실험연구 : 마우스에서 Procaine 투여에 의한 Cisplatin 신독성의 완화 기전

안도환 ( Do Whan Ahn ),김상래 ( Sang Rae Kim ),하동호 ( Dong Ho Ha ),김세환 ( Se Hwan Kim ) 대한마취과학회 2007 Korean Journal of Anesthesiology Vol.52 No.3

Background: Procaine binds to DNA and reduces cisplatin nephrotoxicity, but the mechanism is poorly understood. We explored whether procaine amelioration of cisplatin nephrotoxicity was related to down-and/or up-regulation of inflammatory response gene tumor necrosis factor-α (TNF-α), oxidative stress indicator gene heme oxygenase-1 (HO-1) or cell cycle inhibitor gene p21. Methods: Cisplatin and procaine were intraperitoneally injected to mice at a single dosage of 16 and 80 mg/kg, respectively. Renal evaluation was performed 72 hours after cisplatin administration. The expression of transcripts and proteins was analyzed using real time RT-PCR and Western blot, respectively. Results: Procaine treatment moderately attenuated necrotic changes of renal proximal tubules and increases in BUN and creatinine concentration by cisplatin administration. Kidney platinum level between the cisplatin (cis) group and the cisplatin + procaine (CisPro) group was not different. Although the level of TNF-α mRNA increased 4-fold higher in the Cis group than in the control, this increase was not attenuated by procaine treatment. Gene expression of p21 and HO-1 was elevated 175 and 4-times higher in the Cis group than in the control, respectively. But their expression was no further elevated, rather significantly reduced in the CisPro group compared to the Cis group. Protein abundance of p21 and HO-1 was paralleled by their respective mRNA expression. Conclusions: Procaine amelioration of cisplatin nephrotoxicity is likely to be achieved through processes other than the regulation of TNF-α, HO-1 or p21 gene expression. (Korean J Anesthesiol 2007; 52: 318~27)

토끼 신장의 근위곱슬세관과 근위곧은세관에 존재하는 Na-α-ketoglutarate Cotransport System의 생화학적 특성

안도환 ( Do Whan Ahn ),김경룡 ( Kyoung Ryong Kim ),박희석 ( Hee Seok Park ),박양생 ( Yang Saeng Park ) 대한신장학회 2007 Kidney Research and Clinical Practice Vol.26 No.1

Purpose: alpha-Ketoglutarate (αKG), a Krebs cycle intermediate, is extensively used in the kidney as a fuel substrate and as a counter anion for organic acid secretion. It is known to be taken up by the proximal tubule cells via the brush-border as well as basolateral membranes. We explored biochemical characteristics of the brush-border and basolateral αKG transport systems in pars convoluta and pars recta of the proximal tubule, respectively. Methods: Brush-border and basolateral membrane vesicles (BBMV and BLMV) were isolated from rabbit renal outer cortex and outer medulla by Percoll gradient centrifugation. Vesicular uptake of αKG was determined by rapid Millipore filtration method using α-14[C]KG as a substrate. Results: Both BBMV and BLMV showed a Na-gradient dependent uphill transport of αKG. The systems in both membranes were similarly inhibited by Li and activated by Na (Hill coefficient of 1.4). Kinetic analyses indicated that the Na-αKG cotransporters in the BBMV had a lower substrate affinity as compared with those in the BLMV. The transport systems in BLMVs showed a similar Km but different Vmax between the outer cortex (Km: 34 uM, Vmax: 3.3 nmol/mg protein/10s) and outer medulla (Km: 37, Vmax: 1.8). On the other hand, the systems in BBMVs were different in both Km and Vmax between the outer cortex (Km: 194, Vmax: 3.3) and outer medulla (Km: 89, Vmax: 1.7). Conclusion: The findings suggest that both axial and apical to basolateral heterogeneity of the Na-αKG cotransport system in proximal tubules may be due to a physiological adaptation to efficiently utilize αKG in the kidney.

