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Egypt. J. Histol. Vol. 33, No. 2, June, 2010: 213 - 223 (ISSN: 1110 - 0559) Original Article A Histological Study on the Effect of Diclofenac Sodium (Declophen) Administration on Thyroid Follicular Cells of Albino Rats Nadia M. El-Rouby Histology Department, Faculty of Medicine, Cairo University ABSTRACT Introduction: Diclofenac Sodium (declophen) is a nonsteroidal anti-inflammatory drug recently categorized as thyroid antagonist. Aim of the Work: To determine the histo-physiological changes that might occur in the thyroid follicular cells after declophen administration and evaluate their reversibility. Materials and Methods: Thirty adult male albino rats were randomly divided into three groups; Group I (Control group), Group II received declophen (8 mg/kg /day) for one week then sacrificed. Group III received declophen as group II and then sacrificed one week after drug withdrawal. Blood samples were collected to estimate T3, T4 and TSH. Thyroid glands were processed for microscopic examination. Mean follicular diameter and mean height of thyrocytes were morphometerically evaluated and statistically analyzed. Results: Declophen significantly decreased T3 level which did not return back to normal in group III. Histologically, group II showed considerable light microscopic changes as focal distension of follicles that were lined with flattened epithelium. Their colloid revealed minimal peripheral vacuolations. Some follicles showed shedded epithelial lining. Others appeared irregular with disrupted epithelium. That epithelium had irregular dark pyknotic nuclei and vacuolated cytoplasm. Some follicles had apparently little colloid. Ultrastructural changes included dilated rER with electron lucid material and many darkly stained lysosomes. Group III showed some changes compared with Control. Statistically, mean follicular diameter of group II increased significantly while mean epithelial height decreased significantly compared to Control. Group III revealed neither significant increase in mean follicular diameter or decrease in epithelial height in comparison with the control. Conclusion: Declophen has toxic effect on the thyroid follicular cells which was incompletely recovered. So it should be discontinued prior to evaluation of thyroid functions. Key Words: Declophen, follicular cells, thyroid gland, Corresponding Author: Nadia M. El-Rouby rats. Tel.: 0111675532 E-mail: [email protected] INTRODUCTION Many factors including drugs can influence thyroid function in experimental animals and humans. Studies in humans reported significant effects of non steroidal anti-inflammatory drugs (NSAIDs) on thyroid tests, which can lead to misinterpretation of the result and inappropriate therapeutic decision1-3. and anti-pyretic actions. It is commonly used as a pain killer in many cases including rheumatoid arthritis, dysmenorrhea, artheralgia, tooth ache and postoperative pain7. Sometimes the drug may be used in high doses due to severe agonizing pain or by mistake. As NSAIDs are used more and more frequently in human, it is important to know to what extent they can influence the thyroid gland. Reviewing the literature showed that the influence of NSAIDs, including declophen, on the concentration of thyroid hormones has been established. However, there have been very few previous studies demonstrating the effect of declophen on the histological architecture of thyroid gland8. So the objective of the present study was to evaluate the declophen-induced alterations in the histological structure and function of the thyroid follicular cells. It was also aimed to evaluate the reversibility of such changes. The role of NSAIDs in causing thyroid dysfunction is quite often forgotten. These drugs might influence thyroid hormone homeostasis at any level from their synthesis, secretion, and transport or end-organ action as it might interfere with thyroid hormone binding sites (competitive inhibition). They might also alter the synthesis and secretion of thyrotropin (Thyroid Stimulating Hormone: TSH)4,5. Some cases, however, showed clinically apparent thyroid disease6. Diclofenac Sodium (Declophen) is a potent NSAID. It has anti-rheumatic, anti-inflammatory, analgesic 20 (1185-2010) 213 Nadia M. El-Rouby were fixed in 10% formol saline and processed for paraffin blocks. Sections of 5 micrometer thickness were cut and stained with hematoxylin and eosin (H&E). MATERIALS AND METHODS A- Drug: The drug used was Diclofenac Sodium (declophen), presented as ampoules of 75 mg/3ml (Pharco Pharmaceutical, Alexandria, Egypt). The dose used for rats was 8 mg/kg body weight9. The drug was given daily for a week by intramuscular (I.M) route. The control rats were given physiological saline I.M. in the same volume daily. 2. For Transmission Electron Microscopy (TEM): Thyroid tissues were rinsed in phosphate buffer (PH 7.4) then fixed in 2.5% gluteraldehyde, post fixed in 1% osmium tetroxide and embedded in epoxy resin. Semithin sections of 1 micrometer thickness were cut on an LKB ultramicrotome and stained with toluidine blue. Ultrathin sections were stained with uranyl acetate and lead citrate10 to be examined under Joel EM 100 S TEM using an accelerated voltage of 60 KV. B- Animals: Thirty adult male albino rats were used in this study. They were obtained from Kaser El -Aini animal house. Their weights ranged between 150-200 g. They were kept under good hygienic conditions, fed ad libitum and allowed free water supply. Rats were divided randomly into three equal groups as follows: Morphometric Analysis of Both Thyroid Follicular Diameter and Follicular Epithelium Height: Leica Qwin 500 Ltd, image analysis system was used to determine the diameter and the follicular epithelium height of thyroid follicles. The two parameters were measured in micrometers using low power (X 100) and high power (X 400) magnification, respectively. The internal follicular diameters of 50 follicles, in 10 random fields, of the thyroid gland of each rat of the different groups were measured. They were measured on diagonal axes. The mean of these two readings was then taken as the internal diameter of the follicle. To measure the epithelial height, 50 follicles from each rat in 10 random fields were chosen. The epithelial height was measured at two points on the same axis of each follicle. The mean of these two readings was then taken as the epithelial height. The mean value and Standard Deviation (SD) of the data obtained for each group were calculated. Group I: (Control Group): Rats were given physiological saline daily in a similar manner to other groups. Five rats were sacrificed with animals of group II and the other five rats were sacrificed with animals of group III. Group II: (Experimental Group): Rats were given declophen as an I.M. daily dose of 8 mg/kg body weight 9 for one week. After that week, the rats were sacrificed. Group III: (Recovery Group): Rats received declophen as in group II. The rats were kept alive for another week without treatment. On the 15th day of the beginning of the experiment these rats were sacrificed. Statistical Analysis: Statistical analysis was performed on IBM/PC using SPSS (Version 11) / PC program. Comparison of significance between the different groups was made using student “T” test11. Results were considered significant when probability (p) ≤ 0.05. Methods: At the end of the experimental period, the animals were sacrificed under ether anaesthesia. Blood samples, from all animals, were collected for hormonal assay. Then, the thyroid gland of each rat was excised. RESULTS During the experiment, three rats died; two from the experimental group (Group II) and one from the recovery group (Group III). Hormonal Assay: Hormonal assay was carried out at the Clinical Pathology Department, Faculty of Medicine, Cairo University, to determine the serum levels of T3, T4 and TSH. Hormonal Assay: