Congmujingnosides B-G, triterpene saponins from the stem of Aralia chinensis and their protective effects against H2O2-induced myocardial cell injury

Article abstract of DOI:10.1080/14786419.2017.1399384

Phytochemical investigation of the stem of Aralia chinensis yielded six new oleanane-type triterpene saponins named as congmujingnosides B-G (1–6). The new ones were elucidated on the basis of the chemical and spectroscopic analysis. Protective effects of compounds 1?6 were tested against H₂O₂-induced H9c2 cardiomyocyte injury, and the data showed that compounds 1 and 5 had significant cell-protective effects. No significant DPPH radical scavenging activity was observed for compounds 1?6.

Full text of DOI:10.1080/14786419.2017.1399384

Natural Product Research
Formerly Natural Product Letters
ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: http://www.tandfonline.com/loi/gnpl20
Congmujingnosides B-G, triterpene saponins from
the stem of Aralia chinensis and their protective
effects against H O -induced myocardial cell injury
2
2
Wen Zhang, Nailiang Zhu, Meigeng Hu, Shichun Yu, Zhonghao Sun, Haifeng
Wu, Pengfei Li, Junshan Yang, Guoxu Ma & Xudong Xu
To cite this article: Wen Zhang, Nailiang Zhu, Meigeng Hu, Shichun Yu, Zhonghao Sun, Haifeng
Wu, Pengfei Li, Junshan Yang, Guoxu Ma & Xudong Xu (2017): Congmujingnosides B-G,
triterpene saponins from the stem of Aralia chinensis and their protective effects against H O -
2
2
induced myocardial cell injury, Natural Product Research, DOI: 10.1080/14786419.2017.1399384
To link to this article: http://dx.doi.org/10.1080/14786419.2017.1399384
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Date: 09 November 2017, At: 23:15

