Abstract
Frankincense (Gum Olibanum), made from resins of Burseraceae family, grows in Somalia, India and Yemen. Many years ago the oldest doctors used this plant for treatment of many diseases. This study identifies frankincense impact by biochemical analysis and histological examination on rats. In this study, forty male Wister Albino rats weighing 70–100 g were maintained in clean cages. The rats were divided into 2 groups, each group contained 20 rats. Frankincense extract was prepared by heating distilled water (400 ml) to 80 °C and soaking 20 g of herbs for about 60 min. After cooking at room temperature the dose was given orally through special drinking bottles daily. The first group acted as control drinking water. The second group served as treated group and was given frankincense in the drinking water during the whole duration of the experiment. After 15 and 30 days of treatment, the rats were anesthetized with ether, and blood was collected from the livers and kidneys; some biochemical analyses were performed including aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and non-bilirubin, urea, uric acid, and creatinine. Rats were killed by cervical decapitation of livers and kidneys. Each group was divided into 2 parts. The first part was used for the determination of glutathione (GSH), glucose 6 phosphate dehydrogenase (G6PDH), xanthine oxidase (XO), malonyldealdehyde (MDA), nitric oxide (NO), and xanthine oxidase (XO). The second part of livers and kidneys was kept in formalin solution (10%) and stained by Hematoxylin and Eosin (H & E), to be used for histological examination. I demonstrated in the biochemical analysis in the serum, tissue and histological examination, different impact between group (B) and group (A), and that frankincense is not absolutely safe and that precautions must be taken during it’s us as a traditional medicine and that increase the awareness with safety and health hazards of many other traditional medicine is critically needed.
Keywords: Frankincense, Boswella Serrata, AST, GSH, MDA, Histology
1. Introduction
Frankincense (Gum Olibanum), extract of Boswella Serrata, is a tropical tree that grows as small trees or shrubs; they are of the botanical family Burseraceae. Their natural growing range in limited, but this has been extended by cultivation and current supplies are adequate to meet the worldwide demand. Today, most of the internationally traded Frankincense is produced in Southern Arabian Peninsula (Oman, Yemen) and northeast Africa (Somalia). Frankincense has had spiritual significance since ancient times and it was also adopted as medicine for physical ailments. When referring to this herb, the western people might immediately think of its historic importance in religion (Hostanska et al., 2002; Pedretti et al., 2009).
The resins obtained from different plants have long been known in the traditional medicine of different countries. The main component of frankincense is an essential oil, being alpha-pinene, boswellic acid and many other compounds (Shi et al., 2002). The medicine is used for anti-inflammatory and anti-proliferative activities toward a variety of malignant cells and as a therapeutic agent in the treatment of non-insulin dependent diabetes mellitus (Park et al., 2002; Kim et al., 2008). The effect of boswellic acid enhanced neurite outgrowth, branching, and tubulin polymerization dynamics (Karima et al., 2010). Frankincense oil might represent an alternative intravesical agent for bladder cancer treatment (Frank et al., 2009).
The aim of this study is to find the impact of frankincense by biochemical analysis and histological examination on rats.
2. Materials and methods
2.1. Materials
2.1.1. Animals
In this study, forty male Wister Albino rats weighing 70–100 g were maintained in clean cages. The rats were fed with commercial pelleted diet obtained from King Fahad Medical Research Center in Jeddah.
2.1.2. Chemicals
Aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP), bilirubin, urea, uric acid, and creatinine - source from Dimension (DAD BEHRING Company, USA). Glutathione (GSH), glucose 6 phosphate dehydrogenase (G6PDH), malonyldealdehyde (MDA), nitric oxide (NO), and xanthine oxidase (XO). Formalin solution and Hematoxylin and Eosin (H & E) - source from (Sigma–Aldrich). Frankincense - source from (Abazer-KSA).
2.2. Methods
The rats were divided into 2 groups, each group contained 20 rats. Frankincense extract was prepared by heating distilled water (400 ml) to 80 °C and soaking 20 g of herbs for about 60 min; after cooking to room temperature the dose was given orally through special drinking bottles daily. The first group acted as control drinking water. The second group served as treated group and was given frankincense in the drinking water during the whole duration of the experiment. After 15 and 30 days of treatment, the rats were anesthetized with ether, and blood was collected from the livers and kidneys; some biochemical analyses were performed such as aspartate aminotransferase (AST) (Saris, 1978), alanine aminotransferase (ALT) (Bergmeyer et al., 1978) and alkaline phosphatase (ALP) (Williams, 1984), bilirubin (Jendrassik and Grof, 1938), urea (Talke and Schubert, 1965), uric acid (Bulgar and Johns, 1941), and creatinine (Larsen, 1972), which were measured by Dimension (DAD BEHRING Company, USA).
Rats were killed by cervical decapitation and the livers and kidneys from each group were removed and divided into 2 parts; the first part was used for the determination of glutathione (GSH) (Bentler et al., 1963), glucose 6 phosphate dehydrogenase (G6PDH) (Bergmeyer et al., 1974), xanthine oxidase (XO) (Fried and Fried, 1974), malonyldealdehyde (MDA) (Buege and Aust, 1978), nitric oxide (NO) (Moshage et al., 1995), and xanthine oxidase (XO) (Fried and Fried, 1974). The second part of livers and kidneys was put in formalin solution (10%) and stained by hematoxylin and eosin (H & E) (Bancroft and Stevens, 1996), to be used for histological examination.
2.3. Statistical analysis
Collected data were calculated by ANOVA using SPSS program version 16.
3. Results and discussion
Results of this experiment revealed that the comparison between frankincense group (B) at day 15 and 30 to control group (B) at the same days.
