Effect of Alcoholic Extract of Celery Seeds Apium graveolens on the Histological Characteristic Ovaries of Swiss Rat ratus

 

ABSTRACT:

The present study was conducted to evaluate the effect alcoholic extract of the celery seeds Apium graveolens in the histological characteristics of Swiss rats ovaries. The results showed that the low concentration of alcoholic extract of the celery seeds 150 mg / kg greatly affected and by histological sections in increasing the numbers and sizes of corpus luteum and developing follicles also the cortex of the ovary was free from blood congestion and there were increasing in the numbers of primary and secondary follicles compared with the control group. The treatment group with high concentrations of alcoholic extract 300 mg/kg of the of celery seeds showed that the ovary cortex contain smaller numbers of primary and secondary follicles and there were a hypertrophy , and congestion in the cortex cells and there were thicken in some cells nuclei in cortex and presence of large numbers of atretic follicles and the ovarian cortex consist on smaller numbers of corpus luteums compared with the treatment group with lower concentration 150 mg / kg. In addition to the appearance of signs of congestion in the cortex and increase the diameters of arteries and veins of ovary medulla which is responsible for the blood supply of the ovary.

 

 

INTRODUCTION:

Celery, which dates back to the genus Apium graveolens division Apiaceae belonging to the family of succulent plants Parsley is around plant has meaty root and a group with long necks vehicle that constitutes commercial celery and has a height of about 806 cm and ends leg steem of white flowering and after maturity bearing double-brown fruit and has a smell aromatic and spicy taste⁽¹⁾

Celery contains many important chemical compounds including Phenols and Flavonoid such as Apiginin and Apinin these two compounds have the ability to collect Free radicals and minimize the impact of toxic substances resulting from pollution and digestion and metabolism in the body^{(2, 3)} where the body is not has the ability to produce these materials in sufficient quantities, but get it by eating food rich with flavono.

 

Also celeroy contains fatty acids such as Palmitic acid, Oleic acid, linoleic acid and linolnic acid^{(4 )}, as well as the seeds and leaves of celery contains other active ingredients, the most important vehicles Phthalids and Coumarins ⁽⁵⁾ also celery leaves contain minerals and salts including calcium, manganese, phosphorus, magnesium, sodium, which may have a role in increasing the acidic PH level in the blood ⁽⁶⁾ also celery contains vitamins such as vitamin C⁽⁷⁾.

 

Because of the reproduction is one of the important characteristics of each organism, and which brought back the same object to the kind that belongs.The importance of reproduction in maintaining the continuity of the survival genus and prevent it from extinction through the transmission of life from one generation to another⁽⁸⁾, also used many medicinal plants to improve fertility and reproduction among these plants is a celery leaf, which is used to enhance sexual capacity and organization of the menstrual cycle of females and to reduce the abortion rate among pregnant women and increased secretion of milk⁽⁹⁾. Also because celery contains fatty acids which gave it a role in increasing the efficiency of the production of sperm in males and reduce the sensitivity of sperm to the environmental pollutants⁽¹⁰⁾. Some studies have pointed to the possibility of using of celery in determining the sex of the baby⁽¹¹⁾. Because celery contain similar compounds to sex hormones called Phytoestrogen Some studies have indicated that eating plants that contain these substances excessive quantities reduces fertility and content of sex hormones^(11-15) while other studies conducted on laboratory animals pointed to the role of the alcoholic extracts of the leaves of celery to increase ovarian activity and increasing the level of sex hormones of the female mice^(16-21) .

 

Because of the absence of a certain study about the impact of celery seeds in the histological characteristics of the ovaries such as the growth and development of ovarian follicles and the formation of corpus luteums, this study was designed to evaluate the effectiveness of the alcoholic extract of celery seeds on the histological characteristics of the ovaries of Swiss rats^(22-26) .

 

MATERIALS  AND METHODS:

Preparations of Laboratory Animals:

The present study was carried out in the laboratory animal house follow for biological department of Girls Education Faculty. Twenty seven Swiss rat Ratus ratis) were obtained from their mother that breeding in the previous mention animal house. The animal put in clean plastic cages in the dimension (20× 21× 45 centimeter  which provided by metal covers. The cages were bedded by wooden waste. The mothers were distributed in the cages in order to their reproduction and obtain on the adequate numbers. The cages were improved by cleaning and disinfectant three time weekly. The animals were under the same conditions from ventilation, light (14) hour and temperature degree (22- 28) C°. the animals supplied with water and ration the ration was obtained from Najaf poultry supplement.