It was clear that T3 was significantly decreased in both the experimental and recovery groups (P< 0.05) compared to the control. On the other hand, there was no significant change of both T4 and TSH (P >0.05) compared to the control (Table 1 and Chart 1). Histological Study: 1. For Light Microscopy: The thyroid tissues 214 A Histological Study on the Effect of Diclofenac Sodium (Declophen) Administration on Thyroid Follicular Cells of Albino Rats Table 1: The mean values of T3 in µg/dL, T4 in µg/dL and TSH in µU/mL ± SD in different groups: Item Control group Experimental group Recovery group T3: Mean ±SD “T” test 3.9685 ±.09540 2.7100 ±.07071 * P1< 0.05 3.0990 ±.02767 * P2< 0.05 T4: Mean ±SD “T” test 2.6270 ±.02830 2.6150 ±.02550 P1 >0.05 2.6000 ±.02944 P2 >0.05 0.0118 ± 0.00301 0.0137 ± 0.00295 P1 >0.05 0.0120 ±.00267 P2 >0.05 TSH: Mean ±SD “T” test P1 = between the experimental group and control group. P2= between the recovery group and control group. * Significant P value. was interrupted (Fig. 4-B). The lining follicular cells of the damaged follicles had darkly stained irregular and pyknotic nuclei. The cytoplasm of the affected thyrocytes was vacuolated (Figs. 5-A and B). The follicles showed variable density of colloid staining. Mast cells were frequently seen in the interstitial tissue close to blood vessels (Fig. 6). Electron microscopy revealed that the nuclei of the affected follicular cells appeared indented and hetrochromatic with dilatation of the peri-nuclear space. The cisternae of rER were dilated and contained amorphous lightly stained material (Figs. 7 and 8). The cytoplasm showed many darkly stained lysosomes and prominent Golgi complex (Fig. 9). The distended follicles were lined with flattened thyrocytes that had flat nuclei (Fig. 10). Chart 1: The mean value of T3, T4 & TSH levels. Group III (Recovery Group): The thyroid follicles of rats of this group started to regain their activity and normal appearance but patchy in distribution. These follicles were lined with cubical epithelium and showed peripheral vacuolations of their colloid content. Many distended follicles were still seen. These follicles were lined with flattened thyrocytes. Their colloid had minimal peripheral vacuolations. Also, involuted and distorted follicles persisted (Fig. 11). The follicular cells had vesicular nuclei. Some mast cells with darkly stained granules were seen in the surrounding connective tissue beside the blood vessels (Fig.12). Electron microscopy revealed that the follicular cells appeared more or less normal compared with the control group. They had more or less regular euchromatic nuclei with prominent nucleoli. The cytoplasm had more or less normal rER cisternae but many vacuoles and many darkly stained lysosomes were still seen in some follicular cells (Figs. 13 and 14). Some follicular cells showed sings of regeneration as they were binucleated (Fig. 15). Histological Results: Group I (Control Group): The thyroid gland of control rats was composed of follicles lined with a single layer of cuboidal follicular cells, and filled with acidophilic homogenous colloid having peripheral vacuolations (Figs. 1-A and B). The thyroid follicles were surrounded by vascular connective tissue stroma (Figs. 1-A and 2-A).The follicular cells had vesicular nuclei and prominent nucleoli (Fig. 2-A). Electron microscopy revealed that the follicular cells had euchromatic nuclei, rough endoplasmic reticulum and mitochondria. The apical cytoplasm revealed secretory vesicles. The colloid showed peripheral vacuolations in the form of electron lucid areas (Fig. 2-B). Group II (Experimental group): Examination of the thyroid gland of rats of the experimental group revealed focal markedly distended thyroid follicles. These follicles were lined mostly with flattened epithelial cells that had flattened nuclei. Their colloid showed nearly absent peripheral vacuolations (Fig. 3-A). Some follicles revealed shedded cells. The interstitial tissue had dilated blood vessels (Fig. 3-B). The thyroids of some animals revealed focal involuted follicles with minimal amount of colloid (Fig. 4-A). The wall of some follicles Morphometric Study and Statistical Analysis: The mean diameter of thyroid follicles of rats of the experimental group was significantly increased (p1< 0.05) compared to the control group while the mean follicular diameter of thyroid follicles of the recovery group was insignificantly increased (p 2 >0.05) in comparison with the control group (Table 2 and Chart 2). 