Sirolimus 사용 후 사구체 기저막 세극막 관련 분자의 변화
The Changes of Slit Diaphragm Molecules After Using Sirolimus
0
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
목 적:최근 신이식 환자들에서 cyclosporine A (CsA)의 대체로 sirolimus를 투여 받은 후 단백뇨가 발생한다는 임상보고가 있으나 단백뇨 발생기전의 정확한 메커니즘에 대한 연구가 없다. 이에 sirolimus에 의한 여러 단백뇨 발생기전 중 sirolimus와 CsA 투여 후 족세포의 세극막 관련 분자 변화를 조사하여 족세포와의 직접적인 영향에 대해 알아보고자 하였다. 방법:생체 외 실험- 마우스 족세포를 완충액, CsA (10 μg/mL) 처리 후 sirolimus (10 μg/mL) 처리군, sirolimus 단독군, CsA와 sirolimus 동시처리한 군으로 나누어 RT-PCR을 이용하여 12, 24, 48시간에 족세포의 세극막 관련 분자 변화를 측정하였다. 생체 내 실험- SPF Wistar 쥐 24마리를 각각 4군(완충액, CsA 2주 투여 후 sirolimus 2주간 투여, sirolimus 4주간 투여, CsA와 sirolimus를 4주간 동시투여)으로 분류하여 하루 걸러서 한번 복강 내 약물을 주입하였다(CsA: 25 mg/kg, sirolimus: 0.5 mg/kg). 모든 쥐는 약물주입 후 4주에 희생되어 병리조직은 오른쪽 신장의 일부분을 이용하고, 나머지 신장은 RT-PCR을 이용하여 세극막 관련 분자의 mRNA 발현 변화를 측정하였다. 결 과:생체 외 실험에서 CsA와 sirolimus 동시투여 또는 CsA 처리 후 sirolimus 처리군에서 nephrin 발현이 의미 있게 감소하였다. 생체 내 실험에서 nephrin 발현은 sirolimus를 사용한 모든 군에서, podocin 발현은 CsA와 sirolimus 동시투여 또는 CsA 처리 후 sirolimus 처리군에서 의미 있게 감소하였다. 광학현미경에서 CsA 투여군은 세뇨관 상피세포에서 공포형성 및 석회화가 관찰되었으며, 면역 조직화학검사에서 사구체 모세혈관의 nephrin, podocin항체 침착은 감소되지 않았다. CsA 투여군의 전자 현미경소견에서 사구체 족돌기의 국소적 융합이 있었으며, sirolimus 단독군에서는 특이소견이 없었다. 결 론:본 연구를 통해 sirolimus로 발생한 단백뇨의 많은 기전 중 하나로 sirolimus가 세극막 관련 분자 중 nephrin 및 podocin의 mRNA 발현을 감소시킬 수 있으며 sirolimus 단독보다는 CsA와 함께 작용했을 때 효과가 더 커질 수 있음을 제시하는 바이다.
Abstract
Purpose: Recently, massive proteinuria has been observed in some transplant patients after switching cyclosporine A (CsA) to sirolimus. To evaluate the pathogenesis of sirolimus-associated proteinuria, we investigated the early changes in slit diaphragm molecules by various administrative conditions of sirolimus and CsA. Methods: In vitro-Mouse podocytes were incubated with buffer (C), sirolimus (10 μg/mL) after CsA (10 μg/mL) (C-S), sirolimus only (S) and CsA and sirolimus simultaneously (C+S) for 12, 24, and 48 hours. In vivo- twenty four SPF female Wistar rats were divided into 4 groups buffer (C), sirolimus after 2 weeks of CsA (C-S), sirolimus only (S) and CsA and sirolimus simultaneously (C+S). All groups were treated by intraperitoneal injection every other day for 4 weeks (CsA: 25 mg/kg, sirolimus: 0.5 mg/kg). The changes in mRNA of slit diaphragm molecules were examined by RT-PCR. Results: The mRNA of nephrin was significantly decreased in group C-S and C+S in vitro. In vivo, the mRNA of nephrin in all groups using sirolimus and the mRNA of podocin in group C-S and C+S were decreased. Microscopically, group C-S and C+S showed small vacuolization and calcification in proximal tubular epithelial cells. Immunohistochemistry using nephrin and podocin antibodies did not show remarkable decrease of staining along the glomerular capillaries. Electron-microscopically, focal fusion of foot processes was seen in group C-S and C+S. Conclusion: This study suggests the decrease of slit diaphragm molecules (nephrin and podocin) in podocyte may be one of the causes of sirolimus associated proteinuria, and podocyte injury by sirolimus may need a primary hit by CsA to develop the proteinuria.