Illustrated from Figs. 1–6, impact frankincense of liver function in biochemical analysis from serum and tissue rats. In serum, no significant reduction in AST activity was observed at 15 days, p = 0.09, but the reduction in AST activity was significant at 30 days p = 0.039. In ALT activity, a significant decrease was observed at 15 and 30 days; p = 0.039 and 0.012, respectively. An increase, but not significant,in the activity of ALP was observed at 15 and 30 days compared with group (A); p = 0.266 and 0.585, respectively. In tissue, a very highly significant decrease in GSH and G6PDH was observed, p = 0.000. Very highly significant increases in the level of XO, p = 0.000, compared with group (A) were observed.
Figure 1.
Impact of frankincense on biochemical analysis of liver function in serum of rats.
Figure 2.
Impact of frankincense on biochemical analysis of liver function in serum of rats.
Figure 3.
Impact of frankincense on biochemical analysis of liver function in serum of rats.
Figure 4.
Impact of frankincense on biochemical analysis in liver tissue of rats.
Figure 5.
Impact of frankincense on biochemical analysis in liver tissue of rats.
Figure 6.
Impact of frankincense on biochemical analysis in liver tissue of rats.
In Figs. 13–20, the histological examination of the liver tissue of control group (A) showing hepatic cells around the central vein, nucleus and blood sinusoid is illustrated and also many hepatocytes were binucleated accompanied by granular degeneration and fatty changes activation of Kupffer cells. Frankincense group (B) showed increase in the number of kupffer cells, necrosis of the nucleus, increase in the number and size of nucleus; also, binucleated hepatic cells and necrosis of the nucleus were found.
Figure 13.
Light micrograph of liver tissue of control group (A) showing hepatic cells around the central vein, nucleus and blood sinusoid. Haematoxylin-eosin stain. 400×.
Figure 14.
Light micrograph of liver tissue of control group (A) showing many hepatocytes were binucleated accompanied by granular degeneration and fatty changes activation of Kupffer cells. Haematoxylin-eosin stain. 400×.
Figure 15.
Light micrograph of liver tissue frankincense group (B) showing increase in the number of kupffer cells and necrosis of the nucleus. Haematoxylin-eosin stain. 400×.
Figure 16.
Light micrograph of liver tissue frankincense group (B) showing increase in the number and size of nucleus, also found binucleated hepatic cells and necrosis of the nucleus. Haematoxylin-eosin stain. 400×.
Figure 17.
Light micrograph of kidney tissue of control group (A) showing widening of tubular lumen, extruded nuclei, shortening, and damage of brush border in proximal tubules. Haematoxylin-eosin stain. 400×.
Figure 18.
Light micrograph of kidney tissue of control group (A) degenerative; necrotic changes in kidney tubules and glomerulus. Nuclear pleomorphism in kidney tubules. Haematoxylin-eosin stain. 400×.
Figure 19.
Light micrograph of kidney tissue of frankincense group (B) showing nearly normal distal convoluted tubule with diluted lumen low and nuclei in some proximal epithelial cells. Haematoxylin-eosin stain. 400×.
Figure 20.
Light micrograph of kidney tissue of frankincense group (B) showing extruded nuclei, shortening, damage of brush border in proximal tubules, and hyalinization of glomerulus. Haematoxylin-eosin stain. 400×.
The results in Figs. 7–12, demonstrate the impact of frankincense on kidney function assessed by biochemical analysis of serum and tissue of rats. The same level of urea was observed at 15 days, p = 0.77, and a highly non-significant increase at 30 days, p = 0.096, was observed. Non-significant decrease in uric acid levels was observed at day 15, p = 0.99, but an increase, also non-significant, at day 30, p = 0.597, was observed. The same levels of creatinine at 15 days, p = 1, and a significant decrease at day 30, p = 0.49, were observed. In tissue, a very highly significant increase in MDA, NO and XO, p = 0.000, compared with group (A) was observed.
Figure 7.
Impact of frankincense on biochemical analysis of kidney function in serum of rats.
Figure 8.
Impact of frankincense on biochemical analysis of kidney function in serum of rats.
Figure 9.
Impact of frankincense on biochemical analysis of kidney function in serum of rats.
Figure 10.
Impact of frankincense on biochemical analysis in kidney tissue of rats.
Figure 11.
Impact of frankincense on biochemical analysis in kidney tissue of rats.
Figure 12.
Impact of frankincense on biochemical analysis in kidney tissue of rats.
In Figs. 13–20, the histological examination in the kidney tissue of control group (A) is illustrated showing the widening of tubular lumen, extruded nuclei, shortening, damage of brush border in proximal tubules, degenerative, necrotic changes in kidney tubules and glomerulus, nuclear pleomorphism in kidney tubules. Frankincense group (B) showed nearly normal distal convoluted tubule with diluted lumen low, nuclei in some proximal epithelial cells, extruded nuclei, shortening, damage of brush border in proximal tubules, and hyalinization of glomerulus.
From these results, it may be inferred that the decrease of AST and ALT in serum is caused by the decrease in amino acid followed by decrease in protein synthesis and then increase in MDA, indicated by lipid peroxidation of kidney tissue. At high level of UA and urea at day 30 in group B due to the breakdown of purine base in DNA due to accumulation of UA in the blood and the toxicity of urea by excretion more and more from the liver. Also the level of creatinine is decreased at 30 days due to the accumulation of creatinine level in the kidney. It may be that frankincense inhibits or decreases the activity of XO causing accumulation or increase of UA. Also, increase in NO level causes activation of free radicals in kidney tissue. All of these demonstrated findings agree with histological examination.
I demonstrated in the biochemical analysis in the serum, tissue and histological examination, different impact between group (B) and group (A), and non-safety for drink frankincense in the period of this experiment in serum and livers, kidneys tissues of rats.
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