Journal of Ethnopharmacology 250 (2020) 112497

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Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jethpharm

Anti-inflammatory mechanism and active ingredients of the Chinese tallow T

tree
Rao Fua, Fang Chena, Yiran Guob,c,∗
a
College of Life Sciences, Sichuan University, Key Laboratory of Bio-resource and Eco-environment of Ministry of Education, Chengdu, China
b
School of Biological Science and Medical Engineering, Beihang University, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beijing, China
c
Beijing Advanced Innovation Centre for Biomedical Engineering, Beihang University, Beijing, China

A R T I C LE I N FO A B S T R A C T

Keywords: Ethnopharmacological relevance: The leaf of the Chinese tallow tree (CTT, Sapium sebiferum (L.) Roxb) has been
Chinese tallow tree used in Traditional Chinese Medicine (TCM) to treat eczema, shingles, edema, swelling, ascites, scabs, and
Anti-inflammatory snakebites.
Increased GSH Aim of this study: The present work aimed to explore the antioxidant-related anti-inflammatory mechanisms of
Active ingredients
CTT leaf and to further investigate their possible active ingredients.
Materials and methods: The anti-inflammatory activities of different fractions were determined using a 12-O-
tetradecanoylphorbol-13-acetate (TPA) induced model of acute edema in mouse ears. The SOD, CAT and GCL
activities and the GSH content of the ear tissue were measured using kits, and the ratio of the treated and control
ears was calculated. The anti-inflammatory activities of each single compound and those of a mixture of the
compounds were also determined using the TPA-induced model.
Results: The anti-inflammatory effects of the three fractions were positively correlated with their increasing GSH
capacities. Although the GSH levels decreased during TPA-induced acute edema, the CTT leaf extract could
recover these levels by increasing the glutamate cysteine ligase activity. The mixture of ellagic acid, isoquercitrin
and astragalin showed an anti-inflammatory effect similar to that of the CTT leaf extract. However, none of these
three individual compounds showed comparable activity alone.
Conclusion: These results demonstrated that increasing GSH is an antioxidant-related anti-inflammatory me-
chanism of CTT leaves. In addition, ellagic acid, isoquercitrin and astragalin were found to be jointly responsible
for this bioactivity.

1. Introduction antioxidant, anti-microbial, anti-inflammatory, anti-allergic, anti-hy-

Chinese tallow tree (CTT, Sapium sebiferum (L.) Roxb and referred to
as “Wu Jiu” in Chinese) originated in China and is cultivated as an
ornamental plant. CTT primarily grows in subtropical regions of the
world, and it has been regarded as an invasive species in the United
States (Pile et al., 2017). CTT is a valuable oilseed-producing plant with
great medicinal value. CTT leaf has long been used as an anti-in-
flammatory herb, with diuretic and parasiticidal properties for the
treatment of eczema, shingles, edema, swelling, ascites, scabs, and
snakebites, as described in Traditional Chinese Medicine (TCM) books,
Tang Materia Medica and Compendium of Materia Medica (Peng et al.,
2008). Modern pharmacological studies suggest that CTT leaf possesses
pertensive, analgesic, and hepatoprotection bioactivities (Hassan et al.,
2011; Hsu et al., 1994; Huang et al., 2004; Hussain et al., 2015; Peng
et al., 2008). In terms of its active compounds, several phenolic mole-
cules, including gallic acid, ellagic acid, shikimic acid, quercetin,
kaempferol, isoquercitrin, hyperin, astragalin, trifolin, and rutin, have
been identified (Fu et al., 2015b; Liu and Kong, 2005; Wang et al.,
2007).
During the inflammatory response, reactive oxygen species (ROS)
will burst, which damages the antioxidant defense system (Park et al.,
2016). Theoretically, natural products with antioxidant properties can
directly scavenge radicals at the inflamed area. However, due to the low
bioavailability of natural products, the concentrations in the inflamed

Abbreviations: TCM, Traditional Chinese Medicine; EAF, Ethyl acetate fraction; nBF, n-butyl alcohol fraction; WF, Water residue fraction; SOD, Superoxide dis-
mutase; CAT, Catalase; GSH, Glutathione; TPA, 12-O-tetradecanoylphorbol-13-acetate; DMSO, Dimethylsulfoxide; SEM, Standard error of mean; GCL, Glutamate
cysteine ligase
∗
Corresponding author. No. 37 Xueyuan Road, Haidian District, Beijing, 100191, China.
E-mail address: gyiran@buaa.edu.cn (Y. Guo).

