Journal of Functional Foods 82 (2021) 104514

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Journal of Functional Foods

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Active components and biological functions of royal jelly

Jianying Guo a, 2, Zixu Wang a, Yaoxing Chen a, Jing Cao a, Wenli Tian b, Baochen Ma c, 1,
Yulan Dong a, *
a
Key Laboratory of Precision Nutrition and Food Quality, Ministry of Education, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
b
Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China
c
China Animal Husbandry Group, Beijing 100070, China

A R T I C L E I N F O A B S T R A C T

Keywords: Royal jelly (RJ) is a kind of bee product widely used in cosmetics, medicine and other fields. Not only can RJ
Royal jelly regulate the physiological function of bee population, but also play a specific biological role in many diseases.
Active components This paper overviews the main active ingredients in the functional food RJ, including major royal jelly protein,
Biological functions
fatty acids, phenols, flavonoids, etc, and summarizes the active role of RJ in the maintenance of human health,
such as the regulation of immunity, lifespan, memory, digestive system, blood glucose, obesity, antibacterial and
anti-cancer effect, among which the regulation of memory can be used in the treatment of Alzheimer’s disease.
These findings will benefit for comprehensive understanding and use of RJ, hence making it more effective in
maintaining health.

1. Introduction
Nowadays, more and more natural products are used for health
problems. These products have been proved to reduce the risk of chronic
diseases and reduce the investment in health care (Jenkhetkan, Thi­
tiorul, Jansom, & Ratanavalachai, 2017). In recent years, natural bee
products have been widely used in traditional and modern medicine.
Royal jelly (RJ), as a bee product, is also used to maintain human health
(Pasupuleti, Sammugam, Ramesh, & Gan, 2017). RJ is the creamy
substance secreted by the mandibular and hypopharyngeal glands of
worker bees, which is the food of all honeybee larvae in the first three
days after birth; after three days, worker bee larvae begin to eat worker
jelly, which is mainly composed of honey and pollen. The queen bee
larvae continue to eat RJ (Kayashima, Yamanashi, Sato, Kumazawa, &
Yamakawa-Kobayashi, 2012; Shorter et al., 2015; Yang, Tian, Han, &
Miao, 2017). And royalactin may be a specific factor in RJ to induce bee
larvae to differentiate into queen (Kamakura, 2011). RJ is a kind of
widely used functional food (Ramadan & Al-Ghamdi, 2012) and dietary
supplement, which has many biological activities, such as anti-tumor,
anti-allergy, anti-inflammatory and immune regulation (Pasupuleti
et al., 2017). To understand the biological function of RJ more
comprehensively, we summarized the research in recent years, which

Abbreviations: Ach, acetylcholine; AD, Alzheimer’s disease; ALP, alkaline phosphatase; ApoA/B, apolipoprotein A/B; Bcl-2, B-cell lymphoma-2; β–Lg, β-lacto­
globulin; CAT, catalase; CDDP, Cisplatin; COX-2, cyclooxygenase-2; CP, collagen peptide; DNP-KLH, dinitrophenol-keyhole limpet hemocyanin; EGF, epidermal
growth factor; ERJ, Enzyme-treated RJ; ERK, extracellular regulated protein kinases; GABA, gamma-aminobatyric acid; GR, glutathione reductase; GSH, glutathione;
GST, glutathione S-transferase; GSH-Px, glutathione peroxidase; HCF-1, host cell factor; HPO-DAEE, 4-Hydroperoxy-2-decenoic acid ethyl ester; icv-STZ, strepto­
zotocin; IFN-γ, interferon-gamma; IL-4, interleukin-4; IL-1β, interleukin-1β; LCT, Lambda-cyhalothrin; LDH, lactic dehydrogenase; MAPK, Mitogen activated protein
kinase; MDA, malondialdehyde; MRJPs, major royal jelly proteins; M receptor, muscarinic receptor; MITF, microphthalmia associated transcription factor; NF-κB,
nuclear factor kappa-B; NO, nitric oxide; OVA, ovalbumin; OVX, ovariectomized; PGC-1α, proliferator-activated receptor gamma, coactivator 1 alpha; ROS, Reactive
Oxygen Species; SA, sebacic acid; SOD, superoxide dismutase; SQLE, squalene epoxidase; SLE, systemic lupus erythematosus; T-AOC, total antioxidant capability;
TNF-α, tumor necrosis factor-α; TRP1, TRP2, tyrosinase-related protein1,2; TNBS, 2, 4, 6-trinitrobenzene sulfonic acid; TXL, taxol; WM, working memory; 2-DecDA,
2-decene-1,10-dioic acid; 3-HHDA, 3-hydroxydecanoic acid; 3,10-HDecDA, 3,10-dihydroxydecanedioic acid; 8-HOC, 8-hydroxyoctanoic acid; 9-HDA, 9-hydroxy-2-
decenoic acid; 10-HDA, 10-hydroxy-2-decenoic acid.
* Corresponding author.
E-mail addresses: 1784452025@qq.com (J. Guo), zxwang2007@163.com (Z. Wang), yxchen@cau.edu.cn (Y. Chen), caojing315@126.com (J. Cao), leroytian@
126.com (W. Tian), mabc@cahg.com.cn (B. Ma), ylbcdong@cau.edu.cn (Y. Dong).
1
Co-corresponding author.
2
The first author.

https://doi.org/10.1016/j.jff.2021.104514
Received 17 October 2020; Received in revised form 6 April 2021; Accepted 19 April 2021
Available online 11 May 2021
1756-4646/© 2021 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
J. Guo et al. Journal of Functional Foods 82 (2021) 104514

