Mesenchymal Stem Cells Ameliorate Adriamycin Induced Proteinuric Nephropathy

강희경,박소연,하일수,정해일,최용,Kang, Hee-Gyung,Park, So-Yeon,Ha, Il-Soo,Cheong, Hae-Il,Choi, Yong Korean Society of Pediatric Nephrology 2010 Childhood kidney diseases Vol.14 No.1

목 적 : 사구체신염은 흔히 단백뇨를 보이며 특이 치료법이 없고, 만성 신부전으로 발전하는 경우가 많다. 몇몇 연구에서 중간엽 줄기세포(Mesenchymal stem cell, MSC)를 실험적 사구체신염에 투여하여 단백뇨가 호전된 것을 보고한 바 있으나, 이는 신염을 일으키는 약제와 중간엽 줄기세포를 함께 투여하거나 신장에 직접 투여한 것이었다. 본 연구에서는 실험적 신병증에서 단백뇨가 발현된 시점에서 정주 요법으로 MSC를 투여함으로써 MSC의 임상적인 적용 가능성을 탐색하였다. 방 법 : 실험용 생쥐에 Adriamycin을 투여하여 신병증(ADR-GN)을 유발한 후, 2주 후에 대량의 단백뇨를 확인하고 MSC를 생쥐 꼬리의 정맥에 주사하였다. MSC에 의한 질병 완화의 기전을 확인하기 위한 in vitro 실험으로 mixed lymphocyte culture(MLC)에 MSC를 투여하였을 때의 염증 관련 cytokine인 IFN-$\gamma$ and IL-10의 변화를 측정하였다. 결 과 : 실험용 생쥐에 ADR-GN를 유발하고 단백뇨가 보일 때 MSC를 정주한 군에서는 단백뇨의 소실이 더 먼저 관찰되었다. 또한 MSC를 투여받은 군에서의 생존률이 더 나은 경향이 관찰되었다. MLC 에 MSC를 투여하였을 때, 염증을 유발하는 cytokine인 IFN-$\gamma$ 는 감소하고 염증을 억제하는 cytokine인 IL-10는 증가하였다. 결 론 : 이 연구는 이전의 보고들에서 관찰되었던 사구체신염에서의 MSC의 질병완화 효과가 좀더 임상적으로 적용 가능한 방법으로 투여된 경우, 즉 단백뇨가 있을 때 정주 요법으로 투여한 경우에도 관찰됨을 확인하였다. 이러한 효과의 기전과 임상적용에 요구되는 안전성 등에 대한 확인을 위해서는 추가 연구가 필요하겠다. Purpose : Glomerulonephropathy (GN) often manifests as proteinuria and progresses to chronic renal failure without specific therapy. Mesenchymal stem cell (MSC) has been tried as a therapeutic agent in experimental GN, and previous studies showed that administration of MSC concomitantly to the insult inducing GN or via intra-renal administration ameliorated proteinuria. The purpose of this study was to test the therapeutic potential of MSC administered via intravenous route at the time of clinically evident proteinuria. Methods : MSCs were administered intravenously via tail vain into the mice with adriamycin (ADR) induced nephropathy (ADR-GN), two weeks after ADR injection when massive proteinuria was evident. To test the capacity of MSC modulate the cytokine production in the inflammatory milieu, the concentrations of IFN-$\gamma$ and IL-10 were measured in the supernatant of in vitro mixed lymphocyte culture (MLC) with or without additional MSC. Results : MSCs administered intravenously into the proteinuric mice with ADR-GN accelerated the recovery of this experimental GN with disappearance of proteinuria in two weeks when the saline treated (control) mice still showed significant proteinuria. The mice treated with MSC also had a tendency of better survival. Addition of MSC decreased IFN-$\gamma$ and increased IL-10 in the supernatant of MLC. Conclusion : This study showed that MSC had a therapeutic potential even when administered in a more clinically relevant setting into a proteinuric glomerulonephropathy model. Further study to verify the mechanism and long-term safety of this phenomenon is required.

