Introduction: Mulberry (Morus alba L.) leaves are widely used in traditional Chinese medicine for their antioxidant, anti-inflammatory, antibacterial, anti-obesity, antidiabetic, antiatherosclerotic, and anticancer properties. The current study aimed to investigate the effect of mulberry leaf extract (MLE) on Staphylococcus aureus (S. aureus)-induced conjunctivitis (5 × 109 colony-forming units, 0.5 mL/eye) in a rabbit model. Methods: Rabbits were treated with MLE (5 mL/kg·d−1 and 10 mL/kg·d−1), 0.9% saline, pearl bright eye (PBE) drops, or erythromycin eye ointment (EEO) group for 5 days. The ocular infection symptoms, bacterial negative conversion rate, and conjunctival histopathological changes of rabbits in each group were observed. The expression of caspase-1, apoptosis-associated speck-like protein containing a caspase recruitment domain, NOD-like receptor leucine-rich pyrin domain-containing protein 3 (NLRP3), interleukin (IL)-18, IL-6, IL-1β, TNFα, Keap1, and nuclear factor erythroid 2-related factor 2 (Nrf2) in conjunctival tissue of rabbits were detected by quantitative real-time reverse transcription PCR and/or Western blot analysis. Results: The results showed that MLE treatment significantly reduced the clinical sign scores of conjunctivitis, alleviated clinical signs, and decreased bacterial load, and histological damage in a time- and dose-dependent manner was compared to that in the control group. The antibacterial and anti-inflammatory activities of MLE (10 mL/kg·d−1) were similar to those of the positive control drug PBE and EEO. In addition, MLE significantly decreased the levels of pro-inflammatory cytokines, downregulated the NLRP3 inflammasome, and upregulated the Nrf2 system. Conclusions: MLE is effective in alleviating S. aureus-induced conjunctivitis in rabbits, and this mechanism is associated with the inhibition of the NLRP3 inflammasome and activation of the Nrf2 system to regulate pro-inflammatory signaling.

1.
Liang
QF
,
Du
XH
,
Su
GY
,
Zhang
Y
,
Wang
ZQ
,
Cao
K
.
Genetic typing and susceptibility testing of strains from Staphylococcus aureus keratitis or conjunctivitis patients
.
Zhonghua Yan Ke Za Zhi
.
2018 Oct 11
;
54
(
10
):
767
74
. .
2.
O’Callaghan
RJ
.
The pathogenesis of Staphylococcus aureus eye infections
.
Pathogens
.
2018 Jan 10
;
7
(
1
):
9
. .
3.
Cavuoto
K
,
Zutshi
D
,
Karp
CL
,
Miller
D
,
Feuer
W
.
Update on bacterial conjunctivitis in South Florida
.
Ophthalmology
.
2008 Jan
;
115
(
1
):
51
6
. .
4.
Haas
W
,
Pillar
CM
,
Torres
M
,
Morris
TW
,
Sahm
DF
.
Monitoring antibiotic resistance in ocular microorganisms: results from the Antibiotic Resistance Monitoring in Ocular micRorganisms (ARMOR) 2009 surveillance study
.
Am J Ophthalmol
.
2011 Oct
;
152
(
4
):
567
74.e3
. .
5.
Shu
YS
,
He
D
,
Li
W
,
Wang
ML
,
Zhao
S
,
Liu
LL
,
Hepatoprotective effect of Citrus aurantium L. against APAP-induced liver injury by regulating liver lipid metabolism and apoptosis
.
Int J Biol Sci
.
2020
;
16
(
5
):
752
65
. .
6.
Que
L
,
Yang
G
,
Li
Y
,
Shan
F
,
Chen
M
,
Huang
LQ
.
Overview of revision of the catalogue of the substances traditionally considered as both food and Chinese medicine
.
Chin Pharmaceut J
.
2017
;
52
(
7
):
521
4
. .
7.
El-Sayyad
HI
,
El-Sherbiny
MA
,
Sobh
MA
,
Abou-El-Naga
AM
,
Ibrahim
MA
,
Mousa
SA
.
Protective effects of Morus alba leaves extract on ocular functions of pups from diabetic and hypercholesterolemic mother rats
.