https://doi.org/10.1016/j.jep.2019.112497
Received 17 September 2019; Received in revised form 19 December 2019; Accepted 19 December 2019
Available online 21 December 2019
0378-8741/ © 2019 Elsevier B.V. All rights reserved.
R. Fu, et al.
tissue cannot reach the active levels achieved in vitro. In addition,
natural products might activate the Nrf-2 antioxidant defense system
(Schäfer and Werner, 2015). The anti-dermatitis effect of CTT leaf is
related to its antioxidant activity (Fu et al., 2013, 2015b). However, the
underlying mechanism is unclear. Plant extracts typically possess
complex chemical compositions, and determining which components
contributes to the extract's bioactivity can be challenging. Furthermore,
relatively low concentrations of each single compound might not be
enough to exhibit bioactivity. Thus, synergistic effects of active in-
gredients might contribute to the bioactivity of an extract.
In this context, by comparing the bioactivities of three distinct
chemical composition fractions of CTT ethanol extract, we explored the
antioxidant-related anti-inflammatory mechanisms and possible active
ingredients of CTT leaf. We determined the anti-inflammatory me-
chanism to be an increase in GSH levels through enhanced biosynthesis
enzyme activity. In addition, three main phenolic compounds from CTT
leaf, namely, ellagic acid, isoquercitrin and astragalin, are all re-
sponsible for the anti-dermatitis property of CTT leaf.
2. Materials and methods
2.1. Reagents
12-O-tetradecanoylphorbol-13-acetate (TPA) and indomethacin
were purchased from Sigma-Aldrich Chemical Co. (St Louis, MO, USA).
Protein, superoxide dismutase (SOD), catalase (CAT), glutathione
(GSH), and glutamate cysteine ligase (GCL) measurement kits were
provided by the Institute of Biological Engineering of Nanjing
Jiancheng (Nanjing, China). Ellagic acid, isoquercitrin and astragalin
were obtained from Chengdu Herbpurify Co., Ltd. (Chengdu, China).
All the other chemicals and reagents used in the present study were of
analytical grade.
2.2. Preparation of plant extracts
The plant material was collected in April 2012 in Renshou County
(Sichuan Province, China). A voucher specimen (No. 00721412) was
identified by associate professor Jie Bai, School of Life Science, Sichuan
University, and deposited in the Sichuan University Herbarium. The
three fractions of the ethanol extract of CTT leaves were prepared using
the method described in our previous study (Fu et al., 2015a). Briefly,
the leaves were dried, ground, weighed, and then extracted twice with
95% ethanol in a Soxhlet apparatus. The two extracts were mixed and
concentrated using a vacuum rotary evaporator. The residue was re-
suspended in water and partitioned with petroleum (Boiling range:
30–60 °C) (10 times), ethyl acetate (8 times) and n-butyl alcohol (6
times) using liquid-liquid partition. The fractions were concentrated
and lyophilized. The petroleum fraction was not included due to pig-
ment enrichment and low bioactivity. All fractions were stored at
−20 °C and dissolved in absolute ethanol to perform the experiments.
2.3. Animals
Five-week-old male Kunming mice were purchased from Chengdu
DOSSY Biotechnology Co., Ltd. (Chengdu, China). The mice were
housed under standard conditions (22 ± 2 °C and a 12/12 light/dark
cycle) and given free access to food and water. All the experiments were
approved by the Sichuan University Animal Experimentation Ethics
Committee (Permit No. 20170727) and were in complete compliance
with the National Institutes of Health Guide for the Care and Use of
Laboratory Animals.
2.4. Anti-inflammatory effects of the three fractions
All the measurements were based on a comparison between the
right (treated) and left (untreated) ears of mice. The TPA-induced acute