배양된 토끼 각막간질세포의 Candida albicans 에 대한 탐식능과 Acid Phosphatase 활성도 변화

노주헌(Joo Heon Roh),한영호(Young Ho Hahn),손승완(Seung Wan Sohn),안도환(Do Whan Ahn),원인건(In Gun Won) 대한안과학회 2000 대한안과학회지 Vol.41 No.2

마우스 신장의 피질과 내측수질에서 Real-time RT-PCR을 이용한 Endothelin-1 mRNA와Endothelin Receptor B mRNA의 정량

안도환 대한신장학회 2004 대한신장학회잡지 Vol.23 No.2

카드뮴중독 시 쥐신장의 인산염 이동계의 변화

안도환,박양생 高神大學 醫學部 1991 高神大學醫學部 論文集 Vol.7 No.1

카드뮴중독 시 쥐신장의 인산염 이동계의 변화

안도환,박양생 고신대학교 의학부 1991 高神大學校 醫學部 論文集 Vol.7 No.1

카드뮴이 신장에 존재하는 인산염 이동계에 미치는 영향을 조사하기 위하여 2주간 체중 kg당 2 mg의 카드뮴을 쥐에게 매일 피하주사하였다. 그결과 카드뮴 만성중독 시 나타나는 다뇨, 단백뇨, 당뇨, 인산뇨 등이 관찰되었다. 카드뮴 중독을 일으킨 쥐에서 분리된 세뇨관 강막소포(BBMV)를 이용하여 인산염의 이동을 조사한 바 Na^+dependent phosphate 이동량이 시마게 감소되었다. 따라서 카드뮴 중독은 신장의 세뇨강막에 존재하는 Na^(-)-phosphate cotransport system에 손상을 입혀 인산뇨를 유발함을 알 수 있었다. Effect of cadmium intoxication on the renal cortical phosphate transport system was studied in adult Sprague-Dawley male rats. Subcutaneous injections of CdCl_(2) at a dose of 2 mg Cd/kg body weight per day for 2 weeks induced marked polyuria, glycosuria, proteinuria and phosphaturia, which are the characteristics of chronic cadmium intoxication. In the renal cortical brush-border membrane vesicles prepared from cadmium-intoxicated rats, the Na^(+)-dependent phosphate uptake was markedly attenuated, whereas the Na^(+)-independent uptake was not apparently altered. These results indicate that cadmium intoxication impairs the Na^(+)-dependent phosphate cotransport system in the brush-border membrane of renal proximal tubular cells, which leads to phosphaturia in intact animals.

Cisplatin에 의한 마우스의 급성 신부전 완화에 대한 Endothelin A 수용체의 차단 효과

김일두,안도환 고신대학교의과대학 2007 고신대학교 의과대학 학술지 Vol.22 No.1

Background : Acute renal failure (ARF) can be pathophysiologically classified to hemodynamically-mediated or nephrotoxic type, and in either case, endothelin (ET)-1 expression is enhanced in renal tissues. Hemodynamically-mediated ARF is known to be mitigated by ETA receptor (ETAR) blocker but not ETB receptor (ETBR) blocker. There is poorly understood on mitigation of renal injuries by ETAR or ETBR blockade in nephrotoxic ARF. Therefore, This study examined whether nephrotoxic ARF induced by cisplatin is functionally and/or histologically ameliorated by ETAR blocker BQ123. Materials and Methods : Male C57BL/6 mice are intraperitoneally injected with vehicle (saline), cisplatin or cisplatin+BQ123. Cisplatin was single administrated with a dose of 16 mg/kg/day and BQ123 with a dose of 24 mg/kg/day once a day for 4 consecutive days including a baseline day. Urine flow, excretion of electrolytes and protein, and creatinine clearance were measured. Histological changes were observed through periodic acid-Schiff stain. Real time RT-PCR was performed to evaluate expression of ET-1 and ETAR mRNA. Distribution of ET-1 peptide within renal cortex was examined by immunohistochemistry. Results : Body weight, urine flow, excretion of protein, Na, K, and creatinine clearance tended to decrease by cisplatin administration compared to vehicle treatment, but these parameters did not return to the normal level by combined administration of cisplatin and BQ123. Necroses of several proximal tubules were observed in the cisplatin group, indicating that mild nephrotoxicity occured. In the cisplatin+BQ123 group, this change seemed to increase, rather than decrease. Expression of ET-1 and ETAR mRNA was 6 and 2-fold higher in the cisplatin group than the saline group, respectively. BQ123 treatment reversed ETAR mRNA expression to the basal level but not ET-1 mRNA. Immnuoreactive ET-1 was mainly distributed within proximal tubule cells in all three groups. Immunoreactivity of ET-1 was enhanced in both the cisplatin group and the cisplatin+BQ123 group compared to the saline group, but was similar between the cisplatin group and the cisplatin+BQ123 group. Conclusion : Administration of a ETAR blocker BQ123 in cisplatin-induced nephrotoxic ARF is not likely to be effective in amelioration of renal injuries.

