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ORIGINAL ARTICLE
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Protective effect of pycnogenol against gamma radiation-induced lung injury in rat: DNA damage, lipid peroxidation, antioxidant levels, and histopathological changes


1 Department of Biophysics, Erciyes University, Kayseri, Turkey
2 Department of Histology and Embryology, Erciyes University, Kayseri, Turkey
3 Radiation-Oncology, Faculty of Medicine, Erciyes University, Kayseri, Turkey
4 Genome and Stem Cell Center, Erciyes University, Kayseri, Turkey

Date of Submission28-Jul-2020
Date of Decision15-Jan-2021
Date of Acceptance22-Jan-2021
Date of Web Publication30-Jul-2021

Correspondence Address:
Fazile Canturk Tan,
Department of Biophysics, Faculty of Medicine, Erciyes University, Kayseri 38039
Turkey
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jcrt.JCRT_1062_20

 > Abstract 


Background and Objective: The study aims to evaluate the histopathological changes, enzymatic alterations, and DNA damage in rat lungs induced by whole-body gamma irradiation as well as evaluation of the protective effect of pycnogenol.
Materials and Methods: A hundred adult male rats were equally divided into ten groups including control, four antioxidants, γ-irradiation, four antioxidant + γ-irradiations. This study began the day before radiation treatment and continued for 3 days. The pycnogenol was dissolved 5% dimethyl sulfoxide and then administered orally through a gastric tube at a dose of 37.5 mg/kg, 75 mg/kg, 150 mg/kg, and 300 mg/kg in 24, 48, and 72 h before irradiation. Irradiation was applied with a whole-body irradiation dose of 900 cGy in one fraction. DNA damage, histopathological changes, catalase (CAT), and superoxide dismutase (SOD) activities and malondialdehyde (MDA) levels in lung tissue of rats were evaluated 3 days after irradiation.
Results: CAT and SOD activities were found to be significantly lower in the irradiation group than control (P < 0.001). CAT and SOD activities were higher in the antioxidant + γ-irradiation group than both irradiation and control groups. MDA levels were significantly higher in the irradiation group compared to control (P < 0.001), whereas MDA levels decreased in the antioxidant + γ-irradiation group compared to the irradiation group. The antioxidant groups were significantly increased comet parameters depend on pycnogenol doses compared to control. The antioxidant + γ-irradiation was decreased comet parameter compared to γ-irradiation. As a result of the histopathologically, the antioxidant groups were different than the control group that in the areas of alveolar sacs and connective tissue areas were seen hemorrhage areas similar to the irradiation group.
Conclusion: We demonstrate that 300 mg/kg of pycnogenol might provide significant protection against deleterious effects from whole-body ionizing radiation on the lung tissue. P300+ γ-ray group was significantly reduced radiation-induced lung injury and was possible to observe significantly preservation.

Keywords: Antioxidant activity, ionizing radiation, lung toxicity, protective effect, pycnogenol



How to cite this URL:
Tan FC, YAY AH, Yildiz OG, Kaan D. Protective effect of pycnogenol against gamma radiation-induced lung injury in rat: DNA damage, lipid peroxidation, antioxidant levels, and histopathological changes. J Can Res Ther [Epub ahead of print] [cited 2021 Dec 5]. Available from: https://www.cancerjournal.net/preprintarticle.asp?id=322704




 > Introduction Top


Cancer treatment is mostly based on a multimodal strategy, which includes radiotherapy (RT), chemotherapy, and surgery. The second plays an important role, as more than half of cancer patients will undergo RT[1] with variable efficacy. For nearly two decades, major research has been done in clinical trials to optimize RT methods (density modulation, stereotaxy, and altered fractionation) and limit the damage to normal tissues in the surrounding area, while allowing dose increases in the targeted tumor. On the other hand, recent advances in our understanding of the biological basis of cells and tissue response to ionizing radiation have highlighted some important molecular markers that can predict the effectiveness or toxicity of RT.[2]