215 Nadia M. El-Rouby Table 2: The mean follicular diameter in µm ±SD of thyroid follicles of different animal groups: Mean follicular diameter in µm “T” test Control group Experimental group Recovery group 35.7±9.6 51.9±13 * P1< 0.05 36.6±9.9 P2>0.05 P1 = between the experimental group and control group. P2= between the recovery group and control group. * Significant P value. Chart 2: The mean follicular diameter of thyroid follicles of the different groups. The mean epithelial height of thyroid follicles of the experimental group was significantly decreased (p1< 0.05) compared to the control group while the mean epithelial height of the recovery group was insignificantly decreased ( p2 >0.05) compared with the control group (Table 3 and Chart 3). Table 3: The mean value of epithelial height of thyroid follicles in µm ± SD in different groups: Mean epithelial height in µm “T” test Control group Experimental group Recovery group 9.2±1.6 6.8±1.4 * P1< 0.05 8.9±1.9 P2>0.05 P1 = between the experimental group and control group. P2= between the recovery group and control group. * Significant P value. Chart 3: The mean epithelial height of thyroid follicles in the different groups. 216 A Histological Study on the Effect of Diclofenac Sodium (Declophen) Administration on Thyroid Follicular Cells of Albino Rats Fig. 1: Photomicrographs of sections in the thyroid gland of a control rat (group I) showing: a. Normal thyroid follicles lined with simple cuboidal epithelium and filled with colloid (C). Note blood vessels (BV) in the surrounding connective tissue stroma. H&E X 200. b. The colloid (C) has peripheral vacuolations (V). H & E X 400. Fig. 2: Photomicrographs of: a. Semithin section in the thyroid gland of a control rat. It shows that the follicles are of variable sizes and lined with cuboidal follicular cells that have vesicular nuclei (N). The follicles are filled with colloid (C). Note that the stroma has many blood capillaries. Toluidine blue X 1000. b. TEM section of a control rat thyroid showing a part of thyrocyte. The colloid (C) shows two electron lucid areas (V) of peripheral colloid vaculations. Cisternae of RER and mitochondria (m) are scattered throughout the cytoplasm. Many secretory vesicles (S) in the apical cytoplasm and part of the nucleus (N) are seen. Orig mag X 2000. 217 Fig. 3: Photomicrographs of sections in the thyroid gland of rats from the experimental group (Group II): a. Reveals markedly distended thyroid follicles. They are lined mainly by flat cells with flat nuclei (arrow) and few low cuboidal cells with apparently rounded nuclei (arrow heads). The colloid (C) shows apparently no peripheral vacuolations. H&E X 400. b. Shows that some follicles have shedded epithelial lining (*). The thyrocytes of most of the follicles have vacuolated cytoplasm (arrow). Note the dilated blood vessel (b.v). H&E X 400. Fig. 4: Photomicrographs of sections in the thyroid gland of rats from the experimental group (Group II): a. Shows thyroid follicles of variable activity as some follicles are markedly distended (1) and other follicles appear involuted (2). These follicles have minimal amount of colloid. H&E X 200. b. Reveals a follicle with interrupted follicular wall (arrow). The adjacent follicles showing thyrocytes with vacuolated cytoplasm (curved arrow). H&E X 400. Nadia M. El-Rouby Fig. 5: Photomicrographs of semithin sections in the thyroid gland