단백질 아르기닌 메틸전이효소 5(PRMT5)에 의한 3T3L-1 세포의 지방세포 분화 조절

장민준(Min Jung Jang),양지혜(Ji HyeYang),김은주(Eun-Joo Kim) 한국생명과학회 2018 생명과학회지 Vol.28 No.7

PPARγ는 지방세포의 분화를 조절하는 핵심적인 전사 인자로서 이를 조절하는 후성유전학적 조절 기전이 비만억제 연구에서 중요하게 주목 받고 있다. 선행연구에서 CACUL1이 PPARγ의 전사 활성 및 지방세포의 분화를 억제하는 corepressor로서 작용함을 밝힌 바 있으며 본 연구에서는 CACUL1의 새로운 결합 단백질로 발굴된 protein arginine methyltransferase 5 (PRMT5)의 PPARγ 조절 기능을 분석하였다. PRMT5가 CACUL1과 결합함을 immunoprecipitation assay in vivo와 GST-pull down assay in vitro를 통하여 확인하였다. Luciferase reporter assay 결과로 두 단백질이 상호 협력하여 PPARγ의 전사 활성을 억제함을 확인하였다. PRMT5가 안정적으로 과발현 또는 knockdown되는 3T3-L1 세포주를 제작하여 지방세포 분화에 미치는 영향을 분석한 결과, PRMT5가 3T3-L1세포의 지방세포 분화를 억제함을 증명하였다. 같은 맥락으로 PRMT5는 PPARγ의 타겟 유전자인 Lpl과 aP2의 발현을 억제하는 것을 RT-qPCR로 확인하였다. 이상의 연구 결과로 PRMT5이 CACUL1과 결합하여 PPARγ의 전사 활성을 방해, 나아가 지방세포의 분화를 억제하는 기존에 알려지지 않은 분자적 기전을 처음으로 밝혔다. 따라서, PRMT5 효소 활성의 조절은 비만 억제를 위한 약물 개발에 단서를 제공할 것이다. Peroxisome proliferator-activated receptor gamma (PPARγ) is a key transcription factor that regulates adipogenesis, and epigenetic control of PPARγ is of great interest in obesity-inhibition research. Our previous study showed that CACUL1 (CDK2-associated cullin domain 1) acts as a corepressor that inhibits PPARγ transcriptional activity and adipocyte differentiation. Here, we investigated the roles of protein arginine methyltransferase 5 (PRMT5), a novel binding partner of CACUL1, in regulating PPARγ. The interaction between PRMT5 and CACUL1 was shown by immunoprecipitation assay in vivo and GST pulldown assay in vitro. As shown by luciferase reporter assay, PRMT5 and CACUL1 cooperated to inhibit the transcriptional activity of PPARγ. The suppressive role of PRMT5 in adipogenesis was examined by Oil Red O staining using 3T3-L1 cells, which stably overexpress or deplete PRMT5. Overexpression of PRMT5 suppresses PPARγ-mediated adipogenesis, whereas PRMT5 knockdown increases lipid accumulation in 3T3-L1 cells. Consistently, PRMT5 attenuates the expression of Lpl and aP2, the target genes of PPARγ, as demonstrated by RT-qPCR analysis. Overall, these results suggest that PRMT5 interacts with CACUL1 to impair the transcriptional activity of PPARγ, leading to the inhibition of adipocyte differentiation. Therefore, the regulation of PRMT5 enzymatic activity may provide a clue to develop an anti-obesity drug.

