Chinese Journal of Integrative Medicine

, Volume 20, Issue 7, pp 503–509 | Cite as

Effects of Liuwei Dihuang Granule (六味地黄颗粒) on the outcomes of in vitro fertilization pre-embryo transfer in infertility women with Kidney-yin deficiency syndrome and the proteome expressions in the follicular fluid

  • Fang Lian (连 方)Email author
  • Hai-cui Wu (吴海萃)
  • Zhen-gao Sun (孙振高)
  • Ying Guo (郭 颖)
  • Lei Shi (石 垒)
  • Ming-yue Xue (薛明玥)
Original Article



To observe the effects of Liuwei Dihuang Granule (六味地黄颗粒, LDG) for tonifying Kidney (Shen) on the outcomes of in vitro 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.


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 Western 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.


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).


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.


tonify Kidney Kidney-yin deficiency infertility Liuwei Dihuang Granule protein expression 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Feng YJ, ed. Obstetrics and gynecology. Beijing: People’s Medical Publishing House; 2001:394.Google Scholar
  2. 2.
    Obstetrics and Gynecology Committee of Chinese Association of the Integration of Traditional and Western Medicine. Diagnosis and treatment criteria of traditional and Western medicine for endometriosis, gestational hypertension and female infertility. Chin J Integr Tradit West Med (Chin) 1991;11:376–378.Google Scholar
  3. 3.
    Zhang ZY, ed. Gynecology of Chinese medicine. 1st ed. Beijing: Chinese Medicine Press; 2002:323–324.Google Scholar
  4. 4.
    State Bureau of Technical Supervision. Clinic terminology diagnosis and treatment for Chinese syndrome. Beijing: China Standards Press;1997.Google Scholar
  5. 5.
    Ma BZ, ed. Gynecology of Chinese medicine. Shanghai: Shanghai Science and Technology Press; 1998:206–210.Google Scholar
  6. 6.
    Lu HL, Lu GX, eds. Human reproduction and reproductive engineering. Zhengzhou: Henan Science and Technology Press; 2001:109–110.Google Scholar
  7. 7.
    Puissant F, van Rysselberge F, Barlow P, Deweze J, Leroy F. Embryo scoring as a prognostic tool in IVF treatment. Hum Reprod 1987;2:705–708.PubMedGoogle Scholar
  8. 8.
    Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248–254.PubMedCrossRefGoogle Scholar
  9. 9.
    Wang BQ, Qin ZY, Bai SM, Liu FJ, Li Y, Liu SL. Comparison of different treatments on human serum albumin samples through two-dimensional electrophoresis. J Shandong Univ (Health Sci, Chin) 2009;47:89–94.Google Scholar
  10. 10.
    Qin YJ, Liu SL, Yang YR, Bai SM, Zhang XH, Deng XM. Study of proteomics in cerebrospinal fluid on central nervous system demyelinating diseases. J Shandong Univ (Health Sci, Chin) 2008;46:9–14.Google Scholar
  11. 11.
    Guo HM, ed. Ovary function and its periodical change. 2nd ed. Beijing: People’s Medical Publishing House; 1980:18–20.Google Scholar
  12. 12.
    Artini PG, Battaglia C, D’Ambrogio G, Barreca A, Droghini F, Volpe A, et al. Relationship between human oocyte maturity, fertilization and follicular fluid growth factors. Hum Reprod 1994;9:902–906.PubMedGoogle Scholar
  13. 13.
    Durán Reyes G, Rosales AM, Hicks Gómez JJ. Participation of the follicular fluid in follicular development, oocyte maturation and spermatic function. Ginecol Obstet Mex 1997;65:349–356.PubMedGoogle Scholar
  14. 14.
    Kim YS, Kim MS, Lee SH, Choi BC, Lim JM, Cha KY, et al. Proteomic analysis of recurrent spontaneous bortion: identification of an inadequately expressed set of proteins in human follicular flui. Proteomics 2006;6:3445–3454.PubMedCrossRefGoogle Scholar
  15. 15.
    Lian F, Teng YL, Zhang JW, Sun ZG, Liu YH, Ma FM, et al. Effects of Ergui Tianzhi Granule on leukaemia inhibitory factor in the human follicular fluid and oocyte quality in patients undergoing in vitro fertilization and embryo transfer. Chin J Integr Tradit West Med (Chin) 2007;27:976–979.Google Scholar
  16. 16.
    Chen MH. Management of Liuwei Dihuang Pill and its clinical application. Chin Med J (Chin) 2005;3:340–341.Google Scholar
  17. 17.
    Yang LY. Experimental study of Liuwei Dihuang Pill in treating premature ovarian failure. J Shanxi CoL Tradit Chin Med (Chin) 2002;3:21–23.Google Scholar
