E L S E VI E R

Journal of Ethnopharmacology 54 (1996) 69-75

Journal of
E-THNO-
I ~ M A C ~
Recovery of the hematopoietic system by Si-Jun-Zi-Tang in
whole body irradiated mice
Hs u e - Yi n Hs u a, J e n q - J e r Ya n g b, S h i - L o n g L i a n a, Ya u - Hu i Ho a, Ch u n - Ch i n g L i n b' *
aSehool of Technology for Medical Sciences, Kaohsiung Medical College, Kaohsiung, Taiwan, R.O.C.
bSchool of Pharmacy, Kaohsiung Medical College, Kaohsiung, Taiwan, R.O.C.
Received 23 February 1996; revised 13 June 1996; accepted 14 June 1996
Abstract
The herbal formulation Si-Jun-Zi-Tang reduced the decrease of leukocytes, erythrocytes, thrombocytes and
hematocrit in irradiated mice. In general, its protection was more effective in leukocytes and thrombocytes than other
hematocytes. Protection of bone marrow stem cells by Si-Jun-Zi-Tang was markedly enhanced by increased
radiotolerance under the dose ranging from 0 to 5 Gy. This increased radiotolerance led to a prolonged shoulder in
the survival curve but did not influence the D O value. Si-Jun-Zi-Tang exerted a beneficial effect on clinical syndromes
such as anemia. From the results in this study, we concluded preliminarily that the most effective concentration with
least toxicity was about 20 mg/20 g body weight. At this dose, levels of leukocytes as well as thrombocytes were
enhanced significantly after )~-irradiation. Elevation of erythrocytes and hematocrits could also be found but was not
significant.
Keywords: Radiation; Si-Jun-Zi-Tang; Spleen colony-forming unit (CFUs); Hematocytes; Hematocrit
1. I ntroducti on
Our previ ous investigations have shown t hat
t he i nt raperi t oneal i nj ect i on of crude ext ract s
f r om Kuei - Pi - Tang and Jen-Sheng-Yang-Yung-
Tang before or af t er i rradi at i on can reduce hema-
t opoi et i c injuries ( Hsu et al., 1991, 1993). Several
t radi t i onal Chi nese herbal medi ci nal prescri pt i ons
* Corresponding author.
such as Bu- Zhong- Yi - Qi - Tang, Xi ao- Chai - Hu-
Tang and Shi - Quan- Da- Bu- Tang have also been
f ound t o have r adi opr ot ect i ve activities ( Hoso-
kawa, 1986). These results indicate t hat ext ract s
of some t radi t i onal Chinese medi ci nal prescrip-
tions can diminish r adi at i on- i nduced damage. The
st rat egy of reduci ng r adi at i on i nj ury t o nor mal
tissues will be of significant benefit, allowing t he
use of l arger r adi at i on doses in radi odi agnosi s or
r adi ot her apy. The search f or mor e effective radi o-
prot ect i ve agents displaying di fferent pr ot ect i ons
0378-8741/96/$15.00 1996 Elsevier Science Ireland Ltd. All rights reserved
PH S0378-8741(96)01450-5
70 H.- Y. Hsu et al. / J ournal o f Et hnop harmacol ogy 54 ( 1996) 6 9 - 7 5
against radiation-induced damage is therefore war-
ranted.
Survival brought about by radioprotectors after
potentially lethal irradiation is thought to be due
primarily to their effect on hematopoietic cells
(Floersheim et al., 1988). When measuring bone
marrow hematopoietic stem cell colony-forming
units (Harris and Phillips, 1971), and peripheral
blood hematocytes counts (Smith et al., 1965), it
has been shown repeatedly that some radioprotec-
tors protect hematopoietic cells against radiation
damage. Accordingly, we used peripheral blood
cell counts as indicators of bone marrow function
in order to assess the radioprotection of normal
tissue in this study.
The purpose of the experiments reported here
was to study whether Si-Jun-Zi-Tang, which was
considered to have some effects on promoting
recovery from anemia (Lin, 1986), could act as a
radioprotective agent.