열충격에 의한 조골세포 사멸에 대한 인산염의 상승작용

박경록,안도환 고신대학교의과대학 2007 고신대학교 의과대학 학술지 Vol.22 No.2

Background : Heat and inorganic phosphate (Pi), a major component of bone matrix, are released during implant, where osteoblast survival plays a critical role for successful osseointegration of implant. Because Pi as well as heat stress are known to induce death of osteoblasts, the aim of this study is to investigate the effect of combined treatment of Pi with heat stress on osteoblast viability. Methods : Confluent MC3T3 osteoblasts were exposed to various temperature and time points. They were also incubated with Pi at 37℃ and 48℃. Cell viability was assayed by a MTT method and apoptotic cell death was observed through a TUNEL assay. Results : Heat stress reduced osteoblast survival in a temperature- and time-dependent manner. Increasing Pi concentration did not change cell viability at 37℃, but when osteoblasts were treated at 48℃ in combination with Pi, Pi caused a 4-fold reduction in viability than that by heat stress alone. TUNEL staining showed that some cells were found to die through apoptosis. Addition of phosphonoformic acid, a inhibitor of Pi transporter, completely reversed the Pi-induced cell death without affecting cell death caused by heat stress. Conclusion : Pi potentiates heat stress-induced osteoblastic death, which may reduce the stability and osseointegration of implant.

