Relationship of anti-fertility effects of Andrographis paniculata and hormonal assay in female rats

Aims: This study was aimed to find out anti-fertility effects of Andrographis paniculata (AP) plant. Study design & Methodology: A prospective case control animal study with 85 female and 30 male rats (Total 115 rats) was done in the Department of Pharmacology of Dhaka Medical College and Bangladesh Medical College, Dhaka, Bangladesh from July 2002 to December 2003. The total 85 female rats were grouped into case study or exposure group (N=60 female rats) exposed to water extract of AP & control or non exposure group (N=25 female rats, 30 male rats also considered as control group) Exposure group (water extract) of female were again subdivided in 3 groups according to duration of (A.P) exposure (N=20 in each group) e.g. 4 weeks, 6 weeks and 8 weeks. Mating schedules were done after the completion of scheduled duration of exposure with A.P (Dose was 1 gm/kg). Results: In group I percentage of infertility was 33.33% in 4 weeks exposure, 50% in 6 weeks exposure & 100% in 8 weeks exposure respectively. In control group (Group II) percentage of infertility was 0%. In case study group of female rats the value of FSH was 1.20 1U/L (4 weeks), 1.12 1U/L (6 weeks), 1.00 1U/L (8 weeks), LH 0.78 1U/L (4 weeks), 0.70 1U/L (6 weeks), 0.64 1U/L (8 weeks), Estrogen 45.30 pg/ml (4 weeks), 44.80 pg/ml (6 weeks), 44.20 pg/ml (8 weeks) and Progesterone 4.84 nmol/L (4 weeks), 4.72 nmol/L (6 weeks) and 3.80 nmol/L (8 weeks). In non exposure group the value of FSH was 1.23 1U/L, LH 0.80 1U/L, Estrogen 47.05 pg/L and Progesterone 5.50 nmol/L. In exposure group all the values were lesser than the normal hormonal value. Conclusion: This study suggests that due to lower level of hormone, female rats have promising percentage of infertility with AP. Further study is needed with rat as well as clinical trial with human being.


Introduction
The scientists all over the world are putting much emphasis on herbs and plants in the fertility control.Herbal drugs are usually effective, inexpensive & afforded by people of rural area 1,2,3,4,5 .Bangladesh is rich in natural resources of herbs.Andrographis Paniculata (AP) also known as king of bitters is a member of the plant, family Acanthaceae and has been used for centuries in Asia to treat various ailments including GI tract and upper respiratory infections, fever, sore throat and a variety of other chronic and infectious diseases for prevention of contraception 6,7,8,9 .The leaves contain the highest amount of androdrapholide (2.39%), most active component with very bitter taste & colorless crystalline in appearance 10,11,12,13,14 .According to recent research in human being Andrographis are highly bioavailable and readily absorbed from G.I.T. 80% of andrographolides are excreted by kidney within 8 hours 15,16,17,18 .In animals there is no acute or short time toxicity in very high doses (1 -10 gm/kg body weight) & no effects on major organs 19,20,21 .The antifertility effect has been evaluated in the screening programme done by Zoha et al in 1989 with both male and female mice & confirmed the antifertility effect 22,23 .One study also done by Jean barilla in 1999 with female mice and also shown the promising anti fertility effect & significantly lowers all the hormonal value such as follicle stimulating hormone (FSH), leutinizing hormone (LH), estrogen, progesterone & testosterone 24,25,26 .We have proposed the study to investigate the antifertility effect of Andrographis paniculata with an aim to find out a safe, acceptable and effective method of contraception of female rats.

Materials and Methods
This study was planned to design as a prospective case control study and was carried out to find out any association between Andrographis paniculata plant and hormonal assay related to antifertility effects in female rat and was done in the After completion of exposure with AP (4 weeks, 6 weeks and 8 weeks) female rats were kept into different cages and labeled properly and allowed to take normal diet.In group-I 20 treated female rats from each group and 6 untreated male rats from control group were taken.In group-II 25 untreated female rats and 12 untreated male rats were taken.Then mating outcome has been observed.They have kept together until 3 times birth of litters of control group & in this experiment they have kept together for 3 months because usually rats gave birth of litter 3 times within the 3 months.Then number and percentage of pregnancy and percentage of infertility (%) have been observed.The evidence of pregnancy was observed daily for a period of one week.Hormonal assay such as LH, FSH, Estrogen and Progesterone for female rats in exposure groups (4weeks, 6weeks & 8 weeks) & also in non exposure group were done to observe the anti-fertility effects.

Results
Table I

Table II -
Distribution of study population according to duration of exposure of Andrographis paniculata (4 weeks, 6 weeks and 8 weeks) -Distribution of study population Sakila S. et al.

Table III -
Mating schedule and rat population according to duration of exposure (both male and female rat)

Table IV -
Outcome of mating behavior (percentage of infertility)Relationship of anti-fertility effects of Andrographis paniculata and hormonal assay in female rats

Table V -
Hormonal assay (FSH, LH, Estrogen and Progesterone) of female rat of both exposed and non exposed group (Water Extract)