2. Mat eri al s and methods
2.1. Animals
ICR strain male mice 6- 7 weeks old were pur-
chased from the Animal Center, College of
Medicine, National Taiwan University. These mice
were selected and divided randomly into several
groups and housed 10 to a cage at 24 + IC and
60 __+10% of relative humidity. They were adminis-
tered with nutritional chow and water ad libitum
and, about 2 weeks later, had gained a body weight
between 18 and 23 g when treated with radiation
and Si-Jun-Zi-Tang.
2.2. Preparation of ext ract s
An extract of Si-Jun-Zi-Tang was prepared by
decocting the dried prescription of herbs with
boiling water. The duration of decoction was
about 90 min. The decoction was filtered and
concentrated to 1 g/ml and then stored at 4C
before administration. The ingredients of 16.5 g
Si-Jun-Zi-Tang include 4 g of Ginseng Radix, 4 g
of Atractylodis Rhizoma, 4 g of Poria (sclerotium),
1.5 g of Glycyrrhizae Radix, 1.5 g of Zingiberis
Rhizoma and 1.5 g of Zizyphi Fructus. These
ingredients correspond to parts of the following
plants: Panax ginseng C.A. Meyer (Araliaceae),
At ract ylodes ovata Thunb. (Asteraceae), Poria co-
cos (Schw.) Wol f ( = Pachyma hoelen Rumphius)
(Polyporaceae), Glycyrrhiza uralensis Fischer et
DC. (Febaceae), Zingiber officinale Roscoe (Zin-
giberaceae) and Zizyphus jujuba Mill. var. inermis
(Bunge.) Rehd. (Rhamnaceae), respectively. These
plant materials were obtained from Ko Da Phar-
maceutical Co. (Taoyuan Hsien, Taiwan) and
identified by C.C. Lin, School of Pharmacy, Kaoh-
slung Medical College, and their voucher speci-
mens have been deposited at the Herbarium of the
School of Pharmacy, Kaohsiung Medical College.
2.3. Irradiation and administration
Irradiation with ;(-ray was carried out by 4-MV
linear accelerator (80 cm FSD; 20 x 20 field size;
200 cGy/min dose rate), by total body irradiation.
Before irradiation, 10 mice from each group were
put together in a round plastic box (20 cm in
diameter) and the radiation exposure was such that
each mouse received the same total body irradiated
dose. Stored Si-Jun-Zi-Tang solution was in-
traperitoneally injected daily for seven consecutive
days before irradiation. Control mice were injected
with 0.9% sodium chloride solution.
2.4. Analysis of dose related effect
The dose-related radioprotective effect of Si-
Jun-Zi-Tang was determined by the mean survival
time, mortality rate and increase in life span within
60 days after z-irradiation with 8 Gy. Mice were
injected i.p. for seven consecutive days with the
administered dose, 10, 20, 30, 50, 80 or 100 mg/20
g body weight, once a day, before whole body
z-irradiation. Data of treated samples were calcu-
lated relative to untreated controls. Twenty mice
were used in each treated group.
2.5. Measurement of blood forming st em cells
(spleen colony forming unit, CFUs)
The colony forming unit assay method of Till
and McCulloch (1961) was carried out to measure
H.-Y. Hsu et al. / Journal of Ethnopharmacology 54 (1996) 69- 75
Table 1
Dose effects of Si-Jun-Zi-Tang with or without irradiation on mice
71
Treatment No. of mice MST60 (days) MR60 (%) ILS60 a (%)
Radiation(Gy) Si-Jun-Zi-Tang (mg/20 g body wt.)
