| Objective: The purpose of this study was to investigate the expression of AMHR2 in the endometrium of normal adult female SD rats during estrous cycle and periimplantation period.Methods: A total of 100 female SD rats of 8 weeks were selected,100 rats were divided into two groups,First group of rats,numbered 01-40 sequentially,After a week of adaptive feeding,vaginal smear method was used to identify the period of the estrous cycle.First,select female rats that have a normal regular estrus cycle.On the same day as the vaginal smear on the day of acquisition,the rats were divided into 4 groups: pre-estrus(P),estrus(E),late estrus(M),and inter-temporal(DI)according to the smear results.After taking blood from the eyeball,the rat was killed by breaking the neck and the uterus was taken for cryopreservation and paraffin sectioning.To study the distribution and changes of AMHR2 in the uterus of rats during each estrus cycle,and observe the morphological and histological changes of rat uterus.Radioimmunoassay was used to measure serum sex hormone levels in rats.Next,the second group of rats is numbered in the order of 41-100.After one week of adaptive rearing,vaginal smears were used in the same manner.Female rats with normal regular estrous cycle were also selected.In the P,E phase with the normal maturation of SD male rats mated,the next day microscopic examination found that sperm or naked eye found the vaginal suppository,record pregnancy D1,The pregnant mice were sacrificed on D2-D8 days of pregnancy and were included in the D2-D8 seven groups.After taking blood from the eyeball,the rat was killed by breaking the neck,and the uterus was also taken for cryopreservation and paraffin sectioning.To study the distribution of AMHR2 in uterine endometrium of periimplantation rats.At the same time,the morphological and histological changes of the uterus were observed.Serum sex hormone levels in rats were measured by radioimmunoassay.Results: In each estrous cycle: Radioimmunoassay results showed that the hormone in SD rats changed with the estrous cycle.Immunohistochemistry showed that AMHR2 mainly localized in endometrial stromal cells.q PCR showed that the relative expression of AMHR2 gene level had periodic changes from Proestrus(P)to Diestrus(DI),with the lowest AMHR2 gene level in Estrus(E)and the highest in DI.There was a statistical significance difference in relative expression of AMHR2 gene level between E-phase and Diestrus(DI)(P=0.04,P<0.05).AMHR2 gene and AMH gene have the same tendency in the endometrium of different estrus cycles.Western-blot results further demonstrated that the expression trends of AMHR2 and AMH proteins are consistent with gene expression trends.The AMHR2 protein showed a decreasing trend from P to E phase,with the lowest expression in E phase,gradually increased in Metestrus(M)phase,and highest in DI phase.Compared with P phase,the expression of AMHR2 was increased in M phase,and the difference was statistically significant(P phase vs M phase,P=0.004,P<0.01).AMHR2 expression was also increased in DI phase,and the difference was highly statistically significant(P phase vs DI phase,P=0.000,P<0.001).Compared with E phase,the expression of AMHR2 protein was increased in M phase and DI phase,and the differences were highly statistically significant(E phase vs M phase,P=0.003,P<0.01;E phase vs D1 phase,P=0.000,P<0.001).Compared with DI,the expression of AMHR2 protein in M phase decreased,and the difference was statistically significant(M phase versus DI phase,P=0.003,P<0.01).The AMH protein expression trend was the same as the AMHR2 protein expression trend,and the expression was lowest in the E phase.Compared with P phase,the expression of AMH protein in M phase was increased,and the difference was statistically significant(P phase vs M phase,P=0.006,P<0.01).The expression of AMH in DI phase was also increased,and the difference was highly statistically significant(P phase versus DI phase,P=0.001,P<0.01).Compared with E phase,the expression of AMH protein in both M phase and DI phase was increased,and the differences were highly statistically significant(E phase versus M phase,P=0.002,P<0.01;E phase vs DI phase,P=0.000 P<0.001).In the peri-implantation period of rats: Immunohistochemistry showed that AMHR2 was also localized to mesenchymal cells in the endometrium of pregnant rats D2-D8.The expression changes of AMHR2 protein immunopositive products in the endometrium of D2-D8 were as follows: D2-D5 showed a decreasing trend,D5 was shallowest,and D6-D8 gradually became darker.This suggests that AMHR2 may be associated with the appearance of a planting window,which may be related to the receptivity of the endometrium.q PCR showed that the AMHR2 gene of D2-D8 at the peri-planting stage first decreased and then increased,D2-D4 showed a downward trend,D5-D8 showed an upward trend,and compared with the D8 group,D3 group,D4 group,D5 The expression of AMHR2 gene in group D6 was statistically different(D3 vs.D8,P=0.015,P<0.05;D4 vs D8,P=0.015,P<0.05;D5 vs D8,P=0.014,P<0.05;D6 group vs D8 group,P=0.016,P<0.05).The AMH gene detection results also showed a trend of decreasing first and then increasing.D2-D4 showed a downward trend,and D5-D8 showed an upward trend.Compared with the D8 group,the AMH gene expression in the D3 group,D4 group,D5,and D7 groups had Statistical differences(D3 vs.D8,P=0.014,P<0.05;D4 vs D8,P=0.014,P<0.05;D5 vs D8,P=0.014,P<0.05;D7 vs D8 Group,P=0.018,P<0.05).Western-blotting results further demonstrated that the expression trends of AMHR2 and AMH proteins are consistent with gene expression trends.The AMHR2 protein first decreased and then rose in the D2-D8 group.Among them,compared with the D2 group,the expression of AMHR2 in the D4 group decreased;the expression of the AMHR2 protein in the D8 group increased,with statistical differences(D2 vs.D4,P=0.006,P<0.01;D2 vs.D8,P= 0.002,P < 0.01).Compared with group D3,the expression of AMHR2 protein in D6,D7,and D8 groups was significantly higher than that in D3 group(P<0.01);<0.01;D3 vs.D6,P=0.000,P<0.001).Compared with D4 group and D5 group,the expression of AMHR2 protein in D6,D7,and D8 groups were all increased,and there were highly significant statistical differences(D4 group vs D6 group,P=0.000,P<0.001;D4 group vs D7 group,P = 0.000,P <0.01;D4 group vs D8 group,P = 0.000,P <0.001;D5 group vs D6 group,P = 0.002,P <0.01;D5 group vs D7 group,P = 0.000,P <0.01;D5 group vs D8 group,P=0.000,P<0.001).AMH protein results showed that compared with D7 group,the expression of AMHR2 protein in D2,D3,and D4 groups was decreased,and the difference was statistically significant(D7 group vs D2 group,P=0.04,P<0.05;D7 group vs D2 group,P= 0.04,P<0.05;D7 vs.D2,P=0.04,P<0.05).HOXA-10 protein results showed that compared with D4 group,the expression of HOXA-10 protein in D5,D6,and D7 groups increased,and the difference was statistically significant(D4 group vs D5 group,P=0.000,P<0.001;D4 group Vs D6 group,P=0.000,P<0.01;D4 group vs D7 group,P=0.000,P<0.001).Conclision: AMHR2 expression in endometrium of rats during different estrous cycles and peri-implantation period.AMHR2 was located in the endometrial stromal cells.The AMH protein expression trend was consistent with the expression of AMHR2 protein,suggesting that there may be an AMH-AMHR2 autocrine system in the endometrium.The trend of AMHR2 protein expression was consistent with that of HOXA-10 protein,suggesting that the expression of AMHR2 protein may be related to the appearance of the window of implantation.AMHR2 is also may associated with endometrial receptivity. |
PDF Full Text Request