Two new compounds from Semen celosiae and their protective effects against CCl4-induced hepatotoxicity

Article abstract of DOI:10.1080/14786410902833948

Two new oleanolic acid saponins, namely celosin A (1) and celosin B (2), together with six known compounds, stigmasterol (3), β-sitosterol (4),-daucosterol (5), hexacosoic acid (6), palmitic acid (7) and stearic acid (8), were isolated from the ethanolic extract of Semen celosiae. The structures of celosin A (1) and celosin B (2) were determined by spectral analysis (including 1D-and 2D-NMR). The hepatoprotective activity of 1 and 2 with oral doses 1.0, 2.0 and 4.0 mg kg^-1 were investigated by carbon tetrachloride (CCl₄)-induced hepatotoxicity in mice. The results indicate that they have significant hepatoprotective effects, and that these hepatoprotective effects may be due to the antioxidant capability.

Full text of DOI:10.1080/14786410902833948

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Two new compounds from Semen
celosiae and their protective effects
against CCl4-induced hepatotoxicity
Qian Xue a , Zhen-Liang Sun a , Mei-Li Guo a , Ying Wang b , Ge
Zhang a & Xiao-Kang Wang b
a Pharmacognosy Department , School of Pharmacy, Second
Military Medical University , 325 Guohe Road, Shanghai 200433,
P.R. China
b Pharmacognosy Department , College of Chinese Materia
Medica, Shenyang Pharmaceutical University , 103 Wenhua Road,
Shenyang 110016, P.R. China
Published online: 18 Mar 2010.
To cite this article: Qian Xue , Zhen-Liang Sun , Mei-Li Guo , Ying Wang , Ge Zhang & Xiao-Kang
Wang (2011) Two new compounds from Semen celosiae and their protective effects against CCl4-
induced hepatotoxicity, Natural Product Research: Formerly Natural Product Letters, 25:8, 772-780,
DOI: 10.1080/14786410902833948
To link to this article: http://dx.doi.org/10.1080/14786410902833948
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Natural Product Research
Vol. 25, No. 8, April 2011, 772–780
Two new compounds from Semen celosiae and their
protective effects against CCl4-induced hepatotoxicity
a
a
a
b
a
*
Qian Xue , Zhen-Liang Sun , Mei-Li Guo , Ying Wang , Ge Zhang
and Xiao-Kang Wangb
aPharmacognosy Department, School of Pharmacy, Second Military Medical University, 325
Guohe Road, Shanghai 200433, P.R. China; Pharmacognosy Department, College of
b
Chinese Materia Medica, Shenyang Pharmaceutical University, 103 Wenhua Road,
Shenyang 110016, P.R. China
(Received 29 October 2008; final version received 3 December 2008)
Two new oleanolic acid saponins, namely celosin A (1) and celosin B (2),
together with six known compounds, stigmasterol (3), ꢀ-sitosterol (4),
ꢀ-daucosterol (5), hexacosoic acid (6), palmitic acid (7) and stearic acid (8),
were isolated from the ethanolic extract of Semen celosiae. The structures of
celosin A (1) and celosin B (2) were determined by spectral analysis
(including 1D- and 2D-NMR). The hepatoprotective activity of 1 and 2
with oral doses 1.0, 2.0 and 4.0 mg kgꢀ1 were investigated by carbon
tetrachloride (CCl4)-induced hepatotoxicity in mice. The results indicate
that they have significant hepatoprotective effects, and that these
hepatoprotective effects may be due to the antioxidant capability.
Keywords: Celosia argentea L.; Semen celosiae; celosin A; celosin B;
hepatoprotective effect
1. Introduction
Semen celosiae (Amaranthaceae), the seeds of Celosia argentea L., is widely
distributed in southern China and other subtropical and tropical regions of the
world, such as Sri Lanka, South Asia, Africa and America (Kirtikar & Basu, 1995).
In folk practice, Semen celosiae has been traditionally used for diabetes, cataracts,
keratitis, iridocyclitis, caligo corneae and sarcoptidosis (Anonymous, 1992).