will be helpful to promote the use and further research on the function of
RJ.
2. Active components
RJ contains 60–70% water, 12–15% crude protein, 10–16% total
sugar, 3–6% lipids, vitamins, salts and free amino acids (Chen & Chen,
1995; Howe, Dimick, & Benton, 1985; Schmitzová et al., 1998)
(Table 1). The pH value of RJ varies from 3.6 to 4.2 (Kunugi &
Mohammed Ali, 2019).
2.1. Proteins and free amino acids
More than 80% of the proteins in RJ are major royal jelly proteins
(MRJPs), which mainly include MRJP1-9 (Xin et al., 2016), and the
amino acid sequence homology among the proteins is 60–70% (Oka­
moto et al., 2003). The relative contents of MRJP1-3 and 5 in RJ protein
are 31%, 16%, 26% and 9% respectively, accounting for 82% of the total
RJ protein (Schmitzová et al., 1998). MRJP1 exists in two different
forms: oligomer and monomer. The size of MRJP1 monomer is 57 kDa,
which is called royalactin (Kamakura, 2011), and it is a weak acid
glycoprotein (Xin et al., 2016). MRJP1 oligomer is formed by apisimin
and MRJP1 monomer, and its size is 280–420 kDa. Moreover, different
oligomerization processes seem to depend on the pH value, forming
280/90 kDa oligomers at pH 6 and 7 and 340 kDa oligomers at pH 8
(Cruz et al., 2011; Simúth, 2001; Tamura, Kono, Harada, Yamaguchi, &
Moriyama, 2009). A recent study identified and reported a 16 molecular
structure of natural MRJP 1 oligomer, which contains four MRJP 1
monomer, four apisimin and eight 24-methylenecholesterol molecules,
with a resolution of 2.65 Å, forming the MRJP14-apisimin4-Osl8 complex
(Tian et al., 2018). When the storage temperature and time increase, RJ
will spoil and lose its biological activity. The degradation of MRJP1 in
RJ is positively correlated with both storage temperature and storage
period. Thus MRJP1 is an important index to evaluate RJ quality and
freshness (Shen et al., 2015). MRJP2 is a monomer glycoprotein with
eight forms and molecular weight of 50–59 kDa (Santos et al., 2005).
Abu Serie & Habashy (2019) purified MRJP2 and its isomer X1, their
molecular weights were about 49.95 kDa and 53.12 kDa, respectively.
Then I-TASSER was used to predict the three-dimensional structure of
the two purified proteins. The results showed that the structure of
MRJP2 and its isomer X1 had high similarity. The hydrophilic and hy­
drophobic residues of MRJP2 and its isomer X1 were 182&183 and
160&159, respectively, with 110 neutral residues. The surface charges
of the two three-dimensional structures were zero and (− 2) (Abu-Serie
& Habashy, 2019). MRJP3 is a glycoprotein with the size of 60–70 kDa
(Xin et al., 2016), and there are many subtypes. Santos et al (Santos
et al., 2005) found that there are five subtypes of MRJP3, with molecular
weights ranging from 80.59 kDa to 87 kDa. Sano et al. (2004) found 24
and 10 different subtypes in RJ of honeybees in Europe and Africa, and
these subtypes may be formed by MRJP3 degradation in storage. MRJP4
and MRJP 5 are glycoproteins of 60 kDa and 80 kDa, respectively.
MRJP2-5 are basic proteins (Xin et al., 2016). Post-translational modi­
fication of MRJPs in RJ includes methylation, phosphorylation and
glycosylation. The N-glycosylation of proteins in RJ is related to
developmental regulation, metabolism and immunity. Methylated
modification of proteins plays a key role in the change of biochemical
characteristics. Most of MRJPs are methylated, and methylation in RJ
can promote glycosylation of their proteins. Phosphorylation of MRJP1,
MRJP2 and apolipoprotein III like proteins has been found (Feng et al.,
2015; Zhang et al., 2012).
In addition to MRJPs, RJ also contains a small amount of royalsin,
jelleines, aspirin, royalactina (Fratini, Cilia, Mancini, & Felicioli, 2016).
Royalsin has a molecular weight of 5.5 kDa (Bílikova, Huang, Lin,
Šimuth, & Peng, 2015). Jelleines include jelleine-I (PFKLSLHL-NH2),
jelleine-II (TPFKLSLHL-NH2), jelleine-III (EPFKLSLHL-NH2), jelleine-IV
(TPFKLSLH-NH2). Royalsin and jelleines are common antimicrobial
peptides. A lipid binding protein, apolipophorin-III-like protein, was
also found in RJ, which played an antibacterial role by forming a
protein-lipid complex to bring lipids into the water environment (Han
et al., 2011; Kunugi & Mohammed Ali, 2019).
RJ also contains a lot of free amino acids, among which lysine is the
most abundant (62.43 mg/100 g); followed by proline (58.76 mg/100
g), cystine (21.76 mg/100 g) and aspartic acid (17.33 mg/100 g), amino
acids containing less than 5 mg/100 g are also included, such as valine,
glutamic acid, serine, glycine, cysteine, threonine, alanine, tyrosine,
phenylalanine, hydroxyproline, leucine, isoleucine and glutamine
(Kanbur et al., 2009; Ozcan & Senyuva, 2006).
2.2. Sugars
The main sugars in RJ are glucose, fructose and sucrose. The average
total amount of these three sugars is 13.9%. Fructose and glucose
constitute 90% of the total sugar fraction of RJ, whereas sucrose ac­
counts for 0.8–3.6%. The content of glucose is higher than that of
fructose, and the concentration of sucrose usually changes a lot. RJ also
contains a small amount of maltose, trehalose, etc. The presence or
absence of lactose is uncertain and needs further study and analysis.
Compared to the worker’s food WJ, the content of sugar in RJ is higher
(Kunugi & Mohammed Ali, 2019; Kolayli et al., 2016).

Table 1
Key active components of royal jelly.
Key Active components References

Proteins MRJPs (MRJP1-9); royalsin, jelleines (JelleineI-IV), aspirin, royalactina, apolipophorin-III- (Fratini et al., 2016; Han et al., 2011; Kunugi & Mohammed Ali,
like protein 2019; Xin et al., 2016)
Free amino Gysine, proline, cystine and aspartic acid (Kanbur et al., 2009; Ozcan & Senyuva, 2006)
acids
Sugars Glucose, fructose and sucrose (Kolayli et al., 2016)
Lipids 10-HDA, SA, 3,10-HDecDA (Kocot et al., 2018; Kolayli et al., 2016; Makino et al., 2016)
8-HOC, 9-HAD, 3-HHDA, 2-DecDA
Phenols Pinobanksin, octanoic acid, dodecanoic acid, 1,2-benzenedicarboxylic acid and their esters (Kunugi & Mohammed Ali, 2019)
Flavonoids Naringenin (flavanones), chrysin, acacetin (flavones), isorhamnetin glycosides (flavonols), (López-Gutiérrez et al., 2014)
formononetin (isoflavonoids), etc.
Vitamins Biotin, folate, inositol, niacin, vitamin B6, pantothenic acid, riboflavin, thiamine and vitamin (Kanbur et al., 2009)
E
Minerals K, P, Ca, Mg, Fe, Na, Zn, Cu, Cr, Cd (Adaškevičiūtė et al., 2019)
Other Adenosine monophosphate and its N1 oxide, Ach, testosterone, flavonoids, prolactin and (Hattori et al., 2007; Pasupuleti et al., 2017)
ingredients estradiol

Note: MRJPs, major royal jelly proteins; 2-DecDA, 2-decene-1, 10-dioic acid; 3-HHDA, 3-hydroxydecanoic acid; 3, 10-HDecDA, 3, 10-dihydroxydecanedioic acid; 8-
HOC, 8-hydroxyoctanoic acid; 9-HDA, 9-hydroxy-2-decenoic acid; SA, sebacic acid; 10-HDA, 10-hydroxy-2-decenoic acid; Ach, acetylcholine.

