全 文 ：天然产物研究与开发 Nat Prod Ｒes Dev 2016，28:195-201，227
文章编号:1001-6880(2016)2-0195-08
Ｒeceived:August 1，2015 Accepted:January 5，2016
Fundation Item:Quanzhou Science and Technology Project (2012Z76);Fu-
jian Provincial Education Department Project (JA13394)
* Corresponding author Tel:86-595-22138267;E-mail:qiudanying@ ya-
hoo． com
兖州卷柏水提物对四氯化碳致小鼠肝
损伤的保护作用和抗氧化活性
谢永华1，温扬敏1，2，韩萍萍3，邱丹缨1*
1泉州医学高等专科学校，泉州 362000;2 福建师范大学，福州 350007;3 昆士兰大学，澳大利亚 4072
摘 要:研究兖州卷柏对四氯化碳(CCl4)致小鼠肝损伤的保护效果。昆明小鼠随机分为 6 组(n = 8):正常组、
CCl4 模型组、水飞蓟素对照组、兖州卷柏水提物(高、中、低)剂量组(400、200、100 mg /kg)。各组连续灌胃给药 9
d后，除正常组外，其余各组用 CCl4 溶液(2 mL /kg)灌胃。结果显示兖州卷柏能显著的抑制肝损伤血清中
SGOT、SGPT、ALP、LDH、胆固醇和胆红素升高(P ＜ 0． 01)。兖州卷柏能降低损伤肝组织的脂肪含量，局灶性坏
死，中心静脉充血和正弦空间拥塞。体外抗氧化实验表明兖州卷柏抗氧化活性指标 T-AOC、LPO、T-SOD 和 NO
的 IC50值明显高于维生素 C(P ＜ 0． 05)。提示兖州卷柏水提物可能通过其抗氧化活性对四氯化碳致小鼠肝损伤
产生保护效果。
关键词:兖州卷柏;四氯化碳;肝损伤;抗氧化活性
中图分类号:Ｒ282． 71 文献标识码:A DOI:10． 16333 / j． 1001-6880． 2016． 2． 005
Hepatoprotective and Antioxidant Activity of Selaginella involvens Aqueous
Extract on Carbon Tetrachloride-Induced Hepatic Damage in Mice
XIE Yong-hua1，WEN Yang-min 1，2，HAN Ping-ping 3，QIU Dan-ying 1*
1Microbiology Lab，Quanzhou Medical College，Quanzhou 362000，China;2College
of Life Sciences，Fujian Normal University，Fuzhou 350007，China;3 Tissue Engineering and
Microfluidics Laboratory，University of Queensland，St． Lucia，QLD 4072，Australia
Abstract:In this study，the hepatoprotective and antioxidant activity of Selaginella involventis aqueous extract was evalu-
ated using carbon tetrachloride (CCl4)-induced hepatic damaged mice model． KM mice was randomly divided into 6
groups (n = 8)，including normal，CCl4-model，Silymarin-control groups and three different dosages (400，200，100 mg /
kg)of S． involventis aqueous extract in experimental groups． After oral gavage for 9 days with drug，KM mice were treated
with CCl4(2 mL /kg)by oral gavage，except for normal group． The results showed that administration of S． involventis a-
queous extracts significantly reduced the level of SGOT，SGPT，ALP，Lactic dehydrogenase (LDH)，cholesterol and bili-
rubin when compared to that of hepatotoxin treated mice． More importantly，histopathological data showed that S． invol-
ventis aqueous extracts reduced the fatty contents，focal necrosis，congestion in central vein and congestion in sinusoidal
spaces in CCl4-induced hepatic mice． In addition，the in vitro antioxidant assays demonstrated that the IC50 values of T-
AOC，LPO，T-SOD and NO radical scavenging activities were increased when compared with the level of vitamin C． All
these experimental results suggested that S． involventis aqueous extract may protect hepatic activity through antioxidant
activities in CCl4-induced mice．
Key words:Selaginella involventis;carbon tetrachloride;hepatic damage;antioxidant activity
Introduction
Liver is one of the few organs with highly specialized
function which can undergo an astonishing degree of re-
generation and drug metabolism［1］． However，a few
known risk factors，such as hepatic virus，inflammation，
obesity and alcohol consumption，may lead to hepatic
injury，which is one of the most serious diseases affect-
ing human health［2］． In the process of physiological liv-
er metabolism，reactive oxygen species (ＲOS)includ-
ed three states including free radicals，peroxides and
excited states． When the liver underwent hepatic inju-
ry，elevated generation of ＲOS and decreased capacity
of antioxidant could result in a state of redox imbal-
ance，which may induce loss of cell viability and
death［3］． Then，oxidative stress would trigger an irre-
versible situation that cannot be repaired by antioxidant