Furthermore, the crude EtOH extract from the seeds possessed antipyretic,
antispasmodic, anticancer, diuretic and antibacterial activities. They were also
reported to be useful in the therapeutics of jaundice, inflammation, metrrorhagia,
gonorrhoea, and the healing of wounds and injuries (Anonymous, 1992; Hase,
Kadota, & Basnet, 1997; Vetrichelvan & Jegadeesan, 2000). Previous research
reported that of twelve traditional drugs, the water extract of Semen celosiae
processed the most significant protective effect on CCl4-induced liver injury in rats
(Hase, Kadota, Basnet, Takahashi, & Namba, 1996a). Furthermore, an acidic
polysaccharide compound isolated from the water extract of S. celosiae was an active
*Corresponding author. Email: mlguo@smmu.edu.cn
ISSN 1478–6419 print/ISSN 1029–2349 online
ß 2011 Taylor & Francis
DOI: 10.1080/14786410902833948
http://www.informaworld.com

Natural Product Research
773
component in protection against chemical and immunological hepatitis (Hase,
Kadota, Basnet, Takahashi, & Namba, 1996b). However, to date, little research into
the structure of saponins in S. celosiae has been reported. In the present article,
chemical investigations on the fractions of 30% and 60% ethanol led to the isolation
of eight compounds. Based on spectral analysis, the structures of two new
compounds were elucidated and named celosin A (1) and celosin B (2), respectively.
This article reports the isolation and structure elucidation of two new oleanolic
acid saponins, celosin A and celosin B, using spectroscopic methods including 1D-,
2D-NMR, as well as the testing of their inhibitory effects on the CCl4-induced
hepatotoxicity in mice.
2. Results and discussion
Celosin A (1) was positive to a Liebermann–Burchard reaction and had
a molecular formula of C41H62O15 as determined from 13C-NMR data and
a quasi-molecular ion peak [M þ Na]þ at m/z 793.6762 in the positive-ion HR-
FAB-MS. There were 41 carbon signals in the 13C-NMR spectrum, among which
30 carbons were assigned to the aglycon, with another 11 carbons belonging to
the sugar moiety. 1H-NMR spectrum indicated that the aglycon contained six
methyl protons (s 0.95, 0.98, 1.02, 1.29, 1.45, 1.66), one olefinic proton (s 5.48)
and an active proton (s 9.76). Correspondingly, six methyl carbons (ꢁ 32.18, 22.68,
16.34, 25.13, 15.36, 10.39), two olefinic carbons (ꢁ 121.36, 143.71) and a carbonyl
carbon (ꢁ 207.42) were exhibited in the 13C-NMR spectrum, respectively (Table 1).
In the HMBC spectrum, the methylene proton at ꢁ H 4.19 (H-3) correlated with a
carbonyl carbon (ꢁ 207.42) and a methyl carbon at ꢁ 10.39. Furthermore, the
methylene proton also correlated with an anomeric carbon (ꢁ 102.42) of the sugar
moiety. All of this evidence suggested that celosin A (1) was a 2-hydroxy-23-
ldehyde-12-en-3-oic acid-type triterpenoid (Institute of Medical Plant
Development of the Chinese Academy of Medical Sciences, 1984).
Acid hydrolysis of 1 yielded 2-hydroxy-23-carboxy-oleanolic acid, and the
released monosaccharide units identified as D-glucuronic acid (GlcA) and L-
arabinose (Ara) were detected by co-TLC and GC-MS with standard sugars. The
disaccharide nature of 1 was deduced from the presence of two anomeric proton
1
signals at ꢁ 4.94 (d, J ¼ 7.8 Hz) and ꢁ 5.22 (d, J ¼ 7.2 Hz) in the H-NMR spectrum
and were assigned to ꢀ-glucuronic acid and ꢂ-arabinose units, respectively. A
1H–1H COSY experiment allowed analysis of their spin systems and assignments
of their proton resonances. The assignment of their corresponding carbons, made
by a HMQC spectrum, indicated that arabinose was a terminal unit. The Aha-
(1 ! 2)-GlcA structure of the disaccharide moiety at C-3 of the oleanolic acid
residue was deduced from the HMBC correlations between C-3 (ꢁ 82.27) and H-1
(ꢁ 4.19) of the GlcA unit and between C-2 (ꢁ 84.03) of the GlcA unit and H-1
(ꢁ 5.22) of the Aha unit. The relative stereochemistry of each monosaccharide
were determined as ꢀ-D-glucuronopyranose and ꢂ-L-arabinopyranose based on
the characteristic JH-1,H-2 coupling constants and 13C-NMR data. Therefore, 1
was assigned the structure 2-hydroxy-23-aldehyde-3-O-[ꢂ-L-arabinopyranosyl-
(1 ! 2)-ꢀ-D-glucuronopyranosyl]-oleanolic acid (Figure 1).

