全 文 ：Effect of polysaccharide FⅢ-2 from Ganoderma tsugae on
proinflammatory cytokine production by THP-1 cells and
human peripheral blood mononuclear cells
GONG Yu-Jing1 , FEI Xiao-Fang3 , GAO Xiao-Xia1 , ZHANG Jie2 , WANG Ben-Xiang4 ,
MINAMI Mutsuhiko
5 , NAGATA Toshi6 , IKEJIMA Takashi1＊
(1.China-Japan Research Institute of Medical Sciences and 2.Department of Traditional Chinese Pharmacy , Changchun
College of Traditional Chinese Medicine , Changchun　130021 , China;3.College of Life Sciences , Jilin
University , Changchun　130012 , China;4.Jilin Institute of Natural Medicine , Changchun
130021 , China;5.Department of Parasitology , School of Medicine , Yokohama City
University , Yokohama 236-0004 , Japan;6.Department of Microbiology ,
Hamamatsu Medical University , Hamamatsu , 431-3192 , Japan)
Abstract:Regulatory effects of water insoluble polysac-
charide F Ⅲ-2 (F Ⅲ-2)and its pyridine-chlorosulfonic
acid modified water-soluble polysaccharide F Ⅲ-2-S
(FⅢ-2-S)on the production of proinflammatory cytokines
were investigated.FⅢ-2 and FⅢ-2-S down-regulated IL-
1αproduction by THP-1 cells stimulated by higher con-
centration of lipopolysaccharide(LPS)and phorbol myris-
tate acetate (PMA), whereas they up-regulated IL-8 se-
cretion without stimulants or at weaker stimulatory condi-
tions.F Ⅲ-2 itself induced TNFαproduction by THP-1
cells and at relatively weak stimulatory conditions with
LPS 10mg·L-1 and PMA 200 nmol·L-1 significantly up-
regulated TNFαproduction in a dose dependent manner.
However , at higher concentration of LPS (100 mg·L-1)
and PMA(200 nmol·L-1), FⅢ-2 down-regulated TNFα
production of stimulated THP-1.Human peripheral blood
mononuclear cells(PMBC)responded to FⅢ-2 and FⅢ-
2-S in IL-1α, IL-8 and TNFαproduction in a fashion sim-
ilar to THP-1 cell responses.FⅢ-2 and FⅢ-2-S regulat-
ed mRNA expression for IL-1αand TNFα.Production of
TNFαprotein reflected quantity of TNFαmRNA , howev-
er , IL-1αmRNA expression did not coincide with IL-1α
protein production.In conclusion , the polysaccharides of
Ganoderma tsugae mycelium have immunomodulatory
　
　　Received date:2000-10-18　Accepted date:2001-06-19
　　Foundation item:The project supported by National Natural
Science Foundation of China(39077847);and the Fund of Japan-
China Medical Association(1998)
　　Biography:GONG Yu-Jing(1973 -), female , native of
Tonghua, Jilin Province , main research field is immunopharmaco-
logy.
　　＊Corresponding author.Tel:(0431)5634444 , Fax:(0431)
5634444 , E-mail:ikejimat@sina.com
effects on the production of proinflammatory cytokines.
Chemical modification changes the intensity and reverses
the direction of regulatory effects of the polysaccharides on
proinflammatory cytokine production.
