Original Articles Korean Circulation J 2000;30 1 :56-65 협심증환자의혈장염증성지표및 T- 림프구의활성화에관한연구 이원하 2 이윤 2 김종란 1 이성윤 1 정진옥 1 권현철 1 박승우 1 김준수 1 김덕경 1,2 이상훈 1 홍경

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1 Original Articles Korean Circulation J 2000;301:56-65 협심증환자의혈장염증성지표및 T- 림프구의활성화에관한연구 이원하 2 이윤 2 김종란 1 이성윤 1 정진옥 1 권현철 1 박승우 1 김준수 1 김덕경 1,2 이상훈 1 홍경표 1 박정의 1,2 서정돈 1,2 이원로 1 Plasma Inflammatory Markers and T-Lymphocytes Are Activated in Angina Pectoris Won-Ha Lee, PhD 2, Yoon Lee, MS 2, Jong-Ran Kim, BS 1, Sung-Youn Lee, MD 1, Jin-Ok Jung, MD 1, Hyeon-Cheol Gwon, MD 1, Seung Woo Park, MD 1, June Soo Kim, MD 1, Duk-Kyung Kim, MD 1,2, Sang-Hoon Lee, MD 1, Kyung Pyo Hong, MD 1, Jeong-Euy Park, MD 1,2, Jung Don Seo, MD 1,2 and Won Ro Lee, MD 1 1 Division of Cardiology, Department of Medicine, Sungkyunkwan University, School of Medicine, Cardiac and Vascular Center, Samsung Medical Center, 2 Clinical Research Center, Samsung Biomedical Research Institute, Seoul, Korea ABSTRACT Background and ObjectivesInflammation and activation of immune cells play important roles in the pathogenesis of atherosclerosis. We investigated the activation status of plasma inflammatory markers and immune cells in angina patients. MethodsWe analyzed the plasma level of C-reactive protein CRP as a marker of inflammation in 24 patients with angina pectoris 12 unstable angina, 12 stable angina, and 12 normal subjects. The degree of activation of peripheral blood monocytes was assessed by Northern analysis of pro-atherogenic cytokines and the activation status of T-lymphocytes was measured by flow-cytometric analysis of HLA-DR expression on T-cells. ResultsPlasma level of CRP was highest in unstable angina patients mg/l and lowest in the control subjects mg/lp0.03. We also observed a high correlation between CRP level and the occurrence of minor and major coronary events during 6 months of follow-up. The percentage of HLA-DR positive T-lymphocyte was significantly increased in the unstable angina patients % compared with that in the control %p When baseline levels of cytokine mrna were measured in monocytes, the percentages of the patients expressing higher than normal levels of IL-8, IL-1b, MCP-1, and TF mrnas was 37.5, 29.2, 33.3, and 37.5%, respectively p , , , and , respectively. Basal mrna levels of interleukin IL-8, tissue factor TF, IL-1b and monocyte chemoattractant protein-1 MCP-1 showed a strong correlation with each other 56

2 p0.01 in all combination but not with tumor necrosis factor TNF-or transforming growth factor TGF-1. ConclusionWe observed increase in plasma CRP levels and activation of T-lymphocytes in angina pa-tients. These results may help further classification of angina patients according to the activation of inflammatory markers and under-standing the prognosis of the disease. Korean Circulation J 2000;301:56-65 KEY WORDSAngina Monocytes T-lymphocytes Inflammation C-reactive protein. 서론 대상및방법 대상 57

3 혈장에서의 CRP 및사이토카인의농도측정 단구세포의분리 Northern blot 분석 Table 1. Characteristics of the study subjects Group Unstable angina n12 Stable angina n12 Normal n12 Age yrs Sex male/female 9 / 3 11 / 1 10 / 2 Body mass index kg/m Total cholesterol mg/dl HDL mg/dl LDL mg/dl Smoking current/ex/non-smoker 6 / 3 / 3 4 / 6 / 2 3 / 2 / 7 Hypertension Diabetes mellitus Previous medications Acetylsalicylic acid HMG-CoA reductase inhibitor blockers Calcium antagonists Nitrates ACE inhibitor Age, body mass index, total cholesterol, HDL and LDL levels are expressed as meansd 58 Korean Circulation J 2000;301:56-65

4 활성화된 T-림프구의측정 통계처리 결 Fig. 1. Northern analysis of cytokine mrna levels in peripheral blood monocytes isolated from LPS treated THP-1 cell line, two unstable angina patients UA-1 and UA-2, two stable angina patients SA-1 and SA- 2, and two control subjects N-1 and N-2 are shown. Total RNA was extracted from monocytes before and after LPS treatment for the analysis. One representative blot out of 6 blots with same arrangement is shown. 과 59

5 Table 2. Cytokine mrna levels in monocytes isolated from angina patients and normal subjects Normal Unstable angina Stable angina n Basal level MCP TF IL IL TNF TGF After activation with LPS MCP *** * * TF *** *** *** IL *** *** *** IL *** * ** TNF *** *** *** Data are expressed as means.e. of the cytokine signal/gapdh ratio. Each values were adjusted to set the mean value of normal subjects before activationas 1 *p0.005, **p0.001, and ***p compared with the values before and after LPS stimulation 고찰 60 Korean Circulation J 2000;301:56-65

Fig. 2. Comparison of basal level of cytokines mrnas between normal subjects and angina patients. Individual points represent the cytokine/gapdh signal ratio.

Numbers represent Spearman correlation coefficient. Numbers in the parentheses represent p values IL-8 MCP-1 TF IL-1 TNF- TGF-1 n 36 36 36 33 36 36 IL-8 0.68634 0.7119 0.73188 0.

6 Fig. 2. Comparison of basal level of cytokines mrnas between normal subjects and angina patients. Individual points represent the cytokine/gapdh signal ratio. Shaded areas indicate normal range. *p0.05. Table 3. The correlation among the basal cytokines levels. Calculation was based on Spearman correlation analysis. Numbers represent Spearman correlation coefficient. Numbers in the parentheses represent p values IL-8 MCP-1 TF IL-1 TNF- TGF-1 n IL MCP TF IL TNF TGF

7 62 Fig. 3. Correlation between basal mrna levels of IL-8 and MCP-1 A, TF B, IL-1 C, TNF- D, or TGF-1 E. Monocytes isolated from angina patients and control subjects were tested for the basal level of cytokine mrna by Northern analysis. Individual points represent the cytokine/gapdh signal ratio. Numbers represent Spearman correlation coefficients. Numbers in the parenheses represent p values. Korean Circulation J 2000;301:56-65

8 Fig. 4. Comparison of the percentage of HLA-DR positive T-cells A and B-cells B in patients with unstable angina UA, stable angina SA and normal subjects N. Expression levels of the markers were analyzed by flow cytometry. The horizontal lines of each group indicate mean values. 63

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Pharmacotherapeutics Application of New Pathogenesis on the Drug Treatment of Diabetes Young Seol Kim, M.D. Department of Endocrinology Kyung Hee Univ

Pharmacotherapeutics Application of New Pathogenesis on the Drug Treatment of Diabetes Young Seol Kim, M.D. Department of Endocrinology Kyung Hee Univ Application of New Pathogenesis on the Drug Treatment of Diabetes Young Seol Kim, M.D. Department of Endocrinology Kyung Hee University College of Medicine & Hospital E mail : ycell2@yahoo.co.kr Abstract