and biosynthetic pathways on human pathology ［4］．
However，current treatment is still far from optimal to
cure liver disease． Therefore，it is of great importance to
develop the drugs to protect the liver with induced hep-
atotoxicity and antioxidant properties． In this study，we
investigated the potential of traditional Chinese medi-
cine for this purpose［5］．
A great variety of Chinese traditional drugs have been
used in folk medicine to treat liver diseases and boost
liver functions［6］． Among them，Selaginella involven is
abundantly distributed in south China，especially in Fu-
jian Province． This fern contains a variety of fla-
vonoids，alkaloids，phenols，organic acid and carbohy-
drate compounds． It has been used as a folk herbal
medicine to treat some diseases as its hemostasis，acute
icteric hepatitis，hepatocirrhosis and pneumonia［7］． It is
used in ethnomedical practices in certain remote villa-
ges with the belief that it had some activities，such as
antitumor，antibacterial，antiviral and anti-inflamma-
tion［8］． Ｒecent findings demonstrated that S． involventis
aqueous extract has certain antioxidant abilities and
may inhibit the growth of human esophageal carcinoma
cells in vitro［9］． Its alkaloids may inhibit the growth of
hepatocellular carcinoma cell H22 of mouse in
vitro［10］． However，limited studies exist regarding the
effect of S． involventis onconventional scientific studies，
particularly in hepatoprotective effects area．
The purpose of this study was to evaluate the potential
effect of S． involventis aqueousextract on carbon tetra-
chloride (CCl4)-induced hepatic damage in mice mod-
el． Ｒeactive oxygen species (ＲOC)were the main
causes of carbon tetrachloride (CCl4)-induced acute
liver injury，which results to the mice model marked el-
evation of alanine aminotransferase and aspartate amin-
otransferase．
Materials and Methods
Preparation of S． involventis extract
The matured plants of S． involventis were collected from
valleys in Sanming City，Fujian Province，China． The
specimen samples were immediately sent to Prof SU Q
of the Medicine Ｒesearch Institute of Quanzhou for au-
thentication． The voucher specimen (AUOCAS031)
was subsequently maintained in the herbarium of
Quanzhou Medical College． The herb was washed twice
with distilled water to remove the contaminants，and ov-
en-dried at 60 ℃ for 3 days，and then grinded into
powder．
The powdered sample of S． involventis (500 g)was ex-
tracted with sterile water (5，000 mL)at 80 ℃ for 20
min． Then the solvent was precipitated in vacuum at 60
℃ using a rotary evaporator (Eyela，Japan)． The ex-
tract powder was dissolved with sterile distilled water
and administered orally to mice in final concentrations
of 400，200 and 100 mg /kg body weight．
Animal model and oral treatment with S． involven-
tis extract
Kunming male mice weighed at 18-20 g were obtained
from Slac Laboratory Animal Ltd． (animal certification
number was 2012-0002，Shanghai，China)． The animals
were housed in colony cages at an ambient temperature
of 25 ± 2 ℃ with a 12 h light and 12 h dark circle．
All the animal experiments were conducted in accord-
ance with the guidelines of the experimental animal
ethical committee of Quanzhou Medical College．
In this experiment，the animals were randomly divided
into six groups，with eight mice per group，and treated
as follows:Group 1，Oral gavage with ddH2O (5 mL /kg
·bw)daily for 9 days (normal control group，n = 8)．
Group 2，Oral gavage with sterile distilled water (5
mL /kg· bw)daily for 9 days + single dose of (2
mL /kg·bw，ip)CCl4 with liquid paraffin (1 ∶ 1)for
