Katalinić V, Radić S, Ropac D, Mulić R, Katalinić A.
Odjel sanitarne kemije i toksikologije, Institut pomorske medicine HRM, Split, Hrvatska.
AIM: The aim of this study was to determine the antioxidative activity of propolis from ecologically clean parts of Dalmatia.
METHODS: Phenol concentration in ethanolic propolis extracts was determined by Folin-Ciocalteu reagent using gallic acid as the standard. Flavonoid phenolic compounds were analyzed after precipitation with formaldehyde. The residual non-flavonoid phenolics were also determined by Folin-Ciocalteu method. By determining the change of peroxide number (PN), of tiobarbiture acid reactive species (TBARS), and of DPPH-radical activity, antioxidative efficiency of propolis was tested and compared with well known and widely used synthetic antioxidants. Values of PN and TBARS were determined at 60 degrees C in samples of trigyceride substrate (lard) without and with the addition of antioxidants. Compared was the efficiency of three antioxidants: propolis (alcoholic extract), vitamin E, and (+)-catechin in a concentration of 1%. PN was monitored during 50 days. By the method of Sedlacek, TBARS were measured during 30 days. Antioxidative activity of propolis extract was also measured in terms of hydrogen donating ability using stable radical alpha,alpha-diphenyl-beta-picril hidrazyl (DPPH*) and compared with commercial synthetic antioxidants of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and (+)-cathecin. Inhibition degree of DPPH* was calculated by the formula of Yen and Duh.
RESULTS: Total phenol content, expressed as gallic acid, in propolis extracts varied from 75.2 to 90.2 g/kg propolis. The proportion of flavonoids in total phenols ranged from 62% to 65%. Values of TBARS were not increased only in samples with added propolis. The inhibition of DPPH-radical by propolis extracts ranged from 93% to 96%, by catechin 95%, by BHT 49%, and by BHA 64%. Compared to BHT and BHA, propolis extracts showed greater reducing activity against DPPH-radical.
DISCUSSION: The chemical composition of propolis, and thus its biological activity depend on the plant from which it has been collected, and on the macro- and microclimatic conditions. Many compounds in propolis exert antioxidative activity. A belief was expressed that the biological activity of propolis is very probably based mostly on its antioxidative efficiency. Dalmatian propolis showed high efficiency in the prevention of oxidative processes. This could be explained by the high proportion of polyphenol constituents, especially flavonoids. A very low and equal degree of increase of PN, as a measure of oxidative processes, was noticed in the samples of triglyceride substrate with the addition of propolis and (+)-catechin. The greatest rise of TBARS was measured in the samples of pure lard. There was no increase of TBARS only in the samples with added propolis. Propolis and (+)-catechin showed great efficiency in the inhibition of DPPH-radical, greater than BHT and BHA, which are widely used in food industry.
CONCLUSION: The results indicate that Dalmatian propolis could be an efficient protective agent against oxidative processes in food. The high antioxidative activity of propolis, its natural origin, and present knowledge about its biological properties, make it a very promising nutritional additive for human diet.
Sobočanec S, Šverko V, Balog T, Šaric A, Rusak G, Likić S, Kušić B, Katalinić V, Radić S, Marotti T.
Division of Molecular Medicine, Ruder Boskovic Institute, Zagreb.
Native propolis was defined as propolis powder collected from the continental part of Croatia and prepared according to a patented process that preserves all the propolis natural nutritional and organoleptic qualities.
Nine phenolic compounds (out of thirteen tested) in propolis sample were detected by high performance liquid chromatography (HPLC) analysis. Among them chrysin was the most abundant (2478.5 microg/g propolis). Contrary to moderate antioxidant activity of propolis examined in vitro (ferric reduction antioxidant power; FRAP-assay), propolis as a food supplement modulated antioxidant enzymes (AOE) and significantly decreased lipid peroxidation processes (LPO) in plasma, liver, lungs, and brain of mice. The effect was dose- and tissue-dependent. The lower dose (100 mg/kg bw) protected plasma from oxidation, whereas the higher dose (300 mg/kg bw) was pro-oxidative. Hyperoxia (long-term normobaric 100% oxygen) increased LPO in all three organs tested.
The highest vulnerability to oxidative stress was observed in lungs where hyperoxia was not associated with augmentation of AOE. Propolis protected lungs from hyperoxia by increased catalase (CAT) activity. This is of special importance for lungs since lungs of adult animals are highly vulnerable to oxidative stress because of their inability to augment AOE activity. Because of its strong antioxidant and scavenging abilities, native propolis might be used as a strong plant-based antioxidant effective not only in physiological conditions but also in cases that require prolonged high concentration of oxygen.