 

RESULTS:

Preparation of alcoholic Celery seeds extract :

The alcoholic extract of Celery plant seeds was prepared according to⁽¹⁰⁾ So take (20) gram from dry seeds, the active ingredients were extracted via soxholate apparatus,after that, the extract materials were concentrated by rotary evaporator at temperature degree (45- 40) C° after obtained on the dry material, it weighted in order to prepare the stock, from was made up tow concentrations (150 and 300 milligram/ kilogram of body weight for the present study was conducted.

 

Animal Injection:

After the new borne females became at age three months, they divided in to equal three groups, were included control group and two treated groups, in number nine females for each group. The control group was injected normal saline (0.9 % NaCl . While the treated groups was injected at different doses (150 & 300 milligram/ kilogram B.W from alcoholic extract of Celery plant seeds at respectively, once/ daily for complete month period Intra peritoneal injection.

 

Animals scarified and specimens:

Collecting: after the injection period was complete, the animals were anaethesed by using the diethyl ether [(C₂H₂O₄)]. The abdominal cavities of animals were opened, their ovaries were removed for each animal after the adipose tissue and other tissues were attached to ovaries were pushed out, these organs were transformed into formalin (10 %) in order to fixation the tissues for period (48) hr. after that the routine histological preparations were carried out according to reference^(14,15) which involvement the following steps; washing by tap water, dehydration by series of ascending graduate ethyl alcohol (70 %, 80 %, 90 %, 100 %) and clearing by xylole and infiltration and embedding by paraffin wax and made up blocks, then mounting by Canada- balsam and cover slides. The histological slides were examined by light microscope (type Olympus, Japan(

 

 

Fig. (1): Shows histological section in control group through ovary cortex (white arrow) shows epithelial layer (black arrow) secondary follicle (yellow arrow) corpus luteum (red arrow)(H αE(40 X)

 

Fig. (2):Shows the effect of celery seeds extract (low concentration 150 mg/ kg) on rat ovary shows increasing in numbers and volumes of corpus luteums( red arrow).Also there were increasing in numbers of primary follicles (yellow arrow). The section also shows ovary cortex, ( white arrow) and medulla( green arrow) which seems clear from congestion (H α E 40 X)

 

 

Fig. (3):Shows the effect of celery seeds extract (high concentration 300 mg/ kg) on rat ovary. The section shows ovary cortex (black arrow) which is consist of less numbers and volumes of corpus luteums (red arrow )compared with other group treated with low concentration of extract. The section also shows medulla (white arrow) which is consisted of many congestion spots (blue arrow) also there were increasing in diameter of arteries and veins yellow arrow .(H α E 40 X)

 

 

Fig.(4):Shows the effect of (low concentration of celery seeds extract 150 mg/ kg) shows primary follicle yellow arrow α secondary follicle red arrow growing in a good α normal shape (l00 H α E 40 X)

 

Fig.(5):Shows the effect of celery seeds extract (low concentration 150 mg/ kg) shows good α normal secondary follicle red arrow surrounded with corona radiate (white arrow) and granulose cells (yellow arrow) also the section shows antrum (blue arrow) (H α E 400 X)

 

Fig.(6):Shows the effect of celery seeds extract (high concentration 300 mg/ kg). this section shows ovary cortex which consist numbers of atretic follicles at different stages of growth (red arrow) compared with other treatment group (150 mg/ kg) . (H α E 400 X)

 

 

Fig.(7):Shows the effect of celery seeds extract (low concentration 150 mg/ kg). This section shows ovary cortex which consist of more numbers of primordial follicles (red arrow) compared with other group treated by high concentration (300 mg/ kg) of the extract and control group. This section also shows the ovary cortex which is containing normal cells and nuclei (black arrow) (H α E 400 X)

 

 

Fig. (8,9):Shows the effect of(high concentration of celery seeds extract 300 mg/ kg) on ovary cortex. This section shows ovary cortex which is consist of less numbers of primordial follicles (red arrow) This section also shows the interstitial cells tissue of the ovary cortex the cells look in a hyper trophy appearance (blue arrow). The nuclei look pyknotic (yellow arrow) also there is a congestion in ovary cortex (black arrow) (H α E 400 X)

 

Fig.(10):Shows the effect of(high concentration of celery seeds extract 300 mg/ kg). This section shows a congestion in medulla ovary and there are increasing in numbers α diameters of arteries and veins (blue arrow) and increasing in blood supplies also the section shows pyknotic in nuclei of medulla cells (white arrow). (H α E 100 X)

DISCUSSION:

