全 文 ：·药理实验与临床观察·
Effect of protein and anthraquinone glucosides from Semen Cassiae
on hemorrheology of hyperlipidemic rats
LI Xu-e, YANG Shui-yun
* , ZHAO Wen-ming
* , GUO Bao-jiang

　　 ( Institute of Bio lo gica l Technolog y o f South China Normal Univ e rsity , Guang zhou Guangdong , 510631, China)
　　 Abstract: Object　 To study the effect of pro tein and anthraquinone g lucosides f rom Semen
Cassiae on hemor rheolog y of hyperlipidemic ra ts. Methods　 Rat hyperlipidemic model w as pre-
pared by ig lipid emulsion. Af ter o ral administra tion of the pro tein 0. 25 o r 1 mg /kg , the an-
thraquinone g lucosides 5 or 20 mg /kg , and the protein 0. 25 mg /kg plus the anthraquinone gluco-
sides 5 mg /kg, the blood visco si ty o f high shea r rate (ηH) , blood viscosity of low shea r rate
(ηL) , reduced viscosi ty ( RV ) , plasma viscosi ty (ηP) , pla telet adhesion ratio ( PAR) , hema tocri t
( HCT) , and ery throcy te sedimenta tion rate ( ESR) of the hyperlipidemic rats w ere determined to
assess the ef fect o f the said drugs on these hemo rrheo logic parameters. Results　 The pro tein 1
mg /kg and the anthraquinone g luco sides 20 mg /kg , as w ell as the protein 0. 25 mg /kg plus the
anth raquinone gluco sides 5 mg /kg , could reduce the elev atedηH, ηL, RV , ηP, and PAR of hy-
perlipidemic rats ( P < 0. 05) . Conclusion　 Protein and anthraquinone gluco sides from Semen Cas-
siae could decreaseηH, ηL, RV , ηP, and PAR o f hyperlipidemic ra ts.
Key words: Semen Cassiae; pro tein; anth raquinone gluco sides; hemor rheolog y
决明子蛋白质和蒽醌苷对高脂血症大鼠血液流变学的影响
李续娥 ,杨水云* ,赵文明* ,郭宝江
(华南师范大学生物技术研究所 ,广东 广州　 510631)
摘　要: 目的　探讨决明子蛋白质和蒽醌苷对高脂血症大鼠血液流变学的影响。方法　将大鼠制成高脂血症模型。
测定决明子蛋白质和蒽醌苷的小、大剂量 ,以及两者小剂量合用后对高脂血症大鼠的全血粘度 (高切变率下的ηH,
低切变率下的ηL) ,全血还原粘度 ( RV ) ,血浆粘度 (ηP) ,血小板粘附率 ( PAR) ,红细胞压积 ( HC T )及血沉 ( ESR)的
影响。 结果　决明子蛋白质大剂量 ( 1 mg /kg ) ,蒽醌苷大剂量 ( 20 mg /kg )以及两者的小剂量 (蛋白质 0. 25 mg /kg ,
蒽醌苷 5 mg /kg )合用 ,均可使高脂血症大鼠的 ηH,ηL , RV ,ηP和 PAR显著降低 (P < 0. 05)。 结论　决明子蛋白
质和蒽醌苷皆可降低高脂血症大鼠的 ηH,ηL, RV ,ηP和 PAR。
关键词: 决明子 ; 蛋白质 ; 蒽醌苷 ; 血液流变学
中图分类号: R286. 2　　　文献标识码: A　　　文章编号: 0253 2670( 2002) 05 0429 03
　　Semen Cassiae from Cassia obtusi folia L. o r C.
tora L. w as known to be of therapeutic value in re-
ducing fever, improv ing eyesigh t, nourishing in-
testine and relaxing bow el. Over recent years, it
has been found that this drug is also ef fectiv e to
decrease blood pressure and serum lipid
[1～ 2 ] , a nd
therefo re a tt racted much public interest. In our
previous study, pro tein and anthraquinone gluco-
sides from Semen Cassiae could low er the elev ated
serum to tal cholesterol , trig lyceride, and low den-
·429·中草药　 Chinese T raditional and Herbal Drug s　 2002年第 33卷第 5期
Received Date: 2001-10-06
Fund Item: Project supported by Natu ral Science Foundation of Shanxi Province ( No. 99 SM74) .