Ciglitazone, in Combination with All trans Retinoic Acid, Synergistically Induces PTEN Expression in HL-60 Cells

Lee, Seung-ho,Park, Chul-Hong,Kim, Byeong-Su 충남대학교 형질전환복제돼지연구센터 2007 논문집 Vol. No.10

Peroxisome proliferator-activated receptor-gamma(PPARγ)는 DNA와 결합하기 위해 retinoid-X receptor(RXR)와 heterodimer를 형성해야만 한다. 그리고 전사에 대한 최대활성은 수용체에 대한 리간드 특이성에 의하는 것으로 생각되고 있다. 활성화된 PPARγ와 PPARγ 리간드는 종양억제 PTEN의 조절을 통해 종양세포의 성장에 영향으 끼치게 된다. 본 연구의 목적은 PPARγligand, ciglitazone그리고 RXR ligand로 동시에 자극하였을 때 급성전골수성백혈병(APL)세포에 대해 이들이 함께 PTEN upregulate를 조절할 수 있는지를 결정하기 위함이다. 그리고 이들 세포의 성장과 분화주기에 대해 강력한 억제 능이 있는지를 결정하고자 하였다. 즉, 사람의 백혈병세포주인 HL-60세포에 all-trans-retinol과 ciglutazone을 노출시킨 뒤 PTEN 발현에 대한 측정을 위해 RT-PCR법으로 PTEN mRNA 발현 정도를 확인하고 western blot으로 분석하였다. 세포주기의 분석은 propidium iodide(PI) 염색법과 FACScan으로 분석하였고, HL-60 cells에서 PPARγligand, ciglitazone, 그리고 RXR lignd, retinoic acid 그리고 upregulated PTEN 발현에 대한 time-and dose-dependent 방법으로 각각 확인하였던 바 ciglitazone과 retinoic acid를 동시 조합하여 처치하였을 때 유의적인 효과를 인정할 수 있었다. 더욱이 이들 혼합 물질은 세포의 성장과 G₁phase를 동시 억제하는 능력이 있었다. 그러므로 PPARγ의 활성에 있어 RXR heterodimer가 사람의 백혈병세포에 대한 조절 경로로서 존재하며, PTEN의 upregulation을 통해 백혈병을 조절하기 때문에 백혈병의 예방 및 치료 접근에 PPARγ와 RXR ligands가 중요한 역할을 할 것이다. Peroxisome proliferator-activated receptor-gamma (PPARγ) must form a heterodimer with the retinoid-X receptor (RXR) to bind DNA, and its transcriptional activity is thought to be maximized by ligands specific for either receptor. Activated PPARγ and PPARγ ligands may influence tumor growth through regulation of the tumor suppressor PTEN. Our aim in this study was to determine whether co-stimulation with the PPARγ ligand, ciglitazone, and RXR ligand can synergistically upregulate PTEN in human acute promyelocytic leukemia (APL) cells and consequently potentate the inhibition of cell growth and cell cycle progression of these cells. Human leukemia cell line, HL-60 cells were exposed to all-trans-retinol and ciglutazone. The PTEN expression was measured as the level of PTEN mRNA expression by RT-PCR and as the level of PTEN expression by western blot analysis. Cell cycle analysis was carried out by a propidium iodide (PI) staining method and analyzed with a FACScan. The PPARγ ligand, ciglitazone, and the RXR ligand, retinoic acid, upregulated PTEN expression by HL-60 cells in time- and dose-dependent manners, respectively. This was significantly enhanced by a combination of both ciglitazone and retinoic acid. Moreover, these compounds synergistically induced arrests of both cell growth and the G₁phase of the cell cycle. Thus, the activation of the PPARγ: RXR heterodimer may represent a regulatory pathway for human leukemia cells and there may be important roles for PPARγ and RXR ligands in prophylactic and therapeutic approaches for controlling leukemia through the upregulation of PTEN.