Int J Biol Sci
.
2011 Jun
;
7
(
6
):
715
28
. .
8.
Mahmoud
AM
,
El-Twab
SMA
,
Abdel-Reheim
ES
.
Consumption of polyphenol-rich Morus alba leaves extract attenuates early diabetic retinopathy: the underlying mechanism
.
Eur J Nutr
.
2017 Jun
;
56
(
4
):
1671
84
. .
9.
Assiri
AMA
,
El-Beeh
ME
,
Amin
AH
,
Ramadan
MF
.
Ameliorative impact of Morus alba leaves’ aqueous extract against embryonic ophthalmic tissue malformation in streptozotocin-induced diabetic rats
.
Biomed Pharmacother
.
2017 Nov
;
95
:
1072
81
. .
10.
Ministry of Public Health
..
Traditional Chinese medicine prescription preparation
.
1998
;
17
:
189
.
11.
Committee SP
.
Pharmacopoeia of the People’s Republic of China
.
China Medical Science and Technology Press
;
2015 Aug 21
.
Vol. 1
; p.
297
310
.
12.
Du
XH
,
Zheng
C
,
Gu
XP
,
Qu
CH
,
Ni
J
.
Study on effect and mechanism of Zhenzhu Mingmu eye drops protected HCEC against hypermeability-induced injury
.
Chin Tradit Herb Drugs
.
2017
;
48
(
16
):
3425
30
. .
13.
Da Fonseca
DV
,
Da Silva Maia Bezerra Filho
C
,
Lima
TC
,
de Almeida
RN
,
de Sousa
DP
.
Anticonvulsant essential oils and their relationship with oxidative stress in epilepsy
.
Biomolecules
.
2019 Dec 6
;
9
(
12
):
835
. .
14.
Kulkarni
M
,
Sawant
N
,
Kolapkar
A
,
Huprikar
A
,
Desai
N
.
Borneol: a promising monoterpenoid in enhancing drug delivery across various physiological barriers
.
AAPS PharmSciTech
.
2021 Apr 28
;
22
(
4
):
145
. .
15.
Song
LN
,
Li
HR
,
Wang
HY
,
Li
Z
,
Zhao
WC
,
Lu
FZ
,
Preparation and properties of thermosensitive in situ gel for ophthalmic formulation containing pearl hydrolyzate
.
Acta Pharm Sin
.
2012
;
51
(
10
):
1622
8
. .
16.
Artlett
CM
.
Inflammasomes in wound healing and fibrosis
.
J Pathol
.
2013 Jan
;
229
(
2
):
157
67
. .
17.
Ahmed
SM
,
Luo
L
,
Namani
A
,
Wang
XJ
,
Tang
X
.
Nrf2 signaling pathway: pivotal roles in inflammation
.
Biochim Biophys Acta Mol Basis Dis
.
2017 Feb
;
1863
(
2
):
585
97
. .
18.
Wakamatsu
TH
,
Dogru
M
,
Ayako
I
,
Takano
Y
,
Matsumoto
Y
,
Ibrahim
OMA
,
Evaluation of lipid oxidative stress status and inflammation in topic ocular surface disease
.
Mol Vis
.
2010 Nov
;
16
:
2465
75
. .
19.
Hennig
P
,
Garstkiewicz
M
,
Grossi
S
,
Di Filippo
M
,
French
LE
,
Beer
HD
.
The crosstalk between Nrf2 and inflammasomes
.
Int J Mol Sci
.
2018 Feb 13
;
19
(
2
):
562
. .
20.
Kim
M
,
Nam
DG
,
Ju
WT
,
Choe
JS
,
Choi
AJ
.
Response surface methodology for optimization of process parameters and antioxidant properties of mulberry (Morus alba L.) leaves by extrusion
.
Molecules
.
2020 Nov 10
;
25
(
22
):
5231
. .
21.
Zhou
WW
,
Liang
X
,
Dai
PB
,
Chen
Y
,
Zhang
YX
,
Zhang
M
,
Alteration of phenolic composition in lettuce (Lactuca sativa L.) by reducing nitrogen supply enhances its anti-proliferative effects on colorectal cancer cells
.
