Protective and curative activity of celery seeds on oxidative liver damage induced by cisplatin in male albino rats

Protective and curative activity of celery seeds on oxidative liver damage induced by cisplatin in male albino rats

 

Amira M. El-Moslemany

 

Nutrition and Food Science Department, Faculty of Home Economics, Al-Azhar University, Egypt

 

Abstract

 

This study was carried out to investigate the effect of celery seeds powder on oxidative damage of liver induced by cisplatin in male albino rats. The biological experimental was designed using forty two rats divided into seven main groups including both curative and protective groups (six rats of each) .Curative groups: Group (1) Negative control group was fed on basal diet. Group (2) Positive control group was fed on basal diet and injected with Cisplatin (5mg/kg body wt.) from the first day to induce hepatotoxicity. Group (3) and (4) were injected with Cisplatin and fed on diet containing 5% and 10% celery seeds respectively. Protective groups: Group (5) Positive control group was fed on basal diet and injected with Cisplatin (5mg/kg body wt.) after 28 days. Group (6) and (7) were fed on diet containing 5% and 10% celery seeds powder and injected with Cisplatin (5mg/kg body wt.) after 28 days. At the end of the experimental period rats were fasted overnight and sacrificed; blood samples were collected from the aorta to determine biochemical parameters. Besides, nutritional and biological parameters were recorded.  The results showed that the chemical composition of celery seeds indicates the presence of high amounts of protein, lipids, crude fiber, ash and total carbohydrates, and also low moisture content, respectively. At the end of the biological experimental the results showed that celery seeds powder at 5% and 10%  intake in curative and protective groups improved feed intake, the body weight gain , feed efficiency ratio, liver function, serum lipid levels and antioxidant enzymes compared with positive control groups (injected with cisplatin).It can be concluded that celery seed was fortified in the basal diet at 5 and 10%, improved the activities of liver functions, lipid profile and also improvement of antioxidant enzymes against cisplatin.

 

Introduction

 

Use of medicinal plants to treat common ailments has been prevalent since ancient times and different parts of the plants were used for public health. The use of natural treatments is cost-effective (Tang and Halliwell, 2010).  Since ancient times, plants have been important in reducing pain and today the focus is on their role and ability in healing and their treatment properties for various diseases (Ghasemiboron et al., 2014 and Noori et al., 2016).  

 

Many studies have shown the positive effects of various herbs and medicinal plants on infertility ( Kooti et al., 2014), hormone disorders ( Kooti et al., 2014b), liver disorders (Lone  et al., 2015), anemia (Mansouri et al., 2015),  renal diseases (Wu et al., 2014) and neurologic and mental disorders(Saki et al., 2014).

 

Celery (ApiumGraveolense) is a medical herb used as a food and also in traditional medicine. Celery contains aromatic substances in the stem, leaves and roots. The healing properties of celery are due to its essential oil and flavonoids (Li et al., 2014).

Celery is used as an effective remedy for various ailments such as lower blood pressure, bronchitis, liver and spleen disease, arthritic pain and this natural holistic approach to health is becoming more and more popular now a days. Celery stimulates healthy and normal functioning of kidney by helping in the elimination of the body toxins. It also prevents kidney stones (Kolarovic et al., 2010).

 

Celery seeds contain several substances including volatile oils; flavonoids, antioxidants that give plants their colors and may protect cells from damage; coumarins, chemicals that help thin the blood; and linoleic acid, an omega-6 fatty acid. Celery seed is used mostly as a diuretic, meaning it helps your body get rid of too much water by increasing urine output. Celery seed is also sometimes used for treating arthritis and gout, and to help reduce muscle spasms, calm the nerves, and reduce inflammation (Cheung et al., 2008).

 

Celery (Apium graveolens; family, Apiaceae) is used in Indian system of medicine owing to its richness in flavonoids and antioxidant property for the treatment for liver ailments. Studies have indicated that celery lowers blood pressure, regulates heart function and reduces complications of diabetes (Shivashri et al., 2013). The healing property of celery is due to its bioactive compounds like rutein, quercetin, luteolin, kaempherol, apigenin and myricetin (Mimica-Dukic and Popovic, 2007).