of rats from the experimental group (Group II): a. Shows that the follicles have vacuolated colloid (C). The thyrocytes have vacuolated cytoplasm (V) and indented darkly stained pyknotic nuclei (arrow). Toluidine blue X 1000. b. Shows that the destroyed follicles reveal shedded cells into their lumina (encircled).The nuclei of the shedded cells appear irregular and darkly stained. The thyrocytes reveal vacuolated cytoplasm (V). Toluidine blue X 1000. Fig. 6: A Photomicrograph of semithin section in the thyroid gland of a rat from the experimental group (group II). It shows a distorted follicle with shedded cells (S) into its vacuolated colloid (C). These shedded cells have irregular indented darkly stained nuclei. The colloid (C) reveals variable density of staining. The thyrocytes show vacuolated cytoplasm (V). The surrounding stroma is very vascular. Mast cell (M) is seen close to interstitial blood vessel. Toluidine blue X 1000. Fig. 7: A Photomicrograph of TEM section showing two thyrocytes of two adjacent follicles of a rat from the experimental group, the upper left thyrocyte is of a damaged follicle showing dilated rER (d rER). Its nucleus is hetrochromatic and indented (arrows). The lower thyrocyte is related to more or less normal follicle with normal cisternae of rER but with apparently more darkly stained lysosome (L). Note the projecting microvilli (Mv). Original mag X 6000. 218 Fig. 8: A Photomicrograph of TEM section showing two thyrocytes of a thyroid follicle of a rat from the experimental group. It shows dilated cisternae of rER (d rER) with amorphous material (*). The nuclei (N) appear indented with dilated peri-nuclear space (arrows). The nuclei have prominent nucleoli (Nu) and hetrochromatin (h). Original mag X 6000. Fig. 9: A Photomicrograph of TEM section of a thyrocyte from a rat of the experimental group reveals apical microvilli (Mv) projecting into the lumen that has colloid (C). Many darkly stained lysosomes (L) are seen. The cytoplasm shows prominent Golgi complex (G) and cisternae of rER. Original mag X 8000. Fig. 10: A Photomicrograph of TEM section of the thyroid gland from a rat of the experimental group (group II). It shows two follicular cells with flattened nuclei. The nuclei have hetrochromatin. The cytoplasm shows darkly stained lysosomes (L). The colloid (C) is apparently homogenous. Original mag X 2000. A Histological Study on the Effect of Diclofenac Sodium (Declophen) Administration on Thyroid Follicular Cells of Albino Rats Fig. 14: A photomicrograph of TEM section of the thyroid gland from a rat of the recovery group (Group III) showing two thyrocytes with more or less normal rER. Their nuclei appear euchromatic but with some irregularity. The cytoplasm reveals many vacuoles (V) and mitochondria (M). Many electron dense lysosomes (L) are observed. Original mag X 6000. Fig. 11: A photomicrograph of thyroid gland of a rat from the recovery group (Group III). It reveals some recovered follicles. These follicles show vacuolated colloid (V). Other follicles appear distended with colloid (C) and lined with flat epithelial cells. Note that there are still some distorted follicles (D). A forth group of involuted (I) follicles are seen. H&E X 200. Fig. 12: A photomicrograph of semithin section in the thyroid gland of a rat from the recovery group (Group III). It shows that the follicles are filled with homogenous colloid (C) and lined with cuboidal cells that have vesicular rounded nuclei (curved arrows) and some low cubical cells with flattened nuclei (straight arrows). Some mast cells (m) are seen in the stroma. Toluidine blue X 1000. Fig. 15: A photomicrograph of TEM section of a thyrocyte from a rat of the recovery group (Group III) showing two nuclei with prominent nucleoli. The cytoplasm shows vacuoles (V) and electron dense lysosomes (L). Original mag X 6000. DISCUSSION There is a list of medications that alter thyroid function tests. This list include furosemide, NSAIDs (salicylate, diclofenac sodium), heparin, amiodarone and iodinated contrast media12. As NSAIDs are commonly used drugs so thyroid dysfunction is a potentially forgotten complication of NSAIDs therapy5. Fig. 13: A photomicrograph of TEM section of the thyroid gland of a rat from the recovery group (group III). It shows a follicular cell with more or less euchromatic nucleus (N) and prominent nucleolus (nu). The cytoplasm reveals more or less normal cisternae of rER, many vacuoles (V) and some electron dense lysosomes (L) The lumen shows colloid (C) . Original mag X 6000. 