Evaluation of Antimitotic Activity of Momordica Dioica Fruits on Allium Cepa Root Meristamatic Cells

In today’s world, peoples are suffered from lots of dangerous diseases. Cancer is one of that disease and second leading cause of death in the world. (Cragg and Newman, 2005) Now a day, various new anticancer agents are isolated from the natural sources of drugs. Natural sources of drugs are herbal based drugs consists of about 60-80 % of all other drugs and used as medicine from 1990. (Lee, 2010; Li and Vederas, 2009) The active constituent which is isolated from plants playa vital role in treatment of various diseases and gives special attention such herbal based plants due to its pharmacological activities. (Newman, et al., 2003) There are certain plants which are used in treatment of diseases like cancer. It is found that most of isolated compounds from plants show good anticancer activity (Gonzales and Valerio, 2006; Chopra, 1996). The Momordica dioica is recurring, having distinct male female plants consisting of family curcurbitaceae, which is commonly known as small bitter gourd or spiny gourd throughout the world where as in India it is called as jangle Karela, Kantola or Karola. The plant Momordica dioica shows various pharmacological or medicinal properties like analgesics, anti-tumorogenic, anti-inflammatory and anti-diabetic activity (The useful plants of India, 1992; The wealth of India 1962; Kartika and Basu 1987). The fruit of 10 mg/ml concentration of aqueous extract of Momordica dioica shows good antimitotic activity. (Patil, et al., 2018) From the dichloromethane extract of Momordica dioica roots, five active constituents are isolated which gives good anti-cancer activity on cancerous cell like L1210. At 4 mg/ml it shows good growth inhibitory index about 50 %. (LuoL, et al., 1998) Based on review of literature that no work has to be done on antimitotic activity at concentration of 15 mg/ml of aqueous extracts of fruits of Momordica diocia. Hence in this study aqueous extracts of fruits of Momordica diocia in 15 mg/ml concentration was assessed by using Allium cepa root meristamatic cells. The 15 mg/ ml concentration of aqueous extracts of fruits Momordica diocia was taken because to see the effect of fruits at 15 mg/ ml concentration. Evaluation of Antimitotic Activity of Momordica Dioica Fruits on Allium Cepa Root Meristamatic Cells


Introduction
In today's world, peoples are suffered from lots of dangerous diseases. Cancer is one of that disease and second leading cause of death in the world. (Cragg and Newman, 2005) Now a day, various new anticancer agents are isolated from the natural sources of drugs. Natural sources of drugs are herbal based drugs consists of about 60-80 % of all other drugs and used as medicine from 1990. (Lee, 2010;Li and Vederas, 2009) The active constituent which is isolated from plants playa vital role in treatment of various diseases and gives special attention such herbal based plants due to its pharmacological activities. (Newman, et al., 2003) There are certain plants which are used in treatment of diseases like cancer. It is found that most of isolated compounds from plants show good anticancer activity (Gonzales and Valerio, 2006;Chopra, 1996). The Momordica dioica is recurring, having distinct male female plants consisting of family curcurbitaceae, which is commonly known as small bitter gourd or spiny gourd throughout the world where as in India it is called as jangle Karela, Kantola or Karola. The plant Momordica dioica shows various pharmacological or medicinal properties like analgesics, anti-tumorogenic, anti-inflammatory and anti-diabetic activity (The useful plants of India, 1992; The wealth of India 1962; Kartika and Basu 1987).
The fruit of 10 mg/ml concentration of aqueous extract of Momordica dioica shows good antimitotic activity. (Patil, et al., 2018) From the dichloromethane extract of Momordica dioica roots, five active constituents are isolated which gives good anti-cancer activity on cancerous cell like L1210. At 4 mg/ml it shows good growth inhibitory index about 50 %. (Luo-L, et al., 1998) Based on review of literature that no work has to be done on antimitotic activity at concentration of 15 mg/ml of aqueous extracts of fruits of Momordica diocia. Hence in this study aqueous extracts of fruits of Momordica diocia in 15 mg/ml concentration was assessed by using Allium cepa root meristamatic cells. The 15 mg/ ml concentration of aqueous extracts of fruits Momordica diocia was taken because to see the effect of fruits at 15 mg/ ml concentration.

