Cold Stress가 노화(老化) 흰쥐 심근(心筋)에 미치는 영향(影響)

김정기,정형재,이용덕,박원학,Kim, Jeong-Ki,Chung, Hyeung-Jae,Lee, Yong-Deok,Park, Won-Hark 한국현미경학회 1998 Applied microscopy Vol.28 No.1

The present study was performed to determine the effect of cold stress on myocardium of aging rat. Control groups, which aged 6, 12 and 24 months, were compared with age-matched experimental groups that were exposed to moderate cold stress for a hours daily in a week at laboratory cold room $(4{\pm}1^{\circ}C)$. The histological, histochemical and ultrastructural changes of myocardium were observed. The results were summarized as follow: 1. Age-dependent histological change of control groups was observed the formation of contraction band in 24months aged group. The experimental groups submitted to cold stress showed a similar change pattern as seen in control groups. However, the degree of change in the experimental groups was significantly larger than that of control groups. In the 34 months aged group the formation of hypercontraction band was observed. 2. Regarding age-dependent histochemical changes of control groups, we observed the increase activities of PAS and Masson's trichrome. In experimental groups the activities of PAS and Masson's trichrome were also increased with age. Compare with control group, the activities of PAS was increased but the activities of Masson's trichrome was decreased. 3. Age-dependent ultrastructural changes on vacuolization, lysosome were observed. In control groups the structural changes occur at 12 months. The accumulation of lipofuscin, contraction band, hypercontraction band and a component of connective tissue were observed in 24 months. However, the degree of change in the experimental groups was significantly larger than that of control groups. In contract, the myelin body in intercalated discs was observed in 24 months of experimental groups.

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.

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.

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.

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.

식이중의 Methionine이 흰쥐의 체내 지질과산화와 간 미세구조에 미치는 영향

서정숙,양경미,박원학,정형재,이용덕,Seo, Jung-Sook,Yang, Kyung-Mi,Park, Won-Hark,Chung, Hyeung-Jae,Lee, Yong-Deok 한국현미경학회 1991 Applied microscopy Vol.21 No.1

To study the effects of dietry methionine level on lipid peroxidation of rats, rats were fed vitamin E, selenium and methionine - deficient diet or the same diet supplemented with various levels(0.3%, 0.6%, 0.9%) of methionine for 6 weeks. The biochemincal and mophological changes in the rat liver were investigated. Lipid peroxide levels in plasma and hepatic mitochondrial fraction of MF rats were more increased than those of control rats. However, supplementation with 0.6% methionine modified this increment. Catalase activity was decreased in hepatic mitochondrial fraction from rats fed MF diet. Methionine supplementation did not induce this enzyme. The ultrastructural evidence for lipid peroxidation was found in plasma membranes facing sinusoids. The most striking changes in including disruption and loss of microvilli and development of numerous lipid droplets occurred in rats fed MF diet. These changes were not effectively prevented by the same diet supplemented with 0.3% or 0.9% methionine, but supplementation with 0.6% methionine modulated more or less the changes.

장기간 체중부하 운동훈련이 흰쥐 노화 심근에 미치는 영향

조현국(Hyung Gug Cho),이영실(Young Sil Lee),정형재(Hyeung Jae Chung),이용덕(Yong Deok Lee),박원학(Won-Hark Park) 대한의생명과학회 1998 Biomedical Science Letters Vol.4 No.2

장기간 체중부하운동아 흰쥐의 심근에 미치는 효과를 규명하기 위하여 생후 3개월, 10개월 및 20개월된 흰쥐를 5개월간 운동 시킨 후 각각의 비운동군과 비교하여 심근의 조직학적ㆍ세포학적 변화를 관찰하고, 심근세포내 미세구조적 변화를 입체해석학적으로 비교ㆍ분석하였다. 운동 15개월군부터 노화색소의 양이 현저히 증가되었고 사립체의 변성, 공포화 현상 그리고 간질조직의 증식이 뚜렷하였으며, 25개월군에서는 윤반분리, 근원섬유의 소실 및 과수축대 출현빈도가 증가되었다. Glucose-6-phosphatase의 활성도는 15개월 운동군에서 감소하여 25개월 운동군에서는 활성도가 거의 나타나지 않았다. 업체해석학적 결과에서도 장기간 체중부하운동으로 인해 15개윌, 25개월군에서 모두 사립체와 근원섬유의 체적밀도는 감소되었고, 간질조직의 체적밀도는 증가되었다. 본 연구의 결과로 볼 때 장기간 체중부하운동은 3개월군에 있어서 심장에 미치는 역효과가 없으나 10개월군과 20개월군에서는 심근 기능강화에 역효과를 초래할 우려가 큰 것으로 판단된다. There is considerable interests in the effect of regular, vigorous exercise, in particular weight training as a possible measure in improving myocardial function. The present investigation aimed to examine possible effect of a long-term weight training program on the heart in aging rats. Male rats aged 3-, 10-, and 20-month-old were divided at random into a control (sedentary) and the exercise group. The training group was exercised for 5 days a week by 1 RM of ten times with weight training apparatus. This investigation examined the changes of the heart muscle relative to histological, ultrastructural, cytochemical, and stereo logical studies in rats. Quantity of lipofuscin pigments was clearly increased in the weight training group of 15-month-old rats, and mitochondrial degeneration, vacuolization, and interstitial proliferation were observed as well. In the weight training group of 25-month-old rats the frequency as separated intercalated discs, fraying myofibrils and hypercontraction band increased in number compared with the same 15-month-old group. From the experimental result of glucose-6-phosphatase activity, the enzyme activities decreased in the weight training group of 15-month-old rats, and more decreased in the same 25-month-old group. In stereological study, both 15-and 25-month-old training groups, mitochondrial and myofibrilar volume densities significantly decreased, whereas interstitial volume density significantly increased. From the experimental results obtained in the present study, it is suggested that long-term weight training exercise do not cause any significantly qualitative and quantitative ultrastructural change of the heart muscle in the young. On the contrary, long-term weight training exercise stress may actually induce degenerative changes in the heart muscle in the old age.