219 In the current study, declophen was administered to rats of group II daily for one week to evaluate its effect on the structure and function of the thyroid gland. Group III received declophen for the same duration as group II and this was followed by another week without treatment for recovery to asses the reversibility of these changes. Death of rats that occurred during the experiment was most probably due to liver and/or kidney affection9,13. The present study reinforced the previous recorded results8,14-16 which mentioned that several NSAIDs Nadia M. El-Rouby including diclofenac sodium could lower serum T3 while T4 and TSH were not affected. This might be because NSAIDs block peripheral (tissue) conversion of T4 to T3 and produce a low concentration of T3 as most of T3 is produced from peripheral conversion of T4 to T3 by enzyme 5- deiodinase. The levels of T4 and TSH were not affected. This was most probably because T4 is strongly bound to plasma proteins and its biological half life is long (6-7 days) so subsequently TSH remained without change17. Another cause for insignificant change in T4 level could be because the thyroid gland is a unique gland that stores its hormones extracellular in an amount sufficient for weeks18. NSAIDs including declophen affected the tests of the thyroid gland through alteration in the synthesis, transport, and metabolism of thyroid hormones1,3,5,16. This was proved by both light and electron microscopic findings. At the light microscopic level; some follicles showed degenerative changes in the form of shedded degenerated thyrocytes. The thyrocytes of more affected follicles had vacuolated cytoplasm and small, irregular dark pyknotic nuclei. At the level of TEM; the nuclei of thyrocytes were indented, heterochromatic with widening of the peri-nuclear space. This might be attributed to its connection to dilated cisternae of rER. The rER cisternae were dilated and had intra-luminal electron lucid material of stored unprocessed protein. Also there was apparent increase in dark stained lysosomes. Some follicles revealed variable staining density of colloid. This might indicate variable follicular activities. This finding was supported by Wollman et al.23 who mentioned that density of staining of the accumulated colloid varied from follicle to follicle and correlated the staining reaction of the colloid with the activity of the follicle. Other follicles had vacuolated colloid which might be due to defect in pinocytosis of follicular cell. Thyrocytes of the more affected follicles revealed pyknotic nuclei and vacuolated cytoplasm. This could explain the lowered level of thyroid hormone (T3)14. Ultrastructurally, the follicular cells of the damaged follicles revealed widening of the peri-nuclear space, irregularity and indentation of the nuclei with chromatin condensation, dilatation of rER with intra-luminal electron lucid material, apparently increased darkly stained lysosomes and prominent Golgi complex. Widening of the peri-nuclear space might be due to shrinkage of the nuclei due to the toxic effect of declophen. Also, this might be attributed to its connection to dilated cisternae of rER. The pyknosis of the nuclei and condensation of chromatin observed in thyrocytes of declophen treated rats have been also observed in other tissues as rat liver cells exposed to NSAIDs24. These changes could be degenerative