Ionizing radiation can cause a number of biological consequences such as carcinogenesis, inflammation and death. Damage to normal tissues due to radiation is an important problem during RT or accidental exposure. The purpose of RT is to minimize the damage to normal tissues in the environment while giving a cytotoxic dose to the tumor.[3] RT-induced pneumonitis is one of the most serious dose-limiting toxicities in patients receiving thoracic RT for lung cancer. Therefore, it is important to minimize damage to normal cells by protective intervention before irradiation. Most of the ionizing radiation cytotoxic effects are mediated by reactive oxygen species (ROS) caused by the interaction between ionizing radiation and water molecules or ROS-guided oxidative stress, so the antioxidant system is the key role in treatment-related toxicities.[3] Interest remains in the identification and development of nontoxic and effective radio-protectors that may reduce the effect of ionizing radiation. The identification of radiation-protective agents is an important target for radiation oncologists and basic radiation biologists.

The aim of this study is to investigate a powerful and well-researched antioxidant for protective effects against gamma radiation-induced lung damage. We chose that the dietary supplement Pycnogenol®, extracted from the bark of the French maritime pine (Pinus pinaster) and consisting of procyanidins (US Pharmacopoeia 28) for this study. Pycnogenol® has been shown to be a powerful antioxidant in laboratory tests and clinical studies.[4],[5] Pycnogenol® improves antioxidant capacity, endothelial function, vascular integrity and has significant anti-inflammatory potential in humans.[6] The radical scavenging properties described of Pycnogenol® were assumed to have a potential radioprotective effect. To date, there are no studies evaluating possible radioprotective effects against damage induced by whole-body gamma irradiation.

Therefore, the aim of this study is to test the protective effect of Pycnogenol® on radiation-induced damage in lung tissue with histopathological, DNA damage, and enzymatic evaluations.


 > Materials and Methods Top


Chemicals

All chemicals were obtained from Merck (Darmstadt, Germany). Pycnogenol was donated by Horphag Research Ltd., UK. For chemical and biochemical examinations, ultrapure water received from the two-way water purification system (Purelab ELGA, High Wycombe, UK) was used. All reagents and chemicals were of analytical grade or higher purity.

Animals

A hundred adult male Wistar Albino rats were obtained from the Erciyes University Experimental Research and Application Center (Kayseri, Turkey). Animal experimentation was applied according to The Erciyes University Animal Experiments Local Ethics Committee decision (decision 11/127). Rats were kept in polycarbonate cages under an alternating 12 h light/dark cycle. Animals were maintained on laboratory chow and water ad libitum. Water and food were available from this center.

Irradiation

The animals were placed in a special box for whole-body irradiation with one fraction external acute gamma-rays exposure of 900 cGy delivered by Cobalt 60 Teletherapy (GWXJ80-Co60 Teletherapy Unit) in the RT Unit of the Erciyes University Gevher Nesibe Hospital, Kayseri, Turkey. The fixing boxes contained five rats for each irradiation. The animals were kept in well-ventilated cages and their movements were restricted. Physics calculate radiation doses were used 28 cm × 24 cm of field and 2.5 cm of depth from both back and front and total dose was calculated 900 cGy.[7] While calculating the radiation duration, the tray factor was taken into consideration.

Experimental design

Animals were randomly divided into ten groups, each containing 10 rats. As shown in [Table 1], control group, four antioxidant groups (P37.5, P75, P150, and P300), only irradiation group (γ-ray), and four antioxidant + γ-irradiation groups (P37.5+ γ-ray, P75+ γ-ray, P150+ γ-ray, P300+ γ-ray). The pycnogenol was dissolved in 5% dimethyl sulfoxide (DMSO). Controls and γ-ray groups were received 5% DMSO. Control animals were not irradiated. The γ-ray groups were treated with 900 cGy of gamma-irradiation to the whole-body in one fraction. The antioxidant groups were received 37.5 mg/kg, 75 mg/kg, 150 mg/kg, and 300 mg/kg of pycnogenol, respectively, in 24, 48, and 72 h. Antioxidant + γ-ray groups were received 37.5 mg/kg, 75 mg/kg, 150 mg/kg, and 300 mg/kg of pycnogenol and treated with 900cGy of gamma-irradiation to whole-body in one fraction. All irradiations were carried out between 9:00 a.m. and 11:00 a.m. The 24 h after antioxidant and irradiation applying, the animals were firstly weighed and then killed anesthetized by intramuscular injection ketamine (0.1 mg/kg) and xylazine (0.05 mg/kg). The lungs were quickly removed. Lungs were divided into 3 parts. The first lung was fixed in 10% buffered formal and the second lung was immediately used for comet assay and another lung was stored at −70°C until antioxidant enzymes assay. The tissue was homogenized in 4 volumes of 5 mM phosphate buffer, pH 7.4 and centrifuged at 10,000 × g for 15 min to obtain supernatant which used for the assay of antioxidant profile and protein determination of the animals.[8]
Table 1: Experimental groups of rats treated with pycnogenol and radiation

