| Objective: In this experiment we selected the ovarectomized rats injected with D-gal as the subject to observe the effect of mailuoning,vitamin C+E and traditional Chinese medicine compound recipe on cholinergic receptor , glutamic acid receptor and estrogen receptor .Methods1 Animals and groupsForty female SD rats aged four months were randomly divided into two groups: sham operated group (Sham group),ovariectomized group (OVX group). Sham group was ablated some adipose tissue above ovaries; OVX group was ablated two sides ovaries. Rats in OVX received D-gal (20 mg in 0.5 ml 0.9% saline intraperitoneal injection everyday); rats in Sham received saline (0.5 ml 0.9% saline intraperitoneal injection everyday), after two weeks of operation. The rats in OVX were divided into four groups after six weeks of intraperitoneal injection: one group was administered orally with mailuoning injection at 1.8 ml/kg.d (ODM group); the second group was administed orally with V.C 45 mg/kg.d and V.E 27 mg/kg.d (ODV group); the third group was administed orally with traditional Chinese medicine compound recipe 16.83 g/kg.d (ODZ group); the last group was administed orally with distilled water 1.5 ml/kg.d (OD group). The Sham group was administed orally with distilled water too.2 behavioral testsSix weeks after D-gal injection, beheavioral tests were carried out to evaluate activity and exploratory behavior, learning and memory abilities by using Morris water maze. The same procedure was repeated, after four weeks of medicine administered orally .3 Tissues and data determiningTo anesthetize rats, temporal cortex were quickly separated and deep freezed in liquid Nitrogen. Then tissues were stored at -80℃to extract total RNA. Uterus was quickly separated and weighed by electronic balance. temporal cortex total RNA were extracted by Trizol regent. The relative mRNA contents (M1-5 mAchR,α2-7n?AchR,NMDAR1 and ER)were measured by RT-PCR usingβ-actin as inner standard.Results1 Impairments of behavioral performance Data from Morris water maze showed that the activity and exploratory behavior of the rats was prominently impaired in the OVX+D-gal groups versus the Sham group (P<0.05), Again, this learning impairment was attenuated significantly in the ODM, ODV and ODZ animals (P<0.05).2 The relative expression ofα7n?AchR mRNA in temporal cortex Compared with Sham, the relative expression of ?α7nAchR mRNA of OD was lower (P<0.05). Compared with OD, the ralative espression ofα7nAch receptor mRNA of ODM and ODV was haigher (P<0.05). There was no difference between OD and ODZ (P>0.05).These results above indicated that under the condition of ovariectomized and D-gal injection, mailuoning and V.C+V.E can increase the relative expression ofα7?nAch receptor mRNA .3 The relative expression ofΜ?mAchR mRNA in temporal cortex Compared with Sham, the relative expression of M?mAChR mRNA of OD was higher (P<0.05). Compared with OD , the ralative espression ofΜ1mAchR mRNA of ODZ was lower (P<0.05). There was no difference between Sham and ODM , ODV, ODZ. (P>0.05).These results above indicated that under the condition of ovariectomized and D-gal injection, traditional Chinese medicine compound recipe can increase the relative expression ofΜ?mAchR mRNA.4 The relative expression of M3mAchR mRNA in temporal cortexCompared with Sham, the relative expression of M3mAchR mRNA of OD was lower (P<0.05). Compared with OD, the ralative espression of M3mAchR mRNA of ODM, ODV and ODZ was higher (P<0.05). There was no difference between Sham and ODM, ODV, ODZ. (P>0.05).These results above indicated that under the condition of ovariectomized and D-gal injection, mailuoning,V.C+V.E and traditional Chinese medicine compound recipe can increase the relative expression of M3mAchR mRNA .5 The relative expression of ERαmRNA in tempoal cortex Compared with Sham, the relative expression of ERαmRNA of OD was lower (P<0.05). Compared with OD, the ralative espression of ERαmRNA of ODM was higher (P<0.05). There was no difference between OD and ODV, ODZ. (P>0.05). These results above indicated that under the condition of ovariectomized and D-gal injection, mailuoning can increase the relative expression of E?RαmRNA.6 The relative expression of NMDAR1 mRNA in tempoal cortex Compared with Sham, the relative expression of NMDAR1 mRNA of OD was lower (P<0.05). Compared with OD, the ralative espression of NMDAR1 mRNA of ODM, ODV and ODZ was higher (P<0.05). There was no difference between Sham and ODM, ODV, ODZ. (P>0.05).These results above indicated that under the condition of ovariectomized and D-gal injection, mailuoning,V.C+V.E and traditional Chinese medicine compound recipe can increase the relative expression of NMDAR1 mRNA .Conclusion1 Morris water maze demonstrated that the learning and memory function was deteriorated significantly following long-term OVX combined with D-gal injection. Under this condition, the relative espression ofα7nAChR mRNA,M1 mAchR mRNA,M3 mAchR mRNA,ERαmRNA,NMDAR1 mRNA changed.2 Mailuoning , V.C+V.E and traditional Chinese medicine compound recipe may directly or indirectly regulate some genes expression and therefore compensate for impairment in rats following long-term OVX combined with D-gal injection. The effect of Mailuoning is most significant.
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