2
J. Guo et al.
2.3. Lipids
Lipids are mainly composed of medium chain fatty acids, containing
8–12 carbon atoms, mainly hydroxyl or dicarboxylic acids (Li, Huang, &
Xue, 2013). About 80–90% of RJ’s lipids are rare free fatty acids. The
main fatty acid is 10-hydroxy-2-decenoic acid (10-HDA) (Kocot, Kiełc­
zykowska, Luchowska-Kocot, Kurzepa, & Musik, 2018; Kolayli et al.,
2016), the content of which is vary from 0.771 to 0.928 g/100 g fresh RJ
(Kokotou, Mantzourani, Babaiti, & Kokotos, 2020). 10-HDA can be used
to evaluate the quality of RJ. RJ also includes sebacic acid (SA) (Makino
et al., 2016), 8-hydroxyoctanoic acid (8-HOC), 3,10-dihydroxydecane­
dioic acid (3,10-HDecDA), 9-hydroxy-2-decenoic acid (9-HDA), 3-
hydroxydecanoic acid (3-HHDA), 2-decene-1,10-dioic acid (2-DecDA)
and a small amount of sterols (Ahmad, Campos, Fratini, Altaye, & Li,
2020; Isidorov, Bakier, & Grzech, 2012; Kolayli et al., 2016). The
pharmacological effect of 10-HDA covers immunomodulatory, antidia­
betic, antibacterial and anticancer effects, etc.
2.4. Phenols, flavonoids
RJ includes 1.28 (μg/mg RJ) total flavonoids and 23.3 (μg/mg RJ)
phenolics (Nabas, Haddadin, Haddadin, & Nazer, 2014). Phenolic sub­
stances consist of pinobanksin, organic acids (such as octanoic acid,
dodecanoic acid, 1,2-benzenedicarboxylic acid) and their esters (Kunugi
& Mohammed Ali, 2019). Flavonoids include the following four cate­
gories: flavanones (hesperetin, naringenin, and isosakuranetin), fla­
vones (chrysin, acacetin, and luteolin, apigenin, and its glycoside),
flavonols (kaempferol and isorhamnetin glycosides), and isoflavonoids
(coumestrol, genistein and formononetin) (López-Gutiérrez, Aguilera-
Luiz, Romero-González, Vidal, & Garrido Frenich, 2014). Phenolic
compounds showed certain antioxidant and antibacterial activities
(Viuda-Martos, Ruiz-Navajas, Fernández-López, & Pérez-Álvarez,
2008).
2.5. Vitamins and minerals
The vitamins in RJ are mainly biotin, folate, inositol, niacin, pan­
tothenic acid, vitamin B6, riboflavin, thiamine and vitamin E (Kanbur
et al., 2009). Besides, RJ also has 1.5% mineral salt (Kocot et al., 2018),
including K, P, Ca, Mg, Fe, Na, Zn, Cu, Cr, Cd, Pb and other major and
trace elements, of which the content of K is the most and Cd is the least
(Adaškevičiūtė, Kaškonienė, Kaškonas, Barčauskaitė, & Maruška, 2019).
2.6. Other ingredients
There are also adenosine monophosphate and adenosine mono­
phosphate N1 oxide in RJ. The latter is the product of the oxidation of
adenosine at the N1 position of adenine base moiety. It only exists in RJ
and is not found in other natural products. Adenosine monophosphate
N1 oxide plays an important physiological role (Hattori, Nomoto,
Fukumitsu, Mishima, & Furukawa, 2007). It can activate mitogen-
activated protein kinase (MAPK)/extracellular signal-regulated kinases
1 or 2 (ERK1/2) pathway through A2A Adenosine receptor to promote
neurite outgrowth and induce pheochromocytoma PC12 cell differen­
tiation. (Hattori et al., 2006; Hattori, Nomoto, Fukumitsu, Mishima, &
Furukawa, 2010). The study shows that the adenosine content of pure RJ
is between 27-50 μg/g, while that of RJ as a dietary supplement is be­
tween 2-173 μg/g (Kim & Lee, 2011). In addition, RJ also contains
acetylcholine (ACh) and hormones (testosterone, flavonoids, prolactin
and estradiol) and other bioactive components (Pasupuleti et al., 2017).
3. Biological functions
As a functional food, RJ plays a variety of biological roles. This paper
mainly focuses on the influence of RJ and its active components on
immunity, lifespan, memory, digestive system, blood glucose, obesity,
3
Journal of Functional Foods 82 (2021) 104514
antibacterial, as well as anticancer effects (Table 2 and Table 3).
3.1. Immunoregulatory function
Among the numerous pharmacological activities of RJ, immuno­
modulatory activity is widely studied. Oka, Emori, Kobayashi, Hayashi,
and Nomoto (2001) found that RJ could inhibit mast cells of
dinitrophenol-keyhole limpet hemocyanin (DNP-KLH) mice to produce
antigen-specific IgE and histamine, restore macrophage function and
improve Th1/Th2 cell response. Similarly, in β-lactoglobulin (β-Lg)
allergic mice, RJ could also reduce the levels of anti β-Lg IgE, IgG in
serum and histamine in plasma, and alleviate allergic symptoms
(Guendouz et al., 2017). Moreover, RJ suppressed the occurrence of
atopic dermatitis-like skin lesions in NC/Nga mice treated with picryl
chloride by down-regulating the production of TNP specific IFN-γ and
up-regulating the expression of iNOS (Taniguchi et al., 2003). Oral RJ
could also reduce the level of serum IL-10, anti ssDNA, dsDNA and
erythrocyte autoantibody in New Zealand Black × New Zealand White
F1 mice prone to human systemic lupus erythematosus (SLE), and
reduce the level of splenic autoreactive B cells (Mannoor et al., 2009).
After 3 months of RJ treatment, CD4+ regulatory T cells and CD8+
regulatory T cells increased, CD4+ T cells decreased significantly, and
disease severity decreased in children with SLE (Zahran et al., 2016).
The immunoregulatory activity of RJ against Graves’ disease was mainly
reflected in the decrease of tumor necrosis factor-α (TNF-α) in lym­
phocytes, the increase of IFN-γ and Th1/Th2 cytokine level ratios (Erem,
Deger, Ovali, & Barlak, 2006). The effective uptake of antigens mediated
by specialized M cells of intestinal-associated lymphoid tissues is an
essential step to induce an efficient intestinal mucosal immune response.
The protease-treated RJ can promote antigen-specific mucosal IgA re­
sponses by enhancing the uptake of antigen by M cells in Caco-2 cells,
thus regulating mucosal immune activity (Kai et al., 2013).
Researches showed that the proteins and fatty acids of RJ exerted
some immunoregulatory activity. MRJP3 suppressed the production of
IL-4, IL-2 and IFN-γ by T cells in vitro, and reduced the levels of anti-
OVA (ovalbumin) IgE and IgG1 in serum of OVA/alum-immunized
mice in vivo (Okamoto et al., 2003). Apolipoprotein III like protein
could enhance the immune response after phosphorylation (Han et al.,
2014). However, studies on the immune activity of 10-HDA had shown
that it could bring down the production of IL-6 induced by lipopoly­
saccharide by reducing the expression of IκB ζ in RAW264 cells, and
inhibit the production of NO by inhibiting the activation of NF-κB (nu­
clear factor kappa-B) induced by LPS and IFN-β (Sugiyama et al., 2013).
Furthermore, 10-HDA could restrain the proliferation of T cells (Gasic
et al., 2007); it could also, together with 3, 10-DDA, exert the immu­
nomodulatory effect in vitro by inhibiting the proliferation of allogeneic
T cells and the production of IL-2 dependent on dendritic cells, and
inhibit the antigen-specific response in vivo (Vucevic et al., 2007). It had
been found that RJ also had the function of regulating immune organs,
among which 10-HDA recovered the weight loss in the body, thymus
and spleen of cyclophosphamide-induced mice, improved the thymus/
spleen index decline, affected DNA/RNA/protein activities, then
restored the proliferation of thymus and spleen cells induced by cyclo­
phosphamide, and enhanced the activity of T cells and B cells (Fan et al.,