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Analytical procedures

Antioxidant enzymes assay

The activity of superoxide dismutase (SOD) was measured using a method determined by Sun et al.[9] The activity of CAT was measured using a method determined by Aebi.[10]

Malondialdehyde determination

The concentration of malondialdehyde (MDA) in the homogenate of the liver was measured using a method determined by Ohkawa et al.[11] as thiobarbituric acid reactive substances.

Total protein determination

The total protein concentration was evaluated using a method determined by the Bradford as bovine serum albumin standard.[12]

Assessment of tissue DNA damage using the comet assay

The lung tissue DNA damage was investigated using the comet assay. The comet assay was applied under neutral conditions.[13] The images of 100 chosen nuclei were made at a magnification of 200x using a fluorescent microscope (Olympus, BX51, Tokyo, Japan) and were analyzed using the Comet Assay Software Project (CASP-1.2.2, Windows 2010). We have used two parameters (tail DNA and tail moment TM) to calculate the quantity of DNA damage. The DNA damage was detected by fragmented DNA that migrated from the nuclei of the liver cells, causing a comet figure. However, nuclei without a comet were not evaluated damaged.[14]

Histopathological examination

After euthanasia of the animals at the end of the experimental period, the lung samples from the animals were excised and stored in a 10% formalin solution and then dehydrated and paraffinembedded. Sections of 5 μm thickness were prepared for microtomy and stained with hematoxylin-eosin (H and E), and Masson's trichrome in order to evaluate the morphology of tissue damage. All the sections were examined with a light microscope (Olympus BX51, Tokyo, Japan), and the pieces were photographed.

Statistical analysis

Analysis of comet, SOD, CAT, and MDA data was evaluated using IBM SPSS Statistics 20.0 (IBM Inc., ILL, USA) software. The suitability of the data to normal distribution was evaluated by the Shapiro–Wilk test and variance homogeneity was evaluated by the Levene test. Comparisons between groups were evaluated with one-way analysis of variance and Kruskal–Wallis H tests. Results of ten different rats were expressed as median (25%–75%). The statistical significance was based on P < 0.05. The Student-Newman-Keuls method was used as a multiple comparison test. Data were expressed as mean and standard deviation.


 > Results Top


The tissue enzyme values between groups of lung tissue were given in [Table 2]. To assess the degree of oxidative stress caused by ionizing radiation, the level of lipid peroxidation was measured in the lungs of all rats. The MDA level of the γ-ray group was statistically significantly higher than in other groups and control groups. The MDA level of antioxidant groups was statistically significantly higher than antioxidant + γ-ray groups (P < 0.001). MDA level of the P300 + γ-ray group was statistically significantly lower than the control group and γ-ray group (P < 0.001).
Table 2: The lung malondialdehyde level, catalase and superoxide dismutase activities in the experiment groups

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The CAT activity was measured as an indicator of the oxidant/antioxidant status of the lung. The CAT activity of the control group was statistically significantly higher than in other groups (P < 0.005). The radiation group was statistically significantly lower than other groups (P < 0.005). There were no statistically significant between antioxidant and antioxidant + γ-ray groups (P = 1.000). The CAT activity of the control group was statistically significantly higher than in other groups (P < 0.005). CAT activity of P300+ γ-ray groups was higher than γ-ray group.

