Histoprotective Effect of Aqueous Extract of Cardamom against Cadmium Chloride Induced Toxic Effect in Mice

Muhaned Sachit Al-Delphi¹, Hayder Abdullah Kadhim Al-Sarray ², Israa Jabbar Shamkhi ³, Walaa Fadil Obead⁴

¹ Middle Technical University, Kut Technical Institute, Iraq.

² Middle Technical University, Kut Technical Institute, Iraq.

³ Middle Technical University, Kut Technical Institute, Iraq.

⁴ Kerbala University, Veterinary Medicine college, Department of Anatomy and Histology, Iraq.

Corresponding Author: Dr. Muhaned Sachit Al-Delphi, Middle Technical University, Kut Technical Institute, Iraq. · muhaned-sachit@mtu.edu.iq

Introduction

Elettaria cardamomum Maton is the scientific name of cardamom it is one of the commonly used in spices and is referred to as “the queen of spices” [1]. The family of Cardamom [Zingiberaceae] is one of the most old and costly spices in the world [2]. This plant is appreciated for its dried fruits [3]. The Elettaria genus is a tiny and diminutive natural group originating from the perpetual rainforests of South India and Sri Lanka, via whence they disseminated to other tropical zones [4]. This herb grows all year round, characterized by its foliage, it orginate from  Sri Lanka and  India and is predominantly cultivated in India’s south [5]. The valuable part is a non-perishable herbaceous perennial crop , These seeds are useful in spice mixtures in whole bean or powder, such as  beverages , curries similar as coffee and  team eat items, baked goods, confections, and biscuits flavors that are combined with custards, wines, and liqueurs. In the Middle East, cardamom and coffee are historically combined to make a beverage. Gahwa [6]. The small cardamom seeds are hold a triangular pod at the cross-section the thin papery black shell that covers [7].  The advantageous outcomes of cardamom upon the bacteria Gram positive and Gram negative [8,5], gums and teeth health [9], lung disorders, and finally gastrointestinal disorders [10,11]. The medicinal effects of cardamom stem from its pharmacological qualities, which include antioxidat, antimutagenic, antibacterial, antiinflammatory, antidiabetic, cardioprotective, hepatoprotecte, and chemoprotective attributes. [12-16].

Materials and Methods

Animal management: In this study 15 male mice were obtained from animal house at the pharmaceutical research and control center weighing between 30-45 g. Firstly the mice were given a week to acclimatize to their new surroundings. In this study the animals were divided in to 3 groups 5 male mice in each one. Group 1: administered with 2.5 mg/kg of CdCl2 with normal saline treated orally as control group, Group 2: administered 2.5 mg/kg of CdCl2 and cardamom [200 mg/kg], Group 3: administered the 2.5 mg/kg of CdCl2 with [300 mg/kg] from cardamom for six weeks.

Preparation of Aqueous Extract Solution of Cardamom: The aqueous plant extract prepared by putting dry powder of plant [100 g] and then placed on a conical flask with distilled water about [500 ml] in percentage 1:5 w: v then after make shacked by an electric shaker to two hours and left at ambient temperature. Then was filtered the mixture after 24 hours, by gauze 4 layer and then placed in a container and centrifugation at [2000 rpm/10 min]. Then filter supernatant by filter paper. Crude extract was obtained through concentrating the filtrate mixture for 72 hours in an oven. Before being used, this extract was stored in a dark environment at 4°C, sterile screw-capped bottle [17].

Samples Collected: The mice were sacrificed at the end of experimental period by decapitation under anesthesia and then make incision in abdominal wall therefore make cutting to the liver, heart, kidney and testis. The specimen of these organs were instantly fixed with formalin 10% for histological examination. 

This study was authorized by Scientific Ethical Committee at middle technical university [MEC 136].

Results

In this study show comparative between four organs kidney, testis, liver and heart to explain the protective effect of cardamom on histological change after oral administration by cadmium chloride.

Kidney

The kidney show improvement in histological structure after oral administration of cardamom group [200 and 400] mg/kg compared with group treated by cadmium chloride only. The kidney in cadmium chloride group show necrosis in proximal convoluted tubule [cortex] and its found vacuoles in proximal convoluted tubules also found haemorrheage and degeneration in glomeruli. In cardamom 200 mg group appear significant decrease in necrosis and vacuoles less than cadmium chloride group, degeneration and haemorrheage in medulla [distal convoluted tubule] less than cadmium chloride group. Group with 300 mg/kg cardamom if compare with cadmium chloride group and 200 mg/kg cardamom group find decrease in the necrosis and vacuoles[ less than other group] and medulla is normal [As shown in Figure 1].