2020).
3.2. Antibacterial activity
Jelleines are the family of antimicrobial peptides in RJ. Jelline-I-III
had antimicrobial activities against yeast, Gram-positive and Gram-
negative bacteria. However, no antimicrobial activity was detected in
jelleine-IV (Fontana et al., 2004). Of which, jelleine-I and jelleine-II
were active against Gram-positive Staphylococcus aureus (ATCC 6535),
Staphylococcus saprophyticus and Bacillus subtilis (CCT 2471), while
jelleine-III showed a low activity (Romanelli et al., 2011). The amino
acid sequence contained in jelleine-I is essential for its antimicrobial
J. Guo et al.
Table 2
Biological effects of bioactive compounds in royal jelly.
Bioactive Biological effects
compounds
MRJP1 oligomer Resist intestinal inflammation
Promote the proliferation of IEC-6
cells
MRJP 2 and its Anticancer effect
isomer X1 Promote caspase-dependent
apoptosis and inhibit the
expression of Bcl-2 and p53 in
HepG2 cells.
MRJP3 Immunoregulatory effect
Decrease IL-4, IL-2 and IFN-γ in
vitro and the levels of anti-OVA
IgE and IgG1 in vivo.
MRJPs Improve memory
Improve the spatial memory
ability of the aged rats by
influencing the metabolism of
cysteine, taurine and energy.
Prolong lifespan
By increasing antioxidant activity
and epidermal growth factor
receptor signaling pathway.
Jellines I-III Antibacterial
Inhibit yeast, Gram-positive and
Gram-negative bacteria.
Royalisin Antibacterial
Inhibit Gram-positive bacteria
through damaging cell walls and
cell membranes.
Prolong lifespan
Prolong the lifespan of
Caenorhabditis elegans by
promoting epidermal growth
factor signaling.
Apolipophorin III Immunoregulatory effect
like protein Enhance the immune response
after phosphorylation
10-HDA Immunoregulatory effect
Decrease the production of IL-6 by
reducing the expression of IκB ζ in
RAW264 cells, and inhibit the
production of NO by inhibiting the
activation of NF-κB
Restore the proliferation of
thymus and spleen cells, enhance
the activity of T cells and B cells.
Antibacterial
Suppress Paenibacillus larvae,
animal and human specific
pathogens
10-HDA Prolong lifespan
Prolong the lifespan of
Caenorhabditis elegans.
Antidiabetic effect
Improve hyperglycemia and
insulin resistance by activating the
expression of PGC-1α
Resist intestinal inflammation
Have anti-inflammatory activity
in human colorectal cancer WiDr
cells.
Anticancer effect
Reduce the levels of TRP1, TRP2
and MITF in B16F1 melanoma
cells, inhibit the production of
melanin, and thus inhibited the
pigmentation of mouse skin.
HPO-DAEE (The Anticancer effect
derivative of 10- Promote the apoptosis of human
HDA) lung cancer A549 cells through
ROS-ERK-p38 pathway, through
CHOP pathway partly.
3,10-DDA Immunoregulatory effect
Inhibit the proliferation of
References
(Moriyama et al.,
2015)
(Abu-Serie & Habashy,
2019)
(Okamoto et al., 2003)
(Chen et al., 2017)
(Xin et al., 2016)
(Fontana et al., 2004)
(Shen et al., 2012)
(Detienne, De Haes,
Ernst, Schoofs, &
Temmerman, 2014)
(Han et al., 2014)
(Sugiyama et al.,
2013)
(Fan et al., 2020)
(Šedivá et al., 2018;
Yang et al., 2018)
(Honda et al., 2011)
(Watadani et al., 2017)
(Yang et al., 2018)
(Peng et al., 2017)
(Kamiya et al., 2018)
(Vucevic et al., 2007)
4
Journal of Functional Foods 82 (2021) 104514
Table 2 (continued )
Bioactive Biological effects References
compounds
allogeneic T cells and the
production of IL-2 dependent on
dendritic cells.
ACh Resist intestinal inflammation (Miyauchi-Wakuda
Mediate the excitation of et al., 2019)
intestinal smooth muscle through
M receptor.
AMP-N1 oxide and Anticancer effect (Hattori et al., 2007)
their analogues Promote axonal growth of rat
pheochromocytoma PC12 cells by
activating integrin signaling,
adenosine A2A receptor and
MAPK signaling pathway.
Note: Bcl-2: B-cell lymphoma-2; IL-4: interleukin-4; IFN-γ: interferon-gamma;
OVA: ovalbumin; NF-κB: nuclear factor kappa-B; PGC-1 α, proliferator-
activated receptor gamma, TRP1, TRP2: tyrosinase-related protein 1, 2; MITF:
microphthalmia associated transcription factor; HPO-DAEE: 4-Hydroperoxy-2-
decenoic acid ethyl ester; PGC-1α, proliferator-activated receptor gamma,
coactivator 1 alpha; ROS: Reactive Oxygen Species; ERK: extracellular regulated
protein kinases; M receptor: muscarinic receptor; MAPK: Mitogen activated
protein kinase.
activity, and the presence of the first proline residue is the underlying
reason for the higher efficacy of jelleine-I over other jelleines. N-ter­
minal modification resulted in decreased activity. In fact, the antimi­
crobial activity of jelleine-II, III are lower than jelleine-I (Cabrera, 2014;
Romanelli et al., 2011). Besides, royalisin is a potent antimicrobial
peptide, isolated and purified from the RJ of Asian honeybee Apis cerana
cerana by fusing with glutathione S-transferase (GST) in Escherichia coli
BL21, could inhibit Gram-positive bacteria, Bacillus subtilis, Micrococcus
flavus and Staphyloccocus aureus. It mainly damaged cell walls and cell
membranes. However, it had no antimicrobial activity against Gram-
negative bacteria and fungal strains detected (Shen et al., 2012).
RJ fed to queen bee larvae and added to the jelly of drone and worker
larvae inhibited the growth of European Foulbrood-associated bacteria
by MRJP1 (Vezeteu, Bobiş, Moritz, & Buttstedt, 2017). American foul­
brood is the most serious disease of bee larvae. Food in the larvae is
important for larvae’s resistance to Paenibacillus larvae, which is the
pathogen of American foulbrood. The inhibitory activity of 10-HDA
against Paenibacillus larvae was significantly increased with the
decrease of pH (Šedivá et al., 2018). 10-HDA is also highly effective
against animal or human specific pathogens, including Staphylococcus
aureus, Streptococcus alactolyticus, Staphylococcus intermedius B, Staphy­
lococcus xylosus, Salmonella cholearasuis, Vibro parahaemolyticus and
Escherichia coli (hemolytic) (Yang, Chou, Widowati, Lin, & Peng, 2018).
3.3. Prolong lifespan
Aging is a natural and inevitable life process that is associated with a
gradual decline in physical function (Natarajan et al., 2020). Insulin/
IGF-1 signaling and forkhead box O transcription factor (DAF-16/
FOXO) are major determinants of longevity and are highly conserved in
different species. The nematode Caenorhabditis elegans is an ideal model
organism to study aging and longevity. Increasing the expression of
DAF-16 or decreasing IIS can significantly promote the lifespan and
stress response of Caenorhabditis elegans. HCF-1 (host cell factor) and
SIR-2.1, two cofactors of DAF-16, can interact with DAF-16. Enzyme-
treated RJ (ERJ) is produced by hydrolase digestion of RJ, which breaks
down proteins into amino acids and peptides. It reduces the sensitization
of RJ. It had been shown that both RJ and ERJ can extend the lifespan of
Caenorhabditis elegans by altering the insulin signaling cascade, DAF-16,
and the interactions between DAF-16, HCF-1, SIR-2.1, and FTT-2 (14-3-
3 protein) (Honda et al., 2011; Wang, Cook, Grasso, Cao, & Dong, 2015).
The elderly people also experience weakened immune function during
J. Guo et al. Journal of Functional Foods 82 (2021) 104514