2
Journal of Ethnopharmacology 250 (2020) 112497
inflammation model was established according to the previously de-
scribed method (Fu et al., 2013). Briefly, 2.5 μg TPA (dissolved in 20 μL
DMSO:acetone (v/v) = 1:15) was topically applied to the right ear,
while the left ear received a 20 μL vehicle control (DMSO:acetone (v/
v) = 1:15). The mice in the groups without TPA treatment received the
vehicle on both ears. After 30 min, different fractions were applied to
the right ear at a dose of 0.3 mg/ear. Indomethancin (0.5 mg/ear) was
used as a positive control. The left ears received an equal volume of
ethanol. Mice in the control and model groups received ethanol on both
ears as a reference. The mice were sacrificed at 6 h after TPA treatment,
and thickness was measured using a digital micrometer. To minimize
variation in the technique, a single investigator performed the mea-
surements throughout the experiment. The degree of swelling was
calculated as the thickness of the right ear minus that of the left ear.
Both ears were collected and washed with ethanol (5 times) and water
(5 times) to remove the residue fractions. The cleaned ears were
homogenized in 0.86% saline solution and concentrated at 3500 rpm
for 10 min. The supernatant was used to determine the biochemical
markers including protein, GSH levels and SOD, CAT and GCL activities
with commercial kits according to the manufacturers’ protocols. The
GSH levels and SOD, CAT and GCL activities were normalized to protein
content.
2.5. Kinetic effects of TPA and EAF on mice ear redox parameters
TPA (2.5 μg/ear) or EAF (0.3 mg/ear) was applied to the right ears
as described above, and the mice were sacrificed at different time point
(0, 1, 2, and 6 h post TPA application or 0, 0.5, 1.5, 5.5 and 24 h post
EAF application). All the other procedures were as described above in
section 2.4.
2.6. Identification of active compounds
Mice were divided into seven groups (n = 8) that were treated with
control, model, EAF, ellagic acid, isoquercitrin, astragalin and mixture
of the three single compounds. The EAF dose was 0.3 mg/ear, and the
ellagic acid, isoquercitrin and astragalin doses were 20 μg/ear, which
was calculated from the approximate contents of each single compound
in the EAF according to our previous results (Fu et al., 2015b). All the
other experimental procedures were as described above in section 2.4.
2.7. Statistical analysis
GraphPad Prism (GraphPad Software Inc., San Diego, CA, USA) was
used for statistical analysis of the data. The data are expressed as the
mean ± standard error of the mean (SEM) (n = 8 per group). One-way
analysis of variance (ANOVA) with multiple comparisons was used,
followed by Dunnett's tests. Statistical significance was determined by
P < 0.05.
3. Results and discussion
3.1. Antioxidant-related anti-inflammatory mechanism of CTT leaf
The anti-inflammatory activity of CTT leaf has been studied using
xylene-induced mice ear edema and carrageenan-induced rat paw
edema models (Huang et al., 2004). The results of that study found that
crude ethanol extract had a significant anti-inflammatory effect. Pre-
viously, we prepared a phenolic-enriched fraction from crude ethanol
extract and evaluated its anti-dermatitis activity according to the tra-
ditional use of CTT leaf. Our results found that a phenolic extract of CTT
leaf produces effective anti-dermatitis activity in a TPA-induced acute
mouse ear edema model and a DNFB-induced mouse allergic contact
dermatitis model. In addition, the anti-dermatitis activity of CTT leaf is
related to its antioxidant activity (Fu et al., 2013, 2015b). However, the
underlying mechanism is still unclear. The present study explored this
R. Fu, et al. Journal of Ethnopharmacology 250 (2020) 112497

