Protective effects of major royal jelly proteins on reproductive function of mice during perimenopausal period

doi:10.3785/j.issn.1008-9209.2018.11.123

Journal of Zhejiang University (Agriculture and Life Sciences)

2019, Vol. 45

Issue (6): 751-759 DOI: 10.3785/j.issn.1008-9209.2018.11.123

Animal sciences & veterinary medicine

Protective effects of major royal jelly proteins on reproductive function of mice during perimenopausal period

Xin LIU(

),Yong CHEN,Lirong SHEN(

)

Zhejiang Key Laboratory for Agro-Food Processing, Fuli Institute of Food Science, School of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China

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Abstract

In order to study the effects of the major royal jelly proteins (MRJPs) on the endocrine endocrinology in female mice during the perimenopausal period, an experiment was conducted for the natural aging female ICR mice with the age of 11-13 months. Fifty female mice were randomly divided into five equal groups, including low, middle and high dose groups of MRJPs (administrated with 125, 250, 500 mg/kg MRJPs, respectively), positive control group (administrated with 125 mg/kg casein) and old model group (administrated with physiological saline). Meanwhile, ten female mice with the age of five months were set as the young control group (administrated with physiological saline). Intragastric administrations were conducted for seven weeks once daily continuously. The indexes of uterus and ovary were calculated, and serum levels of estradiol (E₂), progesterone (P), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were detected by radioimmunoassay. The real-time polymerase chain reaction was used to detect the expressions of estrogen receptor α (ERα), estrogen receptor β (ERβ) and progesterone receptor (PR). The histomorphological observation of the uterus and ovary was also carried out under an optical microscope. The results showed that MRJPs supplement in the medium and high dosages could increase the uterus index and ovary index. Serum levels of E₂ and P were significantly elevated in the MRJPs groups, while the levels of FSH and LH were decreased as compared with the old model group. The expressions of ovarian ERα, ERβ and PR were up-regulated significantly in the MRJPs groups. Follicular development was also improved. In conclusion, the above results suggest that MRJPs possess the protective activity in reproduction of mice during the perimenopausal period. It provides scientific evidences of royal jelly and MRJPs in prevention and improvement of perimenopause syndrome.

Key words： major royal jelly proteins mice perimenopause syndrome reproductive function ovary serum hormone estrogenic-like effect

Received: 12 November 2018 Published: 20 January 2020

CLC:

R 151.3

Corresponding Authors: Lirong SHEN E-mail: xinguang0909@163.com;shenlirong@zju.edu.cn

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Cite this article:

Xin LIU,Yong CHEN,Lirong SHEN. Protective effects of major royal jelly proteins on reproductive function of mice during perimenopausal period. Journal of Zhejiang University (Agriculture and Life Sciences), 2019, 45(6): 751-759.

URL:

http://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2018.11.123 OR http://www.zjujournals.com/agr/Y2019/V45/I6/751

蜂王浆主蛋白对围绝经期小鼠生殖功能的保护作用

为研究蜂王浆主蛋白（major royal jelly proteins, MRJPs）对围绝经期小鼠生殖内分泌的影响，选取11～13月龄自然衰老雌性ICR小鼠作为动物模型开展实验。将50只小鼠随机分为蜂王浆主蛋白低、中、高剂量组（分别给予125、250、500 mg/kg MRJPs）、阳性对照组（给予125 mg/kg酪蛋白）和老年模型组（给予生理盐水），另选10只5月龄雌性小鼠作为青年对照组（给予生理盐水），所有小鼠每日灌胃1次，持续7周后，计算小鼠子宫指数和卵巢指数，并通过放射免疫技术检测血清激素雌二醇（estradiol, E₂）、孕酮（progesterone, P）、促卵泡生成素（follicle-stimulating hormone, FSH）和促黄体生成素（luteinizing hormone, LH）含量；此外，采用定量聚合酶链式反应检测小鼠卵巢中激素受体表达量，并在光学显微镜下对卵巢和子宫组织形态进行观察。结果表明：与老年模型组相比，补充中、高剂量的MRJPs可显著提高小鼠卵巢指数和子宫指数，显著提高血清激素E₂和P的水平，降低FSH和LH含量，并提高雌激素受体基因ERα、ERβ和孕激素受体基因PR的表达量，增加子宫内膜平均厚度，改善卵巢的卵泡发育和形态。表明MRJPs具有类雌激素效应，对围绝经期小鼠生殖功能具有一定的保护作用。本研究为蜂王浆和MRJPs应用于预防和改善女性围绝经期综合征提供了科学依据。

