운동훈련이 흰쥐 노화심근에 미치는 영향

박원학(Won-Hark Park),이상선(Sang-Sun Lee),이용덕(Yong-Deok Lee) 대한의생명과학회 1996 Biomedical Science Letters Vol.2 No.1

장기간 반복 주행운동이 흰쥐의 심근에 미치는 효과를 규명하기 위하여 생후 3개월, 10개월 및 20개월된 흰쥐를 운동군과 대조군으로 대별하여 motor driven treadmill을 이용하여 Park등이 사용한 방법에 준해서 5개월만, 주 5일 20분간 운동을 시킨 후 심근의 조직 및 세포학적 변화를 관찰하고 심근 세포 내 미세구조 변화를 입체해석학적으로 비교 분석하였다. 연령 증가에 따라 장기간 반복운동이 흰쥐 심근 미세구조에 미치는 영향은, 8, 15개월의 운동군과 대조군 사이에 뚜렷한 차이를 인정할 수 없으며, 15개월의 운동군에서 대조군에 비하여 변성된 사립체, 리소조옴, 지방적, 공포, 노화색소 등이 증가하는 경향이 있었다. 25개월 운동군은 같은 연령 대조군에 비하여 근원섬유 수축대, 근원섬유 소실, 윤반분리, 세포간질 증식, 핵의 변성, 교원섬유 근섬유내 침입 등 매우 심한 변화를 보였다. 조직상에 나타나는 early lipofusin과 미세구조상에 나타나는 노화색소는 8, 15개월의 운동군과 대조군 사이에 유의한 차이는 없었으며, 25개월 운동군은 같은 연령 대조군에 비하여 유의한 차이로 증가하였다. Glucose-6-phosphatase 활성도 8, 15개월군에서 운동군과 대조군에서 모두 활성이 높았으며 25개월의 대조군과 운동군에서는 모두 활성이 거의 나타나지 않았다. 미세구조 변화를 입체해석학적으로 분석한 결과 8, 15개월의 대조군과 운동군에서 체적 밀도의 모든 항은 양군 사이에 유의한 감소를 나타내었고 근형질세망의 체적밀도는 감소하는 경향을 보였다. 사립체는 대조군에 비하여 유의한 감소를 나타내었고 근형질세망의 체적밀도는 감소하는 경향을 보였다. 사립체와 근원섬유 비는 8개월 운동군에서 유의한 차이는 없지만 대조군에 비하여 높게 나타났으며, 15개월의 운동군과 대조군 사이에서는 차이를 인정할 수 없었다. 그러나 25개월 운동군은 대조군에 비하여 유의한 차이로 감소하였다. 연령증가에 따른 사립체 내막 표면밀도와 사립체수는 대조군과 운동군 사이에는 유의한 변화는 없었다. 본 연구의 성적을 검토한 결과 젊은층(3개월군)과 중령층(10개월군)의 흰쥐에서는 반복된 지구력 운동이 심장에 미치는 역효과를 인정할 수 없었으며 젊은층의 흰쥐에서는 오히려 심장기능 강화를 보이는 경향이 나타났으며, 노화층(20개월군)에서 운동군에서는 스트레스로 작용하여 심장기능의 저하를 초래 하였다고 생각된다. There is considerable current interest in the effect of regular vigorous exercise and in particular endurance-running as a possible measure in improving myocardial function. Some data indicate that the aging heart may actually suffer from vigorous endurance exercise. On the contrary, appropriate exercise in aged animals improves myocardial function and aerobic energy metabolism. So far there is relatively little data to indicate that endurance exercise is in fact beneficial in improving myocardial function or damaging to heart of aged animals. The present investigation aimed to study the possible effect of a long range treadmill training program on the heart in aging rats. Male rats aged 3, 10, and 20 months were divided at random into a control (sedentary) and an exercise group. The training group was exercised for 5 days a week on an automated treadmill for 20minutes at 18m/min over a period of 5 months. The exercise regimen of our experiments did not cause any significant changes in the tissues and ultrastructural as compared with sedentary age-matched control. Tissues and ultrastructures of myocardial cells in trained group aged 8 months are intact and well organized as well as sedentary control group. Age associated tissue and ultrastructural changes of trained group aged 15 months included : an increase in transformed mitochondria, vacuoles, lysosomes, lipid droplets and early lipofuscin. But the trained heart did not show significant difference in tissue and ultrastructural properties from those of sedentary controls. Endurance-trained group aged 25 months showed significant qualitative tissue and ultrastructural difference as compared with age-matched controls. In addition to those found in 25 months control group, focal necrosis, myofibril fraying, hypercontraction band, seperation of intercalated discs, degenerating nucleus and infiltration of collagenous fiber into myocyte were noted in trained 25 months group. The stereological examination of the micrographs disclosed no significant difference in the myofibril, mitochondrion, sarcotubule and interstitium volume density and surface density of mitochondrial cristae and numerical density of mitochondria between trained and control group aged 8 and 15 months. In the trained 25 months group, significant increase in volume density of interstitium, lipofucsin granule were shown as compared to untrained age-matched control. On the other hand, significant decrease in mitochondrion volume density was shown. The myofibril volume density did not differ between trained and control group although trained group showed slight increase. From the data obtained, a reduced mitochondria/myofibrils ratio was found in trained rat heart aged 25 months and there was no difference between trained and control rat aged 15 months. But a slight but not significant increase was found in the trained group aged 8 months as compared with same age control group. Such increase in the ratio in young animals is considered to be of great importance to cardiac pumping and adaptability. Whereas such adaptations don't seem to occur in aged heart muscle. This study proposed that repeated endurance exercise do not cause any significant qualitative and quantitative ultrastructural change of heart muscle in young(3months) and adult(10months) suggesting that the heart is able to adapt to the exercise. On the contrary, the repeated endurance exercise stress may actually induce degenerative changes in the aged heart muscle(20months).