Natural Product research, 2017
ꢁꢁpꢂ://ꢃꢄi.ꢄꢅg/10.1080/14786419.2017.1399384
ꢀ
Congmujingnosides B-G, triterpene saponins from the stem
of Aralia chinensis and their protective effects against H O -
2
2
induced myocardial cell injury
a,b1
a1
a
b
a
Wen Zhang , Nailiang Zhu , Meigeng Hu , Shichun Yu , Zhonghao Sun ,
a
a
a
a
a
Haifeng Wu , Pengfei Li , Junshan Yang , Guoxu Ma and Xudong Xu
ꢆKꢇy lꢆbꢄꢅꢆꢁꢄꢅy ꢄf Biꢄꢆꢈꢁivꢇ sꢉbꢂꢁꢆnꢈꢇꢂ ꢆnꢃ rꢇꢂꢄꢉꢅꢈꢇ uꢁiꢊizꢆꢁiꢄn ꢄf cꢀinꢇꢂꢇ hꢇꢅbꢆꢊ Mꢇꢃiꢈinꢇ, Miniꢂꢁꢅy ꢄf
eꢃꢉꢈꢆꢁiꢄn, Bꢇijing Kꢇy lꢆbꢄꢅꢆꢁꢄꢅy ꢄf Innꢄvꢆꢁivꢇ dꢅꢉg diꢂꢈꢄvꢇꢅy ꢄf tꢅꢆꢃiꢁiꢄnꢆꢊ cꢀinꢇꢂꢇ Mꢇꢃiꢈinꢇ (Nꢆꢁꢉꢅꢆꢊ
Mꢇꢃiꢈinꢇ) ꢆnꢃ tꢅꢆnꢂꢊꢆꢁiꢄnꢆꢊ Mꢇꢃiꢈinꢇ, Kꢇy lꢆbꢄꢅꢆꢁꢄꢅy ꢄf effiꢈꢆꢈy evꢆꢊꢉꢆꢁiꢄn ꢄf cꢀinꢇꢂꢇ Mꢇꢃiꢈinꢇ ꢆgꢆinꢂꢁ
Gꢊyꢈꢄꢊipiꢃ Mꢇꢁꢆbꢄꢊiꢈ diꢂꢄꢅꢃꢇꢅꢂ, sꢁꢆꢁꢇ aꢃminiꢂꢁꢅꢆꢁiꢄn ꢄf tꢅꢆꢃiꢁiꢄnꢆꢊ cꢀinꢇꢂꢇ Mꢇꢃiꢈinꢇ, Inꢂꢁiꢁꢉꢁꢇ ꢄf Mꢇꢃiꢈinꢆꢊ
Pꢊꢆnꢁ dꢇvꢇꢊꢄpmꢇnꢁ, Pꢇking uniꢄn Mꢇꢃiꢈꢆꢊ cꢄꢊꢊꢇgꢇ ꢆnꢃ cꢀinꢇꢂꢇ aꢈꢆꢃꢇmy ꢄf Mꢇꢃiꢈꢆꢊ sꢈiꢇnꢈꢇꢂ, Bꢇijing, cꢀinꢆ;
b
sꢈꢀꢄꢄꢊ ꢄf Pꢀꢆꢅmꢆꢈy, aꢀꢀꢉi univꢇꢅꢂiꢁy ꢄf tꢅꢆꢃiꢁiꢄnꢆꢊ cꢀinꢇꢂꢇ Mꢇꢃiꢈinꢇ, hꢇfꢇi, cꢀinꢆ
ABSTRACT
ARTICLE HISTORY
Phytochemical investigation of the stem of Aralia chinensis yielded six
new oleanane-type triterpene saponins named as congmujingnosides
B-G (1–6). The new ones were elucidated on the basis of the chemical
and spectroscopic analysis. Protective effects of compounds 1−6 were
rꢇꢈꢇivꢇꢃ 2 aꢉgꢉꢂꢁ 2017
aꢈꢈꢇpꢁꢇꢃ 14 oꢈꢁꢄbꢇꢅ 2017
KEYWORDS
Aralia chinensis; ꢁꢅiꢁꢇꢅpꢇnꢇ
ꢂꢆpꢄninꢂ; ꢈꢆꢅꢃiꢄmyꢄꢈyꢁꢇ
ꢄxiꢃꢆꢁivꢇ injꢉꢅy; dPPh ꢅꢆꢃiꢈꢆꢊ
ꢂꢈꢆvꢇnging ꢆꢈꢁiviꢁy
tested against H O -induced H9c2 cardiomyocyte injury, and the
2
2
data showed that compounds 1 and 5 had significant cell-protective
effects. No significant DPPH radical scavenging activity was observed
for compounds 1−6.
CONTACT Gꢉꢄxꢉ Mꢆ
tꢀꢇꢂꢇ ꢆꢉꢁꢀꢄꢅꢂ ꢈꢄnꢁꢅibꢉꢁꢇꢃ ꢇqꢉꢆꢊꢊy ꢁꢄ ꢁꢀiꢂ ꢆꢅꢁiꢈꢊꢇ.
mgxfl8785@163.ꢈꢄm; Xꢉꢃꢄng Xꢉ
xꢃxꢉ@impꢊꢆꢃ.ꢆꢈ.ꢈn
1
sꢉppꢊꢇmꢇnꢁꢆꢊ ꢃꢆꢁꢆ fꢄꢅ ꢁꢀiꢂ ꢆꢅꢁiꢈꢊꢇ ꢈꢆn bꢇ ꢆꢈꢈꢇꢂꢂꢇꢃ ꢆꢁ ꢀꢁꢁpꢂ://ꢃꢄi.ꢄꢅg/10.1080/14786419.2017.1399384.
2017 Infꢄꢅmꢆ uK limiꢁꢇꢃ, ꢁꢅꢆꢃing ꢆꢂ tꢆyꢊꢄꢅ & Fꢅꢆnꢈiꢂ Gꢅꢄꢉp
©