Int J Mol Sci
.
2019 Aug 28
;
20
(
17
):
4205
. .
22.
Sloop
GD
,
Moreau
JM
,
Conerly
LL
,
Dajcs
JJ
,
O’Callaghan
RJ
.
Acute inflammation of the eyelid and cornea in Staphylococcus keratitis in the rabbit
.
Invest Ophthalmol Vis Sci
.
1999 Sep
;
40
(
2
):
385
91
. .
23.
Tang
A
,
Balzli
CL
,
Caballero
AR
,
McCormick
CC
,
Taylor
SD
,
O’Callaghan
RJ
.
Staphylococcus aureus infection of the rabbit cornea following topical administration
.
Curr Eye Res
.
2012 Dec
;
37
(
12
):
1075
83
. .
24.
Zhang
Z
,
Abdel-Razek
O
,
Hawgood
S
,
Wang
G
.
Protective role of surfactant protein D in ocular Staphylococcus aureus infection
.
PLoS One
.
2015 Sep
;
10
(
9
):
e0138597
. .
25.
Darden
JE
,
Scott
EM
,
Arnold
C
,
Scallan
EM
,
Simon
BT
,
Suchodolski
JS
.
Evaluation of the bacterial ocular surface microbiome in clinically normal cats before and after treatment with topical erythromycin
.
PLoS One
.
2019
;
14
(
10
):
e0223859
. .
26.
Magone
MT
,
Chan
CC
,
Rizzo
LV
,
Kozhich
AT
,
Whitcup
SM
.
A novel murine model of allergic conjunctivitis
.
Clin Immunol Immunopathol
.
1998 Apr
;
87
:
75
84
. .
27.
Tanaka
J
,
Tanaka
H
,
Mizuki
N
,
Nomura
E
,
Ito
N
,
Nomura
N
,
Semaphorin 3A controls allergic and inflammatory responses in experimental allergic conjunctivitis
.
Int J Ophthalmol
.
2015 Feb
;
8
(
1
):
1
10
. .
28.
Faggion
CM
 Jr
,
Listl
S
,
Giannakopoulos
NN
.
The methodological quality of systematic reviews of animal studies in dentistry
.
Vet J
.
2012 May
;
192
(
2
):
140
7
. .
29.
He
XR
,
Fang
JC
,
Ruan
YL
,
Wang
XX
,
Sun
Y
,
Wu
N
,
Structures, bioactivities and future prospective of polysaccharides from Morus alba (white mulberry): a review
.
Food Chem
.
2018 Apr 15
;
245
:
899
910
. .
30.
Peng
CH
,
Lin
HT
,
Chung
DJ
,
Huang
CN
,
Wang
CJ
.
Mulberry leaf extracts prevent obesity-induced NAFLD with regulating adipocytokines, inflammation and oxidative stress
.
J Food Drug Anal
.
2018 Apr
;
26
(
2
):
778
87
. .
31.
Gryn-Rynko
A
,
Bazylak
G
,
Olszewska-Slonina
D
.
New potential phytotherapeutics obtained from white mulberry (Morus alba L.) leaves
.
Biomed Pharmacother
.
2016 Sep
;
84
:
628
36
. .
32.
Rodrigues
EL
,
Marcelino
G
,
Silva
GT
,
Figueiredo
PS
,
Garcez
WS
,
Corsino
J
,
Nutraceutical and medicinal potential of the Morus species in metabolic dysfunctions
.
Int J Mol Sci
.
2019 Jan 14
;
20
(
2
):
301
. .
33.
Azari
AA
,
Barney
NP
.
Conjunctivitis: a systematic review of diagnosis and treatment
.
JAMA
.
2013 Oct 23
;
310
(
16
):
1721
9
. .
34.
Cluver
C
,
Novikova
N
,
Eriksson
DO
,
Bengtsson
K
,
Lingman
GK
.
Interventions for treating genital Chlamydia trachomatis infection in pregnancy
.
Cochrane Database Syst Rev
.
2017 Sep 22
;
9
:
CD010485
. .
35.