one day (positive control hepatotoxic group，n = 8)．
Group 3，Oral gavage with silymarin (100 mg /kg·
bw)daily for 9 days + single dose of (2mL /kg·bw，
ip)CCl4 with liquid paraffin (1∶ 1)for one day (well
established standard drug hepatoprotective group，n =
8)． Group 4，Oral gavage with S． involventis aqueousex-
691 Nat Prod Ｒes Dev Vol. 28
tract (400 mg /kg·bw)daily for 9 days + single dose
of (2 mL /kg·bw，ip)CCl4 with liquid paraffin (1∶ 1)
for one day (treatment group，high dose，n = 8)． Group
5，Oral gavage with S． involventis aqueousextract (200
mg /kg·bw)daily for 9 days + single dose of (2mL /
kg·bw，ip)CCl4 with liquid paraffin (1 ∶ 1)for one
day (treatment group，medium dose，n = 8)． Group 6，
Oral gavage with S． involventis aqueousextract (100
mg /kg·bw)daily for 9 days + single dose of (2 mL /
kg·bw，ip)CCl4 with liquid paraffin (1 ∶ 1)for one
day (treatment group，low dose，n = 8)．
Analysis of liver functional enzymes
To investigate the liver function，the enzymes activity
was assessed through the blood samples collected from
each group． On the 10th day，all animals were anesthe-
tized with mild ether and blood sample were collected
by eye bleeding method． The blood serum was separa-
ted from blood clots by centrifugation at room tempera-
ture with 4，000 rpm for 10 min． Then，liver function
tests were estimated with standard kits (Amyjet Scien-
tific Inc，Wuhan China)to detect known biochemical
parameters listed as below:serum glutamic oxaloacetic
transaminase (SGOT)，serum glutamic pyruvic trans-
aminase (SGPT)［11］，lactate dehydrogenase (LDH)，
alkaline phosphatase (ALP) ，total protein，albumin，
sugar，cholesterol and bilirubin．
Followed by the sacrifice for all the animals，the livers
were dissected using a glass rod and a ceramic potter at
4 ℃ with 0． 1 M Tris-HCl (pH 7． 4)． Thereafter，the
supernatant was collected by centrifugation at 4，000
rpm for 10 min for the spectrophotometric tests of lipid
peroxide (LPO) radical scavenging assay，hydroxyl
radical scavenging assay (HＲSA)by using standard
kits (Amyjet Scientific Inc，Wuhan china)．
Various concentrations (2 μg /mL to 600 μg /mL)of
S． involventis aqueous extract was used for the test of to-
tal antioxidant capacity (T-AOC)assay，lipid peroxide
(LPO)radical scavenging assay，hydroxyl radical scav-
enging assay (HＲSA)［12］，nitric oxide radical scaven-
ging assay and total superoxide dismutase (T-SOD)
radical scavenging assay by using standard protocols
with vitamin C (positive control)． All the above activi-
ties were evaluated using standard kits (Amyjet Scien-
tific Inc，Wuhan China)．
Pathological examination and immunohistological
analysis
For histological analysis，the livers were fixed in 10%
formaldehyde for 24 h at room temperature． After rins-
ing with PBS twice，all the specimens were embedded
in paraffin［13］． Then 5 μm thick serial sections were cut
using microtome，stained with haematoxylin-eosin． The
sections were scored as described using a light micro-
scope by Jamshidzadeh et al ［14］． The histological dam-
age was graded on a scale of 0-3 ∶ 0 = no visible cell
damage;1 = focal hepatocytes damage on ＜ 25% -50%
of the tissue;2 = extensive，but focal hepatocytes le-
sion;3 = global hepatocytes necrosis．
To further investigate whether TGFβ1 played role dur-
ing the treatment，the immunohistochemistry staining
was carried out in this study． Briefly，after dewaxing
and dehydration，the slices were incubated with TGFβ1
primary antibody at 4 oC overnight． Followed by incu-
bation at room temperature with a biotinylated universal
secondary antibody (DAKO，CA，USA) for 15 min．