The SOD activity was measured as an indicator of the oxidant/antioxidant status of the lung. The SOD activity of the radiation group was statistically significantly lower than the antioxidant and antioxidant + irradiation groups (P < 0.005). There were no statistically significant between antioxidant and antioxidant + γ-ray groups (P < 0.008). The SOD activity of P300+ γ-ray groups was higher than the other group (P < 0.008).

Exposed to whole-body gamma radiation of the rats were resulted in cellular DNA damage in the lung tissue. The cellular DNA damage values between groups of lung tissue were given in [Table 3]. The comet parameters (except Head DNA) of γ-ray group were increased according to parameters of the other groups [Figure 1]. P < 0.05 was statistically significant.
Table 3: Cellular lung DNA damage in the experiment groups

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Figure 1: The cell of lung tissue (a) Tail DNA of the control group was 2%, (b) Tail DNA of the P37.5 group was 5%, (c) Tail DNA of the P75 group was 12%, (d) Tail DNA of the P150 group was 16%, (e) Tail DNA of the P300 group was 18%, (f) Tail DNA of the γ -group was 21%, (g) Tail DNA of the P37.5+ γ -ray was 17%, (h) Tail DNA of the P75+ γ -ray group was 15%, (ı) Tail DNA of the P150 + γ -ray group was 8%, (i) Tail DNA of P300 + γ-ray group was 7% (Ethidium bromide staining × 200, Olympus, Tokyo, Japan)

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The comet parameters of antioxidant groups (P37.5, P75, P150, P300 groups) were significantly increased comet parameters depend on pycnogenol doses compared to the control group. In the P37.5 + γ-ray, P75 + γ-ray, P150+ γ-ray, P300 + γ-ray groups were decreased comet parameters compared to γ-ray group. P300 + γ-ray groups were possible to observe significantly preservation.

In the lung tissue sections stained using H and E and Masson trichrome staining were evaluated the morphology of tissue damage [Figure 2] and [Figure 3]. The control group had got normal histological apparent. Pulmonary alveolar epithelium is region direct contact with the air. In the lung parenchyma of this group could clearly select blood vessels with terminal and respiratory bronchioles together with alveolar sacs with smooth-appearing single-layer squamous epithelium. When the lung tissue of γ-ray group was examination histological, we were observed increased edema, thickening of alveolar walls, vascular and interstitial hemorrhage in the lung parenchyma. Hemorrhage was evident both on the alveolar surface and in the interstitial connective tissue. In the present study, we were observed that the lung was sensitive to 900 cGy radiation as a result was observed edema and hemorrhage. We were observed that hemorrhage was the similar to radiation group in the connective tissue areas and in the alveolar sacs different from the control group in the antioxidant groups. These bleeding areas were increase depending on antioxidant doses. Just, we were not observed cellular damage or edema in the antioxidant groups. We were not observed protective effect of antioxidant against radiation-induced lung damage in the P37.5 + γ-ray, P75 + γ-ray, P150 + γ-ray groups. We were observed decreased hemorrhage and edema in the P300 + γ-ray groups.
Figure 2: The lung tissue section (a) The control group had normal histological apparent, (b-e) The lung tissue of pycnogenol extract groups was observed that hemorrhage was similar to radiation group in the connective tissue areas and in the alveolar sacs different from the control group. These bleeding areas were increase depending on antioxidant doses. (f) The γ-ray group had observed increased edema, thicking of alveolar walls, vascular and interstitial hemorrhage in the lung paranchyma. Hemorrhage was evident both on the alveolar surface and in the interstitial connective tissue (g-i) The antioxidant + γ-ray groups weren't observed protective effect of antioxidant against radiation-induced lung damage in the P37.5 + γ-ray, P75 + γ-ray, P150 + γ-ray groups. In the P300 + γ-ray groups were observed decreased hemorrhage and edema. *Edema, thick arrow; thickening of alveolar walls, yellow arrow; vascular and interstitial hemorrhage (H and E staining × 400, Olympus, BX51, Tokyo, Japan)