774
Q. Xue et al.
Table 1. 13C-NMR data of celosin A (1) and celosin B (2) (C5D5N þ D2O, TMS, ꢁppm).
Position
1
2
Position
1
2
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
42.54
69.27
82.27
53.43
47.12
19.16
32.01
38.88
46.91
35.04
22.44
121.36
143.71
41.20
26.98
22.68
45.78
40.87
45.43
29.73
33.04
31.33
207.42
10.39
42.64
69.09
86.58
52.39
50.90
19.70
32.19
38.47
47.18
34.96
21.62
120.13
144.76
40.95
26.82
22.53
46.63
41.19
45.96
29.48
33.16
31.35
184.2
13.30
25
26
27
28
29
30
15.56
16.34
25.13
180.57
32.18
22.68
102.42
84.03
75.88
72.70
74.57
174.41
103.70
70.22
73.50
67.71
65.51
15.43
16.13
24.96
184.2
32.19
22.63
102.68
80.92
75.77
72.43
75.12
174.28
glcA-10
glcA-20
glcA-30
glcA-40
glcA-50
glcA-60
ara-100
ara-200
ara-300
ara-400
ara-500
gal-1000
gal-2000
gal-3000
gal-4000
gal-5000
gal-6000
102.34
72.81
73.68
68.88
74.77
60.14
COOH
HO
COOH
O
O
OH
CHO
OH
O
OH
O
OH
HMBC
OH
Figure 1. Chemical structure of compound 1 and its significant HMBC correlations.

Natural Product Research
775
COOH
HO
COOH
OH
O
O
O
COOH
OH
CH2OH
O
OH
HMBC
OH
OH
Figure 2. Chemical structure of compound 2 and its significant HMBC correlations.
Negative-ion HR-FAB-MS of celosin B (2) exhibited a quasimolecular ion at
1
m/z 839.6412, indicating a molecular formula of C42H64O17. The H-NMR and
13C-NMR spectra of 2 were very similar to those of 1, except C-23. Similarly, 2
was also a pentacyclic triterpene saponin. Acid hydrolysis of 2 afforded the
sugar components D-glucuronic acid (GlcA) and D-galactose (Gal) in addition to
1
the aglycon. The H-NMR spectrum of 2 showed the presence of two anomeric
proton signals at ꢁ 5.08 (d, J ¼ 7.2 Hz) and 5.01 (d, J ¼ 7.8 Hz) assignable to
D-glucuronic acid and D-galactose units, respectively. The structure of the
disaccharide moiety at C-3 position was established as Gal (1 ! 2)-GlcA from the
HMBC correlations between galactose H-1 (ꢁ 5.01) and GlcA C-2 (ꢁ 80.92).
The anomeric configurations of the disaccharide moiety were determined to
be D from the J-value of the anomeric proton signals. Therefore, 2 was
assigned the structure 2-hydroxy-23-carboxy-3-O-[ꢀ-D-galactopyranosyl-(1 ! 2)-ꢀ-
D-glucuronopyranosyl] oleanolic acid (Figure 2).
The known compounds were identified by spectroscopic analysis and
comparison with the published data as stigmasterol (3), ꢀ-sitosterol (4), ꢀ-
daucosterol (5), hexacosoic acid (6), palmitic acid (7) and stearic acid (8).