Fig. 1: [A] kidney treated with cadmium chloride only [H &E stain X400], [B] kidney treated with cadmium chloride and 200 mg cardamom [H &E stain X40], [C]  kidney treated with cadmium chloride and 300 mg cardamom [H &E stain X400]

Testis

Testis in normal contain seminiferous tubules, leydig cell  and other histological and anatomical part of testicle in this study treated with cadmium chloride [CdCl2] to make histological damage in the testis then show loss of connective tissue inside the testis lead to the seminiferous tubule appear discontinuous, degeneration in spermatogonia, leydig cell is separated from seminiferous tubules if these results compare with treated group with cardamom [200 & 300 mg/kg] find in 200 mg/kg cardamom no significant differences between cardamom 200 mg/kg group and  CdCl2 group but in group with  300 mg cardamom show recover of connective tissue of testis and return of natural pattern of testis, leydig cell return in normal position between seminiferous tubule. [As shown in Figure 2].

Fig. 2: [A] Testis treated with cadmium chloride only [H &E stain X400], [B] Testis treated with cadmium chloride and 200 mg cardamom [H &E stain X400], [C] Testis treated with cadmium chloride and 300 mg cardamom [H &E stain X400]

Heart

Cardiac muscle tissue is characterized by its appearance is striated, branched cells, and it’s have intercalated discs that allow for coordinated contractions. In group treated with CdCl2 only show the nuclei are longitudinal in shape and cardiac muscle fiber is discontinuous and hemorrhage its present between them.  The compare between treated group with cardamom [200 and 300 mg/kg] with CdCl2 group find the nuclei is longitudinal and circular  and its found in different level in 200 mg/kg group, discontinues and hemorrhage in muscle fiber but less than  from CdCl2 group. In 300 mg/kg group show the significant decrease longitudinal nuclei and return to normal form, normal in cardiac muscle fiber if compare with 200mg/kg cardamom group and CdCl2 group. [As shown in Figure 3].

Fig. 3: [A] Heart treated with cadmium chloride only [H &E stain X400], [B] Heart treated with cadmium chloride and 200 mg cardamom [H &E stain X400], [C] Heart treated with cadmium chloride and 300 mg cardamom [H &E stain X400]

Liver

In group treated with CdCl2 group show degeneration of hepatocyte and find haemrrhage, enlargement in hepatic artery in portal area, haemorrhage in central vein. If compare with 200 mg/kg cardamom treated group show the haemorrheage in hepatic vein less than CdCl2 group. The haemorrhage in central vein is reduce, no arrangement in hepatocyte, reduce in the enlargement of the hepatic artery. Then after group treated with 300 mg/kg cardamom  the haemorrhage in central vein highly decrease compare with CdCl2 group and cardamom 200 mg/kg group, hepatocyte normal strand, hepatic artery is normal and not found  haemorrheage [As shown in Figure 4]

Fig. 4: [A] Liver treated with cadmium chloride only [H &E stain X100], [B]   Liver treated with cadmium chloride and 200 mg cardamom [H &E stain X100], [C] Liver treated with cadmium chloride and 300 mg cardamom [H &E stain X100].

Discussion 

In this study we use the cardamom as protective effect after make histological damage by cadmium chloride. According to properties of cardamom, the different investigations using animals hold presented its antihypertensive, antioxidant, antispasmodic, gastro-protective effect, antimicrobial, against aggregation of platelet as well as anti-cancer features [18]  also cardamom used for medicine to treat oral infection of the gums, teeth, and throat, Additionally, treat the lung congestion and  pulmonary tuberculosis, hypertension, cardiac disease, and digestive tract disorders [19]. The properties of cardamom extracts knew as anti-fungal and antimicrobial properties [20,21]. The anticancer activities of cardamom extract shows in many studies [22]. Other studies show effect of cardamom as anti-proliferative, anti-inflammatory, pro-apoptotic [23] its antioxidant capabilities have been identified as the reasons behind cardamom’s anticancer effects. Cardamom extract has found to be a potent macrophage modulator [24] and as a barrier against the dangers of uranium [25]. These study also agreed by Sameer N et al 2015 [26] that use the cardamom  as  protective substance to myocardial muscle damage proved by fall necrosis in cardiac muscle fiber, edema to minor signs of infection.

Conclusions

This study concluded that use the cardamom as protective effect against histological change by cadmium chloride and can use as antioxidant to prevent histological damage. And finally use the cardamom in 300 mg/kg concentration its more benefit than 200 mg/kg concentration.  

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