In Vitro에서 골 형성과 흡수에 대한 Endothelin-1의 영향

사석진,안도환 고신대학교 의과대학 2010 고신대학교 의과대학 학술지 Vol.25 No.2

Background: Balanced regulation of the receptor activator of nuclear factor-kB (RANK) ligand (RANKL) and osteoprotegerin (OPG) by osteoblasts is important for osteoclastogenesis. Sex hormones, glucocorticoids, and prostaglandin E2 (PGE2) are known to modulate osteoblast proliferation and osteoclast formation. Endothelin (ET)-1 is a mitogen as well as a strong vasoconstrictor.It stimulates proliferation of osteoblasts, but its effects on differentiation are controversial. In addition, little is known about ET-1 regulation of osteoclast formation. Thus, the present study was undertaken to investigate whether ET-1 canregulate the expression of RANKL and OPG genes in osteoblasts and affect RANKL-induced osteoclastogenesis. Methods: Osteoblasts were derived from neonatal calvariae and monocytic preosteoclasts from the bone marrow of adult mice, respectively. Cells were cultured in a-minimum essential medium containing 10 nM ET-1. The gene expressions of RANKL and OPG in osteoblasts and RANK in preosteoclasts were measured by real-time RT-PCR. Mineralization by osteoblasts was determined by Alizarin-red staining. Osteoclastogenesis was examined using tartrate-resistant acid phosphatase (TRAP) staining and resorption pit assay. Results: Osteoblasts expressed both ETA and ETB receptors (ETAR and ETBR). ET-1 (10 nM) increased osteoblast proliferation 1.6-fold compared with the control after 3 days in culture and stimulated differentiation, which was indicated by increased formation of mineralized matrix. Proliferation and differentiation of osteoblasts was blocked by 1 uM BQ123, an ETAR antagonist. ET-1 suppressed RANKL gene expression by 50% but did not affect OPG gene expression, and thus reduced the ratio of RANKL to OPG mRNA. PGE2 production by osteoblasts was increased by ET-1. In preosteoclast cultures, ET-1 had little effect on RANK mRNA expression and suppressed RANKL-induced osteoclastogenesis by decreasing the number of TRAP-positive osteoclasts and the resorbed areas. Conclusions: These findings demonstrate that ET-1 can increase bone formation by stimulating proliferation and differentiation of osteoblasts and decrease bone resorption by suppressing RANKL-induced osteoclastogenesis as well as suppressing RANKL mRNA expression In vitro에서 골 형성과 흡수에 대한 endothelin-1의 영향 배경: 조골세포에서 생성되는 receptor activator of nuclear factor-kB ligand (RANKL)과 osteoprotegerin (OPG) 사이의 균형은 파골세포 형성에 매우 중요하다. Enodothelin-1(ET-1)은 강력한 혈관수축제일뿐만 아니라 다양한 세포에서 세포 증식을 촉진시키는 물질이다. 그러나 ET-1의 조골세포의 분화에 대한 영향은 상반될 뿐만 아니라 ET-1이 조골세포의 RANKL과 OPG의 생성을 변화시켜 간접적으로 파골세포 형성에 영향을 주는지 혹은 파골세포의 분화에 직접적으로 영향을 주는지에 대해서는 연구된 바 없다. 따라서 본 연구에서는 ET-1에 의한 조골세포의 분화와 RANKL 및 OPG 유전자 발현이 파골세포 형성에 미치는 영향을 조사하였다. 재료와 방법: 조골세포는 생쥐의 두개골로부터 그리고 파골전구세포는 골수로부터 각각 분리하였다. 이들 세포들을 10 nM ET-1이 포함된 α-MEM 배지에서 배양하였다. 유전자의 발현은 real-time RT-PCR을 통해 정량적으로 측정하였다. 조골세포의 분화는 alizarin-red 염색으로, 파골세포의 형성과 활성은 tartrate-resistant acid phosphatase 염색과 골 흡수와(resorption pit)의 크기로 평가하였다. 결과: ET-1은 조골세포의 증식과 분화를 촉진시켰으며, 이는 ETA 수용체를 통해 이루어졌다. 조골세포에서 ET-1은 PGE2 생성을 크게 증가시켰을 뿐만 아니라 RANKL 유전자 발현을 대조군에 비해 50% 감소시켰다. 그러나 OPG 유전자의 발현에 영향을 주지 않았다. 파골 전구세포에서 ET-1은 RANK 유전자의 발현에 영향을 주지 않았지만 RANKL에 의해 유도된 파골세포 형성과 활성은 억제시켰다. 결론: ET-1은 조골세포의 증식과 분화를 촉진시키고 RANKL mRNA 발현을 감소시켜 파골세포 형성을 간접적으로 억제한다. 또한 RANKL에 의해 유도되는 파골세포 형성과 활성을 직접적으로 억제한다. 이러한 결과로 미루어 ET-1은 골형성은 촉진하고 골흡수는 억제할 것으로 사료된다.

Peroxisome Proliferator-Activated Receptor Gamma의 골 작용

박경록,안도환 고신대학교의과대학 2008 고신대학교 의과대학 학술지 Vol.23 No.4

Peroxisome proliferator-activated receptor (PPAR) is a class of the nuclear transcription factors that regulates lipid metabolism and cell differentiation. Among three different PPAR isotypes, PPARγis known to be a critical regulator of adipogenesis. Because osteoblasts and adipocytes are originated from common marrow mesenchymal progenitors, PPARγ can also impact on osteogenesis. Recent evidences have shown that PPARγ plays a certain role in bone metabolism and bone turnover. In this review, we summarize its general bone actions of PPARγ including bone cell growth and differentiation, bone formation and loss.