关键词： 蜂王浆主蛋白, 小鼠, 围绝经期综合征, 生殖功能, 卵巢, 血清激素, 类雌激素效应

基因 Gene	引物序列（5 $′$ →3 $′$ ） Primer sequence (5 $′$ →3 $′$ )
β-actin	F：GTGACGTTGACATCCGTAAAGA
β-actin	R：GTAACAGTCCGCCTAGAAGCAC
ERα	F：CAAGGTAAATGTGTGGAAGGCA
ERα	R：ATGGGAAAGAATGAGAAGGAGC
ERβ	F：GACGAAGAGTGCTGTCCCAA
ERβ	R：TCAGCTTCCGGCTACTCTCT
PR	F：GTCACTATGGCGTGCTTACCTG
PR	R：CAACACCGTCAAGGGTTCTCAT

Table 1 Information of primer sequences

Fig. 1 Effects of supplemental MRJPs on body mass of miceYCG: Young control group (administrated with physiological saline); OMG: Old model group (administrated with physiological saline); M125: Low dose group of MRJPs (administrated with 125 mg/kg MRJPs); M250: Medium dose group of MRJPs (administrated with 250 mg/kg MRJPs); M500: High dose group of MRJPs (adminis-trated with 500 mg/kg MRJPs); PCG: Positive control group (adminis-trated with 125 mg/kg casein).

Table 2 Effects of supplemental MRJPs on the ovarian index and uterine index of mice mg/g

Table 3 Effects of supplemental MRJPs on the serum hormone levels of mice

Fig. 2 Effects of supplemental MRJPs on the endometrial thickness of micePlease see the footnote of Fig. 1 for the details of different treatment symbols. Different lowercase letters above the bars represent significant differences at the 0.05 probability level.

Fig. 3 Effects of supplemental MRJPs on the relative expression of ERα, ERβ and PR in ovaries of micePlease see the footnote of Fig. 1 for the details of different treatment symbols. Different lowercase letters above the bars represent significant differences at the 0.05 probability level.

Fig. 4 Histomorphological observation of ovaries in mice by HE staining


[1]	崔清志,田艳君,温海霞,等.益坤宁对围绝经期大鼠卵巢细胞凋亡率及caspase-3表达的影响.中国应用生理学杂志,2010,26(3):318-321. CUI Q Z, TIAN Y J, WEN H X, et al. Effects of Yikunning on the apoptotic rate and expression of caspase-3 in rat ovaries during perimenopausal period. Chinese Journal of Applied Physiology, 2010,26(3):318-321. (in Chinese with English abstract)

[2]	MARNOCHA S K, BERGSTROM M, DEMPSEY L F. The lived experience of perimenopause and menopause. Contem-porary Nurse, 2011,37(2):229-240.

[3]	ROBERTS H, HICKEY M. Managing the menopause: an update. Maturitas, 2016,86:53-58.

[4]	HOLLOWAY D. An overview of the menopause: assessment and management. Nursing Standard, 2011,25(30):47-57.

[5]	WORSLEY R, DAVIS S R, GAVRILIDIS E, et al. Hormonal therapies for new onset and relapsed depression during perimenopause. Maturitas, 2012,73(2):127-133.