Adriamycin이 생쥐 심근 미세구조 및 Glutathione-Glutathione Peroxidase계에 미치는 영향

박원학,정형재,김쌍용,이용덕,최정목,Park, Won-Hark,Chung, Hyeung-Jae,Kim, Ssang-Yong,Lee, Yong-Deok,Choi, Jeung-Mog 한국현미경학회 1989 Applied microscopy Vol.19 No.2

The cardiotoxic effects of acute and chronic administration of adriamycin (ADR) were evaluated in A/J Swiss albino mice. In acute studies, male mice received intravenous ADR, 5mg or 15mg/kg per day for 3 or 1day and were sacrifice 12 hours later. Because the glutathione-glutathione peroxidase system is major pathway for free radical detoxication, glutathione levels and glutathione peroxidase activity was measured. In acute studies, ADR-treated mice exhibited significantly decreased levels(p<0.05) of total glutathione and unchanged levels of oxidized glutathione and percentage of oxidized glutathione. The earliest myocardial fine structural alterations included swelling and degeneration of mitochondria and dilatation of sarcoplasmic reticulum at all dosage of acute models. In chronic studies, mice received 5mg/kg ADR once a week for up to 16 weeks. Levels of total and reduced glutathione were decreased significantly(p<0.01) and oxidized glutathione and percentage of oxidized glutathione were increased significantly (p<0.05). Chronic myocardial lesions included perinuclear vacuolization, seperation of myofibrils and the fasciae adherens of intercalated disc and hypercontraction band within myocyte. Glutathione peroxidase activity reduced significantly (p<0.01) in any group of acute and chronic ADR-treated animals. Test for lipid peroxidation(malondialdehyde) was increased significantly(P<0.01). Thus, we conclude 1) ADR significantly lowers glutathione levels in heart tissue, and 2) cellular damage progress produced by alteration of this system in mouse models of ADR cardiotoxicity. These results suggest that the glutathione-glutathione peroxidase system may be involved in the modulation of ADR-induced cardiotoxicity.

Rat 태생기의 심장성장에 따른 형태측량적 연구

박원학,이용덕,정형재,최정목,Park, Won-Hark,Lee, Yong-Deok,Chung, Hyeung-Jae,Choi, Jeung-Mog 한국현미경학회 1989 Applied microscopy Vol.19 No.2

The ventricular myocardia of 14, 16, 18 and 20-day-old rat fetuses and newborns have been studies by light and electron microscopic morphometrics. The volume density of the myocyte and interstitial compartments as well as volume, surface and numerical density of nuclei were estimated by light microscopic morphometrics. Whereas, the volume density of myofibrils and glycogen granules as well as the volume, surface and numerical density of mitochondria were assessed by electron microscopic morphometrics. The volume density of myocyte compartment of the ventricular myocardia in developing fetuses decreased, but increased in newborn rats. On the other hand, the volume density of the interstitial compartment increased in growing fetuses and decreased in newborns. In all groups the volume, surface and numerical density of nuclei decreased gradually with elongation of myocytes. Conversely, the volume, surface and numerical density of mitochondria and volume density of myofibrils and glycogen granules in ventricular myocytes incresed. The increase in numerical density of mitochondria probably reflects an increase in metabolic activity. Sarcomere length also increased during development.