Compounds 1 and 2 were examined for their inhibitory activities on
CCl4-induced hepatotoxicity. The results indicated that they showed conspicuous
function of prophylaxis, which was relevant to their antioxidation of raising the
values of AST, ALT and ALP induced by CCl4.

776
Q. Xue et al.
3. Experimental
3.1. General experimental procedures
The melting points were measured on a Yanaco micromelting point apparatus
without correction. The IR spectra were obtained on a Nicolet 470 spectrometer.
EI-MS and HR-ESI-MS were measured on a Varian MAT-212 mass spectrometer
and a Micromass Q-TOF mass spectrometer, respectively. Gas chromatography
analysis was operated on an HP-5892 II with FID detector, and an HP-20M
(Carbowx 20M) capillary column (25 m ꢁ 0.3 2 mm ꢁ 0.3 mm) was used. The
1H-NMR, 13C-NMR, DEPT, HMQC and HMBC spectra were recorded on
a Bruker DMX-600 NMR spectrometer with tetramethylsilane (TMS) as an internal
standard. Column chromatography separations were carried out on D101
macroporous resin (Chemical Factory of Nankai University, Tianjin, P.R. China),
silica gel (200–300 mesh, Qingdao Haiyang Chemical Co. Ltd, Qingdao, P.R.
China), ODS (50 mesh, AA12S50, YMC) and Sephadex LH-20 (Pharmacia).
3.2. Plant material
The seeds (Semen celosiae) were collected in Bozhou, Anhui province of China
in October 2004, and were identified by one of the authors (Mei-Li Guo). A voucher
specimen (SMMU 04063) was deposited at the Herbarium of the Second Military
Medical University of China.
3.3. Extraction and isolation
The dried seeds (S. celosiae; 10 kg) were ground to a coarse powder and extracted
with 50% aqueous ethanol at room temperature after being macerated for 24 h. The
extract was concentrated under reduced pressure to give an ethanol extract (3.6 kg).
A part of the ethanol extract (3.0 kg) was subjected to chromatographic separation
over D101 macroporous resin (10 kg) eluted with H2O, 30%, 60% and 95% EtOH–
H2O by turns to give four fractions with the yields 45.3, 50.7, 90.1 and 18.9 g,
respectively. The 30% EtOH–H2O fraction was further isolated on a Si gel column
using a gradient of CHCl3–MeOH (30 : 1–20 : 1–10 : 1–1 : 1, v/v) to give seven
additional fractions; fraction 3 (5.6 g) was separated by a combination of Si gel
column (CHCl3–MeOH–H2O, 65 : 35 : 10) and ODS column chromatography
(MeOH–H2O, 1 : 4) and then by Sephadex LH-20 eluted with 40% MeOH–H2O to
give compounds 1 (50 mg), 2 (50 mg) and 5 (8 mg). The 60% EtOH–H2O fraction
was chromatographed on silica gel column eluted with n-C6H14–EtOAc (25 : 1–15 :
1–5 : 1–1 : 1) and a gradient of CHCl3–MeOH (60 : 1–50 : 1–40 : 1–30 : 1–20 : 1–10 :
1–1 : 1) to afford 15 additional fractions; fraction 12 (5 g) was separated by a Si gel
column (CHCl3–MeOH–H2O, 70 : 30 : 5) and then by Sephadex LH–20 eluted with
40% MeOH–H2O to give compounds 1 (4 g), 2 (60 mg), 3 (20 mg), 4 (25 mg), 6
(6 mg), 7 (10 mg) and 8 (13 mg). Fraction 13 (2 g) was purified on a reversed-phase
column eluted with 10% MeOH–H2O to give compound 2 (80 mg).