Key words:Ganoderma tsugae;polysaccharides;inter-
leukin-1α;interleukin-8;tumor necrosis factorα
CLC number:R285.5
Document code:A
Article ID:1000-3002(2001)06-0401-07
Ganoderma tsugae (G.tsugae), a tradi-
tional Chinese herb has been used as anti-tumor ,
roborant and other remedies in China for cen-
turies[ 1] .The water-insoluble polysaccharide FⅢ-
2(F Ⅲ-2), extracted with alkali from the fruit
body of G.tsugae , inhibited the growth of im-
planted S 180 murine sarcoma cells and increased
the life span of tumor-implanted mice.Three anti-
tumor fractions were obtained by gel filtration of F
Ⅲ-2.They are protein-containing(1-3)-β-D-glu-
cans and the average relative molecular weight is
(l0-190)×103[ 2] .However , the mechanism of
the action of FⅢ-2 is so far unclear.Investigators
have suggested that anti-tumor activity be based on
(l-3)-β-D-glucan of the polysaccharides.In vivo
tumoricidal activity of the polysaccharides might be
related to some extent to the activation of host im-
mune system.In recent years , it has been report-
ed that sulfated polysaccharides have more potent
·401·中国药理学与毒理学杂志　　2001 年 12月;15(6):401-407
anti-tumor and anti-virus activities including
anti-HIV[ 3, 4] .We obtained the water-soluble
polysaccharide FⅢ-2-S(FⅢ-2-S)by sulfation of
water insoluble FⅢ-2 with pyridine-chlorosulfonic
acid.FⅢ-2 showed the stronger anti-tumor activi-
ty than F Ⅲ-2-S.Some cytokines , such as inter-
leukin-1α(IL-1α), interleukin-8(IL-8)and tumor
necrosis factorα(TNFα), are known to play im-
portant roles in the regulation of immune respons-
es.IL-1α, IL-8 and TNFαare produced from a
variety of cells , especially of the macrophage/
monocyte lineage.Therefore , by using human
myelomonocytic leukemic cell line , THP-1 , and
human peripheral blood mononuclear cells
(PBMC), we compared the effects of FⅢ-2 and F
Ⅲ-2-S on proinflammatory cytokine (IL-1α, IL-8
and TNFα)production and expression of mRNA
for these cytokines produced by THP-1 to make
clear the relationship of the polysaccharides and
their immunomodulating and anti-tumor activities.
1　MATERIALS AND METHODS
1.1　Preparation of samples
F Ⅲ-2 was extracted and purified from G.
tsugae fruiting body[ 2] .FⅢ-2-S was prepared by
modification with pyridine-chlorosulfonic acid[ 3] .
After 2mL chlorosulfonic acid was dropped into 10
mL of ice-cold pyridine , F Ⅲ-2 (100 mg)was
added and stirred in the boiling water for 1 h ,
then moved to the ice bath.pH was adjusted to
7.0 with NaOH (3 mol·L-1), then five volumes
of 95% ethanol were added.After centrifugation
at 450 ×g for 20 min , precipitate was dia-
lyzed against distilled water and lyophilized.
133.7 mg F Ⅲ-2-S was finally obtained.To rule
out the possible endotoxin contamination in the
samples(FⅢ-2 and FⅢ-2-S), the Limulus ame-
bocyte lysate (LAL)test was performed by using
assay kit (Seikagaku Kogyo , Japan)[ 5] .The en-
dotoxin levels in the two samples were below l.0
mg·L-1.
1.2　Cell culture
THP-1 cells were stimulated with lipopoly-
saccharide (LPS)(E.coli serotype 0111∶B4 ,
Sigma)at 1 or 10 mg·L-1 and 200 μmol·L-1
phorbol myristate acetate (PMA , Sigma) for in-
duction of IL-1αand IL-8.On the other hand , for
TNFαinduction , 10 mg·L-1 LPS and 200 nmol·
L
-1
PMA were used.Cells were preincubated for
1.5 h prior to administration of various concentra-
tions (4-400 mg·L-1) of F Ⅲ-2 or F Ⅲ-2-
S.THP-1 cells and human PBMC were cultured as
previously reported[ 7 ,8] .
1.3　Quantitation of cytokines
Cytokines (IL-1α, IL-8 and TNFα)were
quantitated by human cytokine specific radioimm-
noassay.Methods for radioimmunoassay for human
cytokines were previously reported
[ 9 , 10] .
1.4　Measurement of IL-1αand TNFαmRNA
expression by THP-1 cells using RT-PCR
method
Before the stimulation with 10 or 100 mg·
L-1 LPS and 200 nmol·L-1 PMA , THP-1 cells
were cultured with various concentrations of FⅢ-2
or FⅢ-2-S.Then , the previously reported meth-
ods were followed[ 7-10] .
1.5　Statistical analysis
Data were expressed as x ±s , and assessed
by unpaired Student′s t test.
