The Effect Of Tiaogeng Yijing Decoction To Patients With Poor Ovarian Response And Kidney Deficiency And Blood Stasis Syndrome Undergoing In Vitro Fertilization-Embryo Transfer

Poor ovarian response(POR)is a pathological state of adverse reactions of ovary to gonadotropin(Gn)stimulation.It is mainly manifested in the development of the eggs in ovarian stimulation cycles,less blood estrogen and low dosage of Gn,the number of eggs,the cycle cancellation rate and clinical pregnancy rate are also low.But nowadays we have no reasonable clinical treatment and the effective strategy to reverse POR.It becomes a difficult problem to be solved in the field of reproduction.Traditional Chinese medicine plays an important role in the treatment of infertility.It can improve reproductive endocrine function and combine modern assisted reproductive technology to create an environmental foundation for POR patients.Therefore,on the basis of earlier animal experiments,we intend to explore the effect of "Tiaogeng Yijing Decoction" on the POR patients with kidney deficiency and blood stasis syndrome undergoing in vitro fertilization-embryo transfer.Cytokines have a very close relationship with reproductive endocrine system.There are many cytokines in human follicular fluid,regulating egg development,maturation and ovulation process;at the same time cytokines can regulate the formation of endometrial receptivity as the intermediary signal system.So we use the most recognized and related cytokines with egg quality and endometrial receptivity as the observation indexes,including: transforming growth factor-?1(TGF-?1),leukemia inhibitory factor(LIF),granulocyte-colony stimulating factor(G-CSF)and vascular endothelial growth factor(VEGF),and the same time we choose growth differentiation factor recognized-9(GDP-9),integrinav?3 as the observation indexes which have been confirmed.At last we analyse the effct of Tiaogeng Yijing decoction improving ovarian reserve and endometrial receptivity,and investigate its mechanisms of improving the outcome of pregnancy.Part 1 Tiaogeng Yijing Decoction Promotes Ovarian Reserve and the Effect of TCM Syndrome of Patients with Poor Ovarian Response and Kidney Deficiency and Blood Stasis Syndrome Undergoing In Vitro Fertilization-Embryo TransferObjective:To explore the effect of ovarian reserve and the kidney and blood stasis syndrome of traditional Chinese medicine(TCM)in poor ovarian response(POR)patients in in-vitro fertilization and embryo transfer(IVF-ET)using Tiaogeng Yijing decoction.From the serum levels of sex hormones,follicular fluid levels of cytokines,the number of eggs,rate of fertilization,high quality embryo rate and so on,we explore the effect of ovarian reserve and provide an effective basis for the treatment of POR in assisted reproductive technology using TCM.Methods:A total of 40 patients who using microstimulation in the last IVF-ET cycle were randomly and equally assigned to treatment group and control group.Two groups of patients were treated with microstimulation using clomiphene(CC)and human menopausal gonadotropin(HMG).Patients in the treatment group received Tiaogeng Yijing decoction for continuous 3 cycles in addition to microstimulation from the seventh day of menstruation taking 14 days,and while patients in the control group had a rest for three months before undergoing microstimulation.The following molecules were measured following treatment.Serum levels of sex hormones,including follicle stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E2)and anti-mullerian hormone(AMH),the antral follicle count(AFC),mean ovarian diameter(MOD);follicular fluid levels of cytokines,including GDF-9,TGF-?1,LIF,G-CSFand VEGF;the change of kidney deficiency and blood stasis syndrome of traditional Chinese medicine of the two groups;comparison of Gonadotropin(Gn)dosage and the Gn days,estradiol(E2)of taking egg day,the number of eggs,rate of fertilization,high quality embryo rate of the two groups.Results:1.Comparison of general clinical data between two groups.There was no significant difference between the two groups,such as age,infertility time,body mass index and sex hormones.