Celosin A (1)
White amorphous powder, m.p.: 259–260ꢂC; [ꢂ]
5
ꢀ5.319 (c 0.019, MeOH); IR
1
D
(KBr) vmax: 3408, 3396, 2948, 2863, 1604, 1094, 1041, 1727, 1691 cmꢀ1; H-NMR

Natural Product Research
777
(C5D5N þ D2O, 600 MHz) ꢁ: 0.95, 0.98, 1.02, 1.29, 1.41, 1.66 (3H each, all
s, H3-29,30, 26,27, 25,24), 3.22 (1H, m, H-18), 4.19 (1H, d, J ¼ 3.0 Hz, H-3), 4.94
(1H, d, J ¼ 7.8 Hz, H-10), 5.22 (1H, d, J ¼ 7.2 Hz, H-100), 5.48 (1H, s, H-12), 9.76
(1H, s, H-23); 13C-NMR data, see Table 1; HR-FAB-MS: m/z ¼ 817.6762 (Calcd
for C41H62O15Na [M þ Na]þ, 817.6755).
Celosin B (2)
White amorphous powder, m.p.: 240–241ꢂC; [ꢂ]
5
ꢀ4.807 (c 0.021, MeOH); IR
D
(KBr) vmax: 3323, 3308, 2947, 2925, 2860, 1617, 1105, 1078, 1772, 1724, 1696 cmꢀ1
;
1H-NMR (C5D5NþD2O, 600 MHz) ꢁ: 0.86, 1.05, 1.09, 1.26, 1.38, 1.80 (3H each,
all s, H3-29, 26, 30, 27, 25, 24), 3.29 (1H, m, H-18), 4.55 (1H, d, J ¼ 7.8 Hz, H-3), 5.01
(1H, d, J ¼ 7.8 Hz, H-1000), 5.08 (1H, d, J ¼ 7.2 Hz, H-10), 5.50 (1H, s, H-12); 13C-NMR
data, see Table 1; HR-FAB-MS: m/z ¼ 863.6412 (Calcd for C42H64O17Na
[M þ Na]þ, 863.6418).
Stigmasterol (3)
White acicular crystal, showed positive to Libermann–Burchard reaction and the
same Rf value with the reference substance, m.p.: 153–155ꢂC; IR (KBr) vmax: 3358,
2934, 2866, 1638, 1459, 1381, 1191, 1133, 1089, 1056, 1022, 969, 837, 800, 591 cmꢀ1
EI-MS m/z: 412 [M]þ (Sadtler Research Laboratories, Inc., 1970).
;
ꢀ-Sitosterol (4)
White acicular crystal, showed the same Rf value with the reference substance, m.p.:
140–142ꢂC; IR (KBr) vmax 3430, 2950, 1640, 1380, 950, 795 cmꢀ1; EI-MS m/z: 414
[M]þ; 1H-NMR (CDC13, 600 MHz) and 13C-NMR (CDCl3, 600 MHz) data:
consistent with the published values (Pei, Zhou, He, Gui, & Jiang 2004).
ꢀ-Daucosterol (5)
White powder, m.p.: 287–289ꢂC; IR (KBr) vmax 3400 2955, 1640, 1075, 1025 cmꢀ1; it
was positive to Liebermann–Burchard and Molish reactions; FAB-MS m/z: 577
[M þ 1]þ; 1H-NMR (CDC13, 600 MHz) and 13C-NMR (CDCl3, 600 MHz) data:
consistent with the published values (Guo, Zhuo, & Hong, 1995).
Hexacosoic acid (6)
White powder, m.p.: 115–117ꢂC; IR (KBr) vmax 3500, 2970, 1710 cmꢀ1; EI-MS
1
m/z: 396 [M]þ, 382, 368, 348, 332, 304, 290; H-NMR (DMSO-d6, 600 MHz) and
13C-NMR (DMSO-d6, 600 MHz): consistent with the published values (Tiriuini &
Manunta, 1991).
Palmitic acid (7)
White crystal fine powder, m.p.: 50–54ꢂC; IR (KBr) vmax 2918, 1703, 1470, 1190, 940,
720 cmꢀ1; EI-MS m/z: 256 [M] þ, 213, 199,185, 171, 157, 143, 129, 115, 101, 87, 73,
69, 57, 43, 41, 29 (Wang, Xiao, & Liang, 2003).

778
Q. Xue et al.
Stearic acid (8)
White crystal, m.p.: 71–72C; IR (KBr) vmax 3200, 2915, 2846, 1700,
1230, 720 cmꢀ1; FAB-MS m/z 284 [M]þ; 1H-NMR (CDC13, 600 MHz) and
13C-NMR (CDCl3, 600 MHz) data: consistent with the published values (Cai &
Wan, 1996).