2　RESULTS
2.1　Effects of FⅢ-2 and FⅢ-2-S on proin-
flammatory cytokine production in THP-1 cells
Tab l illustrates the production of IL-1αand
IL-8 by THP-1 cells with or without FⅢ-2 or FⅢ-
2-S under the stimulation with different concentra-
tions of LPS and PMA.When THP-1 cells were
stimulated with PMA and the higher concentration
of LPS(10mg·L-1), THP-1 cells produced rela-
tively higher amount of IL-1α〔(1.70±0.14)μg·
L-1〕 compared to medium control 〔(0.13±
0.04)μg·L-1〕.F Ⅲ-2 and F Ⅲ-2-S decreased
the production of IL-1αto medium control level.
Untreated THP-1 cells produced considerable
amount of IL-8〔(10±1)μg·L-1〕.Addition of
FⅢ-2 or F Ⅲ-2-S increased IL-8 production.
FⅢ-2 40 mg·L-1 induced 18-fold amount of IL-8
without LPS or PMA.At the same polysaccharide
·402· ChineseJournal of Pharmacology and Toxicology　2001　Dec;15(6)
Tab l.　Effects of water insoluble polysaccharide FⅢ-2 (F Ⅲ-2)and its pyridine-chlorosulfonic acid modified
water-soluble polysaccharide FⅢ-2-S(FⅢ-2-S)on IL-1αand IL-8 production in THP-1 cells
Polysaccharide
/mg·L-1
Interleukin concentration/μg·L-1
Medium
LPS (1 mg·L-1)+
PMA(200 nmol·L-1)
LPS (10 mg·L-1)+
PMA(200 nmol·L-1)
IL-1α
Medium 0.13±0.04 0.17±0.05 1.70±0.14　
FⅢ-2 4 0.16±0.06 0.08±0.06 　0.10±0.03＊＊＊
FⅢ-2 40 0.12±0.07 0.09±0.00 　0.080±0.011＊＊＊
FⅢ-2 400 0.09±0.03 0.32±0.18 　0.12±0.03＊＊＊
FⅢ-2-S 4 0.101±0.022 　0.84±0.22＊＊ 　0.062±0.012＊＊＊
FⅢ-2-S 40 0.082±0.021 0.100±0.023 　0.081±0.012＊＊＊
FⅢ-2-S 400 0.063±0.024 0.093±0.021 　0.17±0.03＊＊＊
IL-8
Medium 　　　　　 10±1 　　　　　　24±5 　　　　　　15±2
FⅢ-2 4 5±1＊＊ 23±6 52±21＊
FⅢ-2 40 180±30＊＊ 58±5＊＊ 21±6
FⅢ-2 400 110±40＊＊＊ 34±1＊ 15±2
FⅢ-2-S 4 13±7 92±8＊＊＊ 50±10＊＊
FⅢ-2-S 40 64±7＊＊＊ 35±10 19±6
FⅢ-2-S 400 42±4＊＊＊ 42±4＊＊ 21±6
LPS:lipopolysaccharides.PMA:phorbol myristate acetate.Polysaccharides were added to THP-1 cell culture 30 min prior to administration of
LPS+PMA. x±s , n=3.＊P<0.05 , ＊＊P<0.01 , ＊＊＊P <0.001 , compared with corresponding medium.
concentration , FⅢ-2-S has similar stimulatory ef-
fect.LPS 1 or 10 mg·L-1 and PMA 200 nmol·
L-1 induced higher amount of IL-8.Addition of F
Ⅲ-2 or F Ⅲ-2-S had similar but much weaker
stimulatory effects at LPS 1 mg·L-1 .