국소성 분절성 사구체 신병증의 동물 모델 (FGS/kist 생쥐) 신 조직의 유전자 발현 양상

강희경,이병섭,이철호,하일수,정해일,최용,Kang, Hee-Gyung,Lee, Byong-Sop,Lee, Chul-Ho,Ha, Il-Soo,Cheong, Hae-Il,Choi, Yong 대한소아신장학회 2011 Childhood kidney diseases Vol.15 No.1

목 적: 국소성 분절성 사구체 경화증(Focal segmental glomerulosclerosis, 이하 FSGS)은 소아신부전의 원인 중 가장 흔한 사구체 질환이다. 일차성 FSGS의 병인은 아직 알려져 있지 않으므로, 저자들은 FSGS의 동물 모델을 대상으로 cDNA 마이크로어레이를 이용한 유전자 발현 양상 분석을 통하여 유전자 발현 수준에서의 FSGS의 질환의 특성을 밝히고자 하였다. 방 법: 사람의 일차성 FSGS와 유사한 질병경과를 보이는 동물모델인 FGS/kist 생쥐의 신피질 조직을 대조군 생쥐(FGS/kist 생쥐의 조상 strain인 RFM/kist 생쥐)와 AB 1700 mouse chip을 이용한 마이크로어레이 실험으로 비교하였다. 결 과: FGS 질병특이 유전자가 62개 추출되었다. 이들은 세포주기/사멸, 면역반응과 지질 대사/혈관 질환과 관련된 유전자들로써, 유전자간 network의 중심유전자가 면역반응(TNF, IL-6/4, IFNg)과 세포사멸 조절 유전자(TP 53), 그리고 지질대사의 중요 유전자인 PPARG이었다. 결 론: 이 연구에서 저자들은 자발적인 FSGS의 임상경과를 보이는 FGS/Kist 생쥐의 신장조직의 유전자 발현의 분석을 통하여 신장세포사멸과 면역반응에 뒤따르는 기질 섬유화, 그리고 지질 대사의 이상과 조기 혈관 질환이 FSGS의 병태생리에 기여할 것임을 다시 확인할 수 있었다. 추가적인 연구가 계속된다면 global transcriptome profiling 기법으로 병인 탐색 및 치료방법 개발 에 의미 있는 결과를 도출할 수 있을 것이다. Purpose: Focal segmental glomerulosclerosis (FSGS) is the most common glomerulopathy causing pediatric renal failure. Since specific treatment targeting the etiology and pathophysiology of primary FSGS is yet elusive, the authors explored the pathophysiology of FSGS by transcriptome analysis of the disease using an animal model. Methods: FGS/kist strain, a mouse model of primary FSGS, and RFM/kist strain, as control and the parent strain of FGS/kist, were used. Kidney tissues were harvested and isolated renal cortex was used to extract mRNA, which was run on AB 1700 mouse microarray chip after reverse transcription to get the transcriptome profile. Results: Sixty two genes were differentially expressed in FGS/kist kidney tissue compared to the control. Those genes were related to cell cycle/cell death, immune reaction, and lipid metabolism/vasculopathy, and the key molecules of their networks were TNF, IL-6/4, IFN${\gamma}$, TP53, and PPAR${\gamma}$. Conclusion: This study confirmed that renal cell death, immune system activation with subsequent fibrosis, and lipid metabolism-related early vasculopathy were involved in the pathophysiology of FSGS. In addition, the relevance of methodology used in this study, namely transcriptome profiling, and Korean animal model of FGS/kist was validated. Further study would reveal novel pathophysiology of FSGS for new therapeutic targets.

Anti-inflammatory Activity of 1-docosanoyl Cafferate Isolated from Rhus verniciflua in LPS-stimulated BV2 Microglial Cells

Lee, Jae-Won,Cheong, Il-Young,Kim, Hae-Sung,Lee, Jae-Jun,Lee, Yong-Suk,Kwon, Yong-Soo,Kim, Myong-Jo,Lee, Hee-Jae,Kim, Sung-Soo,Chun, Wan-Joo The Korean Society of Pharmacology 2011 The Korean Journal of Physiology & Pharmacology Vol.15 No.1

Although various derivatives of caffeic acid have been reported to possess a wide variety of biological activities such as protection of neuronal cells against excitotoxicity, the biological activity of 1-docosanoyl cafferate (DC) has not been examined. The objective of the present study was to evaluate the anti-inflammatory effects of DC, isolated from the stem bark of Rhus verniciflua, on lipopoly-saccharide (LPS)-stimulated BV2 microglial cells. Pretreatment of cells with DC significantly attenuated LPS-induced NO production, and mRNA and protein expression of iNOS in a concentration-dependent manner. DC also significantly suppressed LPS-induced release of cytokines such as TNF-${\alpha}$ and IL-$1{\beta}$. Consistent with the decrease in cytokine release, DC dose-dependently and significantly attenuated LPS-induced mRNA expression of these cytokines. Furthermore, DC significantly suppressed LPS-induced degradation of IKB, which retains NF-kB in the cytoplasm. Therefore, nuclear translocation of NF-kB induced by LPS stimulation was significantly suppressed with DC pretreatment. Taken together, the present study suggests that DC exerts its anti-inflammatory activity through the suppression of NF-kB translocation to the nucleus.