Then，the antibody complexes were visualized by the
addition of a buffered diaminobenzidine (DAB)sub-
strate for 4 min． Mayer’s haematoxylin (HD Scientific
Pty Ltd．)was used for counter staining．
Statistical analyses
All the data were expressed as mean ± standard devia-
tion (SD)． One-way analysis of variance (ANOVA)
was used to calculate statistical significance． Least sig-
nificant difference (LSD)-t test was used for multiple
comparison． Statistical significance was considered as P
＜ 0． 05 using SPSS (version 13． 0)．
Ｒesults
Effect of S． involventis extracts on liver enzymes ac-
tivity
As shown in Table 1，it was noted that hepatotoxin trea-
ted mice had a significantly increased level of SGOT
［(284． 80 ± 14． 33) IU /L］，SGPT ［(212． 85 ±
10. 56)IU /L］，ALP ［(649． 61 ± 43． 14)IU /L］，LDH
［(2664． 17 ± 205． 28)IU /L］，cholesterol ［(9． 34 ±
2． 24)mmol /L］，bilurubin ［(6． 17 ± 1． 72)umol /L］
in Group 2 when compared to those of control group (P
791
Vol. 28 XIE Yong-hua，et al:Hepatoprotective and Antioxidant Activity of Selaginella
involvens Aqueous Extract on Carbon Tetrachloride-Induced Hepatic Damage in Mice
＜ 0． 05)． While，the content of albumin ［(18． 16 ±
2. 15)mmol /L］and total protein ［(31． 84 ± 5． 230．
47)mmol /L］ showed a dramatically decrease (P ＜
0. 05 ) when compared to those of control
group. Interestingly，administration (400 and 200 mg /
kg·bw)of S. involventis aqueous extracts reduced the
level of SGOT，SGPT，ALP，LDH，cholesterol and bili-
rubin (P ＜ 0. 05)when compared with hepatotoxin
treated group. Moreover，it was found that high dosage
of S. involventis (400 mg /kg·bw)extracts had an op-
timal reduction of SGOT ［(164. 19 ± 15. 43)IU /L］，
SGPT ［(79. 91 ± 12. 83)IU /L］，ALP ［(256. 72 ±
31. 69)IU /L］，LDH ［(1444. 02 ± 105. 36)IU /L］，
cholesterol ［(3. 08 ± 0. 61)mmol /L］ and bilurubin
［(5. 18 ± 0. 88)umol /L］. However，the level of albu-
min ［(39. 44 ± 4. 67)mmol /L］ and total protein
［(64. 87 ± 9. 83) mmol /L］ were significantly in-
creased in the high dose group (400 mg /kg·bw) (P
＜ 0. 05)when compared to those of control group (Ta-
ble 1).
Table 1 Effect of S. involventis aqueous extract on the biochemical parameters in CCl4 induced hepatotoxicity in mice
(Mean ± SD)
Parameters Control
Hepatotoxin
group (CCl4)
Silymarin
positive control
S. involventis aqueous extract (mg /kg·bw)
100 200 400
SGOT (IU /L) 126. 27 ± 10. 12 284. 80 ± 14. 33* 145. 73 ± 8. 61* 277. 32 ± 13. 36＊＊ 213. 57 ± 17. 06＊＊ 164. 19 ± 15. 43＊＊
SGPT (IU /L) 52. 41 ± 3. 43 212. 85 ± 10. 56* 68. 30 ± 7. 74* 231. 14 ± 16. 39 139. 62 ± 18. 63＊＊ 79. 91 ± 12. 83＊＊
ALP (IU /L) 129. 51 ± 11. 68 649. 61 ± 43. 14* 147. 05 ± 24. 21 468. 57 ± 34. 14＊＊ 349. 28 ± 53. 17＊＊ 256. 72 ± 31. 69＊＊
LDH (IU /L) 824. 63 ± 114. 32 2664. 17 ± 205. 28* 1182. 83 ± 146. 25* 2787. 31 ± 247. 55 1958. 95 ± 181. 43＊＊1444. 02 ± 105. 36＊＊
CHL (mmol /L) 2. 92 ± 0. 41 9. 34 ± 2. 24* 2. 81 ± 0. 56 10. 22 ± 2. 84＊＊ 5. 42 ± 0. 39＊＊ 3. 08 ± 0. 61＊＊
BIL (μmol /L) 3. 14 ± 0. 24 6. 17 ± 1. 72* 5. 28 ± 0. 62* 6. 86 ± 1. 61 5. 32 ± 1. 28＊＊ 5. 18 ± 0. 88＊＊
ALB (mmol /L) 42. 92 ± 5. 16 18. 68 ± 2. 15* 31. 38 ± 6. 41* 20. 56 ± 2. 72 35. 49 ± 3. 39＊＊ 39. 44 ± 4. 67＊＊
SUG (mmol /L) 5. 29 ± 0. 71 5. 37 ± 0. 73 6. 02 ± 0. 68 5. 97 ± 0. 83 5. 55 ± 0. 69 5. 23 ± 0. 82
TPN (mmol /L) 65. 46 ± 11. 07 31. 84 ± 5. 23* 53. 64 ± 11. 42* 48. 72 ± 13. 56＊＊ 45. 61 ± 10. 60＊＊ 64. 87 ± 9. 83＊＊
Note:Data were mean ± standard deviation (SD)values for groups of six animals each. Values were statistically significant，* P ＜ 0. 05 compared with
control at the corresponding time;＊＊ P ＜ 0. 05 compared with CCl4 -treated group at the corresponding time.