- - 20 60.0 + 0.0 0
- - 10 20 60.0 _+0.0 0 0.0
20 20 60.0 _+0.0 0 0.0
- - 30 20 58.4 _+5.6 10 - 2. 8
50 20 50.9 _+16.9"* 25 - 15.2
80 20 15.1 _+8.4** 100 --74.8
100 20 3.7 _+2.8** 100 --93.9
8 20 35.1 _+14.5 90
8 10 20 38.1 +_15.8 85 8.5
8 20 20 44.1 _+16.4" 60 25.6
8 30 20 44.9 _+13.5" 85 27.9
8 50 20 34.5 _+14.7 95 - 1.7
8 80 20 13.3 _+7.8** 100 -62.1
8 100 20 3.5 _+2.8** 100 -90. 0
Data are presented as mean + S.D. from treatments using 20 mice per group as indicated; MST60, mean survival time within 60
days; MR60, mortality rate within 60 days; ILS, increase in life span within 60 days.
T - C
alLS = x 100, where T is the mean survival time of treated mice and C is the mean survival time of untreated or treated
C
control mice [modified from You et al. (1995)].
*Significant difference with P<0.05.
* * Significant difference with P < 0.01.
t he s ur vi va l o f f e mo r a l ma r r o w s t em cel l s. Gr o u p s
o f d o n o r mi ce wer e gi ven Si - J u n - Zi - Ta n g i n t he
o p t i ma l dos e o f 20 mg/ 20 g b o d y wei ght , c a l c ul a t e d
f r om t he r es ul t s o f d o s e - r e l a t e d ef f ect s ( Ta bl e 1),
a n d t hen i r r a d i a t e d wi t h di f f er ent dos e s a c c o r d i n g
t o t he e x p e r i me n t a l des i gn. Bo t h f e mu r a wer e
r e mo v e d i mme d i a t e l y a f t e r v a r i o u s dos e s o f i r r a di -
at i on. Cel l s f r o m t he b o n e ma r r o w wer e a s p i r a t e d
u n d e r as ept i c c ondi t i ons , a n d a s us pe ns i on o f
si ngl e ma r r o w cel l s was p r e p a r e d . Le t h a l l y i r r a di -
a t e d r eci pi ent mi ce r ecei ved a 9 - Gy bone ma r r o w
s t er i l i zi ng dos e o f whol e b o d y i r r a d i a t i o n 24 h
be f or e i n t r a v e n o u s i nj ect i on o f a p p r o p r i a t e n u m-
ber s o f b o n e ma r r o w cel l s i nt o t he t ai l vei n t o
p r o d u c e a b o u t 1 2 - 1 5 c ol oni e s pe r s pl een 10 d a y s
l at er . Fi ve r eci pi ent mi ce wer e p r e p a r e d f or each
d o n o r mous e . Be t we e n five a n d e i ght d o n o r mi ce
wer e us ed f or e a c h poi nt . Af t e r t he i nj ect i on, mi ce
wer e ki l l ed on t he 10t h d a y a n d t he s pl eens wer e
f i xed i n Bo u i n ' s s ol ut i on. The n u mb e r o f ma c r o -
s copi c s pl een c ol oni e s was t he n s cor ed.
2. 6. Me a s u r e me n t o f he mat oc y t e s and hemat ocr i t s
Wh o l e b l o o d was col l ect ed f r om t he t ai l ends
o f mi ce on di f f er ent d a y s a f t e r v a r i o u s t r e a t -
me n t s a nd t he f l uc t ua t i on o f h e ma t o c y t e s i nc l ud-
i ng l e ukoc yt e s , e r y t h r o c y t e s a n d t h r o mb o c y t e s
wer e a u t o ma t i c a l l y c o u n t e d by a h e ma t o c y t e
c ount e r . Bl o o d was al s o t a k e n wi t h h e p a r i n i ze d
mi c r o h e ma t o c r i t t ube s a n d t hen c e nt r i f uge d at
2260 x g f or 20 mi n t o me a s u r e t he va r i a nc e o f
he ma t oc r i t . The b l o o d c o u n t r e s pons e wa s ex-
pr e s s e d as a pe r c e nt a ge o f t he n o r ma l c o u n t de-
t e r mi n e d 1 d a y be f or e i r r a di a t i on. Av e r a g e
val ues f or each g r o u p wer e o b t a i n e d f r o m five
mi ce pe r g r o u p a n d t he s a me mi ce wer e not
s a mp l e d unt i l 10 d a y s l a t e r t o pr e ve nt t he i nf l u-
ence o f i nf ect i on. I f t he h e ma t o c y t e c ha nge i n
s acr i f i ced mi ce c o u l d n o t be c ount e d, t he t r e a t -
me n t was r e p e a t e d t o i nc r e a s e t he n u mb e r o f
mi ce f or t he e x p e r i me n t a l s t a t i s t i c a l anal ys i s .
