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In This Article
   Abstract
   Introduction
   Material and Methods
   Results
   Discussion
   References
   Article Figures
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RESEARCH PAPER
Year : 2006  |  Volume : 38  |  Issue : 2  |  Page : 111-114
 

Antiovulatory and abortifacient potential of the ethanolic extract of roots of Momordica cymbalaria Fenzl in rats


1 Visveswarapura Institute of Pharmaceutical Sciences, Vokkaligara Sangha Campus,K R Road, Bangalore 560004, India
2 Department of Pharmacy, Faculty of Science and Technology, M.S. University of Baroda, Baroda, India

Correspondence Address:
Raju Koneri
Visveswarapura Institute of Pharmaceutical Sciences, Vokkaligara Sangha Campus,K R Road, Bangalore 560004
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0253-7613.24616

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  Abstract 

Objective: To study the antiovulatory and abortifacient activity of the ethanolic extract of roots of Momordica cymbalaria Fenzl. Materials and Methods: Female Wistar albino rats (150 to 200 g) with at least three regular estrous cycles were administered ethanolic extracts of roots of Momordica cymbalaria Fenzl. at two doses 250 and 500 mg/kg orally for 15 days. Control group received vehicle (tween 80 1%, p.o. daily). Animals were sacrificed on 16th day. One ovary was subjected to histopathological studies and the other for biochemical studies. Abortifacient study was done in another set of three groups of animals. The extracts at doses of 250 and 500 mg/kg were administered orally through gastric gavage from the day 6 to day15 of pregnancy (the period of organogenesis). The animals were laparotomised under light ether anesthesia and semi- sterile conditions on day 19th of pregnancy. Both horns of the uterus were observed for the number of implantation sites, resorptions, dead and alive foetus. Results: Highly significant (P<.001) decrease in the duration of estrous cycle and metaestrous phase and increase in proestrous phase was seen, but diestrous phase was unchanged in both 250 and 500 mg treated group when compared to untreated group. Significant decrease in the ovarian weight and a highly significant increase in serum cholesterol with 250 mg/kg dose were seen. Histology of ovary showed an increase in preovulatory and atretic follicles. Ethanolic extract showed a dose dependent abortifacient effect in pregnant rats during organogenesis period. At 250 mg/kg ethanolic extract did not show any abortifacient activity but reduced the number of viable foetus and resorptions with no change in the foetal weight when compared with control group. At 500 mg/kg ethanolic extract showed highly significant (P< 0.001) abortifacient activity. Conclusion: The ethanolic extract at both doses (250 and 500 mg/kg) showed antiovulatory activity. It is abortifacient at 500 mg/kg but not at 250 mg/kg.


Keywords: Antifertility, contraception, ovulation.


How to cite this article:
Koneri R, Balaraman R, Saraswati C D. Antiovulatory and abortifacient potential of the ethanolic extract of roots of Momordica cymbalaria Fenzl in rats. Indian J Pharmacol 2006;38:111-4

How to cite this URL:
Koneri R, Balaraman R, Saraswati C D. Antiovulatory and abortifacient potential of the ethanolic extract of roots of Momordica cymbalaria Fenzl in rats. Indian J Pharmacol [serial online] 2006 [cited 2019 Mar 25];38:111-4. Available from: http://www.ijp-online.com/text.asp?2006/38/2/111/24616



  Introduction Top


Synthetic estrogens and progesterones, in combination or alone, are extensively used as contraceptives. Although they are highly effective, they are associated with high incidence of side effects. Therefore, the search for new antifertility molecules with minimal side effects continues. Many plant preparations are used to control fertility.[1],[2] The extracts and the dried fruits and leaves of Momordica cymbalaria Fenzl were shown to have antidiabetic and hypolipidemic properties. [3],[4],[5] Roots are used by the natives of north interior Karnataka and Andhra Pradesh to treat gynecological ailments and to induce abortions. However, scientific study has not been conducted to validate these effects . Therefore, this study examined antiovulatory and abortifacient activities of the ethanolic extract of roots of Momordica cymbalaria Fenzl.


  Material and Methods Top


Plant collection and preparation of the extract

Fresh roots of Momordica cymbalaria were collected from Gadag district, Karnataka. They were identified and authenticated by a botonist in the Department of Botany, Bangalore University, Bangalore. A specimen sample of the same was preserved at the herbarium of the Department of Botany, with the voucher no. 18122003, for future reference.

The roots were isolated and chopped into small pieces. Next, they were dried under shade at room temperature for seven days. The dried roots were powdered, passed through sieve (coarse 10/44), and extracted with 95% v/v ethanol using Soxhlet extractor.[6] The combined extracts were concentrated at 40o C to obtain dark brownish yellow residue. The yield obtained from this process was found to be 21.5 % w/w.

