ABSTRACT
The effect and underlying mechanism of Bu-Shen-An-Tai recipe on ovarian apoptosis in mice with controlled ovarian hyperstimulation (COH) implantation dysfunction were studied.The COH implantation dysfunction model in mice was established by intraperitoneal injection of 7.5 IU pregnant mare's serum gonadotrophin (PMSG),followed by 7.5 IU human chorionic gonadotrophin (HCG) 48 h later.Then the female mice were mated with male at a ratio of 2:l in the same cage at 6:00 p.m.The female mice from normal group were injected intraperitoneally with normal saline and mated at the corresponding time.Day 1 of pregnancy was recorded by examining its vaginal smears at 8:00 a.m.of the next day.Fifty successfully pregnant mice were equally randomly divided into 5 groups:normal control pregnant group (NC),COH implantation dysfunction model group (COH),low dosage of Bu-Shen-An-Tai recipe group (LOW),middle dosage of Bu-Shen-An-Tai recipe group (MID) and high dosage of Bu-Shen-An-Tai recipe group (HIGH).Then from day 1,the mice in different groups were respectively intragastrically given corresponding treatments at 9:00 a.m.for 5 consecutive days.The concentrations of 17β-estradiol (E2) and progesterone (P4) were determined by radioimmunoassay (RIA).The ultrastructural changes of ovarian tissues were observed by transmission electron microscope (TEM).The histopathological changes of ovarian tissues were observed by HE staining.The number of atretic follicles and pregnant corpus luteum were also recorded.TUNEL was applied to measure apoptotic cells of ovarian tissues.Western blotting was used to detect the protein expression of apoptosis-related factors like Bax,Bcl-2 and cleaved-caspase-3 in ovarian tissue of mice.The results showed that ovarian weight,the concentrations of E2 and P4,the number of atretic follicles and pregnant corpus luteum,as well as the apoptosis of granulosa cells were significantly increased in the COH group.The ultrastructures of ovarian tissues in the COH group showed that chromatin in granulosa cells was increased,agglutinated,aggregated or crescent-shaped.The focal cavitation and the typical apoptotic bodies could be seen in granulosa cells in the late stage of apoptosis.After the treatment with different doses of Bu-Shen-An-Tai recipe,the ultrastructural changes of ovarian granulosa cells apoptosis were dramatically improved and even disappeared under TEM.Visible mitochondria and mitochondrial cristae were increased and vacuoles were significantly reduced.The lipid dropltes were shown in a circluar or oval shape.The protein expression levels of Bax and cleaved-caspase-3 were decreased,and the expression of Bcl-2 protein was increased after treatment.It was concluded that Bu-Shen-An-Tai recipe can inhibit the apoptosis of ovarian granulosa cells,probably by up-regulating the protein expression of Bcl-2 and down-regulating Bax and cleaved-caspase-3,which contributes to the formation and maintenance of ovarian corpus luteum.It's helpful to promote the embryonic implantation,to reduce embryo loss and ultimately to improve the success rate of pregnancy.
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OBJECTIVE:To investigate the effects of Tiaojingzhuyun capsules on endometrial receptivity in mice with implanta-tion dysfunction and corresponding mechanism. METHODS:Pregnant mice were randomized into normal group,model group and the groups of low and high-dose [12,24 g/(kg·d)] Tiaojingzhuyun capsules,with 10 mice in each group. The mice in the drug ad-ministration groups were given corresponding drugs intragastrically,while those in the normal group and the model group were giv-en normal saline intragastrically,for 3 consecutive days from the 1st day of pregnancy. On the 4th day of pregnancy,the mice in all groups except for the normal group were given mifepristone solution subactaneously to establish model of implantation dysfunc-tion. On the 5th day of pregnancy,the mice were sacrificed,and then the change in endometrial morphology was observed and en-dometrial maturity was scored;the protein expression levels of leukaemia inhibitory factor(LIF),homoeobox gene HOXA10,ma-trix metalloproteinase 9(MMP-9)and matrix metalloproteinase inhibitory factor 1(TIMP-1)expression in endometrial tissues were determined. RESULTS:Compared to the normal group,the mice in the model group and the drug administration groups had endo-metrial hypoplasia and obviously lower endometrial maturity score and protein expression levels of LIF,HOXA10,MMP-9 and TIMP-1 in endometrial tissues(P<0.05). Compared to the model group,the mice in the drug administration groups demonstrated endometrial development which improved or tended to be normal and had significantly higher indexes mentioned above(P<0.05);besides,the effect shown in the group of high-dose Tiaojingzhuyun capsules was stronger than that in the group of low-dose Tiao-jingzhuyun capsules (P<0.05). CONCLUSIONS:Tiaojingzhuyun capsules can improve the endometrial receptivity in mice with implantation dysfunction by a mechanism which may be related to upregulating the gene expression of HOXA10 and LIF,and main-taining the balance of MMP-9 and TIMP-1.
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OBJECTIVE:To study the effects of Tiaojing zhuyun capsules on the number of blastocyst implantation,estrogen, progestogen and their receptors(ER,PR)in mice with implantation dysfunction. METHODS:48 pregnant mice were randomly di-vided into normal control group (normal saline),pathological model group (normal saline) and Tiaojing zhuyun capsules group (24 g/kg),with 16 mice in each group. They were given relevant medicine intragastrically,once a day,for consecutive 3 d. On the 4th day,those mice were given mifepristone 0.1 ml subcutaneously to induce implantation dysfunction model except for normal control group. On the 5th day,the mice were sacrificed. The number of blastocyst implantation sites was observed. The serum lev-els of estradiol (E2) and progesterone (P) were determined by radioimmunoassay,and the expression of ER and PR protein were determined by immunohistochemistry. RESULTS:On the 5th day of pregnancy,the number of embryo implantation sites,serum levels of E2 and P and the protein expression of ER and PR in pathological model group were significantly lower than in normal con-trol group(P<0.05);the number of embryo implantation sites,levels of E2 and P,the protein expression of ER and PR in Tiao-jing zhuyun capsules group were significantly higher than in pathological model group (P<0.05). CONCLUSIONS:Tiaojing zhuyun capsules can effectively improve the serum levels of E2 and P,the protein expression of ER and PR,and increase the num-ber of blastocyst implantation sites in mice.
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<0.01). It was suggested Bangderun may favor a shift from Th1-to Th2-type immune response, therefore inhibiting the maternal immune rejection, inducing the immune tolerance and improving the fetal implantation.
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In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor (LIF) in mice with embryonic implantation dysfunction (EID), 120 Kun- ruing mice post coition were randomized into three groups: normal control group, model group and traditional Chinese medicine group (TCM group) (n=40 in each group). Uterus was collected on the pregnancy day (Pd) 4, 5, 6 after an intravenous injection of Evan's blue. The endometrium was dyed by Evan's blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of L1F mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve em- bryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.
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Objective To observe the effect of Bushen Yiqi Hexue Recipe(Recipe to reinforce kidney,replenish qi and harmonize blood)on the endometrial gland apoptosis in blastocyst implantation dysfunctional mice.Methods The mice models of blastocyst implantation dysfunction were made and divided into control,model and treated groups with 30 in each.Pregnancy rate,number of implantation blastocysts,the expression of proliferating cell nuclear antigen(PCNA),bax,bcl-2 and activated Caspase-3 were observed.Results Compared with the control group,pregnancy rate and mean implanted blastocysts in the model group were remarkably reduced.Compared with the model group,pregnancy rate and mean implanted blastocysts in the treated group were significantly improved(P