　　THP-1 cells produced low amount of
TNFα〔(0.22±0.04)μg·L-1〕 without stimu-
lants , then the addition of FⅢ-2 or FⅢ-2-S mod-
erately increased the production of TNFα, except
FⅢ-2-S 400 mg·L-1(Tab 2).For induction of
TNFαin THP-1 , the concentration of LPS(10 or
100 mg·L-1)was at least 10 times higher than
those for induction of IL-1αand IL-8(LPS 1 or 10
mg·L-1).Under the lower stimulation with LPS
10 mg·L-1 and PMA 200 nmol·L-1 , TNFαpro-
duction slightly increased 〔(2.16 ±0.26)μg·
L-1〕.FⅢ-2 , especially at 400 mg·L-1 , further
stimulated the TNFαproduction by seventy four
times 〔(160±11)μg·L-1〕, however , effect of
F Ⅲ-2-S was not obvious.At higher stimulation
by LPS 100 mg·L-1 and PMA 200 nmol·L-1 ,
remarkably high amount of TNFαwas produced by
THP-1 cells 〔(640±25)μg·L-1〕.Addition of
FⅢ-2 4 and 40 mg·L-1 reduced TNFαproduc-
tion , while F Ⅲ-2 400 mg·L-1 augmented the
TNFαproduction by two times.F Ⅲ-2-S reduced
TNFα production in a concentration-dependent
manner(Tab 2).
2.3　Effects of FⅢ-2 and FⅢ-2-S on pro-
inflammatory cytokine production in human
peripheral blood monocyte cells
F Ⅲ-2 and FⅢ-2-S decreased the production
of IL-1α in PBMC while IL-1α production in
PBMC was highly stimulated with LPS〔(92±18)
μg·L-1 , volunteer #1 and (2.4 ±1.4)μg·
L-1 , volunteer #3〕(Tab 3).Lower amount of
·403·中国药理学与毒理学杂志　　2001 年 12月;15(6)
Tab 2.Effect of FⅢ-2 and FⅢ-2-S on tumor necrosis factor α(TNFα)production in THP-1 cells
Polysaccharide
/mg·L-1
TNFα/μg·L-1
Medium
LPS(10 mg·L-1)+
PMA(200 nmol·L-1)
LPS(100 mg·L-1)+
PMA(200 nmol·L-1)
Medium 0.22±0.04 2.16±0.26　 640±25　　
FⅢ-2 4 　0.72±0.22＊ 1.44±0.26＊ 1.42±0.25＊＊＊
FⅢ-2 40 　　2.0±0.3＊＊＊ 　9.6±0.6＊＊＊ 7.2±1.2＊＊＊
FⅢ-2 400 0.6±1.1 160±11＊＊＊ 　1280±130＊＊＊　
FⅢ-2-S 4 　　3.4±1.1＊＊＊ 　0.121±0.012＊＊＊ 800±60＊ 　
FⅢ-2-S 40 　　0.84±0.20＊＊ 0.8±1.2　 352±40＊＊＊　
FⅢ-2-S 400 　　0.08±0.03＊＊ 1.28±0.14＊＊ 0.35±0.08＊＊＊
Polysaccharides were added to THP-1 cell culture 30 min prior to administration of LPS+PMA. x±s , n=3.＊P <0.05 , ＊＊P<0.01 ,
＊＊＊P<0.001 , compared with corresponding medium.
Tab 3.　Effects of FⅢ-2 and FⅢ-2-S on proinflammatory cytokine secretion in human PBMC
Drug/mg·L-1 Concentration/μg·L
-1
Volunteer#1 Volunteer#2 Volunteer#3
IL-1α
Medium 2.2±0.3 0.0083±0.006　 0.31±0.03
FⅢ-2 400 2.1±0.9 0.23±0.09＊ 4.1±2.5
FⅢ-2-S 400 0.9±0.6 　0.31±0.11＊＊ 3.1±1.8
LPS 1 92±18 0.523±0.012　 2.4±1.4
LPS 1+FⅢ-2 400 　　　　1.5±1.2###　　 　0.021±0.013### 1.9±0.5
LPS 1+FⅢ-2-S 400 　　　 0.73±0.15### 　 　0.032±0.004### 0.7±0.3
IL-8
Medium 500±300　 260±200 　　　　1000±400
FⅢ-2 400 2400±1500　 1700±900　 320±30
FⅢ-2-S 400 980±240　 590±80　 1600±500
LPS 1 2100±1200　 230±70　 30000±20000
LPS 1+FⅢ-2 400 　170±130### 600±140# 590±130###
LPS 1+FⅢ-2-S 400 　230±60### 　1900±800## 1600±500###
TNFα
Medium 0.23±0.05 0.48±0.03 0.23±0.05
FⅢ-2 400 　 16±6＊＊＊ 3.4±1.2＊ 　　6±4＊＊＊
FⅢ-2-S 400 　 11±4＊＊＊ 2.8±1.5 3.3±1.2＊
LPS 1 5.1±2.2 19.8±1.8　 0.57±0.13
LPS 1+FⅢ-2 400 　 59±8### 　　5.4±1.1### 　8.9±1.2##
LPS 1+FⅢ-2-S 400 8.7±1.4　 　　0.99±0.14### 1.6±0.4#
PBMC:peripherial blood monocyte cells.Polysaccharides were added to human PBMC culture 30 min prior to administration of LPS. x±s ,
n=3.＊P<0.05 , ＊＊P <0.01 , ＊＊＊P<0.001 , compared with corresponding medium group;#P <0.01 , ##P<0.01 , ###P <
0.001 , compared with corresponding LPS group.