원발성 수산증에서의 신장이식술 1예

김상준,민승기,정해일,최용 大韓移植學會 1993 Korean Journal of Transplantation Vol.7 No.1

소아의 미세변화형 신증후군 및 초점성 분절성 사구체 경화증 환아에서 혈청 및 요의 용해성 인터루킨-2수용체

하일수,정해일,최용,Ha, Il-Soo,Cheong, Hae-Il,Choi, Yong 대한소아신장학회 1999 Childhood kidney diseases Vol.3 No.1

목 적 : 가능한 교란인자들, 즉 연령, 단백뇨, 스테로이드 사용 등의 영향이 배제된 조건에서 소아의 신증후군, 또는 그 중의 어떤 특성이 혈청이나 요의 용해성 인터루킨-2수용체 (sIL-2R)에 영향을 주는지를 알기 위해 이 연구를 시행하였다. 방 법 : 소아의 일차성 신증후군 중 임상적 혹은 병리소견으로 미세변화형 신증후군으로 진단되거나 병리소견상 초점성 분절성 사구체경화증으로 진단된 환아를 대상으로 이들을 연령 (0-l세, 2-4세, 5세 이상), 단백뇨 및 스테로이드 사용 여부 (PU+Tx-, PU+Tx+, PU-Tx+, PU-Tx-)로 구분하였다. 이들과 대조군의 혈청, 요에서 ELISA법으로 각각 sIL-2R를 정량하고, 요에서는 크레아티닌치도 측정하였다. 각 군의 혈청 sIL-2R치와 요 sIL-2R/크레아티닌 비를 계산하여 비교하였다. 결 과 : 혈청 sIL-2R는 환자와 대조군에서 도두 연령이 어릴수록 높았고, 신증후군에서 대조군보다 높지 않았다. 환자군 중에서 재발한 경우에는 높고 스테로이드 투여 시에는 낮은 경향을 보였다. 요 sIL-2R/크레아티닌 비는 특히 단백뇨가 있을 때 연령이 어릴수록 높았고 (P=0.01), 혈청 치와 마찬가지로 재발과 스테로이드의 영향을 받았다. 혈청 sIL-2R치와 요 sIL-2R/크레아티닌 비는 신 병리소견, 스테로이드 반응도에 따른 차이를 보이지 않았다 (P>0.05). 결 론 : 혈청 sIL-2R치는 연령에 따른 차이가 크고, 신증후군에서 대조군에 비해 높지 않았으나, 재발상태의 환자는 완해 상태의 환자보다 높았고, 스테로이드를 투여할 때에 낮았다. 요 sIL-2R/크레아티닌 비는 특히 단백뇨가 있을 때 혈청 sIL-2R치를 잘 반영하였다. Purpose: This study was designed to investigate the changes in soluble interleukin-2 receptor (sIL-2R) level in sera and urines of children with primary nephrotic syndrome, eliminating the confounding effects of age, proteinuria, and steroid treatment. Methods: Soluble IL-2R was measured by ELISA in sera and urines from patients with minimal change nephrotic syndrome or focal segmental glomerulosclerosis as well as from healthy controls. The serum levels and urinary sIL-2R/creatinine ratios were compared between control group and the 12 patient groups divided by their ages (0-1, 2-4, over 5 years), and presence or absence of proteinuria and/or steroid treatment (PU+Tx-, PU+Tx+, PU-Tx+, PU-Tx-). Results: Though the differences were not statistically significant probably because of the small numbers, serum sIL-2R levels seemed to be higher in younger age groups both in patients and control group. Nephrotic children did not show higher serum levels than normal children. Among the patients, proteinuric condition seemed to raise and steroid treatment tended to suppress the serum sIL-2R levels. Urinary sIL-2R/creatinine ratios were higher in younger age groups, more significantly in patients (P<0.001). Proteinuria and steroid treatment affected the urinary sIL-2R/creatinine ratios by the same way as the serum sIL-2R levels. Serum sIL-2R levels and urinary sIL-2R/creatinine ratios were not different between groups of different histologic findings or steroid responsiveness (P>0.05). Conclusion: Serum sIL-2R levels and the urinary sIL-2R/creatinine ratios were higher in younger age, and they were not higher in nephrotic patients compared to control group. The patients in relapse showed higher levels, while the levels were suppressed with steroid treatment. In proteinuric state, urinary sIL-2R/creatinine ratios reflected serum sIL-2R levels.