외적요인으로서의 Chlorambucil 투여가 흰쥐 심근 노화에 미치는 영향 II. 세포화학 및 생화학적 연구

박원학,정형재,Park, Won-Hark,Chung, Hyeung-Jae 한국현미경학회 1992 Applied microscopy Vol.22 No.1

The present study was investigated to elucidate the effects of chlorambucil the heart tissue of various-aged rats. The male rats ranging from 3 to 36 months were used. The cytochemical and biochemical changes in myocardium of the rats were studied in the aspect of free radical roles in aging process. With the goals of evaluating the potential roles of free radicals in aging process, evidence was shought for alterations of myocardial lipid peroxide levels in control and chlorambucil treated rats. The result are summarized as follows: 1. Cytochemical studies showed that the activities of $Mg^{++}$-ATPase and succinic dehydrogenase increased with age. However, these enzyme activities were decreased with treatment of chlorambucil, when compared with control group. Interestingly it was observed that chlorambucil treatment increased the activity of acid phosphatase from 6 months upto 18 months, and decreased after 18 months. 2. The lipid peroxide level in myocatdium was increased with age; chlorambucil-treated group was higher than that of control group. 3. Age-dependent increase in activities of monoamine oxidase, xanthine oxidase and catalase was observed. But the increase of catalase activity was higher than that of monoamine oxidase and xanthine oxidase activity in control group. However, in chlorambucil-treated group, age-dependent decrease of these enzyme activities was observed, and catalase activity was more significant particularly with regard to other enzymes. In consequently, the morphological alterationsof myocardium due to chlorambucil treatment was exclusively observed. We demonstrate that this alteration is occured by lipid peroxidation upon chlorambucil treatment.

노화에 따른 Rat 기관상피의 세포화학적 및 전자현미경적 연구

박원학,최정목,Park, Won-Hark,Choi, Jeung-Mok 한국현미경학회 1994 Applied microscopy Vol.24 No.1

The present studies were designed to determine the feasibility of using the rat tracheal epithelium as models for induction of aging. The ultrastructural and cytochemical changes of tracheal epithelium were investigated in rats at ages of five, twelve and twenty four months. Some major changes in the tracheal epithelium with advancing age were observed by electron microscopy. The results were summarized as fellow: 1. With the advance of age, lysosome, vacuole and multivesicular bodies were increased in number and numerous myelinoid bodies were observed in cytoplasm of ciliated cells. 2. In goblet cell, serous cell and brush cell lysosome and myelinoid bodies were increased in number with the advance of age, and an myelinoid bodies was often found within the secretory granule. 3. Cytochemical studies showed that acid phosphatase activities was observed in multivesicular bodies and lysosome, strong activities with the advance of age. And alkaline phosphatase activity are observed in microvilli, granule and lateral membrane of secretory granule cells, and strong activities with age. Consequently suggest that with the advance of age, tracheal epithelium show ultrastructural and cytochemical alteration of some kind of cell organelles in all kind of cell.

외적요인으로서의 Chlorambucil 투여가 흰쥐 심근 노화에 미치는 영향 I . 미세구조 및 형태측량적 연구

박원학,정형재,이용덕,최정목,박채수,조현국,Park, Won-Hark,Chung, Hyeung-Jae,Lee, Yong-Deok,Choi, Jeung-Mog,Park, Chae-Soo,Cho, Hyun-Gug 한국현미경학회 1990 Applied microscopy Vol.20 No.2

This study was investigated to elucidate the effect of chlorambucil on the heart in aging rats. Male rat ranging in age from 3 to 36 months were used. Each age groups(3, 6, 12, 18, 24, 36 months) included control and chlorambucil treated rats. As a part of the study, the ultrastructural changes in the left ventricular myocardial cells were described by using the qualitative and morphometric techniques. The results are summarized as follows. Age associated ultrastructural changes included: an increase in vacuoles, protrusion of plasma membranes, lipid droplets, and lipofuscins in myocardium of control groups. These changes which begin to occur at 12 months and continue through 36 months. At the 36 months some instance of unusual formation of contraction band and separation of intercalated discs were encountered. Morever, these changes and contents with chlorambucil treatment were remarkably increased in comparison with control groups. Age-dependent changes of control group measured with morphometry were not observed in the volume densities of mitochondria and myofibrils. But there was increase in interstitium. On the other hand decrease in sarcoplasmic reticulum and T-tubule system. In chlorambucil treated groups, volume densities of mitochondria and interstitium were increased in comparison with those of the control groups. But sarcoplasmic reticulum and T-tubule system were remarkably decreased.