Resume of author: LI Xu-e, female, ass ociate Profes sor, Ph. D. studied at Xi an Univ ersi ty of Medical Science and Xi an Jiaotong Uni-
versity respectiv ely; wo rk ed at Tian jin Ins titute of Pharmaceutical Research , Xing ping City Ins ti tu te of Medicines and
Ch emical Reag ents In spection, and School of Ch emical Engineering of Xi an Jiaotong Univ ersi ty respectively. Being a
pos t-d octor s tudent at Insti tute of Biological Tech nology of Sou th China Normal Universi ty at pres ent . Tel: 020-
85211420
* Xi an Jiaotong Universi ty
si ty lipoprotein of hyperlipidemic rat ( P < 0. 05,
P < 0. 01)
[ 3] . Hyperlipidemia could result in hem-
o rrheolo gic abno rmali ty w hich acted as a promo t-
ing facto r, cause and wo rsen the pa thological
change and clinical symptoms of hyperlipidemia
[4 ]
.
Therefore, the aim o f this study w as to investiga te
the ef fect of pro tein and anthraquinone g lucosides
f rom Semen Cassiae on hemo rrheolog y o f hyperlipi-
demic ra t.
1　Materials and Methods
1. 1　 Prepa ra tions of the protein and the an-
thraquinone g lucosides: Semen Cassiae, the dried
and mature seed of C . obtusifolia L. w as co llected
f rom Anhui Province, PRC in autumn. Its identi ty
w as authentica ted by Prof. LI Yang-li , Depar t-
ment of Pharmacy, Xi an Jiao tong Univ ersi ty. The
pro tein was ex t racted w ith buf fer ( K H2 PO4 /
NaO H, pH 8. 0) , then the ex tract w as separated
w ith ammonium sulfa te f ractiona tion. The pro-
tein, precipi tated a t 40% ～ 50% sa turation of am-
monium sulfate, w as collected by centrifug e and
dia lyzed wi th double distilled w ater fo r 48 hours.
The dialy zed protein w as again centri fuged, and
the centrifuga te w as purified on Sephadex G-75
column in double distil led w ater. The eluate that
had an abso rption peak a t 280 nm was co llected
and lyophi li zed to obtain the puri fied pro tein wi th a
content of 88. 7% . Anthraquinone gluco sides and
anth raquinones w ere ex t racted f rom Semen Cassiae
w ith 90% ethanol, and the anthraquinones were
separated w ith chloroform. Anthraquinone g luco-
sides w ere precipitated wi th Pb2+ , and then Pb2+
was removed by hydrogen sulfide and final ly re-
crystalli zed. The content of anthraquinone g luco-
sides was 94. 8% .
1. 2　 Reagents: Duoxikang ( mix ture o f eicosapen-
taenoic acid and doco sahexaenoic acid ) w as pro-
duced by Zhejiang Hailisheng Pharmaceutical Fac-
to ry ( No. 980417) . Tiamazole w as a product of
Guang zhou Shiqiao Pharmaceutical Facto ry ( No.
980120) .
1. 3　 Experimental rat model and trea tment:
SD rats, 200～ 240 g , w ere prov ided by Animal
Experimental Center, Xi an Jiaotong Univ ersi ty
( GradeⅡ , Certificate No. SX-05) . Ra ts w ere di-
vided into eight g roups ( Table 1) . Every mo rning,
rats in g roup Ⅰ ( cont rol ) w ere given solvent
w hich consisted o f 10% poly oxyethylene ( 20) so r-
bitan monoo lea te+ 10% g lycerine+ 80% distilled
w ater, and the rats in o ther seven g roups w ere giv-
en lipid emulsion which consisted o f 20% lard +
10% cholesterol + 2% sodium cho la te + 0. 1%
thiamazo le + 10% poly oxyethylene ( 20) so rbi tan
monoo lea te + 10% g lycerine + 47. 9% distilled
w ater. Every af ternoon ra ts in g roupⅢ toⅧ were
given drug s at di fferent dosages as show n in
Table 1. This procedure lasted 15 days.
