全 文 ：云南大学学报 (自然科学版) 1998 , 20 (3):162 ～ 165 CN 53-1045/N　ISSN 0258-7971
Journal of Yunnan University
Effect of Petroleum Ether Extract of Hibiscus
rosa-sinensis Flowers on Early Pregnancy and Some
Reproductive Hormones in Rats
①
Jiang Yan　Zhao Cuilan　Li Kaiyuan　Zhou Mingdong
(Biology Department of Yunnan University , 650091 , Kunming , PRC)
Abstract　intrauterine injection of petroleum ether ex tract of Hibiscus rosa-sinensis flowers(HR-1)at 3 deffer-
ent dose levels(10 mg , 20 mg and 30 mg in each uterus)was found to affect early pregnancy and serum hormones of
lutropin , fo llitropin , progesterone and estrogen.With an increase in the do sage of the ex tract , the percentage of preg-
nancy termina ted increased.At the dose level of 30 mg for each uterus , the ex tract led to the termination of pregnancy
in about 99%.The effect was accompanied by reduction of LH , FSH , progesterone and estrogen.
Key words　Hibiscus rosa-sinensis flower , termination of pregnancy , LH , FSH , progesterone , estrogen
Hibiscus rosa-sinensis , commonly known as China rosa plant has been quo ted for it s antifertili ty efficacy
and fo r it s use in correcting menst rual disorders[ 1] .Although there are a lot of information about contraceptive
effect of this f low ers[ 2 ,3] , only few at tempts of pregnancy terminated have been made[ 4 ,5] .As the act ive con-
st ituents of antifertili ty of the f low er have not yet been ident if ied , it was decided to submit it to detailed chemi-
cal and pharmacological investig at ion.The present communicat ion deals wi th the effect of petroleum ether ex-
tract of Hibiscus rosa-sinensis f low ers on termination of early pregnancy in rat.
1　Materials and Methods
1.1　Animals　Wistar female rats weighing betw een 250 ～ 270 gm were maintained under natural light and
temperature , and allow ed free access to balanced food and w ater.
1.2　Drugs　Dried Hibiscus rosa-sinensis f low ers w ere collected f rom the south-west of Yunnan.They were
ex tracted w ith ethanol , concentrated , then ex tracted by pet roleum ether.This product named as HR-1 ,was an
analogy of β-sitosterol[ 6] .125　I-LH kit , 125　I-FSH kit , 125　I-progesterone kit and 125　I-est rogen ki t were purchased
from Beijing Institute of Biological Technology.
1.3　Methods　Adult female Wistar rats w ere caged wi th males of proven fertility in the ratio of 2∶1.The fe-
males w ere examined the follow ing morning for evidence of copulation.The day when spermatozoa w ere de-
tected in the vaginal smears w as considered as day-1 of pregnancy.The pregnant rats w ere laparo tomized under
pentobarbital sodium anaesthesia on day-6 of pregnancy and 0.1ml 0.9%salt water o r 10 mg ,20 mg or 30mg
HR-1 w as injected very slowly into the tw o horns of the uterus , then the abdomen w as sutured.On day-12 of
pregnancy , all animals were sacrified and the uterus w ere examined for the embryo numbers and the fetation
conditions.
① The w ork granted by Yunnan Provicial Science Fundat ion.Received 1998-03-02
Befo re laparotomized , the rats blood w as collected by means of cardiopuncture.All serum specimens were
stored at -20 ℃ for assay LH ,FSH ,progesterone and estrogen.
Data were analyzed by student s t test and a probability of P <0.05 was considered as statistically
singnificant .
2　Results
2.1　Effect of HR-1 on early pregnancy 　In control rats , embryos in uterus developed very well , the size of
the embryo was about 0.75 cm.The treated rats ,when laparotomized on day-12 , appeared defferent interrup-
tion of pregnancy .HR-1 resulted in the termination of pregnancy respectively in 81.6%, 92%and 99%, at the
3 defferent dose levels.The size of embryos appeared significant ly smaller in comparison wi th control (Table
1).