Table 3 Aging is associated with movement disorders that decrease quality of

The biological activities of royal jelly. life. The genetically heterogeneous mice, generated from four different
Biological activities Mechanism of action References inbred lines (BALBC, C57BL/6, C3H and DBA/2), have been used to
detect drugs that have the potential to extend lifespan. The use of RJ
Immunoregulatory Enhance mucosal immune (Kai et al., 2013)
effect (intestinal) reduced age-related motor dysfunction in heterogeneous mice (Oku­
Improve IgA response by mura et al., 2018).
enhancing the uptake of
antigen by M cells 3.4. Improve memory
Antiallergy (Guendouz et al., 2017;
(DNP-KLH, β-Lg allergy, Oka et al., 2001; Taniguchi
atopic dermatitis) et al., 2003) Aging is related to the decline of cognitive function, and is also the
Decrease histamine and IgE, most dangerous factor which causes neurodegenerative diseases (Hal­
increase iNOS loran et al., 2012). Alzheimer’s disease (AD) is a progressive neurode­
Treat autoimmune disease (Erem et al., 2006;
generative disease characterized by the loss of situational memory and
(SLE, Graves’disease) Mannoor et al., 2009;
Reduce TNF-α and IL-10, Zahran et al., 2016) working memory (WM) (Kirova, Bays, & Lagalwar, 2015). Experimental
increase IFN-γ results showed that RJ had a protective effect on the spatial memory
Antibacterial Inhibit animal or human (Fontana et al., 2004; Yang ability of intracerebroventricular injection of streptozotocin (icv-STZ)
specific pathogens (Gram- et al., 2018) rats, icv-STZ is a disease model of sporadic AD (Zamani, Reisi, Alaei, &
positive bacteria et al).
Prolong lifespan Have beneficial effects on (Honda et al., 2011;
Pilehvarian, 2012), the mechanism might be associated with the in­
the lifespan of Okumura et al., 2018; Qiu crease of the retention time for working spatial memory, the prolifera­
Caenorhabditis elegans, et al., 2020) tion of new neurons in the hippocampus, the decrease of oxidative stress
Drosophila and other species level and neurodegeneration in hippocampus (Guardia de Souza E Silva
through EGF and DAF-16/
et al., 2020). By the way, AD models also included 10-month-old APP/
FOXO.
Improve memory Decrease GABA-ergic (Guardia de Souza E Silva PS1 mice. RJ significantly improved memory impairment in APP/PS1
transmission, dopamine 5- et al., 2020; Minami et al., mice by stimulating the cAMP/PKA/CREB/BDNF pathway and inhibit­
HT and levels of oxidative 2016; Pan et al., 2019; ing neuronal apoptosis (You et al., 2018).
stress in the brain Pyrzanowska et al., 2014, Postmenopausal estrogen deficiency can lead to memory impair­
2018)
Antidiabetic effect By resisting hyperglycemia (Sivajothi et al., 2007;
ment, depression and increase susceptibility to AD. RJ improved the
Yoshida et al., 2017) memory impairment of ovariectomized (OVX) rats, suggesting that RJ
Restrain obesity Reduce inflammation and (Petelin et al., 2019) could alleviate the neural symptoms of menopause. In OVX cholesterol-
oxidative stress response fed rabbits, RJ regulated oxidative stress and cholinergic neurotrans­
Anticancer effect Enhance antioxidant (Filipič et al., 2015; Zhang
mitter levels by reducing amyloid-beta, acetylcholinesterase and MDA
activity by SOD, T-AOC and et al., 2017)
GR, inhibit the proliferation levels and increasing the activities of choline acetyltransferase and su­
of cancer cells and induce peroxide dismutase (SOD) in the brain, thus improving memory
apoptosis impairment behavior (Minami et al., 2016; Pan et al., 2019). In addition,
Effects on the Improve intestinal (Karaca et al., 2010, 2012; dopamine and 5-HT in the prefrontal cortex of RJ-treated rats decreased,
digestive system inflammation: TNF-α, IL-1β, Karaca, Uz, Demirtas,
NF-κB Karaboga, & Can, 2015)
and the concentration and metabolic rate of metabolites increased, in
Reduce liver damage: (Kamakura et al., 2005) which 5-HT and dopamine can affect the cognitive function of the pre­
antioxidant activity frontal cortex in the process of WM (Pyrzanowska et al., 2014). Long-
Note: DNP-KLH: dinitrophenol-keyhole limpet hemocyanin; β-Lg: β-lactoglob­
term administration of RJ, striatal gamma-aminobatyric acid (GABA)-
ulin; iNOS: inducible nitric oxide synthase; SLE: systemic lupus erythematosus; ergic transmission and GABA concentration of naturally aged Wistar
TNF-α: tumor necrosis factor-α; IL-10: interleukin-10; EGF: epidermal growth male rats was decreased. The decrease of GABA concentration led to the
factor; DAF-16/FOXO: forkhead box O transcription factor; GABA: gamma- increase of dopamine transmission activity and the improvement of
aminobatyric acid; SOD: superoxide dismutase; T-AOC: total antioxidant capa­ spatial memory (Pyrzanowska et al., 2018). Deeply, among the active
bility; GR: glutathione reductase; NO: nitric oxide. components of RJ, MRJPs could improve the spatial memory ability of
the aged rats, and it was realized by influencing the metabolism of
aging (known as immunosenescence), which can make them more sus­ cysteine, taurine and energy (Chen, Liu, Wan, Lai, & Shen, 2017). In
ceptible to pathogens. The survival time of Caenorhabditis elegans on conclusion, RJ showed a strong ability to improve memory.
different bacterial pathogens was prolonged when treated with RJ. This
result suggested that RJ may be helpful in delaying immunesenescence 3.5. Effects on the digestive system
(Natarajan et al., 2020). The active components of RJ, 10-HDA and
royalactin, play a certain role in prolonging the lifespan of Caeno­ 3.5.1. Resist intestinal inflammation
rhabditis elegans. Among them, royalactin acts by promoting epidermal Intestinal diseases, like diarrhea, is one of the highest morbidity rates
growth factor (EGF) signaling in Caenorhabditis elegans (Detienne, De and fatality rates in the world (Kosek, Bern, & Guerrant, 2003). In­
Haes, Ernst, Schoofs, & Temmerman, 2014; Honda et al., 2011). flammatory bowel disease is a chronic intestinal disease that can cause
In addition, Drosophila can also be used to study aging, such as to test diarrhea and abdominal pain, including Crohn’s disease and ulcerative
the anti-aging effects of dietary antioxidants. Recent studies had found colitis (Mowat et al., 2011), which seriously threatens human health and
that the mixture of ERJ and collagen peptide slowed down the aging of even leads to death. It was found that RJ had a certain effect on intestinal
Drosophila, including extending the average lifespan of the natural aging diseases, which is shown in Fig. 1. Karaca et al. (2010, 2012, 2015)
Drosophila, increasing total superoxide dismutase (T-SOD), glutathione found that RJ could improve the artificially induced colitis: in acetic
peroxidase (GSH-Px), catalase (CAT) activity and reducing the peroxides acid-induced colitis, the treatment of RJ could reduce the ulcerative
product malondialdehyde (MDA) and protein carbonyl (Qiu et al., erosion of colon tissue, the increase of mast cells caused by colitis, the
2020). MRJPs is an important part of RJ to prolong the lifespan of increase of CD3+ and CD45+ T cells, while the number of CD68+ mac­
Drosophila. Its action also benefits from the increase of antioxidant ac­ rophages and CD5+T cells did not change significantly; the pretreatment
tivity. By the way, it is also related to the signaling pathway mediated by of RJ in colon of rats with colitis induced by 2, 4, 6-trinitrobenzene
EGFR (Xin et al., 2016). sulfonic acid (TNBS) inhibited CD3+, CD5+, CD8+ and CD45+ T cells,
pro-inflammatory cytokines, IL-1β and TNF-α, increased the production