Meanwhile，hepatotoxin itself revealed a significant (P
＜ 0. 05)decrease in the levels of LPO ［(0. 757 ±
0. 028)μmol /gport) ］and HＲSA ［(80. 37 ± 7. 75 U /
mgport) ］activities on the liver tissues when compared
to that from control. On the other hand，administration
of S. involventis aqueous extracts (400 and 200 mg /kg
·bw)demonstrated a significant increase in the level
of LPO and HＲSA (P ＜ 0. 05)when compared with
hepatotoxin treated mice. In addition，high dose group
(400 mg /kg·bw)reached the peak reduction level
for LPO ［(1. 337 ± 0. 061)μmol /gport) ］and HＲSA
［(154. 71 ± 12. 51)U /mgport) ］(P ＜ 0. 05)when
compared to that from control group (Table 2).
Table 2 Effect of S. involventis aqueous extract on CCl4-induced oxidative stress in liver tissue of mice (Mean ± SD)
Liver homogenate
parameters
(μmol /L)
Control
Hepatotoxin
group
(CCl4)
Silymarin
positive control
S. involventis aqueous extract (mg /kg·bw)
100 200 400
LPO (μmol /gport) 0. 885 ± 0. 035 0. 757 ± 0. 028* 1. 508 ± 0. 068* 0. 749 ± 0. 076 0. 826 ± 0. 044＊＊＊ 1. 337 ± 0. 061＊＊＊
HＲSA (U /mgport)110. 56 ± 12. 14 80. 37 ± 7. 75* 106. 18 ± 14. 32 108. 55 ± 9. 36＊＊ 133. 67 ± 13. 29＊＊ 154. 71 ± 12. 51＊＊
Note:Data were mean ± standard deviation (SD)values for groups of six animals each. Values were statistically significant:* P ＜ 0. 05 compared with
control at the corresponding time;＊＊ P ＜ 0. 05 compared with CCl4 -treated group at the corresponding time.
Effect of S. involventis extracts on histology of liver
tissues
The representative H＆E histopathological staining
showed that hepatotoxin treated mice induced the maxi-
mum fatty changes，focal necrosis，congestion in central
vein and congestion in sinusoidal spaces. While
S. involventis aqueous extract at the concentration of
100 mg /kg· bw and 200 mg /kg· bw treated mice
891 Nat Prod Ｒes Dev Vol. 28
showed the reduction in fatty changes，focal necrosis，
congestion in central vein and congestion in sinusoidal
spaces when compared to control group. However，no
visible changes were observed with high dose of
S. involventis extracts (400 mg /kg· bw)，except for
fatty changes and focal necrosis (Table 3 and Fig. 1).
Table 3 Dose dependant histopathological scores of S. involventisaqueous extract in CCl4 induced hepatotoxicity in mice
Parameters Fattychanges
Focal
necrosis
Congestionin
central vein
Congestionin
sinusoidal
spaces
Hepatocytes
Deformation Total
Hepatotoxin group (CCl4) 3 3 2 2 0 10
Silymarin 1 1 1 0 0 3
S. involventis treated (400 mg /kg·bw) 1 1 0 0 0 2
S. involventis treated (200 mg /kg·bw) 1 1 1 0 0 3
S. involventis treated (100 mg /kg·bw) 2 2 1 1 0 6
Fig. 1 Dose dependent effect of S. involventis aqueous extract on CCl4 induced hepatotoxicity in mice (100 ×)
Hepatotoxin treated mice induced the maximum fatty changes，focal necrosis，congestion in central vein and congestion in sinusoidal spaces.
A:Control group;B:CCl4 treated mice;C:Silymarin treated mice (100 mg /kg·bw);D:100 mg /kg·bw (Low dose)of S. involventis aqueous extract
treated mice;E:200 mg /kg·bw (Medium dose)of S. involventis aqueous extract treated mice;F:400 mg /kg·bw (High dose)of S. involventis aque-
ous extract treated mice.
Fig. 2 Expression pattern of TGFβ1 on liver tissues. Weak expression of TGFβ1 was observed in healthy liver tissues in
picture A and D.
A:Control group;B:CCl4 treated mice;C:Silymarin treated mice (100 mg /kg·bw);D:100 mg /kg·bw (Low dose)of S. involventis aqueous extract
treated mice;E:200 mg /kg·bw (Medium dose)of S. involventis aqueous extract treated mice;F:400 mg /kg·bw (High dose)of S. involventis aque-
ous extract treated mice.