72 H.-Y. Hsuet al. /Journal of Ethnopharmacology54 (1996) 69-75
1 0 2
u. 10j ~ ~
C) 20mg/20g administratlo~
10 0 ? iadiati~nal": , I I , N~l~ ,
0 1 2 3 4 5 6 7 8 9 10
Irradiation Dose (Gy)
Fig. 1. CFU survival as a funct i on of single doses of x-rays
given alone or i mmedi at el y after 20 mg/20 g of Si -Jun-Zi -Tang
admi ni st ered consecutively for 7 days, once a day.
2. 7. St at i st i cal anal ysi s
Data collected were presented by means and
standard deviations. Statistical analysis was per-
formed by the Student' s t-test to express the
difference between two groups.
3. Results
Table 1 shows the mean survival time, mortality
and increase of life span of the mice within 60
days of treatment with various concentrations of
Si-Jun-Zi-Tang alone or combined with 8 Gy
%-irradiation. Mortality rates of g-irradiation
combined with the Si-Jun-Zi-Tang of 10, 20 and
30 mg/20 g body weight administration were
lower than that of 8-Gy irradiation alone. When
compared with the results of Si-Jun-Zi-Tang ad-
ministered alone, a concentration higher than 30
rag/20 g seemed to be toxic and increased the
mortality rate. X-Irradiation combined with 30
mg/20 g Si-Jun-Zi-Tang administration caused a
slightly lower mean survival time than with 20
mg/20 g. The optimal concentration of Si-Jun-Zi-
Tang seems thus to be 20 mg/20 g. Therefore, we
used 20 mg/20 g body weight to be the Si-Jun-Zi-
Tang administered parameter in the following in-
vestigations to evaluate the radioprotective effect
of this drug. Data from our experiments show
that pre-treatment with Si-Jun-Zi-Tang have some
effects on protecting bone marrow stem cells and
peripheral hematocytes against radiation-induced
regression.
Fig. 1 shows the changes of CFUs counted on
the 10th day with different doses of radiation. The
radiotolerance of bone marrow cells was en-
hanced by Si-Jun-Zi-Tang administration of 20
mg/20 g body weight. Similar radiosensitivity is
indicated from the straight line region of survival
curves of the treatment by 20 mg/20 g body
weight as compared with the treatment of %-irra-
diation alone. It also means that the D O value,
which is an expression of radiosensitivity and is
graphically derived from the exponential portion
Table 2
Leukocyte count s of mice after i rradi at i on and t r eat ment with Si -Jun-Zi -Tang ( %F
Treat ment Days aft er i rradi at i on
z- Ray Si -Jun-Zi -Tang b 5 9 12 t 9 26 33
- - - - 104.8_+21.1 101. 4+19. 7 94.3_+19.8 98.6_+20.1 107.7_+18.4 100. 6+21. 5
20 mg/20 g 96.2_+ 14.4 114.7 _+21.8 107.6_+ 19.2 122. 4+ 19.7 115. 6+ 16.9 123.3 _+ 18.4
500 cGy 23.6_+6.6 16.7_+8.3 11.8_+6.0 33.6_+15.3 54.6_+13.3 84.7_+16.3
500 cGy 20 mg/20 g 32.5 _+ 9.7* 25.4 _+ 7.7* 29.6 _+ 9.4** 53.8 -t- 16.0"* 78.3 _+ 15.6"* 97.2 _+ 18.6
~Percentages of leukocytes were calculated from t he pre-i rradi at i on values t aken as 100%.
bAdmi ni st rat i on of Si -Jun-Zi -Tang was by i.p. injection before irradiation.