Tab 1.Effect of HR-1 on early pregnancy in rats
G roup(No.
of rats)
T reatment schedule
(on day 6)
P regnancy of dead and
injured embryos/ %
Average of embryo
size/ cm
Average of embryos
in each uterus(No.)
I 0.9% 0 0.75+0.02 6.1+0.58
(12) salt w ater
I I HR-1 81.6 0.3+0.03 A＊＊ 5.55+0.38
(11) 10 mg/uterus
II I HR-1 92 0.31+0.03 A＊＊ 6.25+0.35
(12) 20 mg/uterus
IV HR-1 99 0.17+0.03 A＊＊ 5.55+0.55
(9) 30 mg/uterus
　　A:comparison wi th I , ＊＊:P<0.01
2.2　Effect of HR-1 on serum ho rmone concentrations　In the ex tract-t reated rats , the peripheral levels of
FSH and estrogen w ere decreased with an increase of the dosage of HR-1.While the serum LH and proges-
terone concentrations were significantly lower only w hen the dose of injection w as 30 mg (Table 2).
Tab 2.Effect of HR-1 on serum hormones o f pregnancy rats
Group(No.
of rats)
T reatment schedule
(on day 6)
Estrogen
pg/ml
Progesterone
ng/ ml
FSH
mIU/ml
LH
mIU/ ml
I 0.9% 202.7+46.5 79+16.3 4+0.93 9.9+1.42
(7) salt water
II HR-1 189.3+38.6 83.7+12.5 2.56+0.5 10.8+1.84
(8) 10 mg/ uterus
I II HR-1 122.1+18.5 68.2+12.4 2.53+1.12 9.5+0.3
(6) 20 mg/ uterus
IV HR-1 130.6+18.7 24.5+12.2 A＊ 0.45+0.38 A＊ 3.95+1.63 A＊
(5) 30 mg/ uterus
　　A:comparison wi th I , ＊:P <0.05
163第 3 期　　　　江　燕等:扶桑花石油醚提取物(HR-1)的抗早孕作用以及对大鼠血清生殖激素的影响　　　　　　　
3　Discussion
Int rauterine injection of HR-1 w as significant ly affect ive for the termination of early pregnancy , and the
actions w ere related to the dosages of ex tract.The mo rphological ex amination of the uteri showed that the ex-
tract-t reated rats had in uteri embryos at dif frent stages of reso rption.In a number of animals pregnancy termi-
nated , and thei r uteri devoid of any t race of implantation si te , while in some others although no degenerative
changes in embryos w ere apperent , size of the fetation appeared comparatively smaller and tiny black spots
w ere lef t on the inner w all of the uteri.The results w ere the same as that in mice[ 7] ,but only the dosage of in-
trauterine injection was smaller than that of oral administ ration(Fig 2).It suggested that HR-1 is the active
consti tuent of pregnancy terminated in Hibiscus rosa-sinensis flow ers.
F ig 1.Embryos of control rats on day-12 of pregnancy Fig 2.Embryos of HR-1 treated rats on day-12 of pregnancy
The exact regulation of progesterone and estrogen is neccensery for the maintaining pregnancy.The inhi-
bi tion of estrogen or progesterone w ould interrupte the gestation.Estrogen has been credited w ith its ability to
produce uterine progesterone recepter synthesis , then promote uterine uptake and reserve progesterone , thus it
makes the pregnant uterus keep relative “dorment” .HR-1 was found to lower the peripheral level of est rogen ,
i t means that the termination of early pegnancy of HR-1 might be related to the decrease of serum est rogen
which results in the inhibition of progesterone utilizat ion in uterus.