changes. Declophen might induce these degenerative changes either directly or indirectly. This finding was in agreement with Boelsterli25 and Moorthy et al.26 who observed that declophen caused injury of liver cells and they referred this to immune mediated response. Many darkly stained lysosomes were seen in the cytoplasm of thyrocytes of rats administered declophen. The apparent increased number of lysosomes could be explained by the accumulation of highly iodinated thyroglobulin that was resistant to proteolysis27. Marked dilatation of rER was an evidence of disturbed protein synthesis. This dilatation could be due to retention of aberrant protein within their cisternae. Therefore, this might be a form of rER storage disease. This protein could not be processed, folded and transported to appropriate sites. Disruption in protein production might prevent synthesis of apoptosis inhibitors and/or loss of essential proteins involved in cellular homeostasis leading to cellular degeneration28,29. The dilated rER might be the cause of nuclear indentation and irregularity as the dilated rER compressed the nucleus causing its indentation and irregularity. Light microscopic examination of thyroid glands of rats of the experimental group (Group II) revealed marked distortion of follicular structure. Some follicles appeared distended and lined by flat follicular cells due to increased their colloid content that had minimal peripheral vacuolations denoting hypoactivity of these follicles. Other follicles appeared small in size with apparently little colloid. These follicles might be newly formed, collapsed or involuted follicles. A third group of follicles had shedded epithelial cells. These changes could be attributed to cellular distension with accumulated colloid which resulted in cellular disruption and collapse with subsequent collapse of the follicles. The degenerated cells had dark stained nuclei and vacuolated cytoplasm. These findings were recorded in cases of thyroiditis19. Many mast cells were observed frequently in the interfollicular connective tissue beside blood vessels. This finding might be due to an allergic reaction occurred in the thyroid gland as these mast cells secrete cytokines and chemical mediators responsible for allergic reaction. This allergic reaction might be because NSAIDs altered plasma proteins. These NSAIDs- altered proteins could become antigenic. By time these antigens would become immunogens20. Former studies reported that mast cells participate in the process of thyroid hormone secretion and in thyroid gland activity. Mast cells might mediate the action of TSH on follicular cells. TSH has been shown to promote the release of serotonin from thyroid mast cells. Serotonin activates thyroid follicular cells by enhancing them to extend pseudopodia and engulf thyroglobulin from the follicular lumen21,22. Using light microscopy, the thyroid gland of the recovery group revealed that the gland needed more time to return back to normal structure. The thyroid follicles revealed patchy recovery. Some follicles were still large in diameter and distended with colloid with minimal peripheral vacuolations which indicated hypoactivity. Other follicles were smaller in diameter, lined with cuboidal cells mainly and their colloid had some peripheral vacuolations i.e. these follicles regain their activity. Ultrastructurally, the thyrocytes of that group 220 A Histological Study on the Effect of Diclofenac Sodium (Declophen) Administration on Thyroid Follicular Cells of Albino Rats appeared more or less normal with slightly indented euchromatic nuclei and prominent nucleoli. The cytoplasmic organelles were more or less normal in appearance except for the presence of apparently more darkly stained lysosomes. Some thyrocytes revealed variable sized vacuoles in their cytoplasm. This indicated that rat thyroid needed more time to recover completely. 4. 5. 