 

Although cisplatin (CIS) is a highly effective anticancer drug, hepatotoxicity is one of the most common adverse effects associated with its use. Recently, reactive oxygen species (ROS) and inflammation are suggested to be key factors in the pathophysiology of CIS-induced acute liver damage (El-Shitany and Eid, 2017).

 

The aim of this investigation was carried out to evaluate the chemical composition of celery seeds; moreover the effects of celery seeds on oxidative liver damage induced by cisplatin.

 

Material and Methods

 

Materials:

Celery plants (Apium graveolens L.) seeds were purchased from Field Crops Research Institute, Agricultural Research Center, Giza- Egypt.

Soybean oil and starch were purchased from the local market.   Casein, cellulose, vitamins, minerals, dextrin, L-cysteine and choline chloride were obtained from the Cairo Company for Chemical Trading, Cairo, Egypt.

Cisplatin was purchased from Al Tarshouby, Pharmacy, Tanta- Egypt.

 

Male albino rats (Sprague Dawley strain) (42 rats) were obtained from the Laboratory Animal Colony, Helwan, Cairo – Egypt, and weighting were approximately between 150-180g.

         Kits were purchased from Egyptian American Company for Laboratory Service and Supplied by Alkan Company.

 

Methods:

Preparation of celery seeds:

Celery seeds were washed dried and crushed using electric blender to a fine powder which was packed in air-tight plastic bags till use for basal diet supplementation.

Chemical composition of celery:

Approximate chemical composition of celery moisture, ash, crude protein, and fat, were determined according to the methods of the AOAC (2010).While total carbohydrates were estimated by subtracting the difference from initial weight of the samples as follows: -

Carbohydrates% = 100 - (%moisture + % protein + % fat +%ash).

 

Experimental design:

Rats were adapted for one week prior to commencement of the experiment. Water was introduced adlibitum. Rats were divided into seven groups including both curative and protective groups (six rats of each). The first group (6 rats) was considerable as negative control and it was fed on basal diet according to Reeves et al., (1993). The remainders of rats (36) were divided into two main groups (18 rats for each main group).

 

The first main group (18 rats) was injected with Cisplatin (5mg/kg body wt.) for one day only in the first experimental considerable as curative groups according to yogesh et al., (2010)and it wasdivided into three subgroups. The first subgroup was considerable as control positive and it was fed on basal diet while the second and third curative groups were fed on basal diet supplemented with 5 and 10 % from celery seeds.

 

The second main group (18 rats) was injected with Cisplatin (5mg/kg body wt.) at the end of experimental period after 28 day considerable as protective groups and it wasdivided into three subgroups. The first subgroup was considerable as control positive and it was fed on basal diet and the second and third protective subgroups were fed on basal diet supplemented with 5 and 10 % from celery seeds.

 

 

Feed intake, body weight gain and feed efficiency ratio were determined according to Chapman et al. (1959). At the end of experimental period, all rats were fasted overnight sacrificed and the blood samples were collected, and centrifuged to obtain the serum. After then liver from each rats was removed, cleaned in saline solution, dried by filter paper and weighted. Liver was rapidly taken on ice bags and frozen at – 18°C till used for assessment of lipid peroxidation and antioxidant activity in liver tissue.

 

Biochemical analysis of serum:

Serum samples were used for determination of total cholesterol according to (Allain et al., 1974), triglycerides(Fossati and Prencipe 1982). Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) Bergmeyer et al. (1986). Albumin (Drupt,  1974) total protein ( Sonnenwirth and Jaret, 1980).

 

Assessment of Oxidant/Antioxidant Activity:

After then liver was removed, homogenized and centrifuged at 10,000 rpm at 0°C for 20 min. the supernatant was used for estimation of different antioxidant level by calorimetric method using spectrophotometer (Merck thermo spectronic, Model NO. UV-1, double beam), Lipid peroxidation by thiobarbuturic acid-reactive substances (TBARS) methods (Uchiyama and  Mihara1978), Superoxide dismutase (SOD) by method developed by Misra and Fridovich, (1972) and Catalase (CAT) by colorimetric assay  (Sinha,1972).