The PPARδ ligand L‐165041 inhibits vegf‐induced angiogenesis, but the antiangiogenic effect is not related to PPARδ

Park, Jin‐,Hee,Lee, Kuy‐,Sook,Lim, Hyun‐,Joung,Kim, Hanna,Kwak, Hyun‐,Jeong,Park, Hyun‐,Young Wiley Subscription Services, Inc., A Wiley Company 2012 Journal of cellular biochemistry Vol.113 No.6

<P><B>Abstract</B></P><P>Peroxisome proliferator‐activated receptor (PPAR)δ is known to be expressed ubiquitously and involved in lipid and glucose metabolism. Recent studies have demonstrated that PPARδ is expressed in endothelial cells (ECs) and plays a potential role in endothelial survival and proliferation. Although PPARα and PPARγ are well recognized to play anti‐inflammatory, antiproliferative, and antiangiogenic roles in ECs, the general effect of PPARδ on angiogenesis in ECs remains unclear. Thus, we investigated the effect of the PPARδ ligand L‐165041 on vascular EC proliferation and angiogenesis in vitro as well as in vivo. Our data show that L‐165041 inhibited VEGF‐induced cell proliferation and migration in human umbilical vein ECs (HUVECs). L‐165041 also inhibited angiogenesis in the Matrigel plug assay and aortic ring assay. Flow cytometric analysis indicated that L‐165041 reduced the number of ECs in the S phase and the expression levels of cell cycle regulatory proteins such as cyclin A, cyclin E, CDK2, and CDK4; phosphorylation of the retinoblastoma protein was suppressed by pretreatment with L‐165041. We confirmed whether these antiangiogenic effects of L‐165041 were PPARδ‐dependent using GW501516 and PPARδ siRNA. GW501516 treatment did not inhibit VEGF‐induced angiogenesis, and transfection of PPARδ siRNA did not reverse this antiangiogenic effect of L‐165041, suggesting that the antiangiogenic effect of L‐165041 on ECs is PPARδ‐independent. Together, these data indicate that the PPARδ ligand L‐165041 inhibits VEGF‐stimulated angiogenesis by suppressing the cell cycle progression independently of PPARδ. This study highlights the therapeutic potential of L‐165041 in the treatment of many disorders related to pathological angiogenesis. J. Cell. Biochem. 113: 1947–1954, 2012. © 2012 Wiley Periodicals, Inc.</P>

치수세포에서 PPARγ의 항 염증작용에 관한 연구

김정희 大韓齒科保存學會 2006 Restorative Dentistry & Endodontics Vol.31 No.3

치수는 상아질로 둘러싸인 간엽조직으로 다양한 세포와 기저 물질들로 구성되어 있으며 혈관과 신경조직이 분포되어 있다. 치수의 염증은 조직의 분해를 야기하며 이는 Matrix Metalloproteinase에 의해 세포 외 기질의 분해가 촉진되어 병적인 과정을 거치게 된다. 이에 Lipopolysaccharide에 의한 MMP와 inflammatory cytokine의 유도와 peroxisome proliferator-activated receptors (PPAR)에 의한 염증매개 물질의 조절에 대해 알아보고자 하였다. 사람의 치수세포를 다양한 LPS농도에 노출시킨 후 24시간째 MMP-2, MMP-9의 변화를 보고 LPS에 의해 자극된 치수세포에서 ICAM-1, VCAM-1, IL-1ß, TNF-α의 분비가 증가됨을 알 수 있었다. 또한 Adenovirus PPARγ (Ad/PPARγ)와 PPARγ agonist인 rosiglitazone를 LPS로 자극된 치수세포에 처리하였을 때 48시간째 MMPs와 Adhesion molecules, cytokines의 감소를 확인하였다. 이로써 사람의 치수세포에서 PPARγ가 가지는 항 염증효과에 대해 지속적인 연구가 필요할 것으로 사료된다. Dental pulp is a loose, mesenchymal tissue almost entirely enclosed in the dentin. It consists of cells, ground substance, and neural and vascular supplies. Damage to the dental pulp by mechanical, chemical, thermal, and microbial irritants can provoke various types of inflammatory response. Pulpal inflammation leads to the tissue degradation, which is mediated in part by Matrix metalloproteinase leads to accelerate extracellular matrix degradation with pathological pathway. We have now investigated the induction of MMPs and inflammatory cytokines by Lipopolysaccharide (LPS) control of inflammatory mediators by peroxisome proliferator-activated receptors (PPARs). Human dental pulp cells exposed to various concetrations of LPS (1-10 ㎍/㎖) revealed elevated levels of MMP-2 and MMP-9 at 24 hrs of culture. LPS also stimulated the production of ICAM-1, VCAM-1, IL-1ß, and TNF-α. Adenovirus PPARγ (Ad/PPARγ) and PPARγ agonist rosiglitazone reduced the synthesis of MMPs, adhesion molecules and pro-inflammatory cytokines. The inhibitory effect of Ad/PPARγ was higher than that of PPARγ agonist. These result offer new insights in regard to the anti-inflammatory potential of PPARγ in human dental pulp cell.

