Phytochemical constituents of bark essential oils of Cinnamomum zeylanicum Blume and effects on liver tissue of rats

— Cinnamomum zeylanicum Blume is an evergreen tropical tree belonging to the Lauraceae family; it is a useful plant in traditional and modern medicines. Essential oils of Cinnamomum zeylanicum were prepared and identified using gas chromatography–mass spectrometry (GC-MS). For histological study 80 adult male rats were employed in this study. It is the same weight (30 ±5 gm) and 75 days in age. The animals were divided into 4 groups, with ten rats in each group. The results indicated that C. zeylanicum bark contained 27 different phytochemical components. The main components included Cinnamaldehyde (2-Propenal, 3-phenyl-) (46.46%), 9-Methoxybicyclo, [6.1.0] nona-2,4,6-triene (31.31%), alpha-, Muurolene (7.14%) and Copaene (1.63%). Sections of livers were examined for histological abnormalities, such as the breakdown of the liver architecture and increased inflammation, in comparison to control samples, some hepatocytes were swollen with pyknotic, large nuclei and dilated sinusoids. As well as in the rats treated with noticed sever congestion of central vein (CV), degeneration and sluggish of endothelial lining larger, Bile ducts hyperplasia and necrotic hepatocytes. It can be deduced that the main component of cinnamon bark oil is cinnamaldehyde and the liver displayed some tissue changes, including hepatocyte necrosis and the existence of steatosis foci. Histological and cellular changes in liver tissue were dramatically attenuated by C. zeylanicum essential oil


I. INTRODUCTION
Cinnamomum zeylanicum (common Cinnamon) is an evergreen tropical tree 50 m high belonging to the family Lauraceae.The bark of trees of C. zeylanicum is a useful plant in traditional and modern medicines.More than 100 species of the genus Cinnamomum are found throughout Asia.Australia, Vietnam, Madagascar, and Mexico.It is widely, cultivated in southern India and Sri Lanka [1,2].
Numerous health benefits of cinnamon have been suggested, including a reduction in triglycerides, LDL cholesterol, total cholesterol, anti-yeast activity [3], antibacterial, antifungal activity, insects [4,5,6,7,8], blood circulation improvement, anti-platelet aggregation, antiinflammatory activity, Enhancing mental function, decreasing colon cancer risk, anti-Parkinson, anti-Alzheimer, anti-angiogenesis, anti-HIV-1 activity, antiinflammatory qualities, antibacterial action, heart disease prevention, and blood sugar regulation.Furthermore, cinnamon's possess an ability to reduce inflammation and its primary ingredients in a different variety of tissues, such as the pancreas, liver, kidney, heart, and brain [9].Essential oils extracted from plants have long been used to enhance the flavor of food and drink [10,11].Oil from the bark of the cinnamon plant is typically utilized in food and drink.It is highly valuable in industry [12,13], Because of their positive health effects.Traditional plants are increasingly being used as supplemental medicines, including their ability to prevent diabetic mellitus and cardiovascular disease, as well as their anti-inflammatory and antioxidant properties [14,15].It was discovered that in healthy adults, Cinnamon burmannii tea significantly decreased maximal glucose levels after meals and variance in maximum concentration [16].

A. Plant material
This study was conducted in the Basrah University, Science College, Biology Department, Iraq.The GC-MS Chromatography carried at the Basrah University, Agriculture College of, Iraq..

B. Preparation of samples
The bark samples of C. zeylanicum were collected from the market of Basrah governorate.One hundred gram of the bark sample was washed with water, dried and ground in a grinder for 20 seconds.The small pieces were homogenized for 3 min to 40-mesh size.The air-dried sample of cinnamon was pulverized to get the powdered form [18].

C. Essential oils Extraction
A Clevenger apparatus was used to extract essential oils using water distillation for four hours at 100 °C, it was collected in sealed container to prevent any evaporation.Furthermore, Anhydrous sodium sulfate was used to dry the recovered oil, which was then placed in a dark glass container for analysis [28].

D. GC-MS analysis
GC-MS analysis was carried out by utilizing a Shimadzu GC-QP 2010 Ultra gas chromatograph.The temperature of the GC oven was set to rise at a rate of 4.3 °C per minute from 40 °C to 250 °C.The carrier gas was helium.The linear velocity was 48.1 cm/sec and the input pressure was 100.0 kPa.Column flow was 1.78 mL/min, Injector temperature: 250 °C; injection mode: split.MS scan conditions: source temperature, 200 °C; interface temperature, 250 °C; Detector Gain, 0.70 kV +0.10 kV; Scan speed, 1666 Start 50 m/z, End 800 m/z.The components of the Cinnamon oil were identified by comparing the spectra with those of known compounds stored in the NIST library (2005) (Figure 1-5).