The use of medicinal plants has credibility and adoption throughout history, especially in the field of pharmacy and pharmaceutical production where they are drawn to the many effective ones vehicles and multiple ways where these compounds is the core in the manufacture of many drugs to get it licenses the price and lack of side effects and toxic decline in comparable chemical compounds by the pharmaceutical companies around the world manufacture⁽¹⁶⁾ celery leaf is one of the most important plants that studies have shown its importance as food contains many of the elements and nutrients important to the health and safety of the human body.
As for the effect of aqueous and alcoholic seed and leaves of celery and its relationship with fertility in females, the studies are rare and scientific research, completely non-existent, so will be discussed current research results based on other, especially those research that dealt with aqueous extract influence or alcohol extract on the reproductive performance of male rats through access to research studies provided celery plant is one of the more plants that give the water extracts and alcoholic identical results sometimes or dissimilar to each other at other times the results of the present study indicated that the use of alcoholic extract of the seeds of the plant celery by injection under the peritoneum low concentrations 150 mg / kg contributed significantly effective in increasing the level of activity of the ovaries and increase the number of primordial follicles and primary as well as increase the number of secondary follicles and the numbers of corpus luteum fig. (2,4,5,7) compared to the control group fig.(1).
But when using the high concentrations of the alcoholic extract 300 mg/kg histological sections showed remarkable changes in ovary cortex such as a decrease in numbers of developing follicles primary and secondary fig. (8,9) in addition to the decrease in the number and sizes of corpus luteum fig. (3) and increase the number of Atretic follicles in different stages of growth fig.. (6) in addition to emergence of changes in cortex of the ovary within the ovarian cortex fig.(8.9) such as inflation in cortex cells and thickening nuclei compared with low concentration treatment 150 mg / kg,also there were a significant changes in cortex area and medulla like vasodilation veins and arteries of the ovary medulla fig. (3,10).

 

These results were in line with that found       researcher^(17,18) where the researcher said that the histological changes that have emerged in testicle tissue of mice depends on the amount of the dose used during the injection . High doses lead to adverse changes in testicular tissue compared with lower doses which is led to an increase in activity of testicular tissue and increase the number of primary cells and increased numbers of mature sperm while the researchers⁽¹⁹⁾  found that the use of oral daily dosing concentration of 1000 and 2000 mg / kg of the alcoholic extract of the leaves of celery led to increased testicular activity in male Swiss mice and increase the number of cells and movement of sperms ⁽¹⁹⁾ found that the use of high concentrations 1000 mg/kg of alcoholic extract of the celery leaves orally dosing for 26 days led to the increase in the number and diameters of ovarian follicles developing and increasing of corpus luteums in the ovaries of female compared to the lower  concentrations 500 Mg/Kg and the control group ⁽²⁰⁾  had mentioned that the use of certain doses and quantities measured 200 mg / kg could give more positive results on the activity and movement of sperm in the males testes.

 

But in females, it works to increase the activity of the ovaries and increase the level of sex hormones due to the presence of chemical compounds most important flavonoids that have the properties of anti-oxidants and it working to remove free radicals ⁽²¹⁾. while ⁽²²⁾  indicated to the role of celery oil role in the removal of toxic substance cadmium accumulated in the tissues of the testicles of male Swiss mice while ^(20,22) pointed to the use of high concentrations of the alcoholic extract of the celery leaves leads to a reduction in the level of the concentration of sex hormones testosterone and the hormone-induced growth of intra-testicular tissue cells FSH and LH.

 

This is due to contain celery leaves on compounds called Phytoestrogen operate additional hormones affect the activity of the sexual organs as high doses are working on the pituitary gland and the sex glands axis and thus lead to a decline in the level of sex hormones that link to sex glands This is consistent with what has shown the current histological study of female ovaries, which showed dropping in the number s of primary and secondary follicles in addition to the decrease numbers of corpus luteum, which confirms the role of hormones in the histological features of ovary.

 

While the researchers⁽²³⁾. found That increasing doses of oral dosage treatment of female with high concentrations 1000 mg / kg compared to the low- concentrations 500 mg / kg lead to an increased level of sex hormones estrogen, progesterone, FSH and LH. The differences between the use of high concentrations of celery seed, among other studies may be due to the percentage of the active substances in the vegetative parts of the plant celery differ from the roots and seeds⁽¹⁶⁾  have said that the percentage of antioxidant and chemical compounds, called Phalids addition Phytoestrogens vary between the parts of the plant are up to the highest proportion in the seed compared to Arial parts and roots as well as celery seeds contain volatile oils and by large while lacking in Arial parts and roots. Therefore, the present results are consistent with that mentioned ^(25,26) of the differences in the method of preparation of the extract and the type of plant and the parts that were extracted effective, including materials and dosages affect significantly the desired results from the use of the extract as that of the digestive tract of an organism's role in acting bio-alcoholic extracts and water plants as the alcoholic extracts is the best because it is the extraction of materials and components more than water extracts.

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Received on 07.10.2017        Modified on 17.11.2017

Accepted on 06.12.2017      © RJPT All right reserved