마우스 Hepa-1c1c7 세포주에서 B형 간염 바이러스에 의한 tumor necrosis factor-a의 발현 유도

예성수,장원희,양영일,이연재,김미성,석대현,박영홍,백계형,Yea Sung Su,Jang Won Hee,Yang Young-Il,Lee Youn Jae,Kim Mi Seong,Seog Dae-Hyun,Park Yeong-Hong,Paik Kye-Hyung 한국생명과학회 2005 생명과학회지 Vol.15 No.1

B형 간염 바이러스(HBV)에 의한 감염은 인류의 보건에 대단히 중요한 문제이며, 따라서 HBV에 대한 많은 연구가 수행되어져 왔다. 그러나 HBV 연구에 있어서의 주된 장애요인은 그 감염이 사람과 일부 영장류에 국한된다는 점이다. 본 연구에서는 마우스 간암 세포주인 Hepa-1c1c7 세포를 이용하여 HBV의 감염성 및 그에 따른 염증성 사이토카인인 TNF-a의 발현의 변화를 측정하였다. HBV의 표면항원(HBsAg)분비는 microparticle enzyme immunoassay를 사용하여 측정하였고, TNF-a mRNA 발현 측정에는 quantitative competitive RT-PCR 방법을 사용하였다. HBV 발현 벡터를 Hepa-1clc7 세포에 도입시켰을 경우뿐만 아니라 HBV 비리온을 갖고 있는 혈청을 사용하여 Hepa-lc1c7 세포를 감염시켰을 때에도 HBV mRNA 발현 및 HBsAg 분비가 측정되었다. 또한 두 상황 모두에서 TNF-a mRNA발현이 증가됨을 알 수 있었다. 이러한 결과는 사람과 영장류에 특이적인 HBV가 마우스 간암 세포주인 Hepa-lclc7 세포도 감염시킬 수 있다는 가능성을 제시한다. 또한 마우스 기원의 Hepa-lclc7 세포에서도 HBV의 유전자 발현에 필요한 여러 인자들이 존재하며, TNF-a와 같은 사이토카인 유전자 발현을 조절하는 세포 내 기전에 HBV가 영향을 미친다고 할 수 있다. 따라서 마우스 간암 세포주인 Hepa-lclc7 세포는 HBV 연구를 위한 시험 관내 모델로서 사용되어질 수 있을 것으로 보인다. Infection with hepatitis B virus (HBV) is a major health problem worldwide. Although a tremendous amount has been known about HBV, there have been obstacles in the study of HBV due to the narrow host range of HBV limited to humans and primates. In the present study, we investigated the susceptibility to HBV infection of mouse hepatoma cell line, Hepa-1c1c7. In addition, based on that human hepatocytes infected by HBV increase the expression of the pro-inflammatory cytokine TNF-a, the inducibility of TNF-a expression by HBV in the cells was determined. HBV surface antigen (HBsAg) secretion was measured by the microparticle enzyme immunoassay and steady state mRNA expression was analyzed by quantitative competitive RT-PCR. Transient transfection of Hepa-1c1c7 cells with HBV expression vector resulted in a dose-dependent induction of TNF-a expression. Infection of Hepa-1c1c7 cells with the serum of HBV carrier also increased TNF-a mRNA expression. Both in the transfected and infected cells, HBV mRNA was expressed and significant HBsAg secretion was detected. There was no significant variation in $\beta-actin$ mRNA expression by HBV. These results demonstrate that HBV is infectious to Hepa-lc1c7 in vitro and the viral infection induces TNF-a expression, which suggests that Hepa-lc1c7, a mouse hepatoma cell line, may be a possible model system for analysis of various molecular aspects of HBV infection.

담낭 선종과 암종에서 β-catenin의 발현 양상

문정호(Jong Ho Moon),최창수(Chang Soo Choi),양영일(Young Il Yang),백계형(Kye-Hyung Paik),최영길(Young Kil Choi) 대한외과학회 2002 Annals of Surgical Treatment and Research(ASRT) Vol.63 No.2

식도정맥류에 대한 비 Shunt 수술

박원진(Won Jin Park),왕희정(Hee Jung Wang),백계형(Kye Hyung Paik),이혁상(Hyucksang Lee) 대한외과학회 1994 Annals of Surgical Treatment and Research Vol.46 No.4