991
Vol. 28 XIE Yong-hua，et al:Hepatoprotective and Antioxidant Activity of Selaginella
involvens Aqueous Extract on Carbon Tetrachloride-Induced Hepatic Damage in Mice
Expression pattern of TGFβ1 on liver tissues
As seen in Fig. 2，weak expression of TGFβ1 was ob-
served in normal and healthy liver tissues，while there
was strong expression of TGFβ on hepatic portal area
and the interstitial collagen in CCl4 induced-
mice. When the mice was treated with S. involventis ex-
tracts at the dosage of 400 mg /kg·bw，it was noted
that TGFβ1 was strongly expressed when compared to
that from normal control group.
In vitro antioxidant activity of S. involventis
The in vitro antioxidant assays showed that the IC50 val-
ues were (67. 44 + 2. 01)μg /mL，(59. 35 + 1. 29)
μg /mL，(102. 16 + 3. 65)μg /mL，(31. 70 + 2. 28)
μg /mL and (33. 63 + 1. 52)μg /mL for T-AOC，LPO，
T-SOD，NO and HＲSA radical scavenging activities，re-
spectively. Moreover，the results were also comparable
with the positive control of vitamin C (Table 4).
Table 4 IC50 values of S. involventisaqueous extract and vitamin C with various antioxidant activities (μg /mL)
S. involventis aqueous extract IC50 Vitamin C IC50
T-AOC radical scavenging 67. 44 + 2. 01* 25. 61 + 1. 37
LPO radical scavenging 59. 35 + 1. 29* 35. 28 + 1. 47
T-SOD radical scavenging 102. 16 + 3. 65* 28. 85 + 1. 07
NO radical scavenging 31. 70 + 2. 28* 8. 33 + 1. 41
HＲSA radical scavenging 33. 63 + 1. 52 30. 91 + 1. 38
Note:Data were mean ± standard deviation (SD)values. Values were statistically significant:* P ＜ 0. 05 compared with vitamin C.
Discussion and Conclusion
Increasing evidences demonstrated that lots of Chinese
traditional herbal medicines could relieve patients’
pains against some liver diseases;however，there are
still little known whether these herbal extracts possess
this capability. Among of them，S. involventis is widely
used in some different therapeutic applications in folk
medicine. In this study，hepatoprotective potential of
this plant was investigated via in vivo model with hepa-
tocellular damage induced by CCl4 in mice，which has
been established by published protocols with significant
elevations of SGOT and SGPT activities［15，16］ . In the
early of liver injury，SGPT was found with in a high
concentration in cytosolic，while SGOT was localized in
cytosol and mitochondria and released into the circula-
tion. Continuous hepatic damage would result in an ele-
vation ALP，LDH and bilirubin activity in the serum
and further lead to severe hepatic dysfunction. In the
present work，oral administration of S. involventis aque-
ous extract can reduce all the elevated biochemical pa-
rameters such as SGOT，SGPT，ALP，LDH and bilirubin
levels.
In hepatotoxin intoxicated mice，the reduction in the
level of total protein and albumin might be due to the
damage produced and localized in the endoplasmic re-
ticulum which leads to its functional failure with de-
crease in protein synthesis and accumulation of triglyc-
erides. Intoxication of CCl4 also results in inhibition of
the synthesis of the bile acids from the cholesterol and
leads to elevated level of cholesterol ［17］. Suppression of
cholesterol level in the serum parameters suggests the
inhibition of synthesis of the bile acids from cholester-
ol，and is reversed by the pre-administration of the
S. involventis aqueous extracts. Ｒeduction in the level of
SGOT，SGPT towards the normal value is an indication
of the stabilization of the plasma membrane as well as
repair of hepatic tissue damage caused by
CCl［18］4 . Ｒeduction in the level of ALP and bilirubin
suggested the stabilization of the biliary function and
the increased level of protein as well as albumin sug-
gested the stabilization of the endoplasmic reticulum
leading to the protein synthesis.
Oxidative stress is the state of imbalance between the
level of antioxidant defense system and production of
the oxygen derived species［19］. The increased level of
oxygen and oxygen derived species such as superoxide
radicals，hydroxyl radicals and peroxide radicals causes
the oxidative stress［20］. The in vitro assays such as T-
AOC radical scavenging assay，LPO radical scavenging
assay，T-SOD radical scavenging assay，NO radical
scavenging assay and HＲSA radical scavenging assay
002 Nat Prod Ｒes Dev Vol. 28
suggested the ability of S. involventis aqueous extract to
reduce the biological oxidative stress. We observed sig-
nificant increases in the liver levels of LPO radical
scavenging and HＲSA radical scavenging in
S. involventis aqueous extract treated mice compared
with CCl4-treated mice. Hence，the hepatoprotective
effect of the aqueous extract may be achieved by the
scavenging free radical activity of the oxidative stress
［21，22］.
