Introduction

Nux  vomica is a member of the Logoniaceae family and the medicine is prepared from dried seeds. It’s raw seeds are called as Poison-nut due to presence of two alkaloids known as Strychina and Bruccia. Strychina is the principle ingredient. Nux vomica  is one of the best remedies with which to commence treatment of causes that have been dragged by various medicines to counteract these chronic effects. It is a polycrest remedy with the bulk of symptoms corresponding in similarity with the most common diseases of modern era. It is useful for those persons who lead a sedentary life, doing much work or those who remain under stress from prolonged office work and business worries.

Sir Hahnemann said Nux vomica  is chiefly successful with persons of an ardent character of an irritable, impatient temperament, disposed to anger spite or deception. Nux vomica  play a very useful role to cure thin, spare patient with irritable and nervous temperament who is prone to indigestion and haemorrhoids.

Some researchers believe that Nux vomica  has anti-inflammatory properties, used to treat conditions that are worsed by inflammation, such as rheumatism as these or haemorrhoids. Some said that Nux vomica  has antioxidant,  that protect against free radical, which are chemicals in your body that can damage your cells.

Due to lot of beneficial uses of Nux vomica , there shall be no side effects occurs according to mode of treatment. In the present investigations we have an idea to assess the overall effects of Nux vomica  in eukaryotic system which is plant system for fast and better observations and their findings we have used.

We are potentially using  Nux vomica  to cure various kind of diseases and disorders. Nux vomica  is used as natural extract of seed, so that there  is  probability to find some abnormal feature or side effects as genotoxic results by the treatment of Nux vomica  as natural extract (Akinboro and Bakare 2007). Nux vomica  may shows mitodepressive effects on normal mitotic division (Badr 1983). According to Badr 1983 a mitodepressive and chromotoxic activities were found by two natural herbicide in A. cepa another material product extract of Hibiscus cannabis also effects as Cytotoxic activities in Allium cepa (Moujr et al 2007).

In the present investigation we were used extract  of Nux vomica  as homeopathic drugs on  a text system that was a plant Vicia faba to assess various cytotoxic and genotoxic effect of Nux on eukaryotic system.

Material and Method

Mother tincture of extract of dried seeds of Nux vomica  were diluted in various concentrate with distilled water. We have used 5%, 10%, 20% and 30% concentrations for root tip treatment of Vicia faba (4 hrs., 8 hrs., 12 hrs., 24 hrs). First the seeds of Vicia faba were sterilized by using 2%  mercuric chloride to 2 min and rinsed with distilled water. Sterilized seeds were soaked in distilled water for 4-5 min, after that the seed were germinated on moist filter paper in sterilized petriplates. When the root tips were 2.3mm long were subjected into 5%, 10%, 20%, 30% solution of Nux vomica  for 8, 12,  and 24hrs treatment. After treatment root tips were washed with water and fixed in Carnoy’s solution (IGAA and Ethyl alcohol) for 6-14 yrs followed by transfer into 70% of alcohol at 4C. Slides have been prepared by acetocarmine squash method & observations were taken under 100x with Oil emersion. Pictures were taken by Olympus  3nm camera using Nova Oraplus 21 Din or 125  ASA.

Observations

There were so many structural,  behavioural and numerical aberrations were observed in microslides of Vicia treated by Nux.Those were as follows:

Mitotic Index

The mitotic index was decreased with increase in concentration   (5-10) and duration of treatment (4-24) hrs. The mean present value of mitotic index were decreased from 11.50 to 11.20%.(Table-1)

Condensed to Sticky Chromatids and Chromosomes

The treatment of Nux vomica  resulted in stickiness of chromosomes at metaphasic plate (Plate-1, fig –A, C, F). This kind of stickiness was observed increasing order in respect to increasing order of  duration and concentration. The mean per cent value was higher through 12.13 to 13.93% in Vicia. (Table-1, fig-F).

Multipolar Spindles and Inactivation

Multipolar spindles were noticed in ascending order in order of conc. and duration of treatment. The mean percentage was 0.52-0.62% (Table-1, fig- E,H,L).

Chromatin Bridges at Anaphase

It was observed that some chromatin bridges were disturbed the separation of chromatin. This abnormality was increased with increase duration  and conc. of treatment of Nux vomica .

The mean percentage value was 4.47-5.14 (Table-1, fig- G,H,K, L).