((P>0.05)2.Comparison of sex hormone levels,antrol follicle count(AFC),average ovarian volume between two groups.The results showed that Tiaogeng Yijing decoction significantly decreased serum levels of FSH?FSH/LH and increased the serum AMH level(P<0.01).and statistically decreased serum levels of E2,increased AFC compared to the control group and before and after treatment of the treatment group(P<0.05).However,no significant differences were found in LH,MOD(P>0.05).3.Comparison of kidney deficiency and blood stasis syndrome and syndrome score between two groups.Kidney deficiency and blood stasis syndrome significantly decrease(P<0.05)after treatment,and syndrome score difference between two groups was very significant(P<0.05)4.Comparison of GDF-9?TGF-?1?LIF?G-CSF?VEGF concentrations in the follicular fluids between two groups.The levels of GDF-9,TGF-?1 and VEGF in the follicular fluid of the treatment group were significantly higher than that in the control group(P<0.01).However,no significant differences were found in LIF,and G-CSF(P>0.05).5.Comparison of the Gn dosage?the Gn days?the number of eggs?E2?fertilization rate?high quality embryo rate between two groups.There was no significant difference of the Gn days(P > 0.05),and the Gn dosage was obviously lower than the control group(P < 0.05),eggs,E2 levels,the number of eggs,fertilization rate,high quality embryo rate of treatment group were significantly higher than control group(P<0.01).Conclusion: The patients with Tiaogeng Yijing Decoction can improve GDF-9,TGF-?1,VEGF levels in follicular fluid of POR patients,improving the kidney deficiency and blood stasis syndrome,thus improving the number of eggs,fertilization rate,high quality embryo rate.Part 2 Tiaogeng Yijing Decoction Promotes The Receptivity of Endometrium of Patients with Poor Ovarian Response and Kidney Deficiency and Blood Stasis Syndrome Undergoing in vitro fertilizationembryo transfer and The Effect of Pregnancy OutcomeObjective: To explore the influence of Tiaogeng Yijing Decoction to the receptivity of endometrium and pregnancy outcome of the poor ovarian response(POR)patients with kidney deficiency and blood stasis syndrome during IVF-ET.From the expression of related cytokines of the endometrium during the implantation window,endometrial thickness and blood flow index,we explore the influence of Tiaogeng Yijing Decoction to the receptivity of endometrium.From implantation rate,pregnancy rate,we investigate the effect of Tiaogeng Yijing Decoction on pregnancy outcome.At last,we discusse the possible function mechanism.Methods: A total of 40 patients who using microstimulation in the last IVF cycle were randomly and equally assigned to treatment group and control group.Two groups of patients were treated with microstimulation using clomiphene(CC)and human menopausal gonadotropin(HMG).Patients in the treatment group received Tiaogeng Yijing decoction for continuous 3 cycles in addition to microstimulation from the seventh day of menstruation taking 14 days,and while patients in the control group have a rest for three months before undergoing microstimulation.Using Western blot to test integrin av beta 3,TGF-?1,LIF,G-CSF,VEGF protein of two groups patients with planting window phase of endometrium.Comparing endometrial thickness in HCG day,proportion of type A endometrium,blood flow index,planting rate and pregnancy rate of two groups.Results:1.Comparison of general clinical data between two groups.There was no significant difference between the two groups,such as age,infertility time,body mass index and sex hormones.((P>0.05)2.Comparison of planting window endometrium relative grey between two groups Endometrium relative grey value of planting window phase between two groups,integrin av?3,TGF-?1,VEGF protein expression of treatment group was significantly higher than that of control group(P<0.05),while LIF,G-CSF protein was no significant difference(P>0.05).3.Comparison of endometrium thickness,blood flow index between two groups Proportion of type A endometrium,the blood flow index,the difference was statistically significant(P<0.05),but the endometrial thickness was no significant difference(P>0.05).4.Comparison of planting rate and pregnancy rate between two groups Clinical pregnancy rate of two groups,the difference was statistically significant(P<0.05)but the planting rate have no significant difference(P>0.05).Conclusion:1.Tiaogeng Yijing Decoction can improve the receptivity of endometrium,improve the expression of integrin av?3,TGF-?1,VEGF protein of patient with POR.2.POR patients with Tiaogeng Yijing Decoction can improve TGF-?1 and VEGF in the follicular fluid and endometrium at the same time.TGF-?1 plays a role in the induction of VEGF in terms of angiogenesis and acts as an upstream cytokine in VEGF to promote angiogenesis,thereby improving egg quality and endometrial blood flow.So it may be the mechanism of Tiaogeng Yijing Decoction improving the pregnancy rate of POR patients.