1. Introduction

he liver is responsible for vital functions in the body. Liver disease and dysfunction cause significant damage to the body, and even if the disease becomes chronic in the liver and liver problems are not treated, it can lead to death. Liver diseases often have symptoms similar to those of other diseases, which can make it more difficult and slower to diagnose them [1].

The liver is the largest gland in the body and plays an important role in the body’s metabolism and detoxification of various substances, so the health of this organ is important [2]. The liver, which plays an important role in the metabolism and storage of glucose as glycogen, plays a role in regulating blood glucose levels [7].

Albumin is the main plasma protein and its main function is to regulate the colloidal osmotic pressure [3]. Biliberin is one of the yellow bile pigments obtained from the breakdown of hemoglobin [4]. The use of medicinal plants has been common since ancient times. They have fewer side effects than chemical drugs, especially when common treatments are unable to control the disease. In recent years, attention to complementary therapies and therapeutic effects of natural compounds of plant origin has increased [5]. One of these plants is Rosa damascena from Rosaceae family [6]. Its main compounds are anthocyanins, cyanidin 3, 5-d-O-glucosides and several compounds such as camphor, quercetin, galactoside, arabinose, and ginnol. Its antioxidants can be helpful in preventing heart disease [8].

This plant is also used in the treatment of other diseases such as respiratory allergies and insomnia [9, 10, 11]. Another plant is Linum usitatissimum with bay leaves. Its seed is rich in various phenylpropanoids, especially lignans, flavonoids and cyanogenic glycosides. It also contains alpha-linolenic acid and omega 3 [12]. It is used to treat many diseases such as irritable bowel syndrome [13, 14]. Since sometimes, these medicinal plants are overused regardless of their side effects or desired dose, in order to determine their effective dose and prevent liver damage by not interfering with liver factors, this study aimed to investigate the effective doses of the hydroalcoholic extracts of Linum usitatissimum and Rosa damascena on liver enzymes, total protein, bilirubin, albumin, and serum glucose in rats.

2. Materials and Methods

This is a non-randomized clinical trial conducted on 42 male Wistar rats in animal Laboratory of Islamic Azad University of Kazeroon branch in 2019. Rats were divided into 6 groups; control group (group 1) received only sufficient water and food, groups 1 and 2 received 300 and 500 mg/ kgB.W Linum usitatissimum extract, groups 3 and 4 received 500 and 1000 mg/ kgB.W Rosa damascena, and group 6 received 100 mg/ kgB.W Linum usitatissimum plus 250 mg/ kgB.W Rosa damascena extracts intraperitoneally for 28 days. After the last injection, the rats were weighed and their blood samples were collected. The study parameters were measured using a colorimetric method by a spectrophotometer, and then were analyzed using ANOVA and Tukey’s test in SPSS V. 25 at a significance level of P<0.05.

3. Results

There was no significant difference between Alanine Aminotransferase (ALT), Alkaline Phosphatase (ALP), total and direct bilirubin levels in the control group in comparison with other groups (P>0.05). In the groups received Rosa damascena extract, there was a significant difference between total protein and albumin levels compared to the control group (P<0.05). Moreover, there was a significant difference between serum glucose and aspartate Aminotransferase (AST) in the control group compared to other groups (P<0.05).

4. Discussion

Consumption of the hydroalcoholic extract of Linum usitatissimum at a dose of 300 mg/kg increased the glucose levels and caused a significant statistical difference between the mean serum glucose levels of rats in this group and the control group (P<0.05). However, 500 mg/kg Linum usitatissimum reduced the glucose level and this difference was not significant (Table 1). Consumption of hydroalcoholic extract of Rosa damascena alone and in combination with Linum usitatissimum reduced the glucose level, but only the effect of 1000 mg/kg Rosa damascena extract was significant which caused a significant statistical difference between the mean serum glucose levels of rats in this group and other groups except with the group received 500 mg/kg Rosa damascena extract (Table 1).