Moreover，the histopathological analysis showed that，
the normal liver architecture was disturbed by the hep-
atotoxin treated mice. But，the liver sections obtained
from the mice treated with the aqueous extract and in-
toxicated with hepatotoxin，the normal cellular architec-
ture retained as compared with silymarin treated mice，
thereby further confirming the protective effect of the
extract.
TGFβ1 has a prominent role in the disease process to
stimulate hepatic stellate cell activation and transforma-
tion into fibroblast and then affecting liver fibrosis posi-
tively. TGFβ1 could increase the expression of plate-
let derived growth factor (PDGF)and its receptor in
platelets，and then promote hepatic stellate cell prolifer-
ation. In this study，immunohistochemistry data showed
increased expression of TGFβ1 in CCl4 induced chronic
liver injury. S. involventis inhibited TGFβ1 protein ex-
pression in liver tissues，indicating that S. involventis
extracts may possess the capability of anti-liver injury
through TGFβ1 protein expression levels.
The results of the present study suggested that
S. involventis aqueous extract might possess hepatopro-
tective properties through its antioxidant activity in
vivo. However，more in-depth studies will be required to
quantitatively and structurally determine the phyto-
chemical principles during these activities.
Ｒeferences
1 Takate SB，Pokharkar ＲD，Chopade VV，et al. Hepato-pro-
tective activity of the aqueous extract of launaea intybacea
(Jacq)Beauv against carbon tetrachloride-induced hepatic
injury in Albino Ｒats. J Pharm Sci Technol，2010，2:247-
251.
2 Ｒavikumar S，Gnanadesigan M. Hepatoprotective and antioxi-
dant activity of a mangrove plant Lumnitzera racemosa. Asian
Pac J Tropic Biomed，2011，1:348-352.
3 Videla LA. Cytoprotective and suicidal signaling in oxidative
stress. Biol Ｒes，2010，43:363-369.
4 Mukherjee S，Gogoi JB. Free radicals in diseases and poten-
tial role of phytoconstituents-A cause with emerging impor-
tance. Current Chem Biol，2011，5:223-232.
5 Iwalokun BA，Efedede BU，Alabi-Sofunde JA，et al. Hepato-
protective and antioxidant activities of Vernonia amygdalina
on acetaminophen-induced hepatic damage in mice. J Med
Food，2006，9:524-530.
6 Dong H，Lu FE，Zhao L. Chinese herbal medicine in the
treatment of nonalcoholic fatty liver disease. Chin J Integr
Med，2012，18):152-160.
7 He W，Zeng ZP，Wang Y. Ｒesearch overview on the plant
medicine from Selaginella. Chin Tradut Herb Drugs，2000，
31:954-955.
8 Wen YM. Advances in Selaginella involvens. Herald Med，
2012，31:341-343.
9 Xie YH，Wen YM，Luo CL. Extraction of active substances
from Selaginellae involventis Sping and inhibitory effects on
human esophageal carcinoma cells. Herald Med，2012，31:
1132-1135.
10 Qiu DY，Wen YM，Xie YH，et al. Effect of ultrasonic-assis-
ted extraction and anti-proliferation activities on hepatocellu-
lar carcinoma cell H22 of alkaloid from Selaginella invol-
vens. Sci Technol Food Ind，2013，34:265-268.
11 Odebiyi OO，Sofowora EA. Phytochemical screening of Nigeri-
a medicinal plants II. J Nat Prod，1978，41:234-246.
12 Patel T，Shirode D，Ｒoy SP，et al. Evaluation of antioxidant
and hepatoprotective effects of 70% ethanolic bark extract of
Albizzia lebbeckin rats. Int J Ｒes Pharm Sci，2010，1:270-
276.
13 Ofusori DA，Adejuwon AO. Histopathological studies of acute
and chronic effects of Calliandra portoricensis leaf extract on
the stomach and pancreas of adult Swiss albino mice. Asian
Pac J Tropic Biomed，2011，1:182-185.
14 Jamshidzadeh A，Fereidooni F，Salehi Z． Hepatoprotectiveac-
tivity of Gundelia tourenfortii． J Ethnopharmacol，2006，101:
233-237.
15 Yanpallewar SU，Sen S，Tapas S，et al. Effect of Azadirachta
indicaon paracetamol-induced hepatic damage in albino
rats. Phytomedicine，2003，10:391-396.