Chen
JY
,
Zhu
GY
,
Su
XH
,
Wang
R
,
Liu
J
,
Liao
K
,
Identification of ethylparaben as the antimicrobial substance produced by Brevibacillus brevis FJAT-0809-GLX
.
Microbiol Res
.
2015 Mar
;
172
:
48
56
. .
36.
Sun
LB
,
Yu
T
,
Guo
JL
,
Zhang
ZB
,
Hu
Y
,
Xiao
X
,
The estrogenicity of methylparaben and ethylparaben at doses close to the acceptable daily intake in immature Sprague-Dawley rats
.
Sci Rep
.
2016 Apr 28
;
6
:
25173
. .
37.
Liu
Y
,
Li
XM
,
Xie
C
,
Luo
XZ
,
Bao
YG
,
Wu
B
,
Prevention effects and possible molecular mechanism of mulberry leaf extract and its formulation on rats with insulin-insensitivity
.
PLoS One
.
2016 Apr
;
11
(
4
):
e0152728
. .
38.
Burton
MJ
,
Ramadhani
A
,
Weiss
HA
,
Hu
V
,
Massae
P
,
Burr
SE
,
Active trachoma is associated with increased conjunctival expression of IL17A and profibrotic cytokines
.
Infect Immun
.
2011 Dec
;
79
(
12
):
4977
83
. .
39.
Eo
H
,
Lim
Y
.
Combined mulberry leaf and fruit extract improved early stage of cutaneous wound healing in high-fat diet-induced obese mice
.
J Med Food
.
2016 Feb
;
19
(
2
):
161
9
. .
40.
Liu
CJ
,
Lin
JY
.
Anti-inflammatory effects of phenolic extracts from strawberry and mulberry fruits on cytokine secretion profiles using mouse primary splenocytes and peritoneal macrophages
.
Int Immunopharmacol
.
2013 Jun
;
16
(
2
):
165
70
. .
41.
Chen
Y
,
Hong
X
.
Effects of carvedilol reduce conjunctivitis through changes in inflammation, NGF and VEGF levels in a rat model
.
Exp Ther Med
.
2016 May
;
11
(
5
):
1987
92
. .
42.
Wei
CC
,
Kung
YJ
,
Chen
CS
,
Chang
CY
,
Lin
CJ
,
Tien
PT
,
Allergic conjunctivitis-induced retinal inflammation promotes myopia progression
.
EBioMedicine
.
2018 Feb
;
28
:
274
86
.
43.
Taniguchi
K
,
Karin
M
.
NF-kappaB, inflammation, immunity and cancer: coming of age
.
Nat Rev Immunol
.
2018 May
;
18
(
5
):
309
24
.
44.
Fink
SL
,
Cookson
BT
.
Apoptosis, pyroptosis, and necrosis: mechanistic description of dead and dying eukaryotic cells
.
Infect Immun
.
2005 Apr
;
73
(
4
):
1907
16
. .
45.
Medvedev
R
,
Ploen
D
,
Spengler
C
,
Elgner
F
,
Ren
H
,
Bunten
S
,
HCV-induced oxidative stress by inhibition of Nrf2 triggers autophagy and favors release of viral particles
.
Free Radic Biol Med
.
2017 Sep
;
110
:
300
15
. .
46.
Staitieh
BS
,
Ding
L
,
Neveu
WA
,
Spearman
P
,
Guidot
DM
,
Fan
X
.
HIV-1 decreases Nrf2/ARE activity and phagocytic function in alveolar macrophages
.
J Leukoc Biol
.
2017 Aug
;
102
(
2
):
517
25
. .
47.
Kobayashi
EH
,
Suzuki
T
,
Funayama
R
,
Nagashima
T
,
Hayashi
M
,
Sekine
H
,
Nrf2 suppresses macrophage inflammatory response by blocking proinflammatory cytokine transcription
.
Nat Commun
.
2016 May 23
;
7
:
11624
.
48.
Suzuki
T
,
Yamamoto
M
.
Stress-sensing mechanisms and the physiological roles of the Keap1-Nrf2 system during cellular stress
.
J Biol Chem
.
2017 Oct 13
;
292
(
41
):
16817
24
. .
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