[6]	BANKS E, BERAL V, BULL D, et al. Breast cancer and hormone-replacement therapy in the Million Women Study. Lancet, 2003,362(9382):419-427.

[7]	GIACALONE P L, LAFFARGUE F. Ovarian cancer and hormone replacement therapy (HRT). Clinical Aspects, 2001,8(5):394-397.

[8]	陶挺,苏松坤,陈盛禄,等.蜂王浆蛋白生物学功能的研究.应用昆虫学报,2008,45(1):33-37. TAO T, SU S K, CHEN S L, et al. Study on the biological function of royal jelly protein. Chinese Journal of Applied Entomology, 2008,45(1):33-37. (in Chinese with English abstract)

[9]	KAMAKURA M, MITANI N, FUKUDA T, et al. Antifatigue effect of fresh royal jelly in mice. Journal of Nutritional Science & Vitaminology, 2001,47(6):394-401.

[10]	SUSILOWATI H, MURAKAMI K, YUMOTO H, et al. Royal jelly inhibits Pseudomonas aeruginosa adherence and reduces excessive inflammatory responses in human epithelial cells. Biomed Research International, 2017(4):3191752.

[11]	CHEN Y F, WANG K, ZHANG Y Z, et al. In vitro anti-inflammatory effects of three fatty acids from royal jelly. Mediators of Inflammation, 2016(2):3583684.

[12]	KHOSHPEY B, DJAZAYERI S, AMIRI F, et al. Effect of royal jelly intake on serum glucose, apolipoprotein A-I (ApoA-I), apolipoprotein B (ApoB) and ApoB/ApoA-I ratios in patients with type 2 diabetes: a randomized, double-blind clinical trial study. Canadian Journal of Diabetes, 2016,40(4):324-328.

[13]	PYRZANOWSKA J, PIECHAL A, BLECHARZ-KLIN K, et al. Long-term administration of Greek royal jelly improves spatial memory and influences the concentration of brain neurotransmitters in naturally aged Wistar male rats. Journal of Ethnopharmacology, 2014,155(1):343-351.

[14]	SILICI S, EKMEKCIOGLU O, KANBUR M, et al. The protective effect of royal jelly against cisplatin-induced renal oxidative stress in rats. World Journal of Urology, 2011,29(1):127-132.

[15]	VUCEVIC D, MELLIOU E, VASILIJIC S, et al. Fatty acids isolated from royal jelly modulate dendritic cell-mediated immune response in vitro. International Immunophar-macology, 2007,7(9):1211-1220.

[16]	沈立荣,张瓅文,丁美会,等.蜂王浆的营养保健功能及分子机理研究进展.中国农业科技导报,2009,11(4):41-47. SHEN L R, ZHANG L W, DING M H, et al. Research progress on nutritional and health-care functions and molecular mechanism of royal jelly. Journal of Agricultural Science and Technology, 2009,11(4):41-47. (in Chinese with English abstract)

[17]	KAMAKURA M. Royalactin induces queen differentiation in honeybees. Nature, 2011,473(7348):478-483.

[18]	于张颖,肖发,柳丹丹,等.蜂王浆主蛋白提取工艺优化及质量分析.食品工业科技,2014,35(19):185-188. YU Z Y, XIAO F, LIU D D, et al. The optimization and quality analysis on extraction of the major royal jelly proteins (MRJPs) from royal jelly. Science and Technology of Food Industry, 2014,35(19):185-188. (in Chinese with English abstract)

[19]	李玫璐,谭量量,裘卫,等.蜂王浆主蛋白(MRJPs)超滤分离工艺优化及产物质量分析.浙江大学学报(农业与生命科学版),2014,40(5):526-532. LI M L, TAN L L, QIU W, et al. Optimization of ultrafiltration separation and quality analysis of the major royal jelly proteins. Journal of Zhejiang University (Agriculture and Life Sciences), 2014,40(5):526-532. (in Chinese with English abstract)

[20]	XIN X X, CHEN Y, CHEN D, et al. Supplementation with major royal-jelly proteins increases lifespan, feeding, and fecundity in Drosophila. Journal of Agricultural & Food Chemistry, 2016,64(29):5803-5812.