3.4. Prevention effect on CCl4-induced hepatotoxicity in mice
Ninety Kunming mice were randomised to nine groups of 10 mice each: a normal
control group, a model control group, a biphenyl dimethylester positive control
group, and test groups of compounds 1 and 2 with doses of 1.0, 2.0 and
4.0 mg kgꢀ1, respectively. The test groups were orally administered to for 3 days.
One hour after administration on the third day all the mice were intraperitoneally
injected with 0.10% CCl4 resolved in edible oil (Lin, Yen, Lo, & Lin, 1998), with
a dose of 10 mL mg kgꢀ1. Eighteen hours later, blood from the eyeball was
collected and centrifugated to give serum for the determinations of AST, ALT
and ALP. The liver samples were prepared by 10% hepatic tissue homogenation
(adding 9 mL of physiological saline to 1 g of hepatic tissue), mixed ultrasonically
to prepare 10% homogenate and centrifugate at 3000 r minꢀ1 for 10 min, then the
contents of MDA, GSH-PX, CAT and SOD of the supernatant were determined.
See Tables 2 and 3 for pharmacodynamical results.
The values of AST, ALT and ALP of the serum were obviously increased in the
mice of the CCl4 model group, which were significantly different to the control
group. After administrations for 3 days, compounds 1 and 2 could prevent
the increase of AST, ALT and ALP caused by CCl4 effectively. There
was a significant difference when compared to the control within the concentrations
of 1.0–4.0 mg kgꢀ1 (Table 2). The value of MDA increased significantly, while those
of GSH-PX, CAT and SOD decreased in the CCl4 model group. Administration
of
1
and
2
could prevent the biochemical change caused by CCl4,
suggesting that such hepatoprotective effects of 1 and 2 may involve their
antioxidant activity.
Table 2. Blood biochemical indicators of CAS protection of hepatic injury caused by CCl4.
Group
Dose (mg kgꢀ1
)
AST (IU Lꢀ1
)
ALT (IU Lꢀ1
)
ALP (IU Lꢀ1
)
Normal
Model
Bifendate
1 (low)
1 (middle)
1 (high)
2 (low)
2 (middle)
2 (high)
–
–
127 ꢃ 22**
410 ꢃ 56
69 ꢃ 15**
241 ꢃ 51
309 ꢃ 19**
461 ꢃ 27
200
1.0
2.0
4.0
1.0
2.0
4.0
299 ꢃ 77**
317 ꢃ 72**
307 ꢃ 65**
306 ꢃ 43**
312 ꢃ 71**
303 ꢃ 63**
293 ꢃ 54**
167 ꢃ 26**
171 ꢃ 36**
168 ꢃ 47**
163 ꢃ 43**
173 ꢃ 46**
169 ꢃ 46**
162 ꢃ 42**
380 ꢃ 72**
373 ꢃ 77**
365 ꢃ 78**
362 ꢃ 77**
372 ꢃ 76**
366 ꢃ 76**
360 ꢃ 75**
Notes: Values presented as the mean ꢃ SD. **Indicates p 5 0.01.

Natural Product Research
779
Table 3. Hepatic tissue biochemical indicators of CAS protection of hepatic injury caused
by CCl4.