IL-1αin non-stimulated PBMC produced , howev- er , addition of F Ⅲ-2 or F Ⅲ-2-S showed some
·404· ChineseJournal of Pharmacology and Toxicology　2001　Dec;15(6)
tendency to induce IL-1αproduction in an individ-
ual 〔from(0.31±0.03)μg·L-1 to (4.1±2.5)
or(3.1±1.8)μg·L-1 , volunteer #3〕.When
LPS induced relatively lower amount of IL-8
〔(230 ±70)μg·L-1〕 in volunteer #2 , both
kinds of polysaccharide samples obviously in-
creased IL-8 production〔(600±140)and(1900
±800)μg·L-1〕, respectively.On the contrary ,
in volunteers #1 and #3 , F Ⅲ-2 and FⅢ-2-S
significantly down-regulated IL-8 production while
LPS-stimulated PBMC produced higher amount
of IL-8 〔(2.1±1.2)and (30±20)mg·L-1 ,
respectively〕.
　　Without LPS , PBMC itself produced low
amount of TNFα.Addition of F Ⅲ-2 or F Ⅲ-2-S
up-regulated TNFαproduction and FⅢ-2′s stimu-
latory effect was more potent than that of F Ⅲ-2-
S′.Stimulation with LPS induced volunteer #2′s
PBMC to produce higher amount TNFα〔(19.8±
1.8)μg·L-1〕, FⅢ-2 and F Ⅲ-2-S decreased
the TNFαprouduction 〔(5.4±1.1), (0.99±
0.14)μg·L-1〕.In volunteer #1 and #3 , LPS
induced low amount of TNFα〔(5.1±2.2)and
(0.57±0.13)μg·L-1 , respectively〕 and FⅢ-2
up-regulated the TNFαproduction.
2.4　Effects of F Ⅲ-2 and FⅢ-2-S on
cytokine mRNA expression in THP-1 cells
In order to investigate the modulatory effect
of FⅢ-2 and F Ⅲ-2-S , specific concentration of
polysaccharides at which cytokine production in
THP-1 cells affected significantly were selected.
Their effects on IL-1αand TNFαmRNA expres-
sion were further examined by RT-PCR.It has
shown that the cDNA bands for IL-1αwas more
obvious at 30 cycles than that at 24 cycles , mean-
ing that the cDNA amplication had not reached
plateau stage at 24 cycles.Therefore , at 24
cycles of cDNA amplification for IL-1α, the effect
of F Ⅲ-2 and F Ⅲ-2-S on the original amount of
mRNA could be compared.F Ⅲ-2 4 mg·L-1
inhibited IL-1α protein production with higher
stimulation , but have on obvious effect on IL-1α
mRNA expression (Tab 1 and Fig 1 , lane D).
FⅢ-2 4 mg·L-1 or F Ⅲ-2-S 4 mg·L-1 them-
selves did not induce IL-1αmRNA expression
Fig 1.　IL-1αmRNA expression.Lane A:THP-1 cell
control;lane B:LPS(10 mg·L-1)+PMA(200 nmol·L-1);lane
C:FⅢ-2 4 mg·L-1;lane D:FⅢ-2 4 mg·L-1+LPS+PMA;
lane E:F Ⅲ-2-S 4 mg·L-1;lane F:FⅢ-2-S 4 mg·L-1+PMA+
LPS.