Sobočanec S, Šverko V, Balog T, Šaric A, Rusak G, Likić S, Kušić B, Katalinić V, Radić S, Marotti T.
Division of Molecular Medicine, Ruder Boskovic Institute, Zagreb.
Native propolis was defined as propolis powder collected from the continental part of Croatia and prepared according to a patented process that preserves all the propolis natural nutritional and organoleptic qualities.
Nine phenolic compounds (out of thirteen tested) in propolis sample were detected by high performance liquid chromatography (HPLC) analysis. Among them chrysin was the most abundant (2478.5 microg/g propolis). Contrary to moderate antioxidant activity of propolis examined in vitro (ferric reduction antioxidant power; FRAP-assay), propolis as a food supplement modulated antioxidant enzymes (AOE) and significantly decreased lipid peroxidation processes (LPO) in plasma, liver, lungs, and brain of mice. The effect was dose- and tissue-dependent. The lower dose (100 mg/kg bw) protected plasma from oxidation, whereas the higher dose (300 mg/kg bw) was pro-oxidative. Hyperoxia (long-term normobaric 100% oxygen) increased LPO in all three organs tested.
The highest vulnerability to oxidative stress was observed in lungs where hyperoxia was not associated with augmentation of AOE. Propolis protected lungs from hyperoxia by increased catalase (CAT) activity. This is of special importance for lungs since lungs of adult animals are highly vulnerable to oxidative stress because of their inability to augment AOE activity. Because of its strong antioxidant and scavenging abilities, native propolis might be used as a strong plant-based antioxidant effective not only in physiological conditions but also in cases that require prolonged high concentration of oxygen.
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Propolis therapy or therapy that combines propolis with medications has been considerably effective in healing inflammation of nasal cavity, throat, sinusitis, inflammation of the vocal cords, bronchitis and pulmonary tuberculosis.
Asthma
A clinical study of patients suffering from mild to medium asthma has shown that propolis reduces the frequency of seizures at night and considerably improves the ventilatory lung functions.
Lung cancer
Tests on animals have proved that propolis prevents forming of bronchial and alveolar cell carcinoma. Propolis treatment or treatment that combines propolis with cytostatics prevents metastasizing to the lungs. Research at Ruđer Bošković Institute has proved that NATIVE PROPOLIS protects the lungs from damage caused by free radicals and oxidative stress and prevents forming of lung cancer because it markedly strengthens the gene the lack of which leads to appearance of tumour. Furthermore, the research showed that NATIVE PROPOLIS stops the growth and kills the tumour cells.
Therefore, it is important to protect the lungs with propolis when inhaling increased quantities of oxygen (hyperbaric oxygen therapy, diving, increased physical exertion – athletes), working in polluted atmosphere (smog, chemical solvents, smoking), when suffering from pulmonary diseases and when at risk for possible hereditary pulmonary diseases.
Propolis has a healing effect on the digestive system.
It stimulates its regular functioning and protects stomach cells from alcohol-caused damage. Experiences with NATIVE PROPOLIS show that it can be effective in constipation prevention.
Gastric ulcer and duodenal ulcer
Due to its anaesthetic and healing properties, propolis has been used in treatment of gastric and duodenal ulcer for a long time. The practice of using NATIVE PROPOLIS has shown rapid healing effect on duodenal ulcers.This is probably due to its anti-inflammatory properties, forming of film over the mucous membrane, reduction of gastric acidity and soothing the smooth musculature cramps.
Large intestine inflammation
Clinical as well as our own experiences in the application of propolis have shown a very beneficial effect on chronic large intestine inflammation (colitis).
Large intestine carcinoma
It prevents lesion of the large intestine that can cause the development of carcinoma. When the carcinoma has already developed it slows down its growth, kills the tumour cells and reduces the risk of metastasizing to the liver. Combined with cytostatics, it diminishes the progress of abdomen carcinoma considerably and the recuperation of white and red blood cells is more rapid, compared to using only cytostatics.
Therefore, it could be used, with a compulsory physician recommendation, in addition to the cytostatic therapy, because it improves the immune system and alleviates the consequences of chemotherapy.
Clinical experiences of local therapy with propolis preparations have shown good results in treating cervical ectopia, erosion, inflammation and vaginal inflammation.