A. Plant Material & Reagents
Momordica diocia fruits were agglomerated from Nandurbar (MS) region and authenticated by Professor S. K. Tayade, Botany Department, College of Art's, Commerce and Science, Shahada, Lonkheda-425409, (MS). Chemicals and reagents are acquired from Rajesh Chemicals, Mumbai.

B. Preparation of Aqueous Extract of Fruits
Collected fruits were washed with distilled water, shade dried and pulverized to a rough form and extraction is doneby maceration methodusing water-chloroform solvent. The filtrate was evaporated by using rotary-evaporator which removed water solvent in vacuum at 40°C. Take small amount of water extract of Momordica diocia fruits and dissolved in to small amount of distilled water to produce test solution. This test solution was used to study different chemical tests such as alkaloids, flavonoids, tannins, carbohydrates and proteins for detection of different chemical constituents present in the aqueous extract (Harborne, 1988;Kokate, et al., 1999). (Abhang, et al., 1999;Willams and Omoh, 1996)  D. Statistical Analysis (Kulkarni, 1993) The experiments were carried out in triplicates and the data were expressed as mean ± SEM. The significance of difference among the various treated cells and control cells were analyzed by means of one-way ANOVA.

Results
From Table No. 1 showed the presence of phytoconstituents such as alkaloids, flavonoids, tannins, carbohydrates and proteins in aqueous extracts of fruits Momordica diocia.   Table No. 2 showed that fruits Momordica diocia (15 mg/ ml) and methotrexate gives antimitotic in Allium cepa root meristamatic cells. The total number of non-dividing cells in controlled group (distilled water), standard drug group (methotrexate 0.1 mg/ml) and aqueous extract (15 mg/ml) was found to be 120, 91 and 104 respectively. The total number of dividing cells in different stages of mitosis such as prophase, metaphase, anaphase and telophase in controlled group (distilled water) was found to be 15, 4, 2 and 4, in standard group (methotrexate) were found to be 6, 4, 1 and 1 and in 15 mg/ml concentration of aqueous extracts was found to be 10, 3, 2 and 1 respectively. The average mitotic index in water (control), methotrexate (standard) and 15 mg/ml concentration of aqueous extracts of Momordica diocia fruits was found to be 17, 11 and 13 respectively.

Discussion
The fruits of Momordica diocia shows wide spectrum of medicinal activities. The plant contains various phytochemical compounds having diverse chemical structure and nature. The main active constituents are found in plants as phenolic, flavonoids and tannins compound. In existing study, Allium cepa root meristamatic cell model is used to study antimitotic property of fruits Momordica diociain 15 mg/ml concentration. (Abhang, et al., 1991;Latha, 1988) In this parameters, division of cells is same as that of division of cancerous cells in human being. Therefore these cells are used to study antimitotic activity in different parts of plants. Allium cepa meristamatic cells assay is fast, quick to detect toxicity in cells and toxicity in cells genetic material. The inhibition of root growth and antimitotic property gives indication of destruction of cells genetic material. The destruction of cells of genetic material assay action on Allium cepa as a plant helpful in studying karyo type of plant and skill to connect with those of mammalian cells in toxic evaluation. The good genotoxic assay performance of Allium cepa as a plant system has been attributed to the easily studied karyo type of plant and the ability to correlate outcomes of assays with those of mammalian cells in the course toxic evaluations. (Fiskesjo, 1985) In Allium cepa assay, aqueous extracts of fruits of Momordica diocia (15 mg/ml) was found to inhibit root growth in Allium cepa root meristamatic cells and decreased mitotic index after treatment.This suggests that the aqueous extract of fruits of Momordica diocia has fairantimitotic potential in concentration of 15 mg/ml. In Allium cepa assay, MI (mitotic index) is considered as an indication of biomarkers cells proliferation in metaphase cell cycle. (Bhattacharya and Haldar, 2012;Periyanayagam, et al., 2013) Earlier it was proved that fruits of Momordica diocia shows good anticancer activity in different cell lines such as PA-I and Hela cell line by using MTT assay (Ahirrao, 2019).

Conclusion
Our finding support that 15 mg/ml concentration of aqueous extracts of fruits Momordica diocia inhibits cell division in Allium cepa meristamatic cells and may helpful in inhibiting abnormal cell growth like cancerous cells. Further experiments are needed, both in vitro and in vivo to obtain detail mechanisms of action. Positive outcomes from study introduce this plant as a new anticancer drug.