GSH-PX
(unit of
activity)
MDA
CAT
SOD
Dose
(nmol mgꢀ1
proteinum)
(U mgꢀ1
proteinum)
(U mgꢀ1
proteinum)
Group
(mg kgꢀ1
)
Control
Model
Bifendate
1 (low)
1 (middle)
1 (high)
2 (low)
2 (middle)
2 (high)
–
–
90 ꢃ 14**
54 ꢃ 13
1.55 ꢃ 0.42**
2.54 ꢃ 0.62
18.83 ꢃ 1.69**
15.02 ꢃ 2.54
362.3 ꢃ 36.8**
279.6 ꢃ 48.8
341.3 ꢃ 33.0*
320.6 ꢃ 37.1*
324.1 ꢃ 25.9*
334.5 ꢃ 35.9*
318.7 ꢃ 34.6
321.5 ꢃ 32.3*
343.1 ꢃ 36.1**
200
1.0
2.0
4.0
1.0
2.0
4.0
72 ꢃ 11**
66 ꢃ 9*
1.80 ꢃ 0.45**
1.93 ꢃ 0.54*
1.84 ꢃ 0.53*
1.78 ꢃ 0.65**
1.89 ꢃ 0.49*
1.78 ꢃ 0.52**
1.69 ꢃ 0.55**
18.66 ꢃ 1.52**
17.28 ꢃ 2.47
73 ꢃ 16**
74 ꢃ 13**
65 ꢃ 10*
74 ꢃ 13**
79 ꢃ 15**
17.73 ꢃ 1.82*
18.70 ꢃ 1.60**
17.63 ꢃ 1.92*
17.71 ꢃ 2.01*
18.74 ꢃ 1.69**
Notes: Values presented as the mean ꢃ SD. *Indicates p 5 0.05 compared with model group;
**indicates p 5 0.01.
Acknowledgements
This study was supported by the Science and Technology Commission Research Projects of
Shanghai Municipality (06DZ19715-1) and the Research and Development Projects of
Traditional Chinese Medicine of PLA (2006172003).
References
Anonymous. (1992). The wealth of India (pp. 414–415). New Delhi: CSIR.
Cai, L.N., & Wan, H.S. (1996). Studies on the chemical constituents of Alisma orientalis
(Sam.) Juzep. Natural Product Research and Development, 8, 5–9.
Guo, X.M., Zhuo, L., & Hong, Y.F. (1995). Studies on the chemical constituents of
Ranunculus ternatus Thunb. Acta Pharmaceutica Sinica, 30, 931–934.
Hase, K., Kadota, S., & Basnet, P. (1997). Immunostimulating activity of celosian, an
antihepatotoxic polysaccharide isolated from Celosia argentea. Planta Medica, 63,
216–219.
Hase, K., Kadota, S., Basnet, P., Takahashi, T., & Namba, T. (1996a). Research article
hepatoprotective effects of traditional medicines. Phytotherapy Research, 10, 387–392.
Hase, K., Kadota, S., Basnet, P., Takahashi, T., & Namba, T. (1996b). Protective effect of
celosian, an acidic polysaccharide, on chemically and immunologically induced liver
injuries. Biological and Pharmaceutical Bulletin, 19, 567–572.
Institute of Medical Plant Development of the Chinese Academy of Medical Sciences. (1984).
Records of Chinese traditional medicine (p. 441). Beijing: People’s Medical Publishing
House.
Kirtikar, K.R., & Basu, B.D. (1995). Indian medicinal plants (pp. 2053–2054). Dehradun:
Bishen Singh & Mahendra Paul Singh.
Lin, C.C., Yen, M.H., Lo, T.S., & Lin, J.M. (1998). Evaluation of the hepatoprotective and
antioxidant activity of Boehmeria nivea var. nivea and B. nivea var. tenacissima. Journal
of Ethnopharmacology, 60, 9–17.
Pei, G., Zhou, P.H., He, G.H., Gui, F.L., & Jiang, D.F. (2004). Studies on liposoluble
constituents of Huperzia crispate China. Natural Product Research and Development, 16,
213–214.

780
Q. Xue et al.
Sadtler Research Laboratories, Inc. (1970). Standard infrared grating spectra. Philadelphia:
Sadtler Research Laboratories, Inc.
Tiriuini, B., & Manunta, A. (1991). Constituents of the essential oil of the fruits of
Zanthoxylum bungeanum. Planta Medica, 57, 90–91.
Vetrichelvan, T., & Jegadeesan, M. (2000). Hepatoprotective effects of traditional medicines.
Isolation of the active constituent from seeds of Celosia argentea. Indian Drugs, 37,
286–288.
Wang, L., Xiao, H.B., & Liang, X.M. (2003). Studies on chemical constituents of Gastrodia
elata. Chinese Traditional and Herbal Drugs, 34, 584–585.