 

Statistical analysis:

The data obtained in the present study was analyzed by ANOVA. For all analyses, when a significant difference (p ≤ 0.05) was detected in some variable, the data means test was applied to evaluate the difference between the samples. The results were analyzed with the aid of the software SAS System for Windows SAS (2008).

 

Results and Discussion

 

Chemical composition of celery seeds powder.

The results in Table (1) showed the chemical composition of celery seeds powder. The mean value of protein, crude fiber, fat content, ash and total carbohydrates were 16.93, 12.11, 10.61, 18.19 and 42.16%, on dry weight, respectively.

 

The present results are in agreement with those Agarwal and Sharma (2013) and confirmed by Doke and Guha (2014) who found that the chemical composition of celery seeds indicates the presence of high amounts of protein (12.47%-15.00%) and lipids (14.00%-28.03%) which indicated that seeds have high food energy, with low moisture content (3.92%-4.14%) which an index of stability quality and increased shelf life of seeds, also, crude fiber (6.75%-16.50%), ash (14.25%- 24.65%), and total carbohydrates (32.87%-54.00 %) respectively.khare, (2007) reported that the concentration of protein and carbohydrate are good in celery seeds.

 

Effect of feeding celery seeds on feed intake, body weight gain and feed efficiency ratio of rats suffering from liver damage

Body weight gain, feed intake and feed efficiency ratio were determined in rats fed on celery seeds against cisplatin‐induced oxidative liver damage and the results are tabulated in Table (2).

 

Data presented could be observed that there were significant decrease in BWG; FI and FER for control positive group                    (-33.75±0.01, 7.43±0.20and-0.260 ±0.001 respectively) after the single dose of Cisplatin (5mg/kg), as compared to the negative control group (26.98±0.01, 10.52±0.15 and0.115±0.15 respectively).  From the results it could be observed that the curative groups which fed on basal diet containing celery seeds powder at 5 and 10% with cisplatin injection at the first one day of experimental period improved the body weight gain%, feed intake and feed efficiency ratio compared with control positive group. Also, results showed that protective groups showed that groups fed on celery seeds powder at 5% and 10% with cisplatin injection at the end of experimental period caused significant increases and nearly to control negative in the levels of body weight gain, food intake and feed efficiency ratio. These results are agreement with Thebaudin et al. (1997) who reported that some dietary fiber ingredients could be desirable for their nutritional properties but also for their functional and technological properties.

 

The fiber in celery is the insoluble type, which means it does not dissolve in water and passes through intestines undigested. Insoluble dietary fiber helps keep bowels moving regularly and has been used is fat replacer, fat reducing agent during frying, volume enhancer, binder, bulking agent and stabilizer. Celery provides an excellent source of vitamin B1, B2, B6& C and fiber. It's a very good source of folic acid, potassium, and calcium. Also this may be due to presence of polyphenols  which present in celery seeds that improve and protect liver cells against damage via increasing both the levels and activities of antioxidant enzymes in liver and kidneys according to(Jachec et al., 2002)leading to improvement the appetite and increasing feed intake.

 

 

Effect of feeding celery seeds on liver enzymes of rats suffering from liver damage

Results in Table (3) revealed that significantly higher (P< 0.05) value of AST, ALT and ALP with positive group in curative groups (90±4.5, 55.30±4.5, and 383±22.5U/L, respectively) and in protective groups (70.50±10.5, 45.30±3.5 and 322±15.5 U/L, respectively) compared with those of negative control group (48.50±2.5, 22.80±2.5 and146±2.5 U/L, respectively). The increased serum level after Cisplatin administration is due to the reaction between nitric oxide and superoxide anion which forms a radical, peroxynitrite. The process was further continued to oxidize the free radicals, it results to damage of cellular structures, moreover and it causes toxic oxidative damage of lipid components, cell components including proteins, lipids and nucleic acids. Our results appeared to be consistent with many previous findings that indicated a hepatotoxic effect of CIS and its association with increased free radical formation and the subsequent oxidative and nitrosative stress (Rehman, et al 2014). In fact, administration of Curative & protective rats fed on basal diet containing celery seeds powder at 5% & 10% caused a significant reduction in the mean values of serum AST , ALT and ALP as compared to the positive control group, it appears from our results that high concentration of celery seeds was safe and improves liver functions.