2
W. ZHANG ET AL.
1
. Introduction
The genus Aralia Linn belongs to the Araliaceae family is mainly distributed in Asia and less
so in the North America. In China, there are approximately 30 species in this genus (He and
Zeng 1978). Aralia chinensis as one species of this genus, its buds are the major source of
wild vegetables, the roots and barks are found as folk medicine for rheumathism, lumbago,
hepatitis, bruise nephritis and oedema (Sakai et al. 1994). Previous phytochemical investi-
gations indicated that Aralia chinensis species contain a rich source of oleanane-type triter-
pene saponins (Saito et al. 1990; Yoshikawa et al. 1994; Sakai et al. 1994; Zou et al. 2001).
Among these triterpene saponins, some displayed significant biological activities such as
cytotoxic (Tomatsu et al. 2003), anti-inflammatory (Suh et al. 2007), live-protecting (Du and
Chi 2005) and antioxidative (Zhang et al. 2006). However, the chemistry of the stems of
A. chinensis Linn. has been little researched. As a result, the phytochemical investigation of
the stems of A. chinensis Linn. was performed, and obtained six new triterpene saponins
(
Figure 1). In this paper, we report the isolation and structural elucidation of the new com-
pounds, as well as the protective effects of the isolates against cardiomyocyte injury induced
by H O . In addition, their radical scavenging activity was evaluated by DPPH assay.
2
2
2
. Results and discussion
2
.1. New compound structure elucidation
2
0
Compound 1 was obtained in the form of a white amorphous powder with [α]D −20.6 (c 0.1,
MeOH), and the Molish and Liebermann–Burchard reactions were positive. The HRESIMS
+
spectrum showed a quasi-molecular ion at m/z 1259.6067 (calcd for C H O Na [M + Na] ,
5
9
96 27
1259.6037), from which in conjunction with NMR data, the molecular formula was established
as C H O . The NMR spectra (Table S1) of compound 1 showed the characteristic signals
5
9
96 27
of a triterpenoid saponin. Seven signals for tertiary methyl groups at δ 0.84, 1.08, 1.07, 0.86,
H
13
1
.24, 1.27 and 0.87 which agreed with the information obtained from the C NMR spectrum
3
(seven sp carbons at δ : 16.1, 16.9, 17.9, 24.1, 26.6, 28.3 and 33.6) as determined by an HSQC
C
technique. The 1D NMR data also revealed one trisubstituted olefinic proton at δ 5.39 (1H,
H
br s, H-12) with two typical olefinic carbon signals at δ 122.3 and 144.6. The downfield
C
chemical shift of C-3 (δ 89.9) and the upfield chemical shift of C-28 (δ 177.0) showed that
C
C
1
was a bisdesmosidic glycoside, and the chemical shift of C-3 (δ 89.9) indicated an α-ori-
C
1
13
entation for H-3 (Liang et al. 2011). The H and C NMR spectra of 1 showed five anomeric
signals at δ 5.47 (1H, d, J = 7.8 Hz), 4.80 (1H, d, J = 7.8 Hz,), 5.36 (1H, d, J = 7.8 Hz), 5.02 (1H,
H
d, J = 7.8 Hz), and 6.25 (1H, d, J = 7.8 Hz) and carbon signals at δ 105.3, 105.9, 105.7, 105.7
C
and 96.1, which further revealed that compound 1 was a oleanane-type glycoside with five
sugars attached to the C-3 and C-28 positions. Through detailed analysis of the 2D NMR
1
1
spectra (HSQC and H– H COSY), the spin systems for the sugars were assigned. In contrast,
the HMBC spectrum (Figure S1) of 1 showed a proton signal at δ 5.47 (glc-H-1′) that corre-
H
lated with the carbon signal at δ 89.9 (C-3) of the aglycone moiety, and the anomeric proton
C
signals at δ 5.02 (xyl-H-1″″), 4.80 (glc-H-1″) and 5.36 (glc-H-1‴) showed correlations with
H
the carbon signals at δ 79.5 (glc-C-2′), 85.3 (glc-C-3′) and 69.8 (glc-C-6″), respectively, of the
C
inner sugars, which suggests glycosylation at C-3 of aglycone with a [glc (1 → 3)-glc (1 → 6)]-
[
xyl (1 → 2)]-glc moiety. In addition an anomeric proton signal at δ 6.25 (glc-H-1″‴) showed
H
correlation with the carbon signal at δ 177.0 (C-28) indicating the presence of a glucose
C

NATURAL PRODUCT RESEARCH
3
Figure 1. cꢀꢇmiꢈꢆꢊ ꢂꢁꢅꢉꢈꢁꢉꢅꢇ ꢄf Congmujingnosides B-G.
unit at C-28. The anomeric configurations of the sugar moieties were determined to be β
because of the JH–H values (7.8 Hz). The absolute configuration of the monosaccharides was
determined to be D through GC analysis of the chiral derivatives of the monosaccharides in
the hydrolysate. Finally, the structure of
1 was established as 3-O-{[β-D-
xylopyranosyl-(1 → 2)]-[β-D-glucopyranosyl- (1 → 6)-β-D-glucopyranosyl-(1 → 3)]-β-D-glu-
copyranosyl} oleanolic acid 28-O-β-D-glucopyranosyl ester and named congmujingnoside
B (Figure 1).
Compound 2 was obtained as a white amorphous powder, and its elemental formula of
+
C H O Na was evident on the basis of positive HRESIMS [M + Na] ion peak at m/z
5
9
96 27
1
259.6034 (calcd. for 1259.6037), which was the same as that of 1. The acid hydrolysis of 2
1
13
liberated D-glucose and D–xylose. The H and C NMR spectra of 2 were similar to those of