[21]	CHEN D, LIU F, WAN J B, et al. Effect of major royal jelly proteins on spatial memory in aged rats: metabolomics analysis in urine. Journal of Agricultural & Food Chemistry, 2017,65(15):3151-3159.

[22]	钱浩诚,蒋晨旻,陈华才,等.王浆主蛋白作为胎牛血清替代物培养中国仓鼠卵巢细胞CHO-K1的效果及作用机制.浙江大学学报(农业与生命科学版),2018,44(1):1-9. QIAN H C, JIANG C M, CHEN H C, et al. Effect and mechanism of major royal jelly proteins as an alternative of fetal bovine serum to culture Chinese hamster ovary cell CHO-K1. Journal of Zhejiang University (Agriculture and Life Sciences), 2018,44(1):1-9. (in Chinese with English abstract)

[23]	SUZUKI K M, ISOHAMA Y, MARUYAMA H, et al. Estrogenic activities of fatty acids and a sterol isolated from royal jelly. Evidence-Based Complementary and Alternative Medicine, 2008,5(3):295-302.

[24]	BYERS S L, WILES M V, DUNN S L, et al. Mouse estrous cycle identification tool and images. PLoS One, 2012,7(4):e35538.

[25]	王亚凡,封纪珍,王文立,等.围绝经期综合征的治疗进展.河北医药,2010,32(2):223-225. WANG Y F, FENG J Z, WANG W L, et al. Progress in the treatment of perimenopausal syndrome. Hebei Medical Journal, 2010,32(2):223-225. (in Chinese)

[26]	SALEM E Z H. Modulatory effects of Trifolium pretense extract and royal jelly on the function of hypothalamic-pituitary-ovarian axis in ovariectomized rats. International Journal of Pharmacy & Pharmaceutical Sciences, 2013,5(3):593-597.

[27]	HIDAKA S, OKAMOTO Y, UCHIYAMA S, et al. Royal jelly prevents osteoporosis in rats: beneficial effects in ovariectomy model and in bone tissue culture model. Evidence-based Complementary and Alternative Medicine, 2006,3(3):339-348.

[28]	郭子侠,涂晓明.动物性保健食品性激素含量调查及评价.中国食品卫生杂志,2000,12(3):29-30. GUO Z X, TU X M. Investigation and evaluation of sex hormone content in animal health food. Chinese Journal of Food Hygiene, 2000,12(3):29-30. (in Chinese with English abstract)

[29]	张荣富,丁杰锋,屠凤娟.围绝经期女性的4种激素检测分析.放射免疫学杂志,2003,16(4):232-233. ZHANG R F, DING J F, TU F J. Analysis of four hormones in perimenopausal women. Journal of Radioimmunology, 2003,16(4):232-233. (in Chinese)

[30]	KAMAKURA M. Signal transduction mechanism leading to enhanced proliferation of primary cultured adult rat hepatocytes treated with royal jelly 57-kDa protein. The Journal of Biochemistry, 2002,132(6):911-919.

[31]	KIMURA M, KIMURA Y, TSUMURA K, et al. 350-kDa royal jelly glycoprotein (apisin), which stimulates proliferation of human monocytes, bears the β1-3-galactosylated N-glycan: analysis of the N-glycosylation site. Bioscience, Biotechnology, and Biochemistry, 2003,67(9):2055-2058.

[32]	桂黎明,王建六.雌激素受体与细胞信号传导通路.医学分子生物学杂志,2001,23(2):103-106. GUI L M, WANG J L. Estrogen receptor and cell signaling pathway. Journal of Medical Molecular Biology, 2001,23(2):103-106. (in Chinese)

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