16 Ｒajkapoor B，Jayakar B，Kavimani S，et al. Effect of dried
fruits of Carica papaya Linn on hepatotoxicity. Biol Pharm
Bull，2002，25:1645-1646.
(下转第 227 页)
102
Vol. 28 XIE Yong-hua，et al:Hepatoprotective and Antioxidant Activity of Selaginella
involvens Aqueous Extract on Carbon Tetrachloride-Induced Hepatic Damage in Mice
5 Hu LH，Sim KY，Sampsoniones A-M，a unique family of
caged polyprenylated benzoylphloroglucinol derivatives，from
Hypericum sampsonii. Tetrahedron，2000，56:1379-1386.
6 Matsuhisa M， Shikishima Y， Takaishi Y， et
al. Benzoylphloroglucinol derivatives from Hypericum sca-
brum. J Nat Prod，2002，65:290-294.
7 Xu G，Kan WLT，Zhou Y，et al. Cytotoxic acylphloroglucinol
derivatives from the twigs of Garcinia cowa. J Nat Prod，
2010，73:104-108.
8 Marti G，Eparvier V，Moretti C，et al. Antiplasmodial benzo-
phenones from the trunk latex of Moronobea coccinea (Clusi-
aceae). Phytochemistry，2009，70:75-85.
9 Chien S. C，Chyu CF，Chang IS，et al. A novel polyprenylated
phloroglucinol，garcinialone，from the roots of Garcinia multi-
flora. Tetrahedron Lett，2008，49:5276-5278.
10 Tsui WY，Brown GD. Sesquiterpenes from Baeckea frutes-
cens. J Nat Prod，1996，59:1084-1086.
11 Zhu H (朱华旭)，Tang YP (唐于平)，Min ZD (闵知大)，
et al. Bioactive constituents from whole herbs of vernonia ci-
nerea (II). Zhongguo Zhongyao Zazhi (中国中药杂志)，
2009，34:2765-2767.
12 Elgamal MHA，Elewa NH，Elkhrisy EAM，et al. 13 C NMＲ
Chemical shifts and carbon-proton coupling constants of some
furocoumarins and furochromones. Phytochemistry，1979，18:
139-143.
13 Zhang YL (张艳丽)，Feng ZY (冯志毅)，Zheng XK (郑晓
珂)，et al. Chemical constituents from leaves of Ｒehmannia
glutinosa Libosch. Zhongguo Yaoxue Zazhi (中国药学杂
志)，2014，49:15-21.
14 Chanotiya C，Sammal S，Mathela C. Composition of a new
chemotype of Tanacetum nubigenum. Indian J Chem B，
2005，44:1922-1926.
15 Cheng XX (成晓霞)，Zhang ZQ (张志琪)，Xiao YP (肖娅
萍)，et al. Chemical components and cytotoxic activity of
Polytrichum commune L. ex Hedw. Nat Prod Ｒes Dev (天然
产物研究与开发)，2014，26:1733-1738.
16 Yuan CM，Tang GH，Zhang Y，et al. Bioactive limonoid and
triterpenoid constituents of Turraea pubescens. J Nat Prod，
2013，76:
櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵
1166-1174.
(上接第 201 页)
17 Sureshkumar SV，Mishra SH. Hepatoprotective activity of ex-
tracts from Pergularia daemia Forsk against carbon tetrachlo-
ride induced toxicity in rats. Pharmacogn Mag，2007，3:187-
191.
18 Kumar Ｒ，Kumar S. Hepatoprotective activity of aerial parts
of Plumbago zeylanica Linn against carbon tetrachloride in-
duced hepatotoxicity in rats. Int J Pharm Pharm Sci，2009，
1:171-175.
19 Ｒosa MP，Gutiérrez VS. Hepatoprotective and inhibition of
oxidative stress of Prostechea michuacana. Ｒec Nat Prod，
2009，3(1):46-51.
20 Zhu YZ，Huang SH，Tan BK，et al. Antioxidants in Chinese
herbal medicines:a biochemical perspective. Nat Prod Ｒep，
2004，21:478-489.
21 Channabasavaraj KP，Badami S. Hepatoprotective and antiox-
idant activity of methanol extract of Ficus glomerata. Nat
Med，2008，62:379-383.
22 Gebhardt Ｒ，Fausel M. Antioxidant and hepatoprotective
effects of artichoke extracts and constituents in cultured rat
hepatocytes. Toxicol In Vitro，1997，11:669-672.
722Vol. 28 范翊民等:木竹子枝叶的化学成分及细胞毒和抗炎活性研究