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Figure 3: The lung tissue section (a) The control group had normal histological apparent, (b-e) The lung tissue of pycnogenol extract groups was observed that hemorrhage was similar to radiation group in the connective tissue areas and in the alveolar sacs different from the control group. These bleeding areas were increase depending on antioxidant doses. (f) The γ -ray group had observed increased edema, thicking of alveolar walls, vascular and interstitial hemorrhage in the lung paranchyma. Hemorrhage was evident both on the alveolar surface and in the interstitial connective tissue (g-i) The antioxidant + γ-ray groups weren't observed protective effect of antioxidant against radiation-induced lung damage in the P37.5 + γ -ray, P75 + γ-ray, P150 + γ-ray groups. In the P300 + γ-ray groups were observed decreased hemorrhage and edema. Yellow arrow; increased connective tissue areas (Masson trichrome staining × 400, Olympus, BX51, Tokyo, Japan)

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 > Discussion Top


The protection of normal cells from radiation-induced damage is a critical issue in radiation therapy and occupational or daily radiation exposure. To achieve optimum results, a reasonable balance is required between the total dose of RT given and the threshold limit of the normal critical issues in the environment. Normal tissues should be protected against radiation damage to achieve better tumor control with a higher dose. Therefore, the role of radioprotective compounds is very important in clinical RT.[15] It is particularly true for the lung because the lung is very sensitive to ionizing radiation due to its large surface area and higher oxygen levels. The radiation pneumonitis is an acute manifestation and is relatively common after RT for chest wall or intrathoracic malignancies. The lung is an organ that limits the dose during whole-body irradiation for the treatment of leukemia and other hematological malignancies.[16] The radiation damage in a cell is enhanced or mitigated due to various factors such as sulfhydryl compounds, oxygen, and the presence of other molecules in the cellular environment.[17],[18] In the presence of oxygen, hydrated electrons and H atoms react with molecular oxygen to produce radicals such as HO2, O2 as well as other aqueous free radicals. In the presence of oxygen, hydrated electrons, and H atoms react with molecular oxygen to produce radicals such as HO2, O2, apart from other aqueous free radicals.[19],[20] Enzymes such as glutathione peroxidase, catalase (CAT), and SOD protect mammalian cells from oxidative radiation damage.[21] The sensitivity of the cells to ionizing radiation was increased in the presence of oxygen. Oxidative damage to cellular genetic material, namely, DNA, plays an important role in mutagenesis and carcinogenesis. Highly reactive oxygen radicals produced by ionizing radiation cause lesions that lead to cell killing and mutations in DNA.

In addition, ionizing radiation can change the DNA structure, producing specific lesions such as base disorder, single-chain breaks (SSB), and double chain breaks (DSB), leading to cell cycle arrest and apoptosis.[22] A 1 Gy dose can produce more than 2000 base damage, 1000 SSB, 40 DSB and 30 DNA cross-links.[23] It is important to note that DSB is the deadliest RT-induced DNA damage. Antioxidants such as vitamins A, C, and E provide radiation protection because radiation damage mimics oxidative stress associated with active oxygen toxicity. Antioxidants such as Vitamins A, C, and E provide radiation protection because radiation damage mimics oxidative stress associated with active oxygen toxicity.[24],[25] Tocopherol monoglucoside, a water-soluble Vitamin E derivative, has been found to be very effective in protecting DNA after oral or intraperitoneal administration of DNA against in vitro as well as gamma radiation.[26] Melatonin (N-acetyl-5-methoxytryptamine), a pineal hormone that plays a role in the regulation of the neuroendocrine axis, is a highly effective free radical scavenger and antioxidant.[27],[28],[29],[30] When administered to mice before exposure to radiation, melatonin offered significant radiation protection, which is assessed by the frequency of chromosomal abnormalities in bone marrow cells, spermatogonia, spermatocytes and micronuclei.[31] This study was performed to evaluate oxidative status, DNA damage and histopathological changes of radiation-induced lung injury. For this purpose, the activities of antioxidant enzymes such as SOD and CAT and the concentration MDA an indicator of lipid peroxidation, single cell DNA damage and histopathological changes were determined in the lung tissue. Findings in the present study showed that the lung was sensitive to whole-body 900 cGy gamma-irradiation.