Nikbakht and Gheitasi [15] reported that blood glucose levels were significantly reduced in consumers of hydroalcoholic extract of Citrullus colocynthis and Rosa damascena compared to controls. The effect of Rosa damascena extract on blood glucose level can be due to the presence of flavonoids and their glycosidic compounds. In one study, the effect of Rosa damascena methanolic extract in comparison with acarbose (an alpha-glucosidase inhibitor) in normal and diabetic rats was investigated and the strong inhibitory effect of alpha glycosidase was confirmed [16]. It seems that the effects of different amounts of glycosidic compounds and glycosides of different plants as well as the amount of glycosides and flavonoid compounds on the glucose level are different; hence, when using Linum usitatissimum extract, the dose of the extract should be determined based on the blood glucose level of the patient so that reducing or increasing glucose does not cause problems for the person. We can even use the proper dose of Linum usitatissimum extract to regulate serum glucose levels [17].

The effect of different doses of hydroalcoholic extracts of Linum usitatissimum and Rosa damascena on ALP level is similar to the effect of these extracts on the glucose level, but to a lesser extent such that 300 mg/kg Linum usitatissimum hydroalcoholic extract increased the average activity of this enzyme; however, the use of other doses of extracts reduced the activity of ALP and the highest reduction was observed after using 1000 mg/kg Rosa damascena hydroalcoholic extract, although these changes are not large enough to cause a statistically significant difference between the mean level of this enzyme in these groups compared to the control group, but a statistically significant difference was observed between the mean levels of ALP in 300 mg/kg Linum usitatissimum group, in the groups received different dose of Rosa damascena, and the group treated with both Linum usitatissimum and Rosa damascena.

This finding can be a warning to the consumers of Linum usitatissimum extract about the harmful effects of this extract on the liver, especially the bile ducts. In high doses of Linum usitatissimum extract and low and high doses of Rosa damascena extract and even the combination of the extracts of both plants, the antioxidant effect of flavonoids was predominant, such that the consumption of these extracts had no significant effect on the main indicators of a healthy liver (ALP, ALT, total and direct bilirubin) [18, 19].

The use of 1000 mg/kg Rosa damascena hydroalcoholic extract significantly increased the AST level. Given that ALT level, which is more specific for the diagnosis of liver disease than AST [20], has not changed and ALP level even decreased in this group, this increase in AST is most likely not due to a liver problems. If ensuring the absence of hemolysis in the tested samples, this increase can be due to muscle injury. In this case, the creatine phosphokinase enzyme level measurement can be very decisive.

The albumin and total protein levels in groups that consumed low and high doses of Rosa damascena extract and the combination of Rosa damascena and Linum usitatissimum extracts showed a significant decrease. Due to the severe diarrhea observed in these groups that consumed Rosa damascena, the decrease in albumin level can be due to its excretion through the gastrointestinal tract. There was a significant statistical difference between the mean levels of albumin and total protein in the groups received Rosa damascena and Linum usitatissimum hydroalcoholic extracts which was then due to the lack of excretion of total protein and albumin from their body.

The changes in albumin and total protein levels in these groups are not related to the process of protein synthesis in the liver. AKbari et al [21] in a study reported that Rosa damascena has the least nephrotoxic and hepatotoxic effects; however, its very high doses (1440 mg/kg body weight) can lead to hepatotoxic effects. Therefore, the hepatic effects of Rosa damascena have been considered to be dose-dependent [21].

5. Conclusion

Linum usitatissimum and Rosa damascena have no negative effect on the liver function. The probability of diarrhea occurrence and the possible effects on the total protein and serum albumin after using Rosa damascena, and the effects of different doses of Linum usitatissimum on the glucose levels should be taken into account.

Ethical Considerations

Compliance with ethical guidelines

All experiments in this study were according to guidelines of Animal Research Ethics Committee of Islamic Azad University of Kazeroon branch (Code: IR.IAU.KAU.REC.1398.048 ).

Funding

The present paper was extracted from the PhD dissertation of the first author, Faculty of Veterinary Medicine, Kazerun Branch, Islamic Azad University, Kazerun.

Authors' contributions

All authors contributed in preparing this article.

Conflicts of interest

The authors declared no conflict of interest.


 

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