5
J. Guo et al.
Fig. 1. The regulation effect of royal jelly on intestinal inflammation. The treatment of RJ inhibited the increase of CD3+, CD5+, CD8+ and CD45+ T cells, pro-
inflammatory cytokines, IL-1β, TNF–α, and the expression of key inflammatory mediators (COX-2 and NF-κB) in the colon of rats with colitis. TNBS: 2, 4, 6-trinitro­
benzene sulfonic acid; IL-1β: interleukin-1β; TNF-α: tumor necrosis factor-α; NF-κB: nuclear factor kappa-B; COX-2: cyclooxygenase-2.
of anti-inflammatory cytokines IL-10. In the same TNBS colitis experi­
ment, RJ was also found to increase the activity of GSH-Px, reduce the
expression of key inflammatory mediators cyclooxygenase-2 (COX-2)
and NF-κB, and reduce the damage caused by tumor necrosis factor, and
show anti-inflammatory and anti-oxidation effects (Manzo, 2015). In
addition, some studies have found that the intestinal microbiome of IBD
patients changed, in which Bacteroidetes increased significantly and
Firmicides decreased significantly (Jones-Hall & Nakatsu, 2016). RJ had
certain antibacterial activity, among which MRJP 2–5 had antibacterial
activity against Gram-negative E. coli (Park et al., 2019). Moreover,
jellenie I-IV are also important antibacterial ingredients in RJ (Ahmad
et al., 2020). However, whether RJ affects the intestinal microbiome of
IBD patients and healthy people is still unknown.
Studies showed that 10-HDA had anti-inflammatory activity in
human colorectal cancer WiDr cells and had antibacterial activity
against animal pathogens in vitro. This showed that 10-HDA, as an anti-
inflammatory agent and bactericidal agent, could well protect human
gastrointestinal tract (Yang et al., 2018). ACh is the main excitatory
neurotransmitter in the intestinal nervous system. It can mediate the
excitation of intestinal smooth muscle through the muscarinic receptor
(M receptor) (Hansen, 2003). The results showed that ACh contained in
RJ could induce contraction of ileal smooth muscle through M receptor,
but single oral administration of RJ was not enough to increase intestinal
motility or improve constipation. Eating regular doses of honey will also
cause abdominal symptoms due to poor carbohydrate absorption.
However, the content of carbohydrates in RJ is lower than that in honey.
In conclusion, RJ will not cause serious symptoms, like diarrhea under
normal circumstances (Ladas, Haritos, & Raptis, 1995; Miyauchi-
Wakuda et al., 2019; Pasupuleti et al., 2017). MRJP1 is the most
important protein in RJ, MRJP1 oligomer is the main proliferation factor
in RJ, which can promote the proliferation of IEC-6 cells and has certain
6
Journal of Functional Foods 82 (2021) 104514
heat resistance, while MRJP2 and MRJP3 have no effect (Moriyama, Ito,
Omote, Miura, & Tsumoto, 2015). In daily life, oral administration is the
most commonly used drug delivery method of RJ, and whether MRJPs
can withstand the digestive effect is very important for RJ to play its
biological role. MRJP2 had high stability during digestion of pepsin,
intestinal trypsin and chymotrypsin, and because the time of passing
through the stomach when taking it on an empty stomach is far shorter
than after eating (Weitschies, Kosch, Mönnikes, & Trahms, 2005), it can
be inferred that MRJPs may reach a better concentration when taking it
on an empty stomach (Mureşan, Schierhorn, & Buttstedt, 2018).
3.5.2. Reduce liver damage
The liver is one of the most important organs in human body (Abu-
Serie & Habashy, 2019), RJ could regulate 267 liver genes in mice, of
which 148 genes were up-regulated and 119 genes were down-
regulated. Among them, RJ could reduce the gene expression of squa­
lene epoxidase (SQLE) and its transcription factor sterol regulatory
element-binding protein-1, and increase the gene expression of low-
density lipoprotein receptor (LDLR) in the liver, while the changes of
SQLE and LDLR can cause the decrease of cholesterol (Kamakura,
Moriyama, & Sakaki, 2006). At the same time, RJ could also up-regulate
the gene expression level of GST and GSH-Px (Kamakura et al., 2005).
Because the liver is the main organ responsible for metabolism in the
body, the intake of toxic chemicals and drugs mainly affects the function
of the liver (Ramakrishnan, Elangovan, & Pari, 2017). And alanine
aminotransferase (ALT) is the specific enzyme when the liver is
damaged, and aspartate aminotransferase (AST) will also increase when
the liver is damaged (Kanbur et al., 2009). Chemical toxicants such as
cadmium, CCl4 and lambda-cyhalothrin (LCT) could reduce glutathione
(GSH) and increase MDA, a product of lipid peroxidation. Fluoride also
reduced CAT and SOD. RJ could restore antioxidant (SOD, CAT, GSH)
J. Guo et al.
and MDA in liver and reduce AST, ALT levels, alleviating liver damage
(Cavuşoğlu, Yapar, Oruç, & Yalçın, 2011; Çavuşoğlu, Yapar, & Yalçin,
2009; Cemek et al., 2010; Cemek et al., 2012; Kanbur, Eraslan, Silici,
and Karabacak, 2009). RJ played a similar role in liver injury induced by
azathioprine and paracetamol (Ahmed, Khalaf, Moselhy, & Safwat,
2014; El-Nekeety et al., 2007). Cisplatin (CDDP) and taxol (TXL) could
not only cause liver injury similar to the above, but also reduce the levels