발달중인 Mongolian gerbil의 전뇌에서 TH 면역반응세포의 분포에 관한 연구

이경열,박일권,권효정,박미선,송치원,김무강,장규태,이강이,김원식,박원학,Lee, Kyoung-youl,park, Il-kwon,Kwon, Hyo-jung,Park, Mi-sun,Song, Chi-won,Kim, Moo-kang,Chang, Kyu-tae,Lee, Kang-iee,Kim, Won-sik,Park, Won-hark 대한수의학회 2001 大韓獸醫學會誌 Vol.41 No.2

The immunocytochemical localization of tyrosine hydroxylase(TH) is examined in the developmental forebrain of mongolian gerbil in order to determine changes in the distribution and cytology of neurons. At each of the different developmental stage, including prenatal(E)14, E16, E18, E20, postnatal(P)0, P2, P4 days, mongolian gerbils were sacrificed. In E14, TH-IR cells were predominantly round or oval in shape and their processes were very short. In olfactory blub, TH-IR cells were begun to appear from E20. In the striatum, we observed only fibers of TH-IR at all ages. From E16, TH-IR perikarya and fiber were present in periventricular nucleus and paraventricular nucleus of hypothalamus. The changes from the early to the late prenatal stages of development appear to be the result of an increase in number of TH-IR perikarya and processes. These results were shown slight differences to other rodents.

흰쥐 골격근의 노화성 위축에 대한 운동훈련의 영향 - II. 장기간에 걸친 체중부하 훈련의 영향 -

박승한,박원학,이용덕,김정기,Park, Sung-Han,Park, Won-Hark,Lee, Yong-Deok,Kim, Jung-Ki 한국현미경학회 1995 Applied microscopy Vol.25 No.4

The present study was designed to examine effect of long term weight-training on aging atrophy in the rat skeletal muscle. Male rats of 8, 15, and 24 month old were used. Each age groups included control and weight-training for 5 months by using body press apparatus. The histo- and cytochemical, ultrastructural and stereological changes in aging skeletal muscles of the rat were observed in the present study. During the training period the body weight and muscular weight in all groups except the rectus femoris and the gastrocnemius in young age groups remained constant, but muscular weights were increased in the rectus femoris and the gastrocnemius muscles in young age groups. In trained rat, the volume density of muscle fiber type IIA and IIB were increased, but those of type IIC was decreased. Type I remained constant in 8 and 15 month old age groups, but reduced in the tibialis anterior and the gastrocnemius muscles in the 24 month old groups. Some histotological and ultrastructural changes associated with age were found: numerical increase of cytiplasmic vacuoles, lysosomes, lipofuscins, and irregularity of myofibrils. At 24 month old groups some unusual formation of contraction band and muscle splitting were observed. After weight-training, ultrastructural degenerative changes occured in the type I muscle fiber, such as splitting of muscle fiber, disorganization of myofilaments, swelling of mitochondria, accumulation of many lipid droplets, appearance of many lysosomes and residual bodies and necrotic fibers, in the old age groups. But, in the type II muscle fibers hypertrophy of muscle fiber appeared without any noticible damage as the type I. The activities of $Mg^{++}$ -ATPase decreased with age and this enzyme activities in the trained rat were significantly decreased with age. Activities of the acid phosphatase were increased with age and significantly in the trained rat. In stereological analysis, volume density of the myofibrils and the tubular system were increased, on the other hand there mitochondrial capacity was decreased. These experimental results suggested that old rats are not susceptible to be affected by weight-training as young rats, and that physical capacity of the rats must be considered when old rats are exercised for training.