자연과학편 : 흰쥐의 장기간 지구성 운동과 골격근 PPARβ/δ 기능 억제가 GLUT4 발현에 미치는 영향

고진호(JinHoKoh),김기진(KiJinKim) 한국체육학회 2015 한국체육학회지 Vol.54 No.4

본 연구는 장기간 지구성 운동과 Peroxisome proliferator-activated receptor beta/delta(PPARβ/δ) 기능 억제가 흰쥐의 골격근 glucose transporter type 4(GLUT4) 발현에 어떠한 영향을 미치는지 확인하는 것이다. 2주간 수영 운동을 실시하기 전 전기 자극 유전자 전이법(electroporation; EPO)을 이용하여 shot-hairpin PPARβ/δ(shPPARβ/δ)와 scramble(Scr) 유전자(control)를 각각 흰쥐의 왼쪽과 오른쪽 epitrochlearis(Epi) 근육에 전이하였다. 2주간 수영 운동 후 PPARβ/δ, nuclear respiratory factor 1(NRF-1), myocyte enhancer factor 2(MEF2A) 그리고 GLUT4 단백질 발현을 분석하기 위하여 Epi 근육을 적출하였다. shPPARβ/δ가 전이된 좌업 그룹의 골격근 PPARβ/δ, NRF-1, MEF2A 그리고 GLUT4 단백질 발현은 좌업 그룹의 Scr 유전자가 처치된 근육보다 50% 이상 유의하게 감소하였다. Scr 유전자를 처치하고 2주간 수영 운동을 실시한 근육의 PPARβ/δ, NRF-1, MEF2A 그리고 GLUT4 단백질 발현은 좌업 그룹의 Scr 유전자 처치 근육보다 2배 이상 유의하게 증가하였다. 그러나 shPPARβ/δ가 전이된 후 2주간 지구성 운동을 실시한 그룹의 골격근 PPARβ/δ, NRF-1, MEF2A 그리고 GLUT4 단백질 발현은 좌업 그룹의 Scr을 처치한 근육보다 증가되지 않았다. 본 연구는 장기간지구성 운동에 의한 골격근 GLUT4의 발현이 PPARβ/δ에 의하여 조절된다는 것을 확인하였다. The aim of this study was to identify the effects of long-term endurance exercise on glucose transporter type 4(GLUT4) expression in rat skeletal muscle that inhibition of Peroxisome proliferator-activated receptor beta/delta(PPARβ/δ) function by shot-hairpin PPARβ/δ(shPPARβ/δ) overexpression. shPPARβ/δ was over-expressed in rat epitrochlearis (Epi) muscle using electrical pulse-mediated gen transfer (electroporation; EPO) method before 2 wks swimming exercise. To evaluate PPAR β/δ, nuclear respiratory factor 1(NRF-1), myocyte enhancer factor 2(MEF2A) and GLUT4 protein expression in rat skeletal muscle, Epi muscle were dissected after 2 wks swimming exercise. PPARβ/δ, NRF-1, MEF2A and GLUT4 protein expression in shPPARβ/δ over-expression sedentary (Sed) skeletal muscle were significantly decreased over 50% when compare to scramble gene treated Sed muscle. PPARβ/δ, NRF-1, MEF2A and GLUT4 protein expression in 2 wks swimming exercise with Scr gene treated muscle were significantly increased over 2 fold when compare to Scr gene treated Sed skeletal muscle, but PPARβ/δ, NRF-1, MEF2A and GLUT4 protein expression in 2wks swimming exercise with shPPARβ/δ over-expression skeletal muscle were not increased when compare to Scr gene treated Sed skeletal muscle. Our results indicate that GLUT4 protein expression in skeletal muscle was controled by PPARβ/δ following long-term endurance exercise.