Significant differences ( *P<0. 05; **P<0. 01) bet ween z- r ay irradiated gr oup and t he gr oup of z- r ay i rradi at ed combi ned with drug
admi ni st rat i on were statistically anal ysed by t he St udent ' s t-test.
H.- Y. Hsu et al. / Journal of Et hnopharmacology 54 (1996) 6 9 - 75
Tabl e 3
Er yt hr ocyt e count s of mi ce aft er i r r adi at i on a nd t r eat ment wi t h Si - J un- Zi - Tang (%)a
73
Tr e a t me nt Da ys aft er i r r adi at i on
z- Ra y Si - J un- Zi - Tang b 5 9 12 19 26 33
- - 96.6 + 19.3 112.5 + 19.6 102.1 __+20.4 98.4 + 18.8 100.3 + 22.5 96.5 + 19.7
- - 20 mg/ 20 g 110.1 + 15.3 132.9 + 18.6 139.7 + 18.7 146.6 + 19.6 130.3 ___17.4 118.6 + 14.2
500 cGy - - 7 1 . 5 +1 6 . 2 5 0 . 7 +1 4 . 3 4 8 . 2 +1 1 . 3 6 0 . 8 +1 8 . 2 82. 2+19. 1 9 1 . 0 +1 6 . 5
500 c Gy 20 mg/ 20 g 77.6 + 18.2 64.3 + 16.0 59.6 + 12.7 83.6 + 19.4" 94.3 + 18.3 107.2 + 17.5
aPercent ages of er yt hr ocyt es were cal cul at ed f r om t he pr e- i r r adi at i on val ues t aken as 100%.
bAdmi ni s t r at i on of Si - J un- Zi - Tang was by i.p. i nj ect i on before i rradi at i on.
*Si gni fi cant di fferences ( P<0. 05) bet ween z- r ay i r r adi at ed gr oup and t he gr oup of z- r ay i r r adi at ed combi ned wi t h dr ug
admi ni s t r at i on were st at i st i cal l y anal ysed by t he St udent ' s t -t est .
of the cell survival curve in these two treatment, is
almost the same (Travis, 1989). The elevated ra-
diotolerance of bone marrow cells by Si-Jun-Zi-
Tang administration was shown in the shoulder of
the survival curve with a radiation dose between
0- 5 Gy.
As can be seen in Table 2, the tested drug
reduced the radiation-induced decrease in periph-
eral leukocytes from the 5th day after irradiation.
On day 12, when the number of leukocytes of the
irradiated controls decreased to 12% of the initial
values, the counts in the treated groups were 30%
with 20 mg/20 g body weight Si-Jun-Zi-Tang ad-
ministration. Significant differences from the irra-
diated controls were seen at days 19 and 26,
namely 54% and 78% with drug administration
compared with the control values of 34% and
55%. A protection by Si-Jun-Zi-Tang, although
statistically not significant, was seen for erythro-
cytes from the 9th day (Table 3). In contrast to
the irradiated control value of 51%, the erythro-
cyte count was 64% with drug administration. At
day 12, Si-Jun-Zi-Tang gave a value of 60% and
at day 19, a value of 84%, as compared with the
control values of 48% and 61%, respectively. As
shown in Table 4, the thrombocyte reduction was
also influenced by Si-Jun-Zi-Tang administration.
Compared with the thrombocyte control value of
8% at day 12, Si-Jun-Zi-Tang administration led
to values of 24%. The effect of Si-Jun-Zi-Tang
administration on day 12, 19 and 26 was signifi-
cantly superior to the protection of other periods
after irradiation. The control drug administration
enhanced the elevations of leukocyte, thrombo-
cyte and especially erythrocytes counts after treat-
ment.