 

The hepatoprotective activity of celery seeds may be due to its antioxidant effect as evidenced by increasing activity of antioxidant enzymes and significantly changed the physiological parameter showing the reduction in increased serum level (Chandel et al., 2018).

The isolated flavonoid, apigenin in celery seeds act as inhibitory agent against tumor formation in the liver cells(Sultana et al., 2005). Moreover, Abd El-Ghany et al. (2012) reported that celery contains a large amount of vitamin C which is antioxidant to prevent the free radical damage that triggers the inflammatory cascade. Thus, it helps reduce the severity of inflammatory conditions

 

Effect of feeding celery seeds on total protein, albumin and globulin of rats suffering from liver damage

The results of the levels of total protein, albumin, and globulin measurements of rats are reported in Table (4).

 

The results showed that the mean values of these parameters decreased in the total protein, albumin and globulin of positive control group (4.07±0.03, 3.03±0.15 and 1.03±0.20 g/dl, respectively) in the curative groups and in the protective groups (4.60±0.04, 3.30±0.17and 1.30±0.25 g/dl, respectively), compared with the negative control group (7.90±0.05, 4.90±0.75 and 3.03±0.03g/dl, respectively).

 

Also rats fed on diet fortified with celery seeds powder at 5, 10% were nearly to the negative healthy rats group. These finding can explain the possible effect of celeryconsumption at 5% and 10% on improving liver function for rats suffering from hepatotoxicity. The lowest level in the total protein and albumin in control positive injection in the first and end experimental may be caused by the disorder in the liver.

 

This observation corroborates earlier findings on cisplatin-induced alterations to hepatic function markers (Mansour et al., 2006). It has been suggested that the ability of cisplatin to cause alterations in the activity of these enzymes could be a secondary event following cisplain -induced liver damage with the consequent leakage from hepatocytes. Conversely, the different fed from celery protects the hepatocytes against cisplain injury as evidenced by improved levels of biochemical markers of acute hepatocellular injury. Abdou et al., (2009)found that the treatment of celery stimulated theprotein biosynthesis to produce antioxidativeenzymes which treat the oxidative stress of CCl4.

 

Effect of feeding celery seeds on triglyceride and total cholesterol of rats suffering from liver damage

Triglyceride and total cholesterol ware determined in rats fed on celery seeds against cisplatin‐induced oxidative liver damage and the results are tabulated in Table (5).

 

Results indicated that there were significant changes in the serum levels of triglyceride (TG) and total Cholesterol (TC) of negative control group when compared with (positive control groups), the values were120.08 ± 2.51 and 165.61 ± 4.35 in negative control group and 195.71 ± 3.16 and 310.28 ± 10.01mg/dl in curative group and 170.56 ± 0.19 and 230.56 ± 5.61mg/dl in protective group. Meanwhile, the different rats group fed on celery in the basal diet were significant improve the triglyceride and total cholesterol.

 

The results of the present work about the effectiveness of celery seeds  agreed with Kim and Shin (1998),who demonstrated that the lipid-lowering action of this natural product may be mediated through inhibition of hepatic cholesterol biosynthesis, increased faucal bile acids excretion, and enhanced plasma lecithin: cholesterol acyltransferase activity, and reduction of lipid absorption in the intestine (Mansi et al., 2009) and celery has also been shown to have some other medicinal features including hyperlipidemic effects as well as antioxidative and hepatoprotective activities. In a study, kooti et al, (2014) evaluated the effects of celery on serum lipids of mice fed a high-fat meals showed that  celery causes a significant decrease in LDL and cholesterol and a significant increase in HDL. In a study, Tsi and Tan, (2000) examined the attributes of anti-hyperlipidemia of the celery in the rat. The results showed significant reduction in the concentration of serum total cholesterol, triglyceride levels and hepatic lipase triacyl glycerol in the treatment group.