of GST and GSH-Px and increase the number of apoptosis in hepatocytes.
TXL could still cause the levels of alkaline phosphatase (ALP) and lactic
dehydrogenase (LDH), as well as the expression of liver growth regu­
latory factor E2f1 to increase, and the expression of liver growth regu­
latory factor c-myc to decrease, which were relieved after the treatment
of RJ (Karadeniz et al., 2011; Malekinejad, Fani, Shafiee-Roodbari,
Delkhosh-Kasmaie, & Rezaei-Golmisheh, 2017). In addition, RJ also
reduced iNOS and IL-1β in the liver, up-regulated nuclear factor E2-
related factor-2 (Nrf2) and anti-apoptotic protein Bcl-2 but down-
regulated caspases-3 and Bax to resist the toxic effect of Cd on the
liver (Almeer et al., 2018). Later studies found that MRJP2 and its
subtype X1 were important active components in RJ to resist CCl4-
induced liver injury (Abu-Serie & Habashy, 2019).
RJ could also improve the oxidative damage of the liver caused by
chronic stresses, like restraint and cold stress. These stresses caused
some damages to the liver, which were mainly manifested in the
decrease of GSH, GPX and SOD activities, as well as the increase of
glutathione reductase (GR) and MDA (Caixeta et al., 2018). In conclu­
sion, RJ resisted liver injury caused by many factors by improving the
antioxidant capacity of the liver (Fig. 2).
3.6. Antidiabetic effect
Diabetes is a disease characterized by elevated blood sugar, while
type 2 diabetes (T2DM) is a chronic metabolic disorder caused by insulin
secretion disorders and insulin resistance (DeFronzo et al., 2015; Maleki
7
Journal of Functional Foods 82 (2021) 104514
et al., 2019). Impaired insulin action in adipose tissue, liver, and skeletal
muscle is a major contributor to type 2 diabetes (Yoshida et al., 2017).
Diabetes can cause liver diseases such as hepatic cirrhosis, hepatocel­
lular carcinoma, hepatitis C, nonalcoholic fatty liver disease, and acute
liver failure (Guven et al., 2006). As an insulin-dependent tissue, the
liver is essential for the homeostasis of blood sugar and lipids (Sivajothi,
Dey, Jayakar, & Rajkapoor, 2007). RJ significantly reduced the serum
fasting blood glucose and serum glycosylated hemoglobin levels, and
increased the insulin concentration in women with type 2 diabetes
(Pourmoradian, Mahdavi, Mobasseri, Faramarzi, & Mobasseri, 2014).
The reduction of serum glycosylated hemoglobin level further reduced
the risk of microvascular and macrovascular complications, which was
beneficial to the prognosis of type 2 diabetes (Woerle et al., 2007). In
addition, RJ also increased the serum apolipoprotein A-I (ApoA-I) and
changed the apolipoprotein B (ApoB)/ApoA-I ratios in patients with
type 2 diabetes (Khoshpey et al., 2016). The STZ destroyed pancreatic
β-cells, and RJ decreased serum fasting blood glucose, AST, ALT, and
ALP levels and increased insulin, albumin, and total protein levels in
STZ-induced diabetic rats (Ghanbari, Nejati, & Khazaei, 2016). RJ also
enhanced serum total antioxidant capacity and cut down homeostasis
model assessment for insulin resistance in diabetic patients (Shidfar
et al., 2015). Long-term administration of RJ inhibited glucose-6-
phosphatase, a key enzyme in hepatic gluconeogenesis, by inducing
the expression of adiponectin and adiponectin receptor 1 mRNA and
phosphorylated AMP-activated protein kinase in abdominal fat, thereby
improving hyperglycemia (Yoshida et al., 2017). However, one experi­
ment pointed out that RJ did not have a beneficial effect on blood
glucose markers, which might be caused by methodological problems
and dietary factors, and further research is still needed (Mahboobi,
Jafarnejad, & Eftekhari, 2019). Besides, RJ enhanced the metabolic
thermogenesis of brown adipose tissue in mice, thereby improving hy­
perglycemia and liver steatosis induced by a high-fat diet (Yoneshiro
et al., 2018). When the proliferator-activated receptor gamma,
Fig. 2. Protective effects of royal jelly on the
damage of the liver caused by toxic chemicals,
drugs and chronic stress. Chemical toxicants
(cadmium, CCl4 and LCT), chronic stress (re­
straint/cold) and cancer drugs (CDDP and TXL)
could cause oxidative damage of the liver. RJ can
resist multifactorial liver damage by increasing
antioxidant capacity and reducing hepatocyte
apoptosis. LCT: Lambda-cyhalothrin; CDDP:
Cisplatin; TXL: Taxol; GST: glutathione S-trans­
ferase; GSH-Px: glutathione peroxidase; GSH:
glutathione; MDA: malondialdehyde; CAT: cata­
lase; SOD: superoxide dismutase; ALP: alkaline
phosphatase; LDH: lactic dehydrogenase. The “↑”
represents an increase or promotion, the “↓”
represents a decrease, and the “ ′′
represents
inhibiting effect.
J. Guo et al.
coactivator 1 alpha (PGC-1α) is moderately increased, it can regulate
glucose transport and insulin sensitivity (Benton et al., 2010), while 10-
HDA could improve hyperglycemia and insulin resistance by activating
the expression of PGC-1α (Watadani et al., 2017).
3.7. Restrain obesity
Obesity is a chronic disease that has been widely attended and can be
caused by a variety of factors. It is characterized by high oxidation state
and chronic activation of macrophages in peripheral tissues. Therefore,