To assess the degree of oxidative stress caused by ionizing radiation, the level of lipid peroxidation was measured in the lungs of all rats. The MDA level of the irradiation group was statistically significantly higher than in other groups and control groups. The MDA level of antioxidant groups was statistically significantly higher than antioxidant + γ-irradiation groups (P < 0.001). The MDA level of P300+ γ-ray group was statistically significantly lower than the control group and γ-ray group (P < 0.001). The radiation-induced increase in MDA concentration in the lung indicates an escalation of lipid peroxidation in lung tissue. The increased MDA levels indicated that radiation causes oxidative damage to the lung. The reduced MDA concentration in the P300 + γ-ray group allow concluding that the lung antioxidant defense system still effectively protects from the action of radiation-induced free radicals. Pycnogenol protected membrane lipid against oxidative damage of radiation in the P300 + γ-ray groups.

The CAT activity was measured as an indicator of the oxidant/antioxidant status of the lung. The CAT activity of control groups was statistically significantly higher than in other groups (P < 0.005). The irradiation group was statistically significant lower than other groups (P < 0.005). There were no statistically significant between antioxidant and antioxidant + γ-ray groups (P = 1.000). The CAT activity of P300+ γ-ray groups was higher than γ-ray group. However, there were no statistically significant between P300+ γ-ray groups and γ-ray groups (P = 0.239). CAT activities were decreased in the γ-ray group and P300+ γ-ray groups compared to the control group in the study showing that the antioxidant system is insufficient against excessive SOR production.

The SOD activity was measured as an indicator of the oxidant/antioxidant status of the lung. The SOD activity of the radiation group was statistically significantly lower than antioxidant and antioxidant + γ-ray groups (P < 0.005). There was no statistically significant between antioxidant and antioxidant + γ-ray groups (P < 0.008). The SOD activity of P300+ γ-ray groups was higher than in other groups (P < 0.008). Results are showing that the antioxidant system is sufficient against excessive SOR production.

CAT and SOD activities were increased in the P300+ γ-ray groups compared to the γ-ray groups. The pycnogenol is sufficient against the production of ROS. CAT and SOD activities were increased in the pycnogenol extract + γ-ray groups. This shows that both antioxidant enzymes potentiate the effects of each other.

Improved peroxidation of lipids in the intracellular and extracellular membranes leads to damage to cells, tissues, and organs. The SOD and CAT are important antioxidant enzymes that protect this process by eliminating ROS. The SOD catalyzes the reaction of superoxide anion radical (O.-2) dismutation to hydrogen peroxide (H2O2), whereas CAT degrades H2O2 into a molecule of oxygen and a molecule of water.[32]

The radiation-induced escalation of lipid peroxidation in the lung might be a consequence of the increased formation of free radicals as well as the inhibition of SOD and CAT activities. The results obtained indicate that at the used level of radiation treatment, the antioxidant defense system in the lung was insufficient to give complete protection and thus the processes of lipid peroxidation escalated. The increased activity of SOD with the simultaneous increase in the activity of CAT in the lung that the lung antioxidant defense system still effectively protects from the action of radiation-induced free radicals.

In our data, as shown in previous studies with oxidant agents, ionizing radiation-induced lung damage may be caused by the indirect effect of radiation due to SOR, as well as by the direct effect of ionizing radiation. Because CAT activity was decreased in the radiation group, whereas SOD activity and MDA levels were increased in the radiation group. Also, although there was a numerical difference between the groups, it was not statistically significant.

Exposed to whole-body gamma radiation of the rats were resulted in cellular DNA damage in the lung tissue. The cellular DNA damage values between groups in the lung tissue were given in [Table 3]. The comet parameters (except Head DNA) of γ-ray group were increased according to parameters of the other groups [Figure 1]. P < 0.05 was statistically significant. The comet parameters of antioxidant groups (P37.5, P75, P150, P300 groups) were significantly increased comet parameters depend on pycnogenol doses compared with to control group. Simultaneously increased comet parameters with antioxidant doses in the antioxidant group showed that pycnogenol was toxic. In the P37.5 + γ-ray, P75 + γ-ray, P150 + γ-ray, P300 + γ-ray groups were decreased comet parameters compared to γ-ray group. P300 + γ-ray groups were possible to observe significantly preservation according to other groups.

