Therapeutic effect of Bushen Huoxue Recipe (补肾活血方) on autoimmune premature ovarian failure mice established by immunization with recombinant porcine zona pellucida 4 antigen

Chuan-ling Tang; Fang Li; Lan Sun; Da-jin Li

doi:10.1007/s11655-012-1025-y

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Therapeutic effect of Bushen Huoxue Recipe (补肾活血方) on autoimmune premature ovarian failure mice established by immunization with recombinant porcine zona pellucida 4 antigen

OriginalPaper | Updated：2021-08-27

- Therapeutic effect of Bushen Huoxue Recipe (补肾活血方) on autoimmune premature ovarian failure mice established by immunization with recombinant porcine zona pellucida 4 antigen
- Therapeutic effect of Bushen Huoxue Recipe (补肾活血方) on autoimmune premature ovarian failure mice established by immunization with recombinant porcine zona pellucida 4 antigen
- 中国结合医学杂志（英文版） 2013年19卷第6期页码：439-445
- Affiliations：
  
  1. Laboratory for Reproductive Immunology, Hospital and Institute of Obstetrics and Gynecology, Fudan University Shanghai Medical College,Shanghai,China
  2. Jiangsu Kanion Pharmaceutical Co., Ltd.,Jiangsu Province,Lianyungang,China
  3. Department of Obstetrics and Gynecology, the Affiliated Hospital, Hainan Medical College,Haikou,China
- Author bio：
- Funds：
  
  Supported by National Key Research Program of China (No. 2006CB944007), National and Shanghai Leading Academic Discipline Project (No. 211XK22), Program for Outstanding Medical Academic Leader (to LI Da-jin)
- DOI：10.1007/s11655-012-1025-y
  中图分类号：
- 纸质出版日期：2013，
  
  网络出版日期：2012-3-14，
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Tang, Cl., Li, F., Sun, L. et al. Therapeutic effect of Bushen Huoxue Recipe (补肾活血方) on autoimmune premature ovarian failure mice established by immunization with recombinant porcine zona pellucida 4 antigen., Chin. J. Integr. Med. 19, 439–445 (2013). https://doi.org/10.1007/s11655-012-1025-y

Chuan-ling Tang, Fang Li, Lan Sun, et al. Therapeutic effect of Bushen Huoxue Recipe (补肾活血方) on autoimmune premature ovarian failure mice established by immunization with recombinant porcine zona pellucida 4 antigen[J]. Chinese Journal of Integrative Medicine, 2013,19(6):439-445.
Tang, Cl., Li, F., Sun, L. et al. Therapeutic effect of Bushen Huoxue Recipe (补肾活血方) on autoimmune premature ovarian failure mice established by immunization with recombinant porcine zona pellucida 4 antigen., Chin. J. Integr. Med. 19, 439–445 (2013). https://doi.org/10.1007/s11655-012-1025-y DOI：

Chuan-ling Tang, Fang Li, Lan Sun, et al. Therapeutic effect of Bushen Huoxue Recipe (补肾活血方) on autoimmune premature ovarian failure mice established by immunization with recombinant porcine zona pellucida 4 antigen[J]. Chinese Journal of Integrative Medicine, 2013,19(6):439-445. DOI： 10.1007/s11655-012-1025-y.

摘要

To investigate the efficacy and mechanism of Bushen Huoxue Recipe (补肾活血方

BHR) in the treatment of murine autoimmune premature ovarian failure (POF). The recombinant porcine zona pellucida 4 (pZP4) was expressed in E. coli BL21 (DE3) strain within prokaryotic plasmid pET28a (+)

purified by Ni-affinity chromatography and verified by Western blot. Murine autoimmune POF model was established by immunization with pZP4 of female BALB/c mice. Fifty POF mice were randomly divided into 5 groups

which were respectively given low (3.75 mg/kg)

moderate (7.5 mg/kg)

and high dose (15.0 mg/kg) of BHR by gastrogavage once daily for 20 days

with 17-β-estradiol (0.13 mg/kg) and normal saline as positive and negative control. Estrous cycles were analyzed through vaginal smears

serum estradiol (E) levels

and anti-pZP4 antibody titers were detected by ELISA. The proliferative responses in vitro of spleen lymphocytes to pZP4 antigen restimulation were measured by [3H]-thymidine incorporation

and the histomorphology changes of ovary were evaluated by optical microscope. The purified pZP4 was visible as a single lane with 14.4 kD in SDS-PAGE and Western blot. The murine POF model with lengthening estrous cycles

decreased levels of serum E2

high titers of serum anti-pZP4 antibody

and reduced ovarian follicles and corpus lutea were established by immunization with recombinant pZP4. Treatment with moderate and high dosage BHR significantly increased ovarian follicles and reduced the proliferation of spleen lymphocytes to the pZP4 antigen of POF mice (P <0.05). However

only the high dosage BHR administration significantly improved the estrous cycles

elevated the serum E levels (P <0.01)

and decreased the serum anti-pZP4 antibody titers of model mice P<0.05). The recombinant pZP4 could evoke the antigen-specific immune response in mice and induce the autoimmune ovarian injury. It has been demonstrated that BHR was able to increase the serum E levels and protect ovarian functions from the autoimmune injury in murine POF model.

Abstract

To investigate the efficacy and mechanism of Bushen Huoxue Recipe (补肾活血方

which were respectively given low (3.75 mg/kg)

moderate (7.5 mg/kg)

and high dose (15.0 mg/kg) of BHR by gastrogavage once daily for 20 days

with 17-β-estradiol (0.13 mg/kg) and normal saline as positive and negative control. Estrous cycles were analyzed through vaginal smears

serum estradiol (E) levels

and anti-pZP4 antibody titers were detected by ELISA. The proliferative responses in vitro of spleen lymphocytes to pZP4 antigen restimulation were measured by [3H]-thymidine incorporation

decreased levels of serum E2

high titers of serum anti-pZP4 antibody

only the high dosage BHR administration significantly improved the estrous cycles

elevated the serum E levels (P <0.01)

关键词

recombinant porcine zona pellucidaBushen Huoxue RecipePremature Ovarian Failureautoimmunity

Keywords

recombinant porcine zona pellucidaBushen Huoxue RecipePremature Ovarian Failureautoimmunity

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