reducing inflammation and oxidative stress response are relatively
effective treatments. Supplementation of RJ for overweight adults
decreased total cholesterol, inflammatory marker C-reactive protein,
increased anti-inflammatory marker adiponectin, serum total antioxi­
dant capacity, endogenous antioxidant bilirubin, leptin, and so on.
Consequently, it has a positive effect on overweight adults (Petelin et al.,
2019). Studies that added RJ to mice on a high-fat diet found improved
inflammation and increased levels of Irisin, metabolic thermogenesis of
brown adipose tissue (Irandoost et al., 2020; Yoneshiro et al., 2018). It
was found that although 10-HDA was an important active ingredient in
RJ, it could not prevent obesity (Watadani et al., 2017).
3.8. Anticancer effect
RJ also played a great role in the treatment of cancer. For example,
RJ could improve the myelosuppression in mice with Ehrlich ascites
tumor, inhibit the hematopoietic function of the spleen, and improve the
survival rate of mice, which might be caused by the decrease of pros­
taglandin E2. Prostaglandin E2, a product of arachidonic acid meta­
bolism, regulates the proliferation of lymphocytes and inhibits the
tumoricidal activity of normal macrophages. It also regulates immune
responses including T lymphocyte mitosis, lymphokine production, and
tumor toxicity mediated by macrophages and natural killer cells (Bin­
coletto, Eberlin, Figueiredo, Luengo, & Queiroz, 2005). Furthermore, RJ
inhibited the proliferation of human breast cancer MCF-7 cells induced
by environmental estrogen bisphenol A (Nakaya et al., 2007). When
human Caco-2 cells were treated with a mixture of RJ and human
interferon α (2:1), the level of glutathione decreased significantly, while
lipid peroxidation increased significantly (Filipič et al., 2015). RJ
treatment of 4T1 breast cancer mice model, especially prophylactic-
therapeutic method, reduced the weight of tumor, increased the activ­
ity of SOD, total antioxidant capacity (T-AOC), GR, and improved the
antioxidant activity of liver, kidney and serum (Zhang, Shao, Geng, &
Su, 2017).
The mixture of RJ and PBS was slowly shaken overnight at 4◦ C and
then centrifuged at 4 ◦ C for 12 000g for 10 min. The supernatant ob­
tained by separation is RJ Extract. It had strong cytotoxicity on human
glioblastoma multiforme (Borawska et al., 2014). RJP30, a component
obtained by precipitation of RJ crude protein extract (RJCP) with 30%
ammonium sulfate, had a cytotoxic effect on HeLa human cervicouterine
carcinoma cells and could reduce the number of cells (Salazar-Olivo &
Paz-González, 2005). The lipophilic extract of RJ could inhibit the
proliferation of human neuroblastoma (SH-SY5Y) cells, showing the
preventive effect on human neuroblastoma (Gismondi, Trionfera, Can­
uti, Di Marco, & Canini, 2017). AMP-N1 oxide and its analogues were
the only active components in RJ that could promote the axonal growth
of rat pheochromocytoma PC12 cells by integrin signaling and adeno­
sine A2A receptor activation. In addition, AMP-N1 oxide and their an­
alogues could also play a role by activating the MAPK signaling pathway
(Hattori et al., 2007). MRJP2 and its subtype X1 could promote caspase-
dependent apoptosis and inhibit the expression of Bcl-2 and p53 in
HepG2 cells (Abu-Serie & Habashy, 2019). 10-HDA could reduce the
levels of tyrosinase-related protein 1, 2 (TRP1, TRP2) and
microphthalmia-associated transcription factor (MITF) in B16F1 mela­
noma cells, and inhibit the production of melanin, then inhibited the
pigmentation of mouse skin (Peng, Sun, Lin, Kuo, & Li, 2017). The
8
Journal of Functional Foods 82 (2021) 104514
derivative of 10-HDA, 4-hydroperoxy-2-decenoic acid ethyl ester (HPO-
DAEE), could promote the apoptosis of human lung cancer A549 cells
through ROS-ERK-p38 pathway, through CHOP pathway partly, which
meaning the promoting effect of HPO-DAEE on apoptosis was partly
dependent on endoplasmic reticulum stress-related pathways (Kamiya
et al., 2018). The biological activity and mechanism of RJ can be seen in
Table 3.
4. Conclusion
As a kind of bee product and functional food, RJ is rich in major
active ingredients such as major RJ proteins, lipids, phenols and flavo­
noids, which play an active role in maintaining human health. Although
the effects of RJ in anti-inflammatory, anti-cancer, immune regulation,
memory regulation and other aspects have been proved, whether there
are some potential functions is still unknown. For example, the inter­
action between RJ and intestinal tract may become more complex due to
the existence of the intestinal flora. In addition, some of the active in­
gredients may not have been found and the roles of the active in­
gredients have not been fully studied, which need to be further explored.
Moreover, more research evidence can also help RJ to be better used and
more profitable.
5. Funding*
This work was supported by the National Natural Science Foundation
of China (grant nos. 31972633, 31972087, 31572476, 31272483) and
National Natural Science Foundation of Beijing (grant no. 6172022).
6. Ethics Statement
None.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
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