Objective: To observe the effects of Liuwei Dihuang Granule (六味地黄颗粒, LDG) for tonifying Kidney (Shen) on the outcomes of in v/tro fertilization pre-embryo transfer (IVF-ET) of infertility women with Kidne...Objective: To observe the effects of Liuwei Dihuang Granule (六味地黄颗粒, LDG) for tonifying Kidney (Shen) on the outcomes of in v/tro fertilization pre-embryo transfer (IVF-ET) of infertility women with Kidneyyin deficiency syndrome and to explore its mechanism by detecting the proteome expression in the follicular fluid. Methods: Sixty-six infertility patients of Kidney-yin deficiency syndrome who would undergo IVF-ET, were randomly assigned to a treatment group and a control group according to a random number table, 33 cases in each group. Another 33 cases of non-Kidney-yin deficiency syndrome was taken as a syndrome-control group. Besides Westem routine therapy, LDG was given 3 menstrual cycles before IVF to the treatment group, and a placebo granule to the control and syndrome-control groups. The scores of Kidney-yin deficiency symptoms (sore waist and knees, dry vagina, dysphoria with feverish sensation in the chest, palms and soles, etc.) were assessed, the number of retrieved oocytes, rates of high quality oocytes and embryos, fertility rate and clinical pregnancy rate were recorded, and the follicular fluid was collected on the day when the ovum was picked up, the differential protein expression was detected using two-dimensional gel electrophoresis, and then, matrix assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOF-MS) was applied to identify the proteins. Results: The syndrome score in the treatment group decreased significantly from 16.09 ± 2.58 to 8.67 ± 2.13, while it changed insignificantly in the control group, with a significant difference in the lowering score between the two groups (P〈0.05); the high quality rates of oocytes and embryos and clinical pregnancy rate were all superior in the treatment group to the control group (82.29% vs 78.08%, 76.76% vs 68.79%, 63.64% vs 36.36%, all P〈0.05). The protein expression map from the follicular fluid showed that compared with the control group, 33 differential protein expressions were found in the syndrome-control group, among which 18 were down-regulated, and 15 up-regulated; in the treatment group 28 differential protein expressions were found, among which 15 were down-regulated, and 13 up-regulated. Through MALDI-TOF-MS, 14 proteins were identified (P〈0.05). Conclusions: For the infertility patients undergoing IVF, LDG could alleviate clinical symptoms, improve rates of high quality oocytes and embryos, so as to raise clinical pregnancy rate. The mechanism may be through regulating proteome expression in the follicular fluid to improve the developmental microenvironment for oocytes which would lead to a successful embryo implantation.展开更多
基金Supported by the National Natural Science Fund of China(No.81072836)the Natural Science Foundation Project of Shandong Science and Technology Agency(No.2009ZRB14306)and the Special Science and Technology Foundation for Doctoral Subject from Ministry of Education of China(No.20093731110006)
文摘Objective: To observe the effects of Liuwei Dihuang Granule (六味地黄颗粒, LDG) for tonifying Kidney (Shen) on the outcomes of in v/tro fertilization pre-embryo transfer (IVF-ET) of infertility women with Kidneyyin deficiency syndrome and to explore its mechanism by detecting the proteome expression in the follicular fluid. Methods: Sixty-six infertility patients of Kidney-yin deficiency syndrome who would undergo IVF-ET, were randomly assigned to a treatment group and a control group according to a random number table, 33 cases in each group. Another 33 cases of non-Kidney-yin deficiency syndrome was taken as a syndrome-control group. Besides Westem routine therapy, LDG was given 3 menstrual cycles before IVF to the treatment group, and a placebo granule to the control and syndrome-control groups. The scores of Kidney-yin deficiency symptoms (sore waist and knees, dry vagina, dysphoria with feverish sensation in the chest, palms and soles, etc.) were assessed, the number of retrieved oocytes, rates of high quality oocytes and embryos, fertility rate and clinical pregnancy rate were recorded, and the follicular fluid was collected on the day when the ovum was picked up, the differential protein expression was detected using two-dimensional gel electrophoresis, and then, matrix assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOF-MS) was applied to identify the proteins. Results: The syndrome score in the treatment group decreased significantly from 16.09 ± 2.58 to 8.67 ± 2.13, while it changed insignificantly in the control group, with a significant difference in the lowering score between the two groups (P〈0.05); the high quality rates of oocytes and embryos and clinical pregnancy rate were all superior in the treatment group to the control group (82.29% vs 78.08%, 76.76% vs 68.79%, 63.64% vs 36.36%, all P〈0.05). The protein expression map from the follicular fluid showed that compared with the control group, 33 differential protein expressions were found in the syndrome-control group, among which 18 were down-regulated, and 15 up-regulated; in the treatment group 28 differential protein expressions were found, among which 15 were down-regulated, and 13 up-regulated. Through MALDI-TOF-MS, 14 proteins were identified (P〈0.05). Conclusions: For the infertility patients undergoing IVF, LDG could alleviate clinical symptoms, improve rates of high quality oocytes and embryos, so as to raise clinical pregnancy rate. The mechanism may be through regulating proteome expression in the follicular fluid to improve the developmental microenvironment for oocytes which would lead to a successful embryo implantation.