참고문헌 (42)
- Meier-Kriesche HU, "Lack of improvement in renal allograft survival despite a marked decrease in acute rejection rates over the most recent era", Am J Transplant, 4, pp.378~383 (2004)
- Hariharan S, "Improved graft survival after renal transplantation in the United States, 1988 to 1996", N Engl J Med, 342, pp.605~612 (2000)
- Weir MR, "Late calcineurin inhibitor withdrawal as a strategy to prevent graft loss in patients with suboptimal kidney transplant function", Am J Nephrol, 24, pp.379~386 (2004)
- Andrassy J, "mTOR inhibition and its effect on cancer in transplantation", Transplantation, 80, pp.S171~174 (2005)
- Morath C, "Sirolimus in renal transplantation", Nephrol Dial Transplant, 22, pp.viii61~viii65 (2007)
- Lee VW, "Sirolimus: its role in nephrology", Nephrology (Carlton), 10, pp.606~614 (2005)
- Sehgal SN, "Sirolimus: its discovery, biological properties, and mechanism of action", Transplant Proc, 35, pp.7S~14S (2003)
- Poon M, "Rapamycin inhibits vascular smooth muscle cell migration", J Clin Invest, 98, pp.2277~2283 (1996)
- He Z, "Conversion from cyclosporin A to sirolimus retards the progression of chronic allograft nephropathy in the long term in a rat kidney transplantation model", J Int Med Res, 37, pp.1396~1410 (2009)
- Sennesael JJ, "Conversion from cyclosporine to sirolimus in stable renal transplant recipients", Transplantation, 80, pp.1578~1585 (2005)
- Stallone G, "Rapamycin for treatment of chronic allograft nephropathy in renal transplant patients", J Am Soc Nephrol, 16, pp.3755~3762 (2005)
- Bumbea V, "Long-term results in renal transplant patients with allograft dysfunction after switching from calcineurin inhibitors to sirolimus", Nephrol Dial Transplant, 20, pp.2517~2523 (2005)
- Gonwa TA, "Improved renal function in sirolimus-treated renal transplant patients after early cyclosporine elimination", Transplantation, 74, pp.1560~1567 (2002)
- Letavernier E, "Proteinuria following a switch from calcineurin inhibitors to sirolimus", Transplantation, 80, pp.1198~1203 (2005)
- Boratyńska M, "Conversion to sirolimus from cyclosporine may induce nephrotic proteinuria and progressive deterioration of renal function in chronic allograft nephropathy patients", Transplant Proc, 38, pp.101~104 (2006)
- Franco AF, "Proteinuria in transplant patients associated with sirolimus", Transplant Proc, 39, pp.449~452 (2007)
- Izzedine H, "Post- transplantation proteinuria and sirolimus", N Engl J Med, 353, pp.2088~2089 (2005)
- Rangan GK, "Sirolimus-associated proteinuria and renal dysfunction", Drug Saf, 29, pp.1153~1161 (2006)
- Letavernier E, "mToR inhibitors-induced proteinuria: mechanisms, significance, and management", Transplant Rev (Orlando), 22, pp.125~130 (2008)
- Han GD, "IFN-inducible protein-10 plays a pivotal role in maintaining slit-diaphragm function by regulating podocyte cell-cycle balance", J Am Soc Nephrol, 17, pp.442~453 (2006)
- Abbate M, "In progressive nephropathies, overload of tubular cells with filtered proteins translates glomerular permeability dysfunction into cellular signals of interstitial inflammation", J Am Soc Nephrol, 9, pp.1213~1224 (1998)