도롱뇽(Hynobius leechii) 정충의 미세구조(Amphibia, Urodela)

김구환,박원학,이영환,Kim, Kgu-Hwan,Park, Won-Hark,Lee, Young-Hwan 한국현미경학회 1995 Applied microscopy Vol.25 No.1

The ultrastructure of spermatozoa in Hynobius leechii was examined with transmission electron microscope and compared with those of other urodeles and anurans. The perforatorium and the tail show most of the common feature of urodeles. However, there were several ultrastructural characteristics in an acrosome, nucleus, neck, axial rod and mitochondria related to the nucleus. The acrosome was trifoliate in transverse sections and the perforatorium consisted of two different concentric parts with a fine sharp point in a subacrosomal lumen. The nucleus consisted of two different regions of chromatin area and nuclear ridge. The nuclear ridge was composed of several bundles of five to ten minute tubular subunits of 19 nm diameter in this species, while in higher urodeles it was well developed in multi layers. The protoplasmic bead was separated by a cytoplasmic canal except the connection with only the distal portion of the nucleus. The neck was a short cylinder and contained pericentriolar material with transverse striations. In Hynobius the ring was not elongated to the tail and the mitochondria were distributed only in the protoplasmic bead around the nucleus as in Cryptobranchus, while in higher groups of urodeles it elongates to the length of the middle piece and the mitochondria follow the ring. The ring elongation may be related to the distribution of mitochondria. Hynobiidae and Cryptobranchidae are closely related based on structure of neck, the axial rod and location of mitochondria, although they are different in the composition of the axial rod.

피부화상으로 유도된 심근손상에서 Protein Kinase C의 역할

문혜정,조현국,박원학,Moon, Hye-Jung,Cho, Hyun-Gug,Park, Won-Hark 한국현미경학회 2003 Applied microscopy Vol.33 No.4

본 연구는 피부화상으로 유도된 심근손상에서 protein kinase C (PKC)의 역할을 알아보고자 하였다. 수컷 흰 쥐(SD계)에 15%의 피부전층화상을 유도한 뒤, PKC activator인 phorbol 12-myristate 13-acetate (PMA)와 PKC inhibitor인 bisindolylmaleimide (BIS)를 투여하여 5시간, 24시간 후에 심장을 적출하여 생화학적 미세구조적 입체해석학적 방법을 실시하였다. 혈청 AST와 creatinine 은 화상 후 5시간군과 화상 후 5시간+BIS 투여군에서 높게 나타났고, KC와 MPO 활성은 PMA 투여군이 BIS 투여군보다 낮게 나타났다. 미세구조적 관찰 결과 PMA 투여군에서는, 화상으로 인한 핵의 분열, 과수축대 형성, 사이원반의 분리 현상이 다소 완화된 형태로 관찰되었고, BIS 투여군에서는 화상 단독군에서 나타나는 형태적 변화 뿐만 아니라 비정상적인 형태의 사립체도 일부 관찰되었다. 전체적으로 5시간에서 24시간군으로 가면서 손상이 완화된 양상을 나타내었다. 입체해석학적 결과에서는 화상으로 인한 근원섬유의 체적밀도 감소가 PMA와 BIS 투여로 인해 증가되었고, 사립체의 체적밀도와 수밀도의 증가는 BIS군에서 가장 높게 나타났다. 결론적으로 PKC의 활성화는 화상으로 인해 손상된 심근에서 염증반응을 감소시켜 심근 손상을 보호한다고 사료된다. The aim of the present study was to assess the role of protein kinase C (PKC) in the development of cardiac injury following scald burn. Sprague-Dawley rats were induced a scald burn a 15% total body surface area. Phorbol 12-myristate 13-acetate (PMA, 2 mg/kg) and bisindolylmaleimide (BIS, 0.05 mg/kg) were immediately administered i.p. after burn injury. 5 h and 24 h later, heart was removed and examined biochemical assay, ultrastructural changes and stereological analysis. The activity of serum aspartate aminotransferase was significantly increased at 5h (p<0.01) and 5h+BIS (p<0.001) after burn compared with that of control. The activity of serum creatinine was significantly decreased in PMA-treated groups after burn compared with postburn 5 h. PMA caused a decrease in MPO activity and induced wavy fibers in cardiac myocytes at postburn 5 and 24h. BIS induced contraction band, separation of intercalated disk and abnormal mitochondria in cardiac myocytes at postburn 5 and 24h. In stereological analysis, treatment of rats with PMA increased volume density of myofibril and mitochondria compared with postburn 5 and 24h. Our data suggest that the activation of PKC in scald burned heart decreases inflammation and protects the myocardium.