(Fig 1 , lane C and E), but F Ⅲ-2-S obviously
reduced IL-1αmRNA expression induced by the
stimulation(Fig 1 , lane F).
cDNA amplification at 24 cycles mRNA ex-
pression for TNFαapparently did not reach the
plateau stage.cDNA bands were too weak to be
compared(Fig 2).Therefore , at both 24 and 30
cycles , we compared the amplified cDNA amount
which might reflect the original quantities of TNFα
mRNA.FⅢ-2 4 mg·L-1 decreased TNFαprotein
production and mRNA expression when THP-1
cells were stimulated with LPS 10 mg·L-1 and
PMA 200 nmol·L-1(Fig 2 , lane D).FⅢ-2 400
mg·L-1 increased TNFαmRNA expression and
TNFα protein production(Fig 2 , lane F).FⅢ-
2-S 4 mg·L-1 decreased TNFαproduction and
mRNA expression at this stimulatory condition(Fig
2 , lane H).Without stimulants , F Ⅲ-2 4 mg·
L-1 and F Ⅲ-2-S 4 mg·L-1 increased TNFα
mRNA expression(Fig 2 , lane C and G).FⅢ-2-
S 400 mg·L-1 inhibited the expression of TNFα
mRNA and the expression of TNFαmRNA were
completely inhibited in the presence of LPS and
PMA(Fig 2 , lane J).However , F Ⅲ-2-S 400
mg·L-1 slightly reduced the protein production
(Fig 2 , lane I).
·405·中国药理学与毒理学杂志　　2001 年 12月;15(6)
Fig 2.　TNFαmRNA expression.Lane A:THP-1 cell
control;lane B:LPS(10 mg·L-1)+PMA(200 nmol·L-1);lane
C:FⅢ-2 4 mg·L-1;lane D:FⅢ-2 4 mg·L-1+LPS+PMA;
lane E:FⅢ-2 400 mg·L-1;lane F:FⅢ-2 400 mg·L-1+LPS+
PMA;lane G:F Ⅲ-2-S 4 mg·L-1;lane H:FⅢ-2-S 4 mg·L-1+
LPS+PMA;lane I:FⅢ-2-S 400 mg·L-1;lane J:FⅢ-2-S 400
mg·L-1+LPS+PMA.
3　DISCUSSION
Although THP-1 cells and PBMC can be easi-
ly induced by bacterial endotoxin to produce proin-
flammatory cytokines including IL-1α, IL-8 or
TNFα, the contaminated endotoxin in our herbal
polysaccharide samples was not enough to disturb
the quantitation of cytokines in our experiments.
　　FⅢ-2 and F Ⅲ-2-S apparently inhibited the
production of IL-1αwhen THP-1 cells or PBMC
produced relatively higher amount of IL-1α at
higher-dose of LPS and PMA.FⅢ-2 and FⅢ-2-S
up-regulated IL-8 production by THP-1 cells with-
out stimulants or with minor stimulation.When
IL-8 production was at low level in PBMC
culture , FⅢ-2 and FⅢ-2-S increased the IL-8 pro-
duction.However , in the cases of high amounts of
IL-8 production , no matter with or without LPS
stimulation , both polysaccharides reduced the IL-8
production.Under the lower stimulation , TNFα
production was at low level and FⅢ-2 up-regulated
the TNFαproduction.On the other hand , under
the higher stimulation , its production reached sig-
nificantly at high level , then addition of FⅢ-2 in-
hibited TNFαproduction.FⅢ-2′s bidirectional reg-
ulation of cytokine production was the most obvious
in TNFα production by THP-1 cells.Chemical
modification of F Ⅲ-2 by pyridine-chlorosulfonic
acid(FⅢ-2-S)generally reversed the direction of
TNFαproduction.Our previous studies showed that
chemical modification of polysaccharides , FI0-b
and FI0-c , sometimes reverse the direction of cy-
tokine production from up to down regulation and
vice versa[ 3 , 8] .