 

Effect of feeding celery seeds on antioxidant SOD, CAT and lipid peroxide MDA of rats suffering from liver damage

Injected rats with cisplatin in the first day and the end of the experiment and treated with celery seeds with two levels are shown in Table (6). MDA is a main product of lipid peroxidation, as a biomarker of oxygen free radicals;  the liver MDA of rats in the positive control group showed marked increase (0.502± 0.01 nMol/mg ) in curative group and also in protective groups (0.401± 0.02 nMol/mg), compared with rats in the negative control group (0.155±0.0nMol/mg). This may be referred into a direct excessive effect of Cisplatin on liver function. However, rats in other groups experienced significantly (p < 0.05) less of rise in the liver MDA level, as compared to positive control group.

 

Treated rats “curative groups” and "protective groups”   caused a significant reduction in the mean values of MDA as compared to the positive control group. The level ofSOD and CAT was significantly decreased (p < 0.05) in the liver of rats in control positive group (0.089±0.0 1U/L and 0.102±0.0 1ng/mg) compared with those of negative control group (0.251±0.0 2U/L and0.243±0.0 2ng/mg,  respectively) in curative groups. Moreover, fed on basal diet containing celery seeds powder at 5% & 10% in “curative & protective groups” caused a significant increase in the mean values of

 

Serum SOD and CAT, as compared to the positive control group. Cisplatin administration triggered lipid peroxide formation and depleted the hepatic GSH and GPx. Oxidative stress was reported to be involved in cisplatin-induced acute hepatic injury (Bentli  et al .,2013).On the other hand  Louvet and Mathurin (2015) mentioned that the depletion of GSH and GPx, which play major roles in the cellular defense against oxidative stress and cellular damage, renders hepatic tissues more susceptible to oxidative stress. Celery has antioxidant effect (Popovic et al., 2006), it contains a large amount of vitamin C which is antioxidant to prevent the free radical damage that triggers the inflammatory cascade. Thus, it helps reduce the severity of inflammatory conditions (Sultana et al., 2005).

 

The strong antioxidant of celery seed may be attributed to the phenolic compounds and the presence of alkaloids and flavonoids in high concentration in extract of celery seed(Al-Howiriny et al., 2010).

 

In addition to Jain et al.,(2009)found that  co-administration of methanolic extract of A. graveolens seeds along with Di-(2- ethylhexyl) phthalate (DEHP) induced hepatotoxicity in rats,signifycantly prevented the rise in TBARS level with a concomitant elevation in the concentration of hepatic glutathione and ascorbic acid suggesting alleviation of oxidative stress and restoration of antioxidant defense system resulting in membrane stabilization.

 

 

 

 

 

Conclusion

 

According to the results obtained in the present study, it appears that dietary intake of celery seeds at 5 and 10% for four weeks would decrease the toxicity associated with oxidative stress and thereby reducing the damage induced by exposure tocisplatin.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table (1): Chemical composition of celery seeds powder on dry weight basis

*Values are means + standard deviation of three determinations (n=3)

 

Table (2): Effect of feeding celery seeds on feed intake, body weight gain and feed efficiency ratio of rats suffering from liver damage

Values denote arithmetic means ± SD of the mean. Means with different letters (in the same column are significantly at (p ≤ 0.05) using one way ANOVA test, while those with similar letters are non-significant.

 

Table (3): Effect of feeding celery seeds on liver enzymes of rats suffering from liver damage

Values denote arithmetic means ± SD of the mean. Means with different letters (in the same column are significantly at (p ≤ 0.05) using one way ANOVA test, while those with similar letters are non-significant.

 

Table (4): Effect of feeding celery seeds on total protein, albumin and globulin of rats suffering from liver damage

Values denote arithmetic means ± SD of the mean. Means with different letters (in the same column are significantly at (p ≤ 0.05) using one way ANOVA test, while those with similar letters are non-significant.

Table (5): Effect of feeding celery seeds on triglyceride and total cholesterol of rats suffering from liver damage

Values denote arithmetic means ± SD of the mean. Means with different letters (in the same column are significantly at (p ≤ 0.05) using one way ANOVA test, while those with similar letters are non-significant.

 

Table (6): Effect of feeding celery seeds on antioxidant SOD, CAT and lipid peroxide MDA of rats suffering from liver damage