Tabl e 4
Thr omboc yt e count s of mi ce af t er i r r adi at i on and t r eat ment wi t h Si - J un- Zi - Tang (%)a
Tr e a t me nt Da ys aft er i r r adi at i on
z- Ra y Si - J un- Zi - Tang b 5 9 12 19 26 33
- - - - 102.3 + 19.6 101.7 + 18.6 98. 4 + 16.5 102.6 + 19.4 92.7 + 21.6 98.3 + 19.0
- - 20 mg/ 20 g 92.4 + 13.7 95.8 __+13.8 107.9 + 16.2 110.4 __. 18.0 104.6 + 15.8 107.4 + 16.1
500 cGy - - 4 0 . 6 +1 4 . 7 2 5 . 5 +1 1 . 2 7 . 9 +3 . 3 1 6 . 9 +8 . 4 6 6 . 2 +1 8 . 7 8 6 . 4 +1 9 . 8
500 cGy 20 mg/ 20 g 4 8 . 7 + 13.2 39. 2+ 12.7" 23.5__+9.0** 3 8 . 8 + 11.3"* 89. 6+ 14.7"* 9 7 . 4 +2 1 . 8
aPercent ages of t hr ombocyt es were cal cul at ed f r om t he pr e- i r r adi at i on val ues t aken as 100%.
bAdmi ni s t r at i on of Si - J un- Zi - Tang was by i.p. i nj ect i on before i rradi at i on.
Si gni fi cant di fferences ( *P < 0.05; **P < 0.01) bet ween x- r ay i r r adi at ed gr oup and t he gr oup of x- r ay i r r adi at ed combi ned wi t h dr ug
admi ni s t r at i on were st at i st i cal l y anal ysed by t he St udent ' s t -t est .
74 H.- Y. Hsu et al. / Journal o f Et hnopharmacology 54 (1996) 69 75
Table 5
Hemat ocri t count s of mice after i rradi at i on and t reat ment with Si -Jun-Zi -Tang (%,)~
Treat ment Days after i rradi at i on
x-Ray Si-Jun-Zi-Tang b 5 9 12 19 26 33
- - 100.8 19.6 95.3 _+ 19.7 97.1 _+20.3 99.4_+ 19.7 104.5 +21. 2 101.0 -+ 19.8
20 rag/20 g 103.2_+15.3 117.6_+16.5 113.4_+15.7 110.6_+16.3 102.5_+14.7 96.4_+18.8
500 cGy 82.5 _+ 19.6 69.8 + 15.7 51.3 -+ 12.9 68.8 -+ 15.8 88.7 2 18.6 101.3 _+21.4
500 cGy 20 mg/20 g 88.7 _+ 14.3 78.4 _+17.3 64.4 _+ 11.5" 86.9 _+ 15.4* 97.3 19.2 109.6 _+ 21.1
~Percentages of hemat ocri t s were calculated from the pre-i rradi at i on values t aken as 100%.
bAdmi ni st rat i on of Si -Jun-Zi -Tang was by i.p. injection before irradiation.
*Significant differences ( P<0. 05) bet ween z-ray irradiated gr oup and the gr oup of x-ray i rradi at ed combi ned with drug
admi ni st rat i on were statistically anal ysed by t he St udent ' s t-test.
Table 5 illustrates t hat the hemat ocri t value
reached a mi ni mum value at day 12 - - 51% in the
irradiated controls compared with 64% after Si-
Jun-Zi -Tang admi ni st rat i on, which was a signifi-
cant difference ( P<0. 05) . Anot her significant
difference was also seen at day 19 after Si-Jun-Zi-
Tang admi ni st rat i on - - 87% versus the control
value of 69%. Hemat ocri t count s of Si-Jun-Zi-
Tang admi ni st rat i on alone showed some eleva-
tions from the 5th day to the 19th day after
t reat ment , and t hen ret urned to the normal range.