6. Morphometeric study of thyroid follicles of rats of the experimental group (Group II) revealed statistically significant increase in the mean follicular diameter and statistically significant decrease in the mean epithelial height in comparison with the control group (Group I) which denoted hypoactivity of the gland. This was proved histologicaly as many follicles were markedly distended with colloid and lined mainly with flat cells. The colloid was homogenous acidophilic and showed minimal peripheral vacuolations. The less active follicles of the thyroid gland were distended by stored colloid and the lining cells appear flattened against the follicular basement membrane17,30. 7. 8. 9. One week after declophen withdrawal (recovery group; group III), the morphometeric analysis of the thyroid follicles showed insignificant increase in the mean follicular diameter of thyroid follicles and also insignificant decrease in the epithelial height of the lining follicular cells in comparison to the control group. This could be attributed to starting reactivity of the thyroid gland as a result of recovery from the toxic effect of declophen and elimination of harmful waste products from the body. 10. 11. 12. 13. CONCLUSION 14. Declophen, a cytotoxic drug, induced variable structural and functional alterations in the thyroid follicular cells of adult male albino rats. These alterations were incompletely recovered. So, it is advisable to avoid using declophen as first pain killer and if it is used, thyroid function tests’ results should be interpreted cautiously in patients on declophen therapy. The drug should be discontinued prior to evaluation of thyroid function. 15. 16. 17. 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May; 125(1):33-37. 222 دراﺳﺔ ﺗﺄﺛﻴﺮ ﻋﻘﺎر اﻟﺪﻳﻜﻠﻮﻓﻴﻦ ﻋﻠﻰ اﻟﺨﻼﻳﺎ اﻟﻤﺒﻄﻨﺔ ﻟﺤﻮﻳﺼﻼت اﻟﻐﺪة اﻟﺪرﻗﻴﺔ ﻓﻲ اﻟﻔﺌﺮان اﻟﺒﻴﻀﺎء ﻧﺎدﻳﺔ ﻣﺤﻤﻮد ﻣﺤﻤﺪ اﻟﺮوﺑﻲ ﻗﺴﻢ اﻟﻬﺴﺘﻮﻟﻮﺟﻴﺎ ،آﻠﻴﺔ ﻃﺐ اﻟﻘﺼﺮ اﻟﻌﻴﻨﻲ ،ﺟﺎﻣﻌﺔ اﻟﻘﺎهﺮة ﻣﻠﺨﺺ اﻟﺒﺤﺚ ﻳﻌﺪ ﻋﻘﺎر اﻟﺪﻳﻜﻠﻮﻓﻴﻦ ﺣﺪﻳﺜﺎ أﺣﺪ اﻷدوﻳﺔ اﻟﺘﻲ ﻗﺪ ﺗﺆﺛﺮ ﻋﻠﻰ اﻟﻐﺪة اﻟﺪرﻗﻴﺔ وﻗﺪ أﺟﺮﻳﺖ هﺬﻩ اﻟﺪراﺳﺔ ﻋﻠﻰ اﻟﻔﺌﺮان ﻟﻤﻌﺮﻓﺔ ﻣﺪى ﺗﺄﺛﻴﺮ هﺬا اﻟﻌﻘﺎر ﻋﻠﻰ اﻟﻐﺪة اﻟﺪرﻗﻴﺔ ﻣﻦ اﻟﻨﺎﺣﻴﺘﻴﻦ اﻟﺘﺮآﻴﺒﻴﺔ واﻟﻮﻇﻴﻔﻴﺔ وﻣﺎ إذا آﺎﻧﺖ هﺬﻩ اﻟﺘﻐﻴﺮات ﻣﺆﻗﺘﺔ أم ﻻ. أﺟﺮﻳﺖ هﺬﻩ اﻟﺪراﺳﺔ ﻋﻠﻰ ﺛﻼﺛﻴﻦ ﻓﺄرا ،ﻗﺴﻤﺖ إﻟﻰ ﺛﻼث ﻣﺠﻤﻮﻋﺎت ﻣﺘﺴﺎوﻳﺔ ،آﺎﻟﺘﺎﻟﻲ: اﻟﻤﺠﻤﻮﻋﺔ اﻷوﻟﻰ )اﻟﻤﺠﻤﻮﻋﺔ اﻟﻀﺎﺑﻄﺔ( واﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻧﻴﺔ )اﻟﻤﺠﻤﻮﻋﺔ اﻟﻤﻌﺎﻟﺠﺔ( ﻗﺪ ﺗﻢ ﺣﻘﻨﻬﺎ ﺑﺎﻟﻌﻘﺎر ﺑﺠﺮﻋﺔ 8 ﻣﺠﻢ /آﺠﻢ ﻣﻦ وزن اﻟﺠﺴﻢ ﻳﻮﻣﻴﺎ وذﻟﻚ ﻟﻤﺪة أﺳﺒﻮع .أﻣﺎ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻟﺜﺔ )اﻟﻤﺠﻤﻮﻋﺔ اﻟﻤﺘﻤﺎﺛﻠﺔ ﻟﻠﺸﻔﺎء( ﻗﺪ ﺗﻢ ﺣﻘﻨﻬﺎ ﺑﺎﻟﻌﻘﺎر ﻣﺜﻞ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻧﻴﺔ ﺛﻢ ﺗﺮآﺖ ﻟﻤﺪة أﺳﺒﻮع أﺧﺮ ﺑﺪون ﻋﻼج. ﺗﻢ ﺗﺸﺮﻳﺢ اﻟﻔﺌﺮان ﻓﻲ اﻷوﻗﺎت اﻟﻤﺤﺪدة ﻟﺬﻟﻚ آﻤﺎ ﺗﻢ أﺧﺬ ﻋﻴﻨﺎت دم ﻣﻨﻬﺎ ﻟﻘﻴﺎس ﻣﺴﺘﻮى هﺮﻣﻮﻧﺎت اﻟﻐﺪة اﻟﺪرﻗﻴﺔ وآﺬﻟﻚ اﻟﻬﺮﻣﻮن اﻟﻤﺤﻔﺰ ﻟﻬﺎ .و ﺗﻢ أﺧﺬ اﻟﻐﺪة اﻟﺪرﻗﻴﺔ ﻣﻦ اﻟﻔﺌﺮان وﺗﺤﻀﻴﺮهﺎ ﻟﻠﻔﺤﺺ اﻟﻤﻴﻜﺮوﺳﻜﻮﺑﻲ ،آﻤﺎ ﺗﻢ ﻗﻴﺎس ﻣﺘﻮﺳﻂ ﻗﻄﺮ ﺣﻮﻳﺼﻼت اﻟﻐﺪة اﻟﺪرﻗﻴﺔ وآﺬﻟﻚ ﻣﺘﻮﺳﻂ ارﺗﻔﺎع اﻟﻨﺴﻴﺞ اﻟﻄﻼﺋﻲ اﻟﻤﺒﻄﻦ ﻟﻬﺎ وﺣﻠﻠﺖ اﻟﻨﺘﺎﺋﺞ اﺣﺼﺎﺋﻴﺎ ، أوﺿﺤﺖ اﻟﺪراﺳﺔ أن اﻟﺪﻳﻜﻠﻮﻓﻴﻦ ﻗﺪ ﺗﺴﺒﺐ ﻓﻲ ﻧﻘﺺ ذو دﻻﻟﺔ اﺣﺼﺎﺋﻴﺔ ﻓﻲ هﺮﻣﻮن اﻟﻐﺪة اﻟﺪرﻗﻴﺔ اﻟﺜﻼﺛﻲ ) (T3وذﻟﻚ ﻓﻲ اﻟﻤﺠﻤﻮﻋﺔ اﻟﻤﻌﺎﻟﺠﺔ ﺑﺎﻟﻤﻘﺎرﻧﺔ ﻣﻊ اﻟﻤﺠﻤﻮﻋﺔ اﻟﻀﺎﺑﻄﺔ. آﻤﺎ أﻇﻬﺮ اﻟﻤﻴﻜﺮﺳﻜﻮب اﻟﻀﻮﺋﻲ ﺑﻌﺾ اﻟﺘﻐﻴﺮات ﻓﻲ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻧﻴﺔ وذﻟﻚ ﻓﻲ ﺻﻮرة اﻧﺘﻔﺎخ ﻓﻲ ﺑﻌﺾ اﻟﺤﻮﻳﺼﻼت اﻟﺘﻲ آﺎن ﻧﺴﻴﺠﻬﺎ اﻟﻄﻼﺋﻲ ﻣﺴﻄﺤﺎ وﻣﻤﺘﻠﺌﺔ ﺑﺎﻟﻬﺮﻣﻮن اﻟﻐﺮوي واﻟﺒﻌﺾ اﺣﺘﻮى ﻋﻠﻰ ﺧﻼﻳﺎ ﻣﺘﺴﺎﻗﻄﺔ وأﻇﻬﺮت اﻟﺤﻮﻳﺼﻼت اﻷآﺜﺮ ﺗﺄﺛﺮًا ﺗﻐﻴﺮات ﻓﻲ ﺧﻼﻳﺎهﺎ اﻟﻤﺒﻄﻨﺔ وذﻟﻚ ﻓﻲ ﺻﻮرة ﻓﺮاﻏﺎت ﺳﻴﺘﻮﺑﻼزﻣﻴﺔ وﺗﺪﻣﻴﺮ ﺑﺄﻧﻮﻳﺘﻬﺎ وﻟﻢ ﻳﻈﻬﺮ اﻟﻬﺮﻣﻮن اﻟﻐﺮوي ﻓﻲ اﻟﺒﻌﺾ ﻣﻦ اﻟﺤﻮﻳﺼﻼت اﻻ ﺑﻜﻤﻴﺎت ﻗﻠﻴﻠﺔ. وأوﺿﺢ اﻟﻤﻴﻜﺮﺳﻜﻮب اﻹﻟﻜﺘﺮوﻧﻲ اﻧﺘﻔﺎخ ﻓﻲ اﻟﺸﺒﻜﺔ اﻷﻧﺪوﺑﻼزﻣﻴﺔ اﻟﺨﺸﻨﺔ ﻣﻊ وﺟﻮد ﻣﻮاد ﻣﺨﺰﻧﺔ ﺑﺘﺠﺎوﻳﻔﻬﺎ وزﻳﺎدة واﺿﺤﺔ ﻓﻲ اﻟﻠﻴﺴﻮزوﻣﺎت ﻣﻊ ﺗﻌﺮﻳﺞ ﻓﻲ ﺷﻜﻞ اﻟﻨﻮاة وﺑﻘﻴﺖ ﺑﻌﺾ هﺬﻩ اﻟﺘﻐﻴﺮات ﻣﻮﺟﻮدة ﻓﻲ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻟﺜﺔ . وأﻇﻬﺮ اﻟﺘﺤﻠﻴﻞ اﻻﺣﺼﺎﺋﻲ ﻟﻠﻨﺘﺎﺋﺞ زﻳﺎدة ذو دﻻﻟﺔ اﺣﺼﺎﺋﻴﺔ ﻓﻲ ﻣﺘﻮﺳﻂ ﻗﻄﺮ اﻟﺤﻮﻳﺼﻼت ﻓﻲ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻧﻴﺔ ﻣﻊ ﻧﻘﺺ ﻓﻲ ﻣﺘﻮﺳﻂ ارﺗﻔﺎع اﻟﻨﺴﻴﺞ اﻟﻄﻼﺋﻲ اﻟﻤﺒﻄﻦ ﻟﻬﺎ ﺑﺎﻟﻤﻘﺎرﻧﺔ ﺑﺎﻟﻤﺠﻤﻮﻋﺔ اﻟﻀﺎﺑﻄﺔ . وﻳﻤﻜﻦ اﻻﺳﺘﻔﺎدة ﻣﻦ هﺬﻩ اﻟﺪراﺳﺔ ﻓﻲ أﺧﺬ اﻟﺤﻴﻄﺔ ﻋﻨﺪ اﺟﺮاء ﺗﺤﻠﻴﻞ اﻟﻐﺪة اﻟﺪرﻗﻴﺔ ﻓﻲ اﻷﺷﺨﺎص اﻟﺨﺎﺿﻌﻴﻦ ﻟﻠﻌﻼج ﺑﻌﻘﺎر اﻟﺪﻳﻜﻠﻮﻓﻴﻦ. 223