack of estrogen. The reason emotional stimulation furthur increasing the the expression of ERβmay be that stress act on nerve-endocrine-immune network system. With the intervention of Kidney Yin Invigorating Pill, the expression of ERβdecreased to an approximate level of the sham group. We speculated that the Kidney Yin Invigorating Pill could increase the level of estrogen and modulate neuroendocrine as a whole.
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