원문 : PPARδ가 생쥐 골격근의 미토콘드리아 생합성에 미치는 영향

고진호 ( Jin Ho Koh ),김상현 ( Sang Hyun Kim ),정수련 ( Su Ryun Jung ),김기진 ( Ki Jin Kim ) 한국운동생리학회(구 한국운동과학회) 2014 운동과학 Vol.23 No.4

본 연구는 골격근에 PPARδ의 과발현이 미토콘드리아 효소의 발현에 어떠한 영향을 미치는지 확인하는 것이다. 전기자극유전자 전이기법을 이용하여 생쥐의 골격근에 PPARδ의 과발현을 유도하였으며, PGC-1α가 증가하기 전과 증가된 시점에서 전자전달계 효소인 NADH dehydrogenase(NADH), cytochrome c oxidase subunit Ⅳ(COXⅣ) 및 succinate-ubiquinone oxidoreductase(SUO)와 구연산회로의 효소인 citrate synthase(CS)의 발현을 분석하였다. 생쥐의 골격근에 PPARδ가 2주 또는 4주간 과발현된 근육은 PPARδ가 대조군보다 각각 2.1배 2.8배 증가하였다. PPARδ가 2주간 과발현된 근육은 PGC-1α, COXⅣ, CS가 대조군과 차이가 없는 것으로 나타났으나, NADH와 SUO는 대조군보다 각각 1.5배 2.4배 증가하였다. PPARδ가 4주간 과발현된 근육은 PGC-1α, COXⅣ, CS, NADH 그리고 SUO가 대조군보다 각각 1.6배, 2.1배, 1.8배, 2배 그리고 2.5배 증가하였다. 본 연구는 PPARδ가 PGC-1α의 단백질 발현을 증가시키거나 또는 다른 기전을 통하여 미토콘드리아 효소의 발현을 증가시킬 수 있음을 증명하였다. The purpose of this study was to evaluate the role of PGC-1α and PPARδ on the mitochondrial biogenesis in mouse skeletal muscle. PPARδ overexpression in mouse skeletal muscle was achieved by electroporation (EPO). To evaluate mitochondrial biogenesis, such as NADH dehydrogenase (NADH), cytochrome c oxidase subunit Ⅳ (COXⅣ), citrate synthase (CS) were analyzed 2 and 4 weeks post PPARδ overexpression by EPO. Two and four weeks after PPARδ EPO, PPARδ expression in muscle was increased 2.1 and 2.8 fold respectively when compared to empty vector (EV) treated muscle. Two weeks after PPARδ EPO in muscle, PGC-1α, COXⅣ, and CS were not increased, but NADH and SUO were increased 1.5 and 2.4 fold respectively when compared to EV treated muscle. Four weeks after PPARδ EPO in muscle, PGC-1α, COXⅣ, CS, NADH, and SUO were increased 1.6, 2.1, 1.8, 2, and 2.5 fold respectively when compared to EV treated muscle. Thus, this study demonstrated that PPARδ can increase PGC-1α by posttranscriptional mechanism causing mitochondrial biogenesis, and PPARδ can affect directly or indirectly mitochodrial biogenesis without PGC-1α.