Table 1
Effects of different fractions on the SOD and CAT activities with and without
TPA treatment.
Groups SOD ratio CAT ratio

-TPA Control 1.013 ± 0.046a 0.970 ± 0.087a

EAF 0.999 ± 0.032a 0.981 ± 0.022a
nBF 0.955 ± 0.040a 0.986 ± 0.060a
WF 0.950 ± 0.081a 0.921 ± 0.056a

+TPA Model 0.746 ± 0.049b 0.592 ± 0.025b

EAF 0.768 ± 0.049b 0.748 ± 0.048b
nBF 0.789 ± 0.029ab 0.735 ± 0.025b
WF 0.557 ± 0.092b 0.601 ± 0.041b
Indo 0.775 ± 0.033b 0.748 ± 0.051b

Data are expressed as the mean ± S.E.M (n = 8). Different lowercase letters
indicate significance at P < 0.05 with each other.

the antioxidant-related mechanism of CTT leaf. To minimize individual

Fig. 1. Anti-inflammatory activities of three different fractions from CTT
leaf. (a) The degree of swelling. Mice were divided into two groups with and
without TPA treatment. In the group without TPA treatment, four subgroups
were created: control, EAF, nBF and WF. In the groups with TPA treatment, five
other sub-groups were created: model, EAF, nBF, WF and positive control in-
domethacin. Various samples were applied to study the therapeutic effects.
After 6 h of TPA application. Both ears were measured, and degree of swelling
was calculated as the thickness of the right ear minus that of the left ear control.
(b) The ratio of the GSH levels between the right and left ears among different
groups. Data are expressed as the mean ± S.E.M (n = 8). The means with
different lowercase letters above the column are significantly different from
each other (P < 0.05).
mechanism by investigating bioactivity of the three distinct chemical
composition fractions.
Three fractions from the CTT ethanol extract including the ethyl
acetate fraction (EAF), n-butyl alcohol fraction (nBF) and water residue
fraction (WF), have shown different in vitro antioxidant and anti-mi-
crobial capacities (Fu et al., 2015a). Therefore, we first used a TPA-
induced acute inflammation model to compare the bioactivities of the
three fractions. Compared with the ear thickness in the control group,
the ear thickness in the model group was elevated to over 0.3 mm after
TPA treatment. At the same doses, EAF and nBF showed significant
inhibition of edema similar to that of the positive control indomethacin
(P < 0.05). However, WF did not exhibit any suppression (Fig. 1a). In
addition, when applied to the ears not treated with TPA, none of the
three fractions exhibited any effect on swelling compared with the
control group. The antioxidant parameters were determined to explore
differences, the ratio of both was calculated. After treatment with TPA
for 6 h, and with increasing edema, the SOD and CAT activities were
significantly decreased (Table 1), and the GSH levels were also sig-
nificantly reduced (Fig. 1b); this finding was consistent with those of
published reports (Chaudhary et al., 2009). EAF and nBF, with swelling
suppression abilities, also exhibited the potential alleviation of SOD and
CAT activities, while WF did not (Table 1). EAF showed the strongest
ability to increase GSH levels relative to nBF and WF (Fig. 1b). Inter-
estingly, the GSH-increasing effect was positively correlated with the
anti-inflammatory activity (r = 0.682). Furthermore, when these three
fractions were applied to normal ears, they did not change the SOD and
CAT ratios (Table 1). However, these three fractions did influence the
GSH ratios, in the order of EAF > nBF > WF (Fig. 1b). It seems that
increasing GSH is the antioxidant-related anti-inflammatory me-
chanism of CTT leaf rather than the recovery of SOD and CAT activities.
GSH, which is composed of glutamate, cysteine and glycine, is a key
regulator that maintains intracellular redox homeostasis and protects
cells against oxidative injury (Zhang et al., 2019). It has been shown
that supplementation with γ-glutamylcysteine, an intermediate dipep-
tide of the GSH biosynthesis pathway, produced anti-inflammatory
activities (Yang et al., 2019). Another GSH prodrug, N-acetyl-l-cysteine,
has been reported to be effective in the treatment of various diseases
(Rushworth and Megson, 2014). These results indicate that CTT leaf can
play an anti-inflammatory role by increasing the GSH levels.
To better understand the antioxidant-related mechanism, we in-
vestigated the dynamic effects of CTT leaf on normal mouse ears. The
most active fraction, EAF, was used. We evaluated the effect at four
timepoints: 0.5, 1.5, 6.5 and 23 h after EAF application. During this
period, EAF did not change the SOD and CAT activities. However, EAF
did significantly increase the GSH levels 0.5 h after application; this
increase lasted for 24 h (Fig. 2). These results demonstrated that CTT
leaf can indeed upregulate the GSH levels. During the development of
TPA-induced inflammation, the GSH levels significantly decreased as
swelling increased (Fig. 3). After the application of TPA for 6 h, edema
reached the highest levels, and GSH level was reduced by half. These
results further support the conclusion that increasing the GSH levels is
the antioxidant-related anti-inflammatory mechanism of CTT leaf ra-
ther than recovery of SOD and CAT activities.
We further measured glutamate cysteine ligase (GCL) activity which
is responsible for the biosynthesis of GSH (Li et al., 2016). Under both
inflamed and uninflamed conditions, EAF was found to significantly
increase the GCL activity (P < 0.05) (Fig. 4). These results demon-
strated that CTT leaf can increase the GCL activity, which subsequently
results in the elevation of the GSH levels and further support of the
antioxidant defense system and anti-inflammatory function.