개의 동소성 간 이식술시 이식 간의 재관류시에 관찰되는 혈행 동태에 관한 연구

황성환,오상훈,백계형,김상효 인제대학교 1996 仁濟醫學 Vol.17 No.2

본 연구의 목적은 개의 동소성 간이식 모델에서 혈역학적 변화를 정확히 측정하여 임상에서 많은 문제를 일으키는 이식간의 재관류 후에 발생하는 혈역학적 변화 양상을 좀 더 정확히 밝혀 보려는 데 있다. Biopump에 의한 정-정맥 우회술을 이용하여 개에서 동소성 간이식을 시행하였으며, 재관류 전(III-5)과 후(III+5)의 혈역학 지수의 평균치 변화를 비교하였다. 10례 중 4례에서 재관류 후 증후군이 나타나 종래의 방법에 의한 개의 간이식 실험에서보다 적은 빈도를 보였으며 간의 허혈 방지와 수술 수기의 향상과 전신 대사 이상의 적극적인 치료 등 종래보다 발전된 간이식으로 재관류 후의 혈행 동태를 안정화시킬 수 있었고, 재관류 후에 페동맥 고혈압은 거의 나타나지 않아 재관류 후에 나타나는 폐혈관 저항의 증가 양상은 재관류 후 증후군의 한 현상이 아닌 별개의 현상으로 생각된다. Objective: To study the hemodynamic changes after reperfusion during orthotopic liver transplantation(OLT). Methods and Materials: During OLT, hemodynamic changes after reperfusion were studdid in 10 mongrel dogs. And then hemodynamic variables at each stage of operation were measured and compared with the stage of 5-minutes before reperfusion and 5 minutes after reperfusion. Results: The mean value of mean arterial pressure was decreased from 90±9.5mmHg to 74.3±27.0 mmHg(p=0.13). In four of ten cases, mean arterial pressure was decreased over 30% and showed post-reperfusion syndrome(PRS). The mean value of systemic vascular resistance was decreased from 3857.2±1900 dyn.sec/cm3 to 2640.4 ±769dyn.sec.cm3(p<0.05). Cardiac output was decreased in 7 cases(p=0.63). The mean value of mean pulmonary arterial pressure was decreased from 14.6±2.27 mmHg to 11.7±2.71mmHg(p<0.05), and pulmonary vascular resistance was decreased from 284.0 ±239 dyn.sec/cm3 to 255.2±199 dyn.sec/cm3(p =0.57). Pulmonary hypertension which could be seen in orthotopic liver transplantation was not noted with this study even though 4 cases showed PRS. Conclusion: Increased pulmonary vascular resistance or pulmonary arterial pressure after reperfusion could be an independent phenomenon rather than a part of PRS.

간이식 수술중 혈전탄성묘사도 변수에 따른 혈액응고관리의 효과 분석

김명호,김종운,우성,이혁상,백계형,김문철,왕희정 대한마취과학회 1997 Korean Journal of Anesthesiology Vol.32 No.4

Introduction: Thromboelastography (TEG) provides an overall assessment of the platelet-coagulation protein cascade interaction. The information generated from the TEG is rapidly obtained and made useful to guide replacement therapy. The purpose of this study was to evaluate the efficacy of the TEG as its guided blood replacement therapy and pharmacological therapy during liver transplantation. Methods: This study was carried out in 13 consecutive patients who were subjected to TEG-guided replacement therapy during liver transplantation. A prepared mixture of blood products used for continuous replacement therapy was a fluid composed of red blood cells(2 units), fresh frozen plasma (2 units), and normal saline(500 ml). The pharmacological therapy was performed by comparing TEG of untreated blood and blood treated with antifibrinolytic and heparin neutralizing agent. Based on the findings of TEG, platelet concentrates were given. The TEG samples were obtained at various intervals. Additional TEG tracing was obtained as needed to see the effect of therapeutic intervention. Results: In all patients the reaction time was kept in an acceptable range in the preanhepatic stage by administration of the mixture of blood products. Heparin-induced anticoagulation was observed in 3 cases in the anhepatic stage and in 11 cases upon reperfusion. Fibrinolysis was seen in all but one patients: 8% in the preanhepatic stage, 41% in the anhepatic stage, 69% at reperfusion, and 2% in the postanhepatic stage. Early and aggressive treatment with epsilon-aminocaproic acid effectively inhibited fibrinolysis without complications. Ten patients needed platelet transfusion in the postanhepatic stage with significant improvement in the TEG. Conclusions: The results of this study suggest that TEG monitoring and TEG-guided replacement and pharmacological therapy are clinically effective in maintaining blood coagulability. (Korean J Anesthesiol 1997; 32: 604∼615)