Animal

Colony bred female adult albino rats of Wistar strain (150-200 gm) were maintained under controlled standard animal house conditions. They had access to standard rat feed and water ad libitum. All the animal procedures were performed according to the CPCSEA norms. The Institutional Animal Ethics Committee approved the experimental procedures. The acute oral toxicity study was performed using the up and down procedure (OPPT guidelines).

Antiovulatory activity

Vaginal smear from each rat was examined daily for 15 days, and those rats exhibited three regular cycles[7] were included in the study. The selected rats were divided into three groups of six animals each. Drugs and vehicle were started in the estrous phase and administered orally, daily for 15 days. Group I received vehicle only (1%Tween 80, p.o. daily) and served as control. Groups II and III received ethanolic extract at 250 and 500 mg/kg, respectively. The 15-day treatment was to cover three regular estrous cycles. Vaginal smear from each animal was observed every morning between 9-10 A.M. On the 16th day, 24 hours after the last treatment, the animals from each group were sacrificed. Ovaries and uteri were dissected out, freed from extra deposition, and weighed on a sensitive balance (Precisa, XB series). One ovary from each animal was processed for biochemical analysis of cholesterol. The other ovary was fixed in 10% formalin buffer for histological study.

Abortifacient activity

The female rats were caged with male rats of known fertility in the ratio of 2:1 in the evening of proestrous. They were examined the following morning for the presence of sperms. Rats exhibiting thick clumps of spermatozoa in the vaginal smear were separated, and that day was designated as day one of pregnancy. The pregnant rats were divided into three groups of six animals each. Group I received vehicle only (1%Tween 80, p.o. daily) and served as control. Group II and group III received ethanolic extract at 250 and 500 mg/kg, p.o. daily, respectively. The extracts were administered orally through gastric gavage from the 6th to the 15th day of pregnancy (period of organogenesis). The animals were laparotomised under light ether anesthesia on the 19th day of pregnancy. Both horns of the uterus were observed for the number of im-plantation sites, resorptions, and dead and alive fetuses.[9],[10],[11]

The results are expressed as mean±SEM. The statistical significance between groups was analysed using the one-way ANOVA test and the Tukey Kramer multiple comparison post-test. P<0.05 was considered significant.


  Results Top


Mortality in the acute toxicity test was not seen in the limit test at a dose of 5000 mg/kg. Therefore, 1/10th & 1/20th of the dose were selected for this study.

Ethanolic extract at 250 and 500 mg/kg caused a significant (P<0.001) decrease in the duration of the estrous and metes-trous phases, no change in the duration of the diestrous phase, and a significant (P<0.001) increase in the duration of the proestrous phase, compared with the control group. [Table - 1]

Ethanolic extract at 250 and 500 mg/kg significantly (P<0.01) decreased the weight of ovaries, compared with the control group. [Table - 2] An increase in preovulatory follicles and atretic follicles was seen in the treated groups [Figure - 2][Figure - 3]. A highly significant (P<0.001) increase in ovarian cholesterol level was observed at 250 mg/kg dose, compared with the control group. [Table - 3]

At 500 mg/kg, ethanolic extract showed a highly significant (P<0.001) abortifacient effect. At 250 mg /kg, ethanolic extract did not show any abortifacient activity, but reduced the number of viable fetuses and resorptions (6.16±1.24). There was no change in the fetal weight, compared with the control group. Table 4


  Discussion Top


Many morphological, histological, physiological, and biochemical changes occur in the ovary during the estrous cycle. During the maturation of preovulatory follicles, ovulation takes place under the combined and balanced influence of ovarian and extra ovarian hormones. Imbalance in these hormones leads to irregularity in the ovarian functions and duration of the estrous cycle.[12],[13],[14]

The estrous cycle in the rats treated with extract (250 and 500 mg/kg) showed a decrease in the duration of estrous and the metestrous phases. It was also characterised by a prolongation of the proestrous phase. The prolongation of the proestrous phase indicates that maturation of the follicle in the preovulatory phase was delayed, leading to non-maturation of graffian follicle. Non-availability of matured graffian follicle was indicated by reduction in the estrous and the metastrous phases. Therefore, ovulation was inhibited. This result was further supported by our histopathological studies [Figure - 2][Figure - 3] in which the transverse section of the ovary showed the presence of primary or developing follicles.

Ovary can be considered an aggregate of three endocrine tissues, the stroma, the follicle and the corpus luteum. The weights of these tissues constitute the net weight of the ovary. During the estrous cycle the weight of the ovarian tissue increases under the influence of gonadotrophic and steroidal hormones. The decrease in the weight of ovaries of the rats treated with extract indicates a decrease in the activity of the stroma, the follicle, and the corpus luteum in the ovary. This decrease may be due to the non-availability of gonadotrophic or steroidal hormones or both.[13]

Atretic follicles are degenerating preovulatory follicles. The degeneration of preovulatory follicles takes place due to non-availability of steroidal hormones (essential for their maturation and differentiation), non-availability of local estrogen produced by granulosa cells, or imbalance in endogenous steroid, protein and hormones. The presence of increased atretic follicles in the rats treated with ethanolic extract, compared with control rats, indicates that the extract promotes the degeneration of preovulatory follicles. Cholesterol is the precursor for the steroidogenesis of ovarian endocrine tissues. The significant increase in ovarian cholesterol in the treated group (250 mg/kg) indicates that cholesterol is not used for steroidogenesis.[13] But 500 mg dose did not show a similar effect indicating that it occurs only at a lower dose.