Regulatory effects of FⅢ-2 and FⅢ-2-S on
the production of proinflammatory cytokines by
THP-1 cells had similar tendency to the cytokine
production by human PBMC.Regardless of indi-
viduals or kinds of cyokines , when cytokine
production was at high levels , F Ⅲ-2 or FⅢ-2-S
down-regulated the production.If the cytokine
production was at low level , these herbal polysac-
charides up-regulated their production.However ,
each blood donor has different genetic background
on production of proinflammatory cytokines.Sen-
sitivities to FⅢ-2 or FⅢ-2-S were not similar
among blood donors.Therefore , we first used
THP-1 cells instead of using human PBMC for
examining the polysaccharides′ effects on the
production of the cytokines.
FⅢ-2 4 mg·L-1 up-regulated IL-1αmRNA
amount under the treatment of LPS and PMA , but
the radioimmunoassay results showed that FⅢ-2
4 mg·L-1 down-regulated IL-1αprotein produc-
tion , indicating that FⅢ-2 4 mg·L-1 affected the
post-transcriptional regulation.However , TNFα
protein production was reflected by its mRNA
quantity.Cytokines′gene expression might be
regulated by FⅢ-2 and FⅢ-2-S at both transcrip-
tional and post-transcriptional levels.
In conclusion , FⅢ-2′s bidirectional regula-
tion of proinflammatory cytokine production was
more significant than FⅢ-2-S , especially on TNFα
production.The pyridine-chlorsulfonic acid modifi-
cation might change the immunological functions.
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铁杉灵芝多糖 FⅢ-2对人组织瘤细胞和人外周血白细胞
产生炎症性细胞因子的影响
宫毓静1 , 费晓方3 , 高晓霞1 , 张　洁2 , 王本祥4 , 南陆彦5 , 永田年6 , 池岛乔1
(长春中医学院 1.附属医院中日医学研究所 , 2.中药系分析化学教研室 , 吉林 长春　130021;3.吉林大学
生命科学院 , 吉林 长春　130012;4.吉林天然药物研究所 , 吉林 长春　130021;5.横滨市立大学
医学部寄生虫学教研室 ,横滨 , 236-0004 ,日本;6.滨松医科大学微生物学教研室 ,
滨松 ,431-3192 ,日本)
摘要:用放射免疫分析法及逆转录聚合酶链反应法 ,
比较研究水不溶性铁杉灵芝多糖(FⅢ-2)及其吡啶-
氯磺酸修饰产物(FⅢ-2-S)对人炎症性细胞因子蛋
白质及其mRNA产生的影响.结果表明 ,FⅢ-2(4 ,40
或400 mg·L-1)显著提高低剂量脂多糖(LPS)10 mg·
L-1协同佛波醇 14烷酰乙酸盐(PMA)200 nmo1·L-1
诱导的人组织瘤 THP-1 细胞产生肿瘤坏死因子 α
(TNFα),然而明显地抑制高剂量 LPS l00 mg·L-1协
同PMA诱导的THP-1细胞产生 TNFα.FⅢ-2-S(4或
40 mg·L-1)提高无刺激剂细胞产生TNFα,高浓度
F Ⅲ-2-S(400mg·L-1)抑制 TNFα产生.FⅢ-2与FⅢ-
2-S 提高白介素-8 的产生 ,降低刺激剂引起的白介
素-1α的产生.F Ⅲ-2 的抗肿瘤作用可能是与 TNFα
产生有关.FⅢ-2对人外周血单核细胞产生各种细
胞因子和对 THP-1细胞产生细胞因子的机理基本上
相同.结果提示 ,松杉灵芝菌丝体水不溶性多糖在
不同的刺激条件下具有双向免疫调节作用.化学修
饰的多糖可改变原多糖对细胞因子产生的调节方
向.
关键词:铁杉灵芝;多糖类;白介素-1α;白介素-8;
肿瘤坏死因子α
　
　　基金项目:国家自然科学基金资助项目(39077847);日
中医学协会助成金(1998)
(本文编辑　乔　虹)
·407·中国药理学与毒理学杂志　　2001 年 12月;15(6)