Fragments

Another kind of abnormality  observed in Vicia, that was fragments of chromatids during anaphasic  separation. The heigher value was 4.85 at 12 hrs treatment.(Table-1,Fig-I,J).

Swelled and Stretched Chromosomes

Some swelled and stretched chromosomes were also observed in Vicia at higher concentration only (fig-D,F).

Figures 1: Showing various chromosomal aberrations during different stages of mitotic cell division in root tips cells of Vicia faba.   Click here to View figure

Fig. A-Condensed and sticky chromosomes, B-Unoriented chromosomes, C-clumped and sticked chromosomes, D- swelled and ring chhromosmes, E-scattered chromosomes and multipolar spindle, F-Sticky and swellon chromosomes, G-chromatin bridges at anaphase, H-multipolar spindle and chromatin bridges, I-fragmented chromatides, J-chromatin fregmentation during separation, K-chromatin bridges at anapase, L-multipolar separations of chromatids.

Table 1: Types and distribution of somatic chromosomal abnormalities in (%) incduced by different conc. and duration of Nux vomica extract (Root-tip treatment) in Vicia Faba

Result and Discussion

The present investigation has suggested that Nux vomica  has potential to damage chromosomes and its content. Various types of cytogenetical abnormalities were appeared by treatment of Nux vomica  on Vicia faba.

The inhibition in MI values are perhaps due to reduction DNA and RNA contents in dividing root meristem cells as observed by Anuradha (1991), through the pesticides treatment on the basis of cytogenetical, marphological and physiological studies in Lens and Vicia. Amato (1942, 1952) suggested that reduced MI  has resulted due to Prophase poisoning and restriction of the division, Kuras et al (2009) found alkaloids from Licaria fomentosa bark were retard on inhibit mitosis and change mitotic phase in A. cepa.

In the present investigation it was clearly showed that appearance of  sticky chromosomes was the most prominent abnormality  observed (Table-1).The differential response of the plant against drugs may be due to diverse correlation in the heterochromatin and euchromatin contents of the plant chromosomes (Evans,H.J.1962). Stickiness  is the most common abnormality. It is probably due to delay in chromosome movement through drug effect or influenced by gene action ( Darlington 1945). In the present study, stickiness of chromosomes may be attributed to the alteration in the chromosomal protein resulting in the physiological changes of surface nucleoprotein configuration or to disturbances in cytochemically balanced reaction, (Salimuddin, B.Ramesh 1994).

Another significant effect of Nux vomica  was partial or entire inactivation of spindles leading to scattering of chromosomes, restitution of nucleus and multipolar spindle. The frequency of multi poles were observed in higher conc. of Nux vomica . The chromosome movement is adversely affected by inactivation and inhibition of spindle formation, due to inhibited ATP synthesis (Freg Wyssling 1938). The present study reveals that there is a tendency towards the arrest of division at metaphase or lengthened  the period of division, similar aberration was also reported by the effect of drug (El-Bayoumi et al 1987).

Chromatin bridges were also observed due to occurrence of chromatin stickiness and subsequent failure of anaphase separation or it may be the result of chromosome breaks and reunion. Grover and Virk (1986). Fragments and breaks of chromatids were seen in some cells, it was may be the result of cytotoxic property of drug disturbed the synthesis and degradation of nucleic acids, proteins, etc. According to Sharma (1985) fragments followed by translocation may lead to heritable phenotypic differences. Kato (1960) observed that the extracts of young pods of A. fistilerium induced fragmentation and chromosomal breakes. These breaks are know to produced chromosomal mutations because of deletion and duplication, which lead to cell death Swaminathan , and Natrajan  (1956) has fragmentation A. sativum. Induced by vegetable oils and edible fats.

Grant W.F. (1978) was reported Laggards same as present observations. It may be attributed by the adhesion of the centromere of a prometaphase chromosomes at the equator. The Lagging chromosomes has been reported earlier by Somashekhar et. al (1984) in Allium.The Laggards usually form micro nuclei able to perform DNA synthesis and mitotic condensation with the main nucleic. They can subsequently be integrated into the main nuclei in the first mitosis . Rao et al (1986) in A. cepa,.

The micro nuclei and multi nuclei were observed in  low  frequency. They may  arise from laggards and chromosomes fragments (Torkoglu et al 2007), or may also be  attributed to the  multiple spindle, or building of nucleus. Micronuclei are true mutagenic effect.( Kirkla D. 1998).