흰쥐 골격근의 노화성 위축에 대한 운동훈련의 영향

박승한(Sung Han Park),박원학(Won-Hark Park),정형재(Hyeung-Jae Chung) 대한의생명과학회 1996 Biomedical Science Letters Vol.2 No.1

본 실험은 골격의 위축이 일어난 노화 흰쥐를 사용하여 단기간동안 트레드밀 훈련과 웨이트 트레이닝을 시켰을 때 골격근에 나타나는 변화들을 고찰하고자 고안하였다. 출생 후 24개월 된 수컷 흰쥐를 사용하여 대조군과 트레드밀 훈련군, 웨이트 트레이닝군으로 구분하여 각 군별로 4주간 훈련시켜 각 군의 골격근은 조직 및 조직화학, 미세구조 및 세포화학적 연구 및 입체해석학적 연구에 적용하였다. 각 훈련 기간동안 개체의 체중과 근육의 무게는 큰 변화가 없었고, 근섬유의 분포양상은 트레드밀 훈련군에서는 ⅡB형과 ⅡC형이 증가를 한 반면 ⅡA형은 감소를 하였고 Ⅰ형의 근섬유 조성은 변화가 없었다. 웨이트 트레이닝 후에는 ⅡA와 ⅡB형이 증가를 한 반면 ⅡC형은 감소하였다. 그러나 모든 근섬유 조성의 변화에는 유의성이 없었다. 노화 대조군의 조직구조 및 미세구조의 특징은 노화색소의 출현, 약한 수축대의 출현, 근섬유들의 분할현상 등이 관찰되었다. 트레드밀 훈련 후에는 손가락형의 돌출부를 포함한 근초의 불규칙성이 나타났으며, 밝은띠의 확장으로 인한 Z-선의 불규칙한 배열, 공포화, 장축과 평행한 근섬유의 분할현상이 나타났으며, 웨이트 트레이닝 후에는 많은 용해소체들과 함께 포식소자를 함유한 자가용해소체들이 나타났으며 심한 수축대가 나타났고 근원섬유들의 배열이 와해되는 현상도 나타났다. 조직화학 및 세포화학적인 측면에서 succinic dehydrogenase와 acid phosphatase는 활성도의 변화가 거의 없었으며, Mg<SUP>++</SUP>-ATPase의 활성은 훈련 후 약간 감소하는 경향이었다. 미세구조의 입체해석학적 분석에서는 측량한 구조물들의 체적밀도와 수밀도는 변화가 거의 없었고 사립체 내막의 표면밀도만 트레드밀 훈련군에서 증가되었다. 이상의 결과에서 볼 때 단기간의 지구력 훈련을 실시할 경우는 노화 골격근은 잘 적응하여 골격근의 위축 방지에 효율적이었으나 웨이트 트레이닝은 노화 골격근의 위축방지에 영향을 주지 못하였다. The present study was designed to examine effect of short term treadmill and weight-training on aging atrophy in the rat skeletal muscle. Male rats of 24 months old were used. Each groups included control, treadmill and weight-training for 4 weeks by using treadmill apparatus and body press apparatus. The histo and cytochemical, ultrastructural and stereological changes in senile skeletal muscles of the rat were observed in the present study. During the training period the body weight and muscular weight in all groups remained constant. The volume density of muscle fiber type ⅡC and ⅡB were increased, that of type ⅡA was decreased, but type Ⅰ remained constant in treadmill-training group. In weight-training rat, the density of type ⅡA and ⅡB were increased, but those of type ⅡC was decreased. But, all changes of muscle fiber type is not significant. Senile control group some usual formation of mild contraction band, lipofuscin pigment and muscular splitting were observed. After treadmill-training, histological and ultrastructural changes occured in the muscle fiber, such as irregularity of the sarcolemma, interfibrillar vacuolization, longitudinal splitting, and widened I-band. After weight-training, the changes occured in the trained muscle fiber, such as apearance of many lysosomes and autophagic vacuoles, severe contraction band, and breakup of myofibrils. Histo and cytochemical studies showed that the activities of succinic dehydrogenase and acid phosphatase remained constant, activities of Mg<SUP>++</SUP>-ATPase decrease with training. Stereo-logical changes were not observed in the volume and numerical density of all subject component, but the surface density of mitochondrial inner membrane was increased with treadmill-training. These experimental results suggested that the endurance training during short-term may result in the adaptible response in senile skeletal muscles. On the other side, weight-training is bad for senile skeletal muscle.