Abortion refers to the premature expulsion of the products of conception from the uterus. Abortion may be due to maternal exposure to chemicals, which can disrupt pregnancy and cause detachment of the embryo.[10] Ethanolic extract at 500 mg/kg showed 100% abortifacient activity, while 250 mg/kg did not show abortifacient activity. However, it reduced the number of viable fetuses.

To conclude a highly significant decrease in the duration of the estrous and the metestrous phases and increase in the duration the proestrous phase was seen. In addition, a highly significant decrease in ovarian weight and increase in cholesterol level, compared with the control group, was noted. These findings indicate that extract Momordica cymbalaria produces temporary inhibition of ovulation. The result of administration of extract to the pregnant rats during organogenesis shows that the extract is abortifacient only at the higher dose of 500 mg. These findings could explain the traditional use of Momordica cymbalaria as abortifacient and antiovulatory[Figure - 1].

 
  References Top

1.Nadkarni AK, Nadkarni KM. Cucurbitaceae. Indian materia medica. 2nd ed. Bombay: Popular Book Depot; 1982.  Back to cited text no. 1    
2.Chopra SR, Badhwar RL, Ghosh S. Cucurbitaceae. Poisonous plants of India. 1st ed. Jaipur: Academic Publishers; 1984.  Back to cited text no. 2    
3.Rao BK, Kesavulu MM, Giri R, Apparao C. Antidiabetic and hypolipidemic effects of Momordica cymbalaria Hook. Fruit powder in alloxan diabetic rats, J Ethnopharmacol 1999;67:103-9.  Back to cited text no. 3    
4.Rao BK, Kesavulu MM, Apparao C. Antihyperglycemic activity of Momordica cymbalaria in alloxan diabetic rats. J Ethnopharmacol 2001;78:67-71.  Back to cited text no. 4  [PUBMED]  [FULLTEXT]
5.Kameswararao B, Kesavulu MM, Apparao C. Evaluation of antidiabetic effect of Momordica cymbalaria fruit in alloxan-diabetic rats. Fitotherapia 2003; 74:7-13.  Back to cited text no. 5  [PUBMED]  [FULLTEXT]
6.Shivalingappa H, Satyanarayan ND, Purohit MG. Antiimplantation and pregnancy interruption efficacy of Rivea hypocrateriformis in the rat. J Ethnopharmacol 2001;74:245-9.  Back to cited text no. 6  [PUBMED]  [FULLTEXT]
7.Hafez ES. Reproduction in breeding techniques for laboratory animals. Philadelphia: Lea and febiger; 1970.  Back to cited text no. 7    
8.Kulkarni SK. Biostatistics in experimental pharmacology. Hand book of experimental pharmacology. 3rd ed. Delhi: Vallabh Prakashan Publishers; 1999.  Back to cited text no. 8    
9.Medeiros RM, Gorniak SL, Guerra JL. Fetotoxicity and reproductive effects of monocrotaline in pregnant rats. J Ethnopharmacol 2000;69:181-8.  Back to cited text no. 9  [PUBMED]  [FULLTEXT]
10.Feranada CG. Almedia, Ione P. Lemonica . The toxic effects of Coleus barbatus B. on the different periods of pregnancy in rats. J Ethnopharmacol 2000;73: 53-60.  Back to cited text no. 10    
11.Nath D, Sethi N, Singh RK, Jain AK. Commonly used Indian abortifacient plants with special reference to teratologic effects in rats. J Ethnopharmacol 1992;36:147-54.  Back to cited text no. 11  [PUBMED]  
12.Prakash AO, Mathur R. Studies on oestrous cycle of albino rats: Response to embelia ribes extracts. Planta Medica 1979;36:131-41.  Back to cited text no. 12    
13.Shivalingappa H, Satyanarayan ND, Purohit MG, Sharanabasappa A, Patil SB. Effect of ethanol extract of Rivea hypocrateriformis on the estrous cycle of the rat. J Ethnopharmacol 2002;82:11-7.  Back to cited text no. 13  [PUBMED]  [FULLTEXT]
14.Circosta C, Sanogo R, Occhiuto F. Effects of Calotropis procera on oestrous cycle and on oestrogenic functionality in rats. Farmaco 2001;56:373-8.  Back to cited text no. 14  [PUBMED]  


    Figures

[Figure - 1], [Figure - 2], [Figure - 3]

    Tables

[Table - 1], [Table - 2], [Table - 3]

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