Conclusion

On the basis of observations and results, present investigation revealed that the action of the Nux vomica  as homeopathy is a very cytogenotoxic with increase in concentrations  and duration of treatment. It has suggested that the dose and duration of given drug Nux vomica  as homeopathy drug should be tested before for it mutagenic an genotoxic properties in various eukaryotic system.

References

1. Akinboro A and Bakare A. A. 2007-Cytotoxic and genotoxic effects of aqueous extracts of five medicinal plants on. Allium cepa journal of Ethenopharmacology. 112:470-475.
2. Anuradha and Sarbhoy.  Inhibitions of Chlorophyll synthesis and leaf senescence by pesticides carbaryl and dimethorate in Lense cullinaris78^th ind.sci.cong.part III.
3. Badr A. 1983. Mitodepressive and chemotoxic activities of two herbicides in A. cepa. Cytologia. 1991;48:451-457.
   CrossRef
4. Amato D.F.  Prophase poisoning by chemical agents. Caryologia. 1949;1:327-328.
   CrossRef
5. Amato D.F. The cytological study of chemical mutagens. Genet. I——–ber. 1952;4:3-20.
6. Darlington C.D. Chromosome chemistry & gene action. Nature. 1942;149:66
   CrossRef
7. El-Bayoumi  A.S,Habib  A.A and Haleem A.S. Mutagnecity of the drug Algaphan using the Allium test.Abstreact 16th annual meeting of European Enviro.Mutagen Society(EEMS), Mutab. 1987.
   CrossRef
8. Evans H.J. Chromosome aberrations induced by ionizing radiation. Int. Rev. Cytol. 1962;13:221-231.
9.  Wyssling F.A.  Submikroskpische morphologie des protoplasma and seiner derivate Berlin, Borntager. 1938.
10. Grant W.F. Chromosomal aberrations in plants as a monitoring system .Environ.With Persp. 1978;27:37-43.
    CrossRef
11. Grover I.S and Virk G.S. A comparative study by gamma rays and some     chemical mutagens on the induction of chromosomal aberrations in mungbean(Vigna radiataL.). Acta Botanica Indica. 1986;14:170-180.
12. Kato Y.  Chromosomal breaks and bridges induced by extracts from spinach and beet root apahyton (Argentina). 1957;8:131-136.
13. Kirkla D. Chromosome aberration testing in genetic toxicity –past , present and future. Mutation Research. 1998;404:173-185.
    CrossRef
14. Kuras M,Pilarski R .b,  Nowakowskac J, Zobel A.d, Krzysztof.
15. Brzost a, Antosiewicz J.e. Krzyszt of Gulewicz b (2009). Effect of Alkaloid-Free and Alkaloid-Rich preparations from Uncaria tomentosa bark on mitotic activity and chromosome morphology evaluated by. Allium Journal of Ethnopharmacology.  2009;121:140–147.
16. Moujir L,Seca A.M.L, Silva A.M.S,Lopez M.R, Padilla N, Cavaleiro J.A and Neto S.
17. P. 2007-Cytotoxic activity of lignans from Hibiscus cannabinus. Fitotererpia. 78:385-387.
18. Rao B.V, Sharma B.S.R and Rao  B.G. Clastogenic and turbagenic effects of organophosphorus pesticides on Allium cepa root meristems. Proc. Ind. Acad. Sci. (Plant sci.). 1986;96:303-309.
19. Salimuddin B.R. Karyotype, nuclear and chromosomal DNA variation in Lense culinaris. Cytologia (Tokyo). 1994;59(1):7-15.
20. Sharma S.  Cytogenetical studies on the effect of some carbamate pesticides on Cyamompsis and Lens.  Ph.D.Thesis Agra Univ. Agra. 1985.
21. Somashekhar and Gowda M.T.G. Effect of a fungicide vitavaxon on. Allium cepa.Cytologiya. 1984;49:177-181.
    CrossRef
22. Swaminathan M.S, Natrajan A.T.  has fragmentation sativum. Induced by vegetable oils and edible fats. 1956.
23. Turkoglu M, Gurer U.S, Akarsu B.G. 2007, Colonic delivery of compression coated nisin  ctin/HPMC polymer mixture. Eur J Pharm Biopharm. 2007 Aug;67(1):202-10.
24. Erden T.M, Ozdemir M. 2008, Isolated right ventricular infarction due to occlusion of the right ventricular branch in the absence of percutaneous coronary intervention. Can J Cardiol. 2008 Oct;24(10):793-4.
    CrossRef