- Straathof-Galema L, "Sirolimus-associated heavy proteinuria in a renal transplant recipient: evidence for a tubular mechanism", Am J Transplant, 6, pp.429~433 (2006)
- Mreich E, "Sirolimus does not reduce receptor-mediated endocytosis of albumin in proximal tubule cells", Transplantation, 83, pp.105~107 (2007)
- Bennett WM, "Chronic cyclosporine nephropathy: the Achilles' heel of immunosuppressive therapy", Kidney Int, 50, pp.1089~1100 (1996)
- Saurina A, "Conversion from calcineurin inhibitors to sirolimus in chronic allograft dysfunction: changes in glomerular haemodynamics and proteinuria", Nephrol Dial Transplant, 21, pp.488~493 (2006)
- van den Akker JM, "Proteinuria following conversion from azathioprine to sirolimus in renal transplant recipients", Kidney Int, 70, pp.1355~1357 (2006)
- Diekmann F, "Influence of sirolimus on proteinuria in de novo kidney transplantation with expanded criteria donors: comparison of two CNI-free protocols", Nephrol Dial Transplant, 22, pp.2316~2321 (2007)
- Letavernier E, "High sirolimus levels may induce focal segmental glomerulosclerosis de novo", Clin J Am Soc Nephrol, 2, pp.326~333 (2007)
- Letavernier E, "Sirolimus interacts with pathways essential for podocyte integrity", Nephrol Dial Transplant, 24, pp.630~638 (2009)
- Pätäri-Sampo A, "Molecular basis of the glomerular filtration: nephrin and the emerging protein complex at the podocyte slit diaphragm", Ann Med, 38, pp.483~492 (2006)
- Kawachi H, "Role of podocyte slit diaphragm as a filtration barrier", Nephrology (Carlton), 11, pp.274~281 (2006)
- Kerjaschki D, "Caught flat-footed: podocyte damage and the molecular bases of focal glomerulosclerosis", J Clin Invest, 108, pp.1583~1587 (2001)
- Kwoh C, "Pathogenesis of nonimmune glomerulopathies", Annu Rev Pathol, 1, pp.349~374 (2006)
- Vollenbröker B, "mTOR regulates expression of slit diaphragm proteins and cytoskeleton structure in podocytes", Am J Physiol Renal Physiol, 296, pp.F418~426 (2009)
- Kahan BD, "Efficacy of sirolimus compared with azathioprine for reduction of acute renal allograft rejection: a randomised multicentre study", Lancet, 356, pp.194~202 (2000)
- Kokui K, "Cyclosporin reduces proteinuria in rats with aminonucleoside nephrosis", J Pathol, 166, pp.297~301 (1992)
- Faul C, "The actin cytoskeleton of kidney podocytes is a direct target of the antiproteinuric effect of cyclosporine A", Nat Med, 14, pp.931~938 (2008)
- Meyrier A, "Antiproteinuric and immunological effects of cyclosporin A in the treatment of glomerular diseases", Nephrol Dial Transplant, 7, pp.S80~84 (1992)
- Nacar A, "Ultrastructural examination of glomerular and tubular changes in renal allografts with cyclosporine toxicity", Ren Fail, 28, pp.543~547 (2006)
- DiJoseph JF, "The effect of rapamycin on kidney function in the Sprague-Dawley rat", Transplantation, 53, pp.507~513 (1992)
- Liew A, "Factors associated with proteinuria in renal transplant recipients treated with sirolimus", Transpl Int 2009;22:313-22., 22, pp.313~322 (2009)
- Ruiz JC, "Early sirolimus use with cyclosporine elimination does not induce progressive proteinuria", Transplant Proc, 39, pp.2151~2152 (2007)