Progesterone appears to play a key role in pregnancy of almost all species.In rats , progesterone mainly
comes from ovarial corpora.Lower serum progesterone results f rom the inhibition of ovarian progesterone syn-
thesis one side , another side it results from the interruption of progesterone secreation or the degeneration of
corpora lutea.HR-1 could decrease the serum levels of progesterone and LH at the dose of 30 mg , it indicates
that synthesis and secreation progesterone in HR-1 treated rats w ere all inhibited , and the lower progesterone
level might be a main facto r of the pregnancy terminated of HR-1.Although the maintenance of corpora lutea
164 　　　　　云南大学学报(自然科学版)　　　　　　　　　　　　　　　　第 20 卷
probably more relyed on prolactin in rat and mouse[ 8] , exogenous PRL was only partially to prevent the termi-
nat ion of pregnancy of the ex tract of Hibiscus rosa-sinensis ,while exogenous HCG was able to completely pre-
vent this effect[ 5] .It may therefo re be assumed that LH might play an impo rtant role in this process.
The result also indicated that HR-1 interfered with the serum level of FSH.Whether FSH participats reg-
ulating the function of corpora lutea of rats remains to be determined.
References
1　Kholkute S D , Mudgal V , Udupa K N.Stdies on the antifertility potentiality o f Hibiscus rosa-sinensis.Planta Medica , 1977 , 31:
35～ 39
2　Singh M P , Singh R H , Udupa K N.Antifertility activity of a benzene extract of Hibiscus rosa-sinensis flowers on female A lbino
rats.Journal of Medicinal Plant Research , 1982 , 44:171～ 174
3　Zhao Cuilan , Jiang Yan , Li Kaiyuan.Antitertility activity of a ethanol ex tract of Hibiscus rosa-sinensis flowers on mise.Yunnan
Journal of T raditional Chinese Medicine and Materia Medica , 1995 , 16(6):57～ 58
4　Zhao Cuilan , Li Kaiyuan , Jiang Yan.Effect of ethanol ex tract of Hibiscus rosa-sinensis flowers on secreatary function of human
chorionic tissues in vitro.Yunnan Journal of T raditional Chinese Medicine , 1994 , 15(1):50～ 51
5　Anita Pakrashi , Bhattacharva K , Syed M K , et al.Flow ers of Hibiscus rosa-sinensis , a po tentail source of contrag estative agent.
I II:Interceptiv e effect of bezene in mouse.Contraception , 1986 , 34(5):524～ 536
6　Zhou M ingdong , Jiang Yan , Zhao Cuilan , et al.Study on compositions of ether pentrolum ex tract of Hibiscus rosa-sinensis and
its antifertility.Yunnan J.Traditional Chinese Medicine and Materia Medica , 1997 , 18(2):33～ 34
7　Tu Yuanquan , Zhao cuilan , Jiang Yan , et al.Studies on the antifertility constituent of Hibiscus rosa-sinensis L.(HR-1).Joural
of Yunnan University , 1998 , 20(3):159 ～ 161
8　Turner C D , Bagnara J T.General Endocrinology(six th edition).Science Publishers , 1983.359 ～ 369
扶桑花石油醚提取物(HR-1)的抗早孕作用以及对大鼠血清生殖激素的影响
江　燕　　赵翠兰　　李开源　　周铭东
(云南大学生物系 , 昆明 , 650091;第一作者 40 岁 ,女 , 副教授)
摘要　观察不同剂量的扶桑花提取物在宫内注射大鼠的抗早孕效果.实验结果显示:(1)每侧子宫给
予 10 mg ,20mg 和 30 mg 的石油醚提取物(HR-1)后 ,子宫内死胚和损伤胚的百分率分别达到 81.6%,92.
0%和99.0%,胚胎平均大小分别为 0.30 cm ,0.31 cm 及0.17 cm ,明显小于对照组的胚胎0.75 cm(P<0.
01).(2)给药各组的血清 FSH()和雌二醇呈下降趋势 ,血清 LH()及孕酮水平仅在 30 mg 剂量组明显低
于对照组(P <0.05).这提示 ,HR-1在大鼠的抗早孕作用与垂体和卵巢激素的降低有关.
关键词　扶桑花 ,抗早孕 ,孕酮 ,雌二醇
分类号　R285
165第 3 期　　　　江　燕等:扶桑花石油醚提取物(HR-1)的抗早孕作用以及对大鼠血清生殖激素的影响