3
R. Fu, et al.
Fig. 2. Effect of EAF on the antioxidant defense system. EAF was applied to
the right ear, and the left ear was used as a control. The mice were sacrificed at
different timepoints according to protocol, and the antioxidant parameters were
determined. The data are expressed as the mean ± S.E.M (n = 8). *P < 0.05
versus 0 h (control).
Fig. 3. Degrees of swelling and ratios of the GSH level after TPA treatment.
The data are expressed as the mean ± S.E.M (n = 8). The means with different
lowercase letters above or below the point are significantly (P < 0.05) with
each other.
Fig. 4. EAF upregulated GCL to increase the GSH level. The data are ex-
pressed as the mean ± S.E.M (n = 8). The means with different lowercase
letters above the column are significantly (P < 0.05) with each other.
3.2. Identification of active compounds
The three fractions of CTT leaf each possess distinct chemical
compositions, of which EAF contains more identified phenolic com-
pounds than nBF and WF (Fu et al., 2015a). Combining the above anti-
4
Journal of Ethnopharmacology 250 (2020) 112497
Fig. 5. Exploring the possible active ingredients of CTT leaf. (a) Anti-in-
flammatory effects of the main chemical components; (b) Effects on the GSH
levels during the anti-inflammatory period. The data are expressed as the
mean ± S.E.M (n = 8). The means with different lowercase letters above the
column are significantly (P < 0.05) with each other.
inflammatory results with the previously reported antioxidant and anti-
bacterial activities of each fraction, they were found to be in the order
of EAF > nBF > WF. These results suggested that the identified
phenolic compounds presented in EAF might be responsible for the
bioactivity of CTT leaf. The main components of EAF are ellagic acid,
isoquercitrin and astragalin in amounts of 52.62 ± 0.05,
74.14 ± 0.01 and 71.21 ± 0.01 mg/g, respectively (Fu et al., 2015b).
Our unpublished results also suggested that isoquercitrin and astragalin
were the major compounds absorbed in the mouse ear model. Thus, we
investigated the anti-inflammatory activities of each single component
and the mixture of all three components. The EAF dose was 0.3 mg/ear,
and the doses of ellagic acid, isoquercitrin and astragalin were 20 μg/
ear according to their quantities in EAF. The anti-inflammatory abilities
of each component are presented in Fig. 5a. EAF exhibited the strongest
inhibitory effect. All three single compounds and the mixture of the
three showed significant inhibition of swelling (P < 0.05). However,
none of three single compounds exhibited effects similar to that of EAF
(P < 0.05). Interestingly, the mixture showed the same effect as EAF
(P > 0.05). The above results demonstrate that increasing the GSH
levels is the antioxidant-related anti-inflammatory mechanism of CTT
leaf. Therefore, the GSH levels among the groups were also measured.
As shown in Fig. 5b, all the samples exhibited potential effects of in-
creasing the GSH levels compared with the levels in the model group.
EAF, isoquercitrin and the mixture of the three exhibited significant
effects (P < 0.05). However, no significant difference was detected
R. Fu, et al.
among different samples. In summary, these results indicated that el-
lagic acid, isoquercitrin and astragalin, all contributed to the bioactivity
of CTT leaf. It is possible that synergistic or additive effects exist among
these compounds.
4. Conclusion
In the present study, by comparing of three distinct chemical com-
position fractions of CTT leaf ethanol extract, we found that increasing
GSH is the underlying antioxidant-related anti-inflammatory me-
chanism of CTT leaf. Furthermore, all three main phenolic compounds,
namely, ellagic acid, isoquercitrin and astragalin, contributed to the
anti-dermatitis properties of CTT leaf. This work provides new insight
into how this antioxidant plays its anti-inflammatory role. A study of
the molecular mechanism by which GSH in increased is ongoing.
Author contributions
R. F., F. C. and Y. G. designed the experiment. R. F. performed ex-
periments. R. F., F. C. and Y. G. wrote the manuscript. All authors re-