  18. 18.
    Yu ZL, Yan YQ, Lv JF, Kou JP. Effects of Liuwei Dihuang Decoction on sperms in mouse injured by tripterygium glycosides. Lishizhen Med Mater Med Res (Chin) 1999;10:81–82.Google Scholar
  19. 19.
    Sun X, Ando Y, Haraoka K, Katsuragi S, Yamashita T, Yamashita S, et al. Role of VLDL/chylomicron in amyloid formation in familial amyloidotic polyneuropathy. Biochem Biophys Res Commun 2003;311:344–350.PubMedCrossRefGoogle Scholar
  20. 20.
    Qiao YF, Pang DZ, Zhang ZL, Jiang YS. Research progress on early diagnostic index for diabetec nephropathy. Foreign Med (Urinary System, Chin) 2005;25:670–672.Google Scholar
  21. 21.
    Takebayashi K, Suetsugu M, Wakabayashi S, Aso Y, Inukai T. Retinol binding protein-4 levels and clinical features of type 2 diabetes patients. J Clin Endocrinol Metab 2007;92:2712–2719.PubMedCrossRefGoogle Scholar
  22. 22.
    Klöting N, Graham TE, Berndt J, Kralisch S, Kovacs P, Wason CJ, et al. Serum retinol-binding protein is more highly expressed in visceral than in subcutaneous adipose tissue and is a marker of intra-abdominal fat mass. Cell Metab 2007;6:79–87.PubMedCrossRefGoogle Scholar
  23. 23.
    Do HQ, Nazih H, Luc G, Arveiler D, Ferrières J, Evans A, et al. Influence of cholesteryl ester transfer protein, peroxisome proliferator-activated receptor alpha, apolipoprotein E, and apolipoprotein A-I polymorphisms on high-density lipoprotein cholesterol, apolipoprotein A-I, lipoprotein A-I, and lipoprotein A-I:A-II concentrations: the Prospective Epidemiological Study of Myocardial Infarction study. Metabolism 2009;58:283–289.PubMedCrossRefGoogle Scholar
  24. 24.
    Eriksson M, Carlson L A, Miettinen TA, Angelin B. Stimulation of fecal steroid excretion after infusion of recombinant proapolipoprotein A-I. Potential reverse cholesterol transport in humans. Circulation 1999;100:594–598.PubMedCrossRefGoogle Scholar
  25. 25.
    Francis GA, Tsujita M, Terry TL. Apolipoprotein A-I efficiently binds to and mediates cholesterol and phospholipid efflux from human but not rat aortic smooth muscle cells. Biochemistry 1999;38:16315–16322.PubMedCrossRefGoogle Scholar
  26. 26.
    Nanjee MN, Cooke CJ, Garvin R, Semeria F, Lewis G, Olszewski WL, et al. Intravenous apoA-I/lecithin discs increase pre-beta-HDL concentration in tissue fluid and stimulate reverse cholesterol transport in humans. J Lipid Res 2001;42:1586–1593.PubMedGoogle Scholar
  27. 27.
    Zhang H, Wen MP, Lou B, Chen PF. Antiendotoxin effects of high-density lipoprotein. J Fudan Univ (Med, Chin) 2003;30:474–476.Google Scholar
  28. 28.
    Furlaneto CJ, Ribeiro FP, Hatanaka E, Souza GM, Cassatella MA, Campa A. Apolipoproteins A-I and A-II downregulate neutrophil functions. Lipids 2002;37:925–928PubMedCrossRefGoogle Scholar
  29. 29.
    Silbum KA, McPhee DA, Maerz AL, Poumbourios P, Whittaker RG, Kirkpatrick A, et al. Efficacy of fusion peptide homologs in blocking cell lysis and HIV-induced fusion. AIDS Res Hum Retroviruses 1998;14:385–392.CrossRefGoogle Scholar
  30. 30.
    Brown S, Hay RV, Schreiber JR. Relationship between serum estrogen and level of apolipoprotein E in human ovarian follicular fluid. Fertil Steril 1989;51:639–643.PubMedGoogle Scholar
  31. 31.
    Zerbinatti CV, Mayer LP, Audet RG, Dyer CA. Apolipoprotein E is a putative autocrine regulator of the rat ovarian theca cell compartment. Biol Reprod 2001;64:1080–1089.PubMedCrossRefGoogle Scholar
  32. 32.
    Drela N, Kozdron E, Szczypiorski P. Moderate exercise may attenuate some aspects of immunosenescence. BMC Geriatr 2004;4:8.PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Chinese Association of the Integration of Traditional and Western Medicine and Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Fang Lian (连 方)
    • 1
    Email author
  • Hai-cui Wu (吴海萃)
    • 1
  • Zhen-gao Sun (孙振高)
    • 1
  • Ying Guo (郭 颖)
    • 1
  • Lei Shi (石 垒)
    • 1
  • Ming-yue Xue (薛明玥)
    • 1
  1. 1.Integrative Medicine Research Centre of Reproduction and Hereditythe Affiliated Hospital of Shandong University of Traditional Chinese MedicineJinanChina

Personalised recommendations