인체 대장암의 진행과 DCC 유전자 및 단백 발현과의 연관성

김종익,정해진,양영일,백계형,윤혜경,홍관희,최경현 대한대장항문학회 2003 Annals of Coloproctolgy Vol.19 No.1

개의 동소성 간이식술에서 혼합정맥혈 산소포화도의 지속적 감시에 의한 산소운반 및 섭취능력 평가

조강희,왕희정,우성,김문철,백계형,이혁상 대한마취과학회 1994 Korean Journal of Anesthesiology Vol.27 No.11

There are many hemodynamic and physiologic changes during liver transplantation much more than other surgical interventions. The oxygen delivery and oxygen consumption are af- fected by depressed hemodynamic and metabolic status during the operation. At the lower levels of oxygen present in venous blood, a linear relationship exists between saturation and tension. The use of fiberoptic oximetry system in conventional pulmonary artery flotation catheters has made the bedside application of this relationship of practical value in the continuous assessment of mixed venous oxygen saturation. This study was performed to determine changes in Sv￣O₂ and other variables of oxygen kinetics during canine OLT and study the correlation between Sv￣O₂ and cardiac output, Sv￣O2 and oxygen consumption and oxygen utilization ratio. The continuous rnixed venous oxygen saturation and cardiac output by SO₂/CO computer were monitored and the oxygen delivery, oxygen consumption and oxygen utilization ratio were calculated by arterial and venous blood gas analysis and modified Fick's equation during orthotopic liver transplantation in 20 dogs. The results were as follow as ; 1. There was no significant difference in tissue oxygen extraction between preoperative control and anhepatic phase, while cardiac output were decreased during anhepatic phase. 2. By utilizing centrifugal pump(venovenous bypass) oxygen delivery and oxygen utilization ratio were well maintained even though suppressed the change of oxygen delivery and oxygen consumption during anhepatic phase. 3. There was a significant decrease in Sv￣O₂ immediately after declamping the suprahepatic vena cava, whereas the oxygen utilization rate and oxygen consumption following reperfusion were significantly increased than just prior to reperfusion of transplanted liver. 4. A Statistically significant correlation was found between Sv￣O₂ and cardiac output, oxygen consumption in all surgical stages except reperfusion(CO;r=0.478, p$lt;0.001, VO₂,r=-0. 272, p=0.004), but their correlations were relatively poor. However, there was highly significant correlation among Sv￣O₂ and oxygen utilization ratio in all surgical stages(O₂UR; r=- 0.834, P$lt;0.001). In conclusion, continuous monitoring mixed venous oxygen via a fiberoptic pulmonary catheter could be used as the index for evaluation of hemodynamics and oxygen kinetics during canine OLT, but further research should be performed to determine whether these measurements indicate viability of the grafted liver.

마우스 간세포암 기원의 Hepa-1c1c7 세포주에서의 B형 간염 바이러스에 의한 TNF-α발현의 유도

예성수,장원희,양영일,이연재,김미성,백계형 인제대학교 2000 仁濟醫學 Vol.21 No.2

Infection with hepatitis B virus (HBV) is a major health problem worldwide. Although a tremendous amount has been known about HBV. there have been obstacles in the study of HBV due to the narrow host range of HBV limited to humans and primates. In the present study, we investigated the susceptibility to HBV infection of mouse hepatoma cell line, Hepa-Iclc7. In addition, based on that human hepatocytes infected by HBV increase the expression of the pro-inflammatory cytokine TNF-α, the inducibility of TNF-α expression by HBV in the cells was determined. HBV surface antigen (HBsAg) secretion was measured by the microparticle enzyme immunoassay and steady state mRNA expression was analyzed by quantitative competitive RT-PCR. Transient transfection of Hepa-1c1c7 cells with HBV expression vector resulted in a dose-dependent induction of TNF-α expression. Infection of Hepa-lclc7 cells with the serum of HBV carrier also increased TNF-α mRNA expression. Both in the transfected and infected cells, HBV mRNA was expressed and significant HBsAg secretion was detected. There was no significant variation in β-actin mRNA expression by HBV. These results demonstrate that HBV is infectious to Hepa-1c1c7 in nitro and the viral infection induces TNF-α expression, which suggests that Hepa- 1c1c7. a mouse hepatoma cell line, may be a possible model system for analysis of various molecular aspects of HBV infection.