화분의 영양생화학적 연구 - Aniline이 유도한 간 독성에 미치는 진달래 화분의 영향 -

권정숙(Chong-Suk Kwon),조수열(Soo-Yeul Cho),정형재(Hyeung-Jae Chung),박종민(Jong-Min Park),허근(Keun Huh) 한국식품영양과학회 1989 한국식품영양과학회지 Vol.18 No.3

진달래 화분의 해독 작용을 연구할 목적으로 aniline을 model 약물로 하여 간 조직에 대한 병태 생리학적 동태를 대사 효소의 활성과 관련지어 검토하였다. 진달래 화분 각 추출물을 투여한 군이 대조군에 비해 간 aniline hydroxylase의 활성이 증가되었으며, 진달래 화분 물 추출물 투여에 의한 간 aniline hydroxylase 활성 증가를 동력학적인 측면에서 관찰하였을 때 Km치는 변하지 않았으나 Vmax가 증가하는 점으로 보아 본 효소의 단백 합성이 촉진될 결과라고 사료된다. 또한 aniline만 투여한 군에 비해 진달래 화분 각 추출물을 투여한 실험군이 aniline의 혈중 농도와 간 손상을 현저히 감소시켰고, 이러한 작용한 물 추출물에서 가장 강하게 나타났다. 이로써 진달래 화분 투여에 의한 aniline의 간독성 예방 효과는 진달래 화분 추출물에 의한 간 aniline hydroxylase의 활성증가와 관련되어 나타난 것으로 사료되어진다. Preventive effect of azalea pollen extracts against aniline-induced hepatic toxicity in mice was investigated in this experiment. When the biochemical and histological changes were measured, preventive effect was more striking by treatment with water extract. After treatment with azalea pollen extracts, hepatic microsomal aniline hydroxylase activity increased as compared to control. Whereas, aniline level in serum and liver significantly decreased. The Vmax value without affecting Km value increased by the water extract treatment, the results obtained suggest that the characteristics of increase in the aniline hydroxylase activity may include induction of enzyme proteins. These data indicate that the observed preventive effects of azalea pollen extracts against hepatotoxicity is due to the induction of aniline metabolizing enzyme.

노화 흰쥐 간에 미치는 비타민 A 급성과다 효과 ㅣ.조직학적 연구와 Alkaline phosphatase 활성

조현국,박원학,정형재 嶺南大學校 基礎科學硏究所 1993 基礎科學硏究 Vol.13 No.-

급성 비타민 A 과다가 노화과정의 진행정도(3, 6, 20개월)에 따라 간장에 미치는 영향을 형태학 및 효소화학적으로 관찰하였다. 대조군에 대한 광학현미경적 변화로 3, 6개월군은 대체로 그 형태가 잘 보존되어 있으나 20개월군에서는 동양혈관의 확장, 불규칙한 간세포삭, 국소적 세포팽창과 염증세포의 침윤이 관찰하였다. 비타민 A 투여 3개월군에서의 광학현미경적 변화로, 간소엽의 주변부변성이 관찰되었으며, 6개월군에서는 중심정맥과 간정맥 내막의 섬유증식, 그리고 섬유모세포와 성상세포의 중식이 국소적으로 형성되었다. 20개월군에서는 소엽 전반에 걸친 동양혈관의 확장, 간정맥 경계부의 섬유중식, 중심정맥과 동양혈관의 심한 섬유증식에 기인된 동양혈관 내막에서의 적혈구 정체 및 세포질의 소실도 관찰되었다. 효소활성의 변화로, 혈액중의 ALT, AST는 변화가 없는 반면, ALP 활성은 비타민 A 투여 20개월군에서 증가하는 경향을 보였다. The effect of acute hypervitaminosis A on liver tissue, obtained from 3,6, 20months of rats, were studied by analyzing morphological changes and serum enzyme activities. In light microscopic results of control groups(3,6 months), the tissue structure is preserved well, whereas from 20 months old group, dilatation of sinosoids, irregular hepatic cell cord, and focal cell swelling and infiltration of inflammatory cell were observed. From the 3 months groups treated with vitamin A, peripheral degeneration in the hepatic lobule was observed. From the 6 months groups, fibrosis of central and portal veins, and formation of focal proferation of fibroblasts and Kupffer cells. The dilatation of sinusoids in the whole liver tissue, fibrosis of continuity of liver tissue in adjacent lobule, and focal stagnated red blood cells(caused from the fibrosis of central vein and sinusoids) with cytoplasmic loss were observed from 20 months group. The activity of serum ALP level in 20 months group treated with vitamin A, compared with 3, 6 months groups, was increased, while the changes of serum ALT and ASt activities were insignificant.