4. Discussion
Survival brought about by radi oprot ect ors after
a potentially lethal i rradi at i on is t hought to be
due primarily to their effect on hemat opoi et i c
cells (Floersheim et al., 1988). When measuring
bone marrow hemat opoi et i c stem cell colony-
formi ng units (Harris and Phillips, 1971), and
peripheral bl ood hemat ocyt es count s (Smith et al.,
1965), it has been shown repeatedly t hat thiols
protect hemat opoi et i c cells against radi at i on dam-
age. The dat a from our experiments show t hat
pre-t reat ment with Si-Jun-Zi-Tang has some effect
on protecting bone marrow stem cells and periph-
eral hemat ocyt es against radi at i on-i nduced regres-
sion. It is generally agreed t hat radi at i on deat h in
the midlethal dose range is due to i mpai rment of
bone marrow hemat opoi et i c funct i on such as the
leukopenia, eryt hropeni a and t hrombocyt openi a
which will ultimately lead to whole body infec-
tion, hemorrhage and even deat h (Floersheim et
al., 1988). Thus, the peripheral bl ood count can
be considered to be a biologically meani ngful
paramet er to demonst rat e the effect of radi at i on
and radi oprot ect i on on a normal tissue which is
critical for survival. This assumpt i on is corrobo-
rated by the fact t hat , as outlined in the results,
Si-Jun-Zi-Tang allowed %-irradiated mice to
mai nt ai n radi ot ol erance to radi at i on doses which
were sublethal or lethal for mice not pre-treated
with Si-Jun-Zi-Tang. In this study, the protective
effect of leukocytes was significant at day 12, 19
and 26 after exposure to ionizing radiation. A
reduction of the radi at i on-i nduced fall in the he-
mat ocri t and numbers of t hrombocyt es and ery-
throcytes after Si-Jun-Zi-Tang admi ni st rat i on was
also seen in our experiments.
As to the question whet her Si-Jun-Zi-Tang af-
fords hematological prot ect i on by preventing the
destruction of bl ood cells or by enhanci ng hema-
topoietic recovery, it seems t hat the protective
effect probabl y involves bot h pathways. As seen
in Fig. 1, the decrease of bone mar r ow stem cells
was reduced or, alternatively, their radiotolerance
was enhanced, under the doses from 1 to 5 Gy.
According to the conclusions of Floersheim et al.
(1988), this could mean t hat bot h circulating
bl ood cells and progeni t or cells are protected.
As for the mechani sm of radi oprot ect i on by
Si-Jun-Zi-Tang, we suggest the possible hypot he-
sis of a stabilization of radi at i on induced cellular
injuries. It has been report ed t hat a single injec-
tion of the partially purified extract of ginseng
H.- Y. Hsu et al. / Journal of Ethnopharmacology 54 (1996) 69 75 75
before or after whole body 2-ray irradiation pro-
tected mice from bone marrow death (Yonezawa
et al., 1981). Likewise, licorice has been found to
have a protective effect against radiation injury
through the accelerated recovery of hematocyte
counts (Hosokawa, 1986). According to previous
investigations, glycyrrhizin which is one of the
components of licorice may stabilize lysosomes by
inhibiting phospholipase A2 activity, which is in-
volved in the lipid metabolism of cell membranes
(Shiki et al., 1983). Since the components of Si-
Jun-Zi-Tang include bot h ginseng and gly-
cyrrhizin, one of its radioprotective effects is
likely afforded through a similar mechanism. That
is, the glycyrrhizin in Si-Jun-Zi-Tang may play an
important role in eliminating the lysis of damaged
cells after irradiation (Hsu et al., 1991; Hsu et al.,
1993). P. cocos, as well as A. ovat a and Z. offici-
nal e have been reported to exert some effects on
enhancing the secretion of cytokines by human
peripheral bl ood mononuclear cells (Chang et al.,
1995; Tseng and Li, 1996). Among those cytoki-
nes, granulocyte-macrophage colony-stimulating
factor (GM-CSF) was originally found to stimu-
late colony formation of granulocytes and
macrophages (Burgess et al., 1977), and was sub-
sequently demonstrated to stimulate the growth
and differentiation of progenitor cells in bone
marrow (Moore, 1991). Therefore, the augmenta-
tive effect of hematopoietic recovery after irradia-
tion was probabl y due to the modulation caused
by P. cocos, A. ovat a and Z. officinale.
Although our investigations might provide an
experimental basis for the use of Si-Jun-Zi-Tang
as a radioprotector of bl ood cells, its effect on
other normal tissues such as the immune system,
gut and kidney should be further examined.
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