Table 1　 Experimental groups and treatment
group diet and drug　　
Ⅰ cont rol ( i g solvent , ip normal saline)
Ⅱ hyperlipidemic model ( ig LE, ip normal saline)
Ⅲ posit ive drug con t rol ( ig L E, ip d uoxikang 5 m L /kg
Ⅳ small dosage P ( ig LE, ip P 0. 25 mg /k g)
Ⅴ large dosage P ( ig LE, ip P 1 mg /kg )
Ⅵ small dosage AG ( ig LE, ip AG 5 mg /kg )
Ⅶ large dosage AG ( ig LE, ip AG 20 mg /kg)
Ⅷ small dosage P+ smal l d osag e AG ( ig L E, ip P 0. 25
mg /kg+ AG 5 mg /k g)
LE: lipid emulsion　 P: protein　 AG: an th rqauinon e g lucosides
1. 4　 Experimenta l methods and sta tistical analy-
sis: After the last t reatment, the ra ts w ere fasted
fo r 12 h, and anesthetized wi th pentobarbi tal sodi-
um ( 30 mg /kg , ip) , and then blood samples w ere
co llected f rom ca ro tid ar tery by intuba tion. The
ηH ( 200 s- 1 ) , ηL ( 40 s- 1 ) and RV were measured
wi th the LG-Ⅲ tube-shaped spin blood viscometer
( Transducer Techno logy Development Co. , Chi-
nese Academy o f Sciences) . The PAR was deter-
mined w ith 9103-3 platelet adhesiometer ( Trans-
ducer Techno logy Development Co. , Chinese A-
cademy o f Sciences) . TheηP w as determined w ith
a falling ball v iscometer ( Huaxing High Tech nolo-
gy Co. ) , a nd the ESR and HCT were taken w ith
ESR and HCT reading board ( Huax ing High Tech-
no logy Co. ) . Results w ere expressed as x± s , a nd
stati stical analysis was carried out using t-test.
2　Result
The changes of ηH, ηL, RV , ηP, PAR,
ESR, and HCT in the eight g roups w ere show n in
Table 2.
·430· 中草药　 Chinese T raditional and Herbal Drug s　 2002年第 33卷第 5期
Table 2　 Changes in hemorrheologic parameters (x± s , n= 10)
group
ηH
(m Pa· s )
ηL
( m Pa· s )
RV
( m Pa· s)
ηP
( m Pa· s )
PAR
(% )
HCT
(% )
ESR
( mm /h )
Ⅰ 3. 77± 0. 09* * 9. 31± 1. 97* * 6. 21± 2. 23* * 2. 85± 0. 60* * 20. 36± 6. 50* 45. 00± 3. 90 1. 63± 1. 54
Ⅱ 6. 00± 1. 26 15. 19± 3. 07 11. 76± 3. 29 4. 26± 0. 90 26. 76± 6. 43 45. 30± 3. 89 2. 60± 2. 40
Ⅲ 4. 04± 1. 08* * 9. 08± 2. 22* * 7. 72± 2. 29* * 3. 29± 0. 75* 18. 20± 8. 19* 44. 80± 5. 40 1. 60± 1. 70
Ⅳ 4. 89± 1. 59 12. 13± 3. 59 8. 74± 2. 88* 3. 60± 1. 06 19. 21± 6. 72* 44. 21± 6. 51 1. 90± 1. 80
Ⅴ 4. 48± 1. 22* 11. 20± 3. 38* 7. 33± 2. 60* 3. 32± 0. 82* 17. 62± 7. 79* 46. 19± 5. 06 1. 30± 0. 90
Ⅵ 5. 49± 1. 23 12. 95± 3. 05 9. 65± 2. 47 4. 00± 0. 81 19. 19± 7. 20* 44. 25± 6. 67 1. 99± 1. 58
Ⅶ 4. 44± 1. 45* 11. 34± 3. 29* 7. 93± 2. 41* 3. 36± 0. 93* 20. 26± 6. 70* 43. 85± 5. 84 1. 60± 1. 10
Ⅷ 4. 46± 1. 54* 10. 83± 4. 05* 8. 02± 4. 26* 3. 35± 1. 05* 17. 48± 3. 71* 45. 10± 4. 30 1. 30± 1. 00
　　　　* P < 0. 05, * * P < 0. 01　 vs groupⅡ