4
W. ZHANG ET AL.
1
, except for the signals due to the upfield chemical shifts of C-3 (δ 79.1), which suggests
C
C-3 without sugar unit. And the sugar linkages were determined from the HMBC spectrum.
The long-range couplings observed between the proton signal at δ 6.24 (glc-H-1′) and the
H
carbon signal at δ 177.0 (C-28) of the aglycone moiety, and the anomeric proton signals at
C
δH 5.02 (xyl-H-1″″), 4.81 (glc-H-1″), 5.58 (glc-H-1″‴), and 5.36 (glc-H-1‴) showed correlations
with the carbon signals at δ 79.8 (glc-C-2′), 90.1 (glc-C-3′), 89.3 (glc-C-3″), and 69.8 (glc-C-6″)
C
of the inner sugar, respectively, confirm the glycosylation at C-28 of the aglycone with a
{
[glc-(1 → 6)-glc-(1 → 3)]-glc-(1 → 3)}-[xyl-(1 → 2)]-glc moiety. Therefore, the structure of 2
was established as [β-D-xylopyranosyl-(1 → 2)]-{[β-D-glucopyranosyl- (1 → 3)]-[β-D-glu-
copyranosyl-(1 → 6)]-β-D-glucopyranosyl-(1 → 3)}28-O-β-D-glucopyranosyl oleanolic acid
ester and named congmujingnoside C. (Figure 1)
Compound 3, which was isolated as a white acicular crystal. In the positive-ion HRFABMS
analysis revealed the molecular formula of 3 to be C H O . By the acid hydrolysis of 3,
5
4
88 23
1
13
D-glucose were identified by TLC analysis. The H and C NMR spectra of 3 were very similar
to those of 2, apart from the lack of signals of a β-D-xylopyranosyl. In contrast, long-range
couplings were observed from a proton signal at δ 6.32 (glc-H-1′) to the carbon signal at
H
δ 176.9 (C-28), δ 5.33 (glc-H-1″″) to δ 80.1 (glc-C-2′), δ 4.83 (glc-H-1″) to δ 89.8 (glc-C-3′),
C
H
C
H
C
and δ 5.57 (glc-H-1‴) to δ 89.1 (glc-C-3″) in the HMBC spectrum of 3, which suggests gly-
H
C
cosylation at C-28 with a{ [glc (1 → 3)]-glc-(1 → 3)]}-[glc(1 → 2)]-glc moiety. This result agrees
with the above observations and suggests that compound 3 is [β-D-glucopyrano-
syl-(1 → 2)]-{[β-D-glucopyranosyl-(1 → 3)]-β-D-glucopyranosyl-(1 → 3)}28-O-β-D-
glucopyranosyl oleanolic acid ester and named congmujingnoside D. (Figure 1).
Compound 4 was obtained as an amorphous powder. The HRESIMS showed a deproto-
+
nated molecular ion peak at m/z 1097.5500 [M + Na] , indicating the molecular formula to
1
13
be C H O . The H and C NMR spectroscopic data (Table S2) were closely related to those
53
86 22
of 3 except for one D-glucose in 3 was replaced by D-xylose in 4. The sugar moieties and
1
3
linkages were the same as 3 on the basis of the similar C NMR and HMBC correlations,
1
1
which were further confirmed by HSQC and H– H COSY experiments. All of the correlations
showed that compound 4 was an oleanolic acid glycoside at C-28 with a {[glc (1 → 3)]-glc-
(
1 → 3)]}-[xyl-(1 → 2)]-glc moiety. Therefore, the structure of 4 was established as [β-D-xy-
lopyranosyl-(1 → 2)]-{[β-D-glucopyranosyl-(1 → 3)]-β-D-glucopyranosyl-(1 → 3)}28-O-β-D-
glucopyranosyl oleanolic acid ester and named congmujingnoside E. (Figure 1).
2
0
Compound 5 was obtained as an amorphous white powder with [α]D −26.3 (c 0.1,
MeOH). It was determined to have the molecular formula of C H O on the basis of the
5
9
96 26
1
3
+
C NMR data and HRESIMS spectrum at m/z 1243.6085 [M + Na] (Calcd for C H O Na,
243.6088). An examination of the H and C NMR data (Table S3) showed the structure
5
9
96 23
1
13
1
of 5 to be similar to that of 1 except for one D-glucose in 1 was replaced by D-quinovose
in 5. Through detailed analysis of the 2D NMR spectra, the spin systems for the sugars
were assigned. In the HMBC spectrum, proton signal at δ 5.47 (glc-H-1′) had correlation
H
with the carbon signal at δ 89.2 (C-3) of the aglycone moiety, and the anomeric proton
C
signals at δ 5.02 (xyl-H-1″″), 4.81 (glc-H-1″), and 6.20 (qui-H-1‴) showed correlations with
H
the carbon signals at δ 79.2 (glc-C-2′), 83.7 (glc-C-3′) and 69.9 (glc-C-6″), respectively, of
C
the inner sugar. In addition an anomeric proton signal at δ 6.25 (glc-H-1″‴) showed corre-
C
lation with the carbon signal at δ 177.0 (C-28), which suggests glycosylation at C-3 of
C
aglycone with a [glc (1 → 3)-qui (1 → 6)]-[xyl (1 → 2)]-glc moiety glycosylation and at C-28
with a glucose. Therefore, taking together with the NOE spectrum, the structure of 5 was