viewed the results and approved the final version of the manuscript.
Declaration of competing interest
All authors report no conflicts of interest and no competing financial
interests exist.
Acknowledgement
Yiran Guo was supported by a grant from the Fundamental Research
Funds for the Central Universities. Rao Fu was supported by China
Postdoctoral Science Foundation Grant (No. 2018M631080).
References
Chaudhary, S.C., Alam, M.S., Siddiqui, M.S., Athar, M., 2009. Chemopreventive effect of
farnesol on DMBA/TPA-induced skin tumorigenesis: involvement of inflammation,
Ras-ERK pathway and apoptosis. Life Sci. 85 (5), 196–205.
Journal of Ethnopharmacology 250 (2020) 112497
Fu, R., Zhang, Y.-T., Guo, Y.-R., Huang, Q.-L., Peng, T., Xu, Y., Tang, L., Chen, F., 2013.
Antioxidant and anti-inflammatory activities of the phenolic extracts of Sapium se-
biferum (L.) Roxb. leaves. J. Ethnopharmacol. 147 (2), 517–524.
Fu, R., Zhang, Y., Guo, Y., Chen, F., 2015a. Chemical composition, antioxidant and an-
timicrobial activity of Chinese tallow tree leaves. Ind. Crops Prod. 76, 374–377.
Fu, R., Zhang, Y., Peng, T., Guo, Y., Chen, F., 2015b. Phenolic composition and effects on
allergic contact dermatitis of phenolic extracts Sapium sebiferum (L.) Roxb. leaves. J.
Ethnopharmacol. 162, 176–180.
Hassan, J.C., Asma, Z., Asghari, B., Fahd, R., Shah, F., Wajid, N., 2011. Antimicrobial
activities of Sapium sebiferum L. belonging to family Euphorbiaceae. J. Med. Plants
Res. 5 (24), 5916–5919.
Hsu, F.-L., Lee, Y.-Y., Cheng, J.-T., 1994. Antihypertensive activity of 6-O-galloyl-D-glu-
cose, a phenolic glycoside from Sapium sebiferum. J. Nat. Prod. 57 (2), 308–312.
Huang, B.-x., Huang, Z.-q., Xu, X.-l., Wei, J.-q., Lai, S., Pan, Y., 2004. Analgesic and anti-
inflammatory effect of extract of Sapium sebiferum leaves on animal model. Chin
Tradition. Patent. Med. 26 (6), 476–479.
Hussain, L., Akash, M.S.H., Tahir, M., Rehman, K., 2015. Hepatoprotective effects of
Sapium sebiferum in paracetamol-induced liver injury. Bangladesh J. Pharmacol. 10
(2), 393–398.
Li, C., Zhang, W.-J., Choi, J., Frei, B., 2016. Quercetin affects glutathione levels and redox
ratio in human aortic endothelial cells not through oxidation but formation and
cellular export of quercetin-glutathione conjugates and upregulation of glutamate-
cysteine ligase. Redox Biology 9, 220–228.
Liu, R.-h., Kong, L.-y., 2005. Reaserch on the phenolic constituent of Sapium sebiferum
leaves. China J. Chin. Mater. Med. 30 (15), 1123–1125.
Park, S.-H., Seo, W., Eun, H.S., Kim, S.Y., Jo, E., Kim, M.-H., Choi, W.-M., Lee, J.-H., Shim,
Y.-R., Cui, C.-h., Kim, S.C., Hwang, C.Y., Jeong, W.-I., 2016. Protective effects of
ginsenoside F2 on 12-O-tetradecanoylphorbol-13-acetate-induced skin inflammation
in mice. Biochem. Biophys. Res. Commun. 478 (4), 1713–1719.
Peng, X., Yi, N., Cheng, T., 2008. Research advances in chemical constituents and phar-
macological effects of Sapium sebiferum. Chin. Wild. Plant. Res. 27 (3) 1,2,11.
Pile, L.S., Wang, G.G., Stovall, J.P., Siemann, E., Wheeler, G.S., Gabler, C.A., 2017.
Mechanisms of Chinese tallow (Triadica sebifera) invasion and their management
implications – a review. For. Ecol. Manage. 404, 1–13.
Rushworth, G.F., Megson, I.L., 2014. Existing and potential therapeutic uses for N-acet-
ylcysteine: the need for conversion to intracellular glutathione for antioxidant ben-
efits. Pharmacol. Ther. 141 (2), 150–159.
Schäfer, M., Werner, S., 2015. Nrf2—a regulator of keratinocyte redox signaling. Free
Radical Biol. Med. 88, 243–252.
Wang, H.-q., Zhao, C.-y., Chen, R.-y., 2007. Studies on chemical constituents from leaves
of Sapium sebiferum. China J. Chin. Mater. Med. 32 (12), 1179–1181.
Yang, Y., Li, L., Hang, Q., Fang, Y., Dong, X., Cao, P., Yin, Z., Luo, L., 2019. γ-gluta-
mylcysteine exhibits anti-inflammatory effects by increasing cellular glutathione
level. Redox Biology 20, 157–166.
Zhang, L., Wang, X., Cueto, R., Effi, C., Zhang, Y., Tan, H., Qin, X., Ji, Y., Yang, X., Wang,
H., 2019. Biochemical basis and metabolic interplay of redox regulation. Redox
Biology 26, 101284.