Anti-inflammatory Activity of 1-docosanoyl Cafferate Isolated from <I>Rhus verniciflua</I> in LPS-stimulated BV2 Microglial Cells

Jae-Won Lee,Il-Young Cheong,Hae-Sung Kim,Jae Jun Lee,Yong-Suk Lee,Yong-Soo Kwon,Myong-Jo Kim,Hee Jae Lee,Sung-Soo Kim,Wanjoo Chun 대한생리학회-대한약리학회 2011 The Korean Journal of Physiology & Pharmacology Vol.15 No.1

Although various derivatives of caffeic acid have been reported to possess a wide variety of biological activities such as protection of neuronal cells against excitotoxicity, the biological activity of 1-docosanoyl cafferate (DC) has not been examined. The objective of the present study was to evaluate the anti-inflammatory effects of DC, isolated from the stem bark of <i>Rhus verniciflua</i>, on lipopoly</I>saccharide (LPS)-stimulated BV2 microglial cells. Pretreatment of cells with DC significantly attenuated LPS-induced NO production, and mRNA and protein expression of iNOS in a concentration- dependent manner. DC also significantly suppressed LPS-induced release of cytokines such as TNF-Ձ and IL-1Ղ. Consistent with the decrease in cytokine release, DC dose-dependently and significantly attenuated LPS-induced mRNA expression of these cytokines. Furthermore, DC significantly suppressed LPS-induced degradation of IKB, which retains NF-kB in the cytoplasm. Therefore, nuclear translocation of NF-kB induced by LPS stimulation was significantly suppressed with DC pretreatment. Taken together, the present study suggests that DC exerts its anti-inflammatory activity through the suppression of NF-kB translocation to the nucleus.

Pseudohypoaldosteronism Type 1

Hae Il Cheong 대한의학유전학회 2013 대한의학유전학회지 Vol.10 No.2

Pseudohypoaldosteronism (PHA), a rare syndrome of systemic or renal mineralocorticoid resistance, is clinically characterized by hyperkalemia, metabolic acidosis, and elevated plasma aldosterone levels with either renal salt wasting or hypertension. PHA is a heterogeneous disorder both clinically and genetically and can be divided into three subgroups; PHA type 1 (PHA1), type 2 (PHA2) and type 3 (PHA3). PHA1 and PHA2 are genetic disorders, and PHA3 is a secondary disease of transient mineralocorticoid resistance mostly associated with urinary tract infections and obstructive uropathies. PHA1 includes two different forms with different severity of the disease and phenotype: a systemic type of disease with autosomal recessive inheritance (caused by mutations of the amiloride-sensitive epithelial sodium channel, ENaC) and a renal form with autosomal dominant inheritance (caused by mutations of the mineralocorticoid receptor, MR). In the kidneys, the distal nephron takes charge of the fine regulation of water absorption and ion handling under the control of aldosterone. Two major intracellular actors necessary for the action of aldosterone are the MR and the ENaC. Impairment of the intracellular aldosterone signal transduction pathway results in resistance to the action of mineralocorticoids, which leads to PHA. Herein, ion handling the distal nephron and the clinico-genetic findings of PHA are reviewed with special emphasis on PHA type 1.

Pseudohypoaldosteronism Type 1

Cheong, Hae Il Korean Society of Medical Genetics and Genomics 2013 대한의학유전학회지 Vol.10 No.2

Pseudohypoaldosteronism (PHA), a rare syndrome of systemic or renal mineralocorticoid resistance, is clinically characterized by hyperkalemia, metabolic acidosis, and elevated plasma aldosterone levels with either renal salt wasting or hypertension. PHA is a heterogeneous disorder both clinically and genetically and can be divided into three subgroups; PHA type 1 (PHA1), type 2 (PHA2) and type 3 (PHA3). PHA1 and PHA2 are genetic disorders, and PHA3 is a secondary disease of transient mineralocorticoid resistance mostly associated with urinary tract infections and obstructive uropathies. PHA1 includes two different forms with different severity of the disease and phenotype: a systemic type of disease with autosomal recessive inheritance (caused by mutations of the amiloride-sensitive epithelial sodium channel, ENaC) and a renal form with autosomal dominant inheritance (caused by mutations of the mineralocorticoid receptor, MR). In the kidneys, the distal nephron takes charge of the fine regulation of water absorption and ion handling under the control of aldosterone. Two major intracellular actors necessary for the action of aldosterone are the MR and the ENaC. Impairment of the intracellular aldosterone signal transduction pathway results in resistance to the action of mineralocorticoids, which leads to PHA. Herein, ion handling the distal nephron and the clinico-genetic findings of PHA are reviewed with special emphasis on PHA type 1.

Molecular Pathogenesis of Nephrogenic Diabetes Insipidus

Hae Il Cheong,Yong Choi 대한전해질학회 2004 대한전해질대사연구회지 Vol.2 No.1

Genetic diagnosis of Alport syndrome

Cheong, Hae Il The Korean Pediatric Society 2019 Clinical and Experimental Pediatrics (CEP) Vol.62 No.5