　　 Compared wi th contro l g roup (Ⅰ ) , ηH, ηL,
RV, ηP, and PAR of hyperlipidemic model g roup
(Ⅱ ) w ere signi ficant ly increased ( P < 0. 05, P <
0. 01) , g roup w ith duoxikang (Ⅲ ) could low er the
above raised indexes ( P < 0. 05, P < 0. 01) . This
manifested that the hyperlipidemic model of rat
w as successfully established by ig lipid emulsion
fo r 15 d.
Results showed that the pro tein 1 mg /kg
(Ⅴ ) , the anthraquinone glucosides 20 mg /kg
( Ⅶ ) , and the pro tein 0. 25 mg /kg plus an-
thraquinone g lucosides 5 mg /kg (Ⅷ ) could de-
creaseηH, ηL, RV andηP of the hyperlipidemic
ra ts ( P < 0. 05) ; but nei ther protein 0. 25 mg /kg
(Ⅳ ) , no r the anth raquinone g lucosides 5 mg /kg
(Ⅵ ) show ed any ef fect (P> 0. 05) . The pro tein 0.
25 (Ⅳ ) and 1 mg /kg (Ⅴ ) , the anthraquinone glu-
cosides (Ⅵ ) 5 and 20 mg /kg (Ⅶ ) , and the protein
0. 25 mg /kg plus the anthraquinone glucosides 5
mg /kg (Ⅷ ) could low er PAR o f the hyperlipidem-
ic ra ts ( P < 0. 05) , but could no t low er ESR and
HCT o f the hyperlipidemic ra ts ( P> 0. 05) .
3　 Discussion
Clinical study indicated that a kind of tea made
from Semen Cassiae could low er serum lipid
[1 ] . In
our previous study, w e found that pro tein and an-
thraquinone gluco sides f rom Semen Cassiae could
low er serum lipid of hyperlipidemic rats. The re-
sul ts of the present study show ed tha t they could
also lower the elev atedηH, ηL, RV ,ηP, and PAR
o f hyperlipidemic rats. Therefo re, pro tein and an-
thraquinone g lucosides f rom Semen Cassiae are not
only beneficial to hyperlipidemia , but also benefi-
cial to hemo rrheolo gic disorders.
It has been repo rted tha t Semen Cassiae has
eyesight-improv ing function and can be used to
cure many kinds o f eye diseases
[2 ] . The severity of
diabetic retinopathy was po si tiv ely co rrela ted w ith
total cho lesterol and low densi ty lipoprotein
[5 ]
.
One o f the main pathogenesis o f retinal v ein occlu-
sion w as due to abno rmali ties of hemo rrheo log y
[6 ] ,
and th e raised blo od viscosity and the change of
pla telet activ ity play ed an impo rtant ro le as the
chief culpri t
[ 7, 8] . Pro tein and anthraquinone gluco-
sides f rom Semen Cassiae could low er to tal choles-
tero l, low density lipoprotein, blood viscosi ty , and
pla telet adhesion ratio, thus they w ere useful fo r
t rea ting diabetic retinopathy and retinal vein occlu-
sion. This might , in pa rt at least , account fo r the
improvement of eyesigh t by Semen Cassiae.
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·431·中草药　 Chinese T raditional and Herbal Drug s　 2002年第 33卷第 5期