Carcinomas
It has been proved at the Faculty of Natural Sciences and Mathematics in Zagreb that Croatian propolis has anti-tumour and anti-metastasizing effects on mammary gland carcinoma.
Research results of Ruđer Bošković Institute have shown that NATIVE PROPOLIS stimulates processes that prevent the formation of carcinomas of reproductive organs. Furthermore, it stops the progression of carcinoma and kills tumour and deformed cells.Cytostatics combined with propolis are more efficient in preventing the progression of carcinoma and its metastasizing and the side effects of chemotherapy are substantially diminished (nausea, weakness, vomiting, and the reduction in the quantity of corpuscles).
Therefore, the usage of propolis is important for protection of persons at increased risks for hereditary reproductive organs diseases and, with a compulsory physician recommendation, it can be used to treat the above mentioned diseases.
KIDNEYS
Antioxidative substances in propolis not only protect the kidneys from acute damage caused by some cytostatics (cisplatin) during chemotherapy, but also prevent forming of carcinoma on the kidneys.
LIVER
Ruđer Bošković Institute has determined that NATIVE PROPOLIS prevents liver damage caused by free radicals and enhances its antioxidative defence.
Tests conducted on mice have shown that, generally, propolis protects the liver from chemical toxins and alcohol, which is attributed to its ability to capture free radicals. Its protective effect is stronger than the well known substance glycyrrhizin, used by official medicine.
Carcinomas
Stops the growth of some kinds of liver tumours and kills cancerogenic cells.
Liver protection is important to chronic patients who must take medications regularly, in cases of liver diseases and to persons prone to alcohol abuse.
NATIVE PROPOLIS prevents lesion and strengthens the defence of the heart muscle from oxidative stress and free radicals that are formed under the influence of cardio-toxic substances.
The protective activity is especially important in cases of heart diseases and persons at risk for hereditary heart diseases.
Blood
Propolis improves the absorption of minerals (iron, calcium, phosphorus, magnesium) from the digestive system into the blood circulatory system.
Anaemia
In combination with pollen (NATIVE PROPOLIS), it increases resorption of iron into the blood and raises the level of haemoglobin in cases of iron deficiency anaemia (ferropenic anaemia).Tests conducted in Ruđer Bošković Institute have shown that the presence of NATIVE PROPOLIS in blood substantially reduces the level of free radicals, increases the antioxidative defence and prevents oxidative stress. Thus, it protects blood corpuscles from free radical damage.
Lymphatic system
It has been determined in laboratory conditions on the model of human leukaemia cells (HL-60) that one of the propolis components kills the tumour cells.
Using the DNA chip method in Ruđer Bošković Institute it has been determined that NATIVE PROPOLIS markedly strengthens the gene the lack of which leads to appearance of lymphatic system carcinoma.
The protective activity of NATIVE PROPOLIS is significant for groups at high-risk for hereditary blood diseases.
Propolis stops the reproduction of bacteria in the oral cavity, heals necrotic stomatitis, aphthae and candidiasis, prevents the forming of dental plaque and dental caries, reduces the painful sensitiveness of teeth, increases tooth enamel hardness, strengthens dental pulp, reduces the acute and chronic inflammation of gums (paradontosis).
Tests on mice have shown that propolis prevents the forming of cataract and enhances the antioxidative defence of blood and lens.
The same antioxidative as well as anti-inflammatory mechanisms protect the cornea from the results of chemical and mechanical injuries.
The effect of propolis on brain cells had been unknown in professional literature. Tests conducted by the Ruđer Bošković Institute have shown that the NATIVE PROPOLIS could protect the brain from damage and atrophy of nerve cells, particularly in cases of nervous system diseases (Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, diabetic neuropathy, neuritis, brain atherosclerosis, epilepsy, depression, schizophrenia, ischaemic-reperfusion brain injury, aging, etc.), because it:
-prevents the brain oxidative stress,
-increases antioxidative defence of the brain tissue,
-neutralizes free radicals in the brain,
-markedly strengthens the gene with a weakened function apparently linked to the damage of nerve cells and an increased risk of Parkinson’s disease,
-repairs the free-radicals induced DNA damage,
-strengthens the gene that aids transmission of nerve impulses
-stimulates the DNA replication in the brain.
In cases of brain tumour (glioma) it stimulates the organism to produce the drug by itself (immunosuppression therapy).
It has been proved that, in cases of brain stroke, propolis in synergy with the bee pollen (NATIVE PROPOLIS) increases the blood supply of the brain and facilitates the more rapid recovery of disrupted and lost functions.
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