NATURAL PRODUCT RESEARCH
5
defined as 3-O-{[β-D-xylopyranosyl-(1 → 2)]-[β-D-quinovopyranosyl-(1 → 6)-β-D-glucopyra-
nosyl-(1 → 3)]-β-D-glucopyranosyl} oleanolic acid 28-O-β-D-glucopyranosyl ester and
named congmujingnoside F.
+
Compound 6 exhibited an [M + Na] ion peak at m/z 1081.5567 using HRESIMS, corre-
1
13
sponding to the molecular formula C H O . The H and C NMR signals (Table S3) were
5
3
86 21
closely related to those of 4 except for the sugar unit. The monosaccharides obtained after
aqueous acidic hydrolysis of compound 6 were identified as glucose, xylose and quinovose
byTLC comparison with authentic samples, which displayed one D-glucose in 4 was replaced
by D-quinovose in 6. The absolute configuration of the monosaccharides was determined
to be D by GC analysis of the chiral derivatives of the monosaccharides in the hydrolysate.
The similar NOEs of 6 and 4 indicated that their absolute configurations were identical.
Therefore, the structure of compound 6 was [β-D-xylopyranosyl-(1 → 2)]-{[β-D-glucopyra-
nosyl-(1 → 3)]-β-D-glucopyranosyl-(1 → 3)}28-O-β-D-glucopyranosyl oleanolic acid ester
and named as congmujingnoside G.
In this study, the protective effects of compounds 1−6 against H O -induced myocardial
2
2
cell injury were tested. As shown in Table S4, compounds 1 and 5 (15−200 μM) significantly
increased the viability of H9c2 induced by H O in a dose-dependent manner, ranging from
2
2
the concentrations at 15–200 μM. Compound 5 was shown to be the most efficient among
the isolates. It increased the cell viability at a low concentration of 30 μM and was more
efficient than vitamin E, which was used as the positive control. Compounds 1 showed
moderate protective effects. They significantly improved cell viability at a concentration of
200 μM. No significant DPPH radical scavenging activity was observed for compounds 1−6.
3
. Conclusion
In summary, Phytochemical investigation of the stem of Aralia chinensis yielded six new
oleanane-type triterpene saponins named as congmujingnosides B-G (1–6). Protective
effects of compounds 1−6 were tested against H O -induced H9c2 cardiomyocyte injury,
2
2
and the data showed that compounds 1 and 5 had significant cell-protective effects. No
significant DPPH radical scavenging activity was observed for compounds 1−6.
Supplementary material
Supplementary material relating to this article is available online, alongside Experimental, Tables S1–S5
and Figures S1–S37.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
This study was financially supported by the technologic large platform for comprehensive research
and development of new drugs in the Twelfth Five-Year“Significant New Drugs Created”Science and
Technology Major Projects [grant number 2012ZX09301-002-001-026], the National Science and
Technology Support Program [grant number 2012BA127B06], and the Innovation Capacity building
in Anhui Science and Technology Agency [grant number 10100027-3].

6
W. ZHANG ET AL.
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