The control group had got normal histological apparent. Pulmonary alveolar epithelium is region direct contact with the air. In the lung parenchyma of this group could clearly select blood vessels with terminal and respiratory bronchioles together with alveolar sacs with smooth-appearing single-layer squamous epithelium. When the lung tissue of γ-ray group was examination histological, we were observed increased edema, thickening of alveolar walls, vascular and interstitial hemorrhage in the lung parenchyma. Hemorrhage was evident both on the alveolar surface and in the interstitial connective tissue. In the present study, we were observed that the lung was sensitive to 900 cGy radiation as a result was observed edema and hemorrhage. We were observed that hemorrhage was similar to the irradiation group in the connective tissue areas and in the alveolar sacs different from the control group in the antioxidantgroups. These bleeding areas were increase depending on antioxidant doses. Just, we weren't observed cellular damage or edema in the antioxidant groups. We were not observed protective effect of antioxidant against radiation-induced lung damage in the P37.5+ γ-ray, P75 + γ-ray, P150 + γ-ray groups. We were observed decreased hemorrhage and edema in the P300+ γ-ray groups [Figure 2] and [Figure 3].

The results of this study supported by some research. Farzipour et al. evaluated total body irradiation at doses of 5 Gy from X-ray source-induced acute lung damage in mice. The biochemical studies were performed at 24 h after irradiation. Results from biochemical analyses demonstrated increased MDA, nitric oxide (NO) and protein carbonyl (PC) levels of lung tissues in only irradiated group. Histopathologic findings also showed an increase in the number of inflammatory cells and the acute lung injury in this group.[33] Haddadi et al. investigated a single dose of 18 Gy γ-rays-induced lung damage in rats. They evaluated acute and chronic histopathological changes lung which 24 h after RT for acute and 8 weeks after RT for chronic. The acute results indicated significant increases in inflammation, lymphocyte, macrophage, and neutrophil frequency in RT group when compared with sham group. The chronic results indicated a majority of factors such as frequency of inflammatory cells and mast cells and the incidence of inflammation, pulmonary fibrosis, alveolar thickness, and vascular thickness increased in irradiated lungs as compared to the sham group.[34] Cantürk Tan et al. evaluated that different doses of Pycnogenol might provide substantial protection against radiation-induced oxidative, DNA damage, and histopathological changes in the liver.[35] Furthermore, Ramos et al. showed that the study shows that both doses of Pycnogenol® provide significant protection against the harmful effects of whole-body ionizing radiation on the intestinal mucosa.[36]

Our results were clearly demonstrated that the pycnogenol was caused oxidative stress, DNA, and tissue damage in rat lungs, which was evidenced by the dose-dependent increase in cellular DNA damage, tissue damage and lipid peroxidation up to 24 h after pycnogenol. We were demonstrated γ-radiation caused oxidative stress, DNA and tissue damage in rat lungs, which was evidenced by an increase in DNA damage, tissue damage and lipid peroxidation up to 24 h after irradiation. Also, we were demonstrated that pycnogenol + γ -ray protective from oxidative stress, DNA and tissue damage in rat lungs, which was evidenced by a decrease in DNA damage, tissue damage, and lipid peroxidation up to 24 h after irradiation. Pycnogenol® was administered orally to rat in a dosage of 37.5, 75, 150, and 300 mg/kg body weight, which is in the same range as in our experiments.


 > Conclusion Top


Our study demonstrates that both applied dosages of Pycnogenol® provide significant protection against deleterious effects from whole-body ionizing radiation on the lung tissue. Furthermore, this study describes for the first time a radioprotective function for Pycnogenol® on lung tissue. It is proposed that pycnogenol may be expected to reduce damage from ionizing radiation. Also, our study was demonstrated that pycnogenol has a toxic effect on normal lung tissue.

Financial support and sponsorship

The authors acknowledge the financial support from Erciyes University—The Scientific Research Projects of Turkey (ERUBAP); Project number: TSD-12-3936.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

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    Tables

  [Table 1], [Table 2], [Table 3]



 

 
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