E. The animals
In total, 80 adult male rats were employed in this study.They were in good health, approximately having the same weight (30 ±5 gm) and 75 days in age having cages with three animals in each group.at a temperature of 18-25 ºC under a 12 h dark-light cycle.The animals were left for one week before starting the treatments.The animals were divided into 4 groups, with ten rats in each group.First, distilled water was given orally to the control group.Diethyl ether was used to anesthetize the animals following each treatment, fixed liver was taken for sectioning.Experimental design was approved by College of Science/ Basrah University.Twenty male adults healthy Two groups of rats were created.: Group 1 represented Control rats (n= 10) no treatment and Group 2: The animals were dosed orally with a concentration of 50mg/kg atenolol for thirty days (n=10).

F. Preparation of tissues
Tissue samples were fixed in equivalent formaldehyde solution (10%), then washed with tap water.The samples were then dehydrated by being passed through serious of ethanol from 30% up to 100%.Xylene was then applied for 30-40 minutes to clear the samples from ethanol.The tissues were subjected to paraffin infiltration using melted paraffin wax inside the oven at 60 ºC for one hour.and the tissue samples were transferred from the oven inside these molds which were then cooled to room temperature.The tissue sections were stained by using hematoxylin for one minute then in alcoholic eosin 1% [29].
Researchers have different estimates of how many chemical components are present in cinnamon essential oil.It was 4% of the essential oil, which is predominantly composed of 60-75% cinnamaldehyde [24].[4] reported that C. zeylanicum consists of 38 components, the majority components (68.41%) was Cinnamaldehyde, followed by benzaldehyde.While [33] discovered that cinnamaldehyde made up about 91.82% of the composition of cinnamon oil extract, with eight compounds having small percentage.Cinnamic aldehyde was discovered to be the primary component in the essential oil extract when GC-MS analysis of C. zeylanicum essential oil was performed [40].Cinnamaldehyde content in C. zeylanicum extracted was 90% [28].While [41] identified thirty compounds.Other constituents include cinnamyl acetate, cinnamyl alcohol, eugenol, linalool, cuminaldehyde, and pinene [42].
Climate, soil, harvest season, drying technique, storage conditions, and even the specific portion of the plant tissue are examples of environmental variables.The amount of essential oil in cinnamon species (Cinnamomum burmannii) differs between the tree age of plant [20].Different agricultural dates, origins, vegetable states, plant growth seasons, and market storage conditions can all affect the composition of essential oils [43,44].Although there were discrepancies between the present study and past studies in the number of essential oils, the analytical results were compatible with earlier publications [45].This may be brought on by various cultivars and extraction techniques.It could be because the microwave's heating effect caused the essential oil's cinnamic aldehyde to change into 2methoxycinnamaldehyde [30].

B. Histopathology of the liver
Examinations of liver sections under a light microscope for histopathology found that when compared to the control group, the rat given doses of the essential oil extract showed a number of alterations.In the section in rat liver from group treated with C. zeylanicum referred to dilated branch of portal duct, hepatocytes with cytoplasmic vacuolation, some hepatocytes were swollen with pyknotic, large nuclei and dilated sinusoids (Figure 6 A).As well as in the rats treated with noticed sever congestion of central vein (CV) (Figure 6 B), degeneration and sluggish of endothelial lining larger, Bile ducts hyperplasia and necrotic hepatocytes (Figure 6 C).A section of the rat liver from the C. zeylanicum-treated group revealed vacuolation in most hepatocytes, congested central vein, dilated sinusoid, complete degeneration of endothelial lining layer of central vein and the parenchyma tissue lost normal structure (Figure 6 D).
The histological sections taken from the livers of animals dosed orally from the volatile oil extract of cinnamon at a concentration of 20 mg/kg showed congestion, inflammations around central vein in treated rat after 30 days and showed congestion around portal triads vein (Figure 6); as well as found necrosis and degenerations liver cells in addition inflammation of cells after 30 days, In addition.One of the mechanisms of plant extracts is the process of inhibiting the synthesis of purine and pyrimidine [46,47].Under a microscope, liver sections were examined for histological alterations such as When compared to the control, there was greater inflammation and damage of the liver architecture [48].

IV. CONCLUSION
From the current investigation, it can be deduced that the main component of cinnamon bark oil is cinnamaldehyde.Consequently, cinnamaldehyde may be thought of as a component used to identify cinnamon bark oil.The liver displayed several tissue changes, including hepatocyte necrosis and the existence of steatosis foci.Histological and cellular changes in liver tissue were dramatically attenuated by C. zeylanicum essential oil.From this investigation, we may infer that essential oil of C. zeylanicum possesses hepatoprotective properties [39].

CONFLICT OF INTEREST
Authors declare that they have no conflict of interest.

Table ( 1
): Compounds Identified By Mass Spectrometry Gas Chromatography (Gc-Ms) In The Essential Oil Of C. Zeylanicum Bark.