생쥐의 지구성 운동에 의한 Tfam 발현이 AMPK, PPARβ/δ 및 PGC-1α의 발현에 미치는 영향

고진호(KohJin-ho),김기진(KimKi-jin) 한국체육학회 2017 한국체육학회지 Vol.56 No.2

Mitocondrial transcription factor A (Tfam)의 과발현이 미토콘드리아 생합성 및 호흡 기능을 증가시키는 것으로 보고되었으나 지구성 운동에 의한 Tfam의 발현이 어떠한 기전으로 핵으로 신호를 보내며 근 적응의 주요 인자들과 상호작용하는지 확인되지 않았다. 본 연구는 지구성 운동에 의한 Tfam의 발현이 어떠한 기전으로 골격근 적응의 주요 요인인 AMP-activated protein kinase (AMPK), Peroxisome proliferator-activated receptor (PPARβ/δ) 및 Peroxisome proliferator activated receptor gamma coactivator-1α (PGC-1α) 발현에 영향을 미치는지 확인하는 것이다. 생쥐들은 2주간 수영 운동을 실시하거나, Tfam이 골격근에 과발현되도록 유전자 전이를 실시하였다. 또한 PPARβ/δ와 Tfam의 관련성을 확인하기 위하여 PGC-1α knockout 생쥐의 골격근에 PPARβ/δ를 EPO 하였다. 2주간 지구성 운동을 실시한 생쥐의 근육에서 Tfam, PPARβ/δ 및 PGC-1α의 발현이 증가하였다. Tfam이 과발현 된 근육의 AMP:ATP 비율, pAMPK, PPARβ/δ 및 PGC-1α가 증가하였다. PGC-1α knokout 생쥐의 근육에 PPARβ/δ의 과발현은 Tfam의 발현을 증가시키는 것으로 나타났다. 본 연구는 지구성 운동에 의한 Tfam의 발현 증가가 세포내 AMP:ATP 비율 증가를 유도하여 AMPK의 활성을 증가시키며, AMPK의 활성은 PPARβ/δ와 PGC-1α의 발현을 증가시킨다는 것을 확인하였다. 이는 Tfam이 AMPK-PPARβ/δ 및 PGC-1α를 통하여 자신의 발현도 조절할 수 있음을 나타낸다. Tfam overexpression has been shown to increase mitochondrial biogenesis and respiratory function, but there is not clear how Tfam signals to the nucleus, and that a key factors associated with muscle adaptation are involved in Tfam overexpression by endurance exercise. The aim of this study was to identify the effects of Tfam expression by endurance exercise on AMPK, PPARβ/δ and PGC-1α that are associated with skeletal muscle adaptive response. Mice were subjected to forced swimming for 2 wk or Tfam was electroporated in tibialis anterior (TA) muscle. To identify PPARβ/δ effect on Tfam, PPARβ/δ was electroporated in PGC-1α knockout mouse muscle. Mice Swimming for 2 wk resulted in an increase Tfam, PPARβ/δ and PGC-1α in TA muscle. Tfam overexpression in mouse TA muscle resulted in an increase AMP:ATP ratio, AMPK phosphorylation, PPARβ/δ and PGC-1α expression. PPARβ/δ overexpression in PGC-1 α knockout muscle in mouse resulted in an increase Tfam expression. Our finding indicate that an increased Tfam expression by endurance exercise increases AMPK activation through controled AMP:ATP ratio that result in an increase PPARβ/δ and PGC-1α expression. These results indicate that Tfam can increase itself through AMPK-PPARβ/δ and/or PGC-1α axis.

지구성 운동에 의한 PPARβ의 발현이 고지방 식이에 의한 골격근 미토콘드리아 H₂O₂의 배출과 GLUT4 발현에 미치는 영향

고진호(KohJin-ho),김기진(KimKi-jin) 한국체육학회 2017 한국체육학회지 Vol.56 No.4

고지방 식이는 골격근 미토콘드리아로부터 hydrogen peroxide (H2O2) 배출을 증가시키며, 과도한 H2O2의 배출은 인슐린 저항성의 원인이다. 본 연구는 지구성 운동에 의한 Peroxisome proliferator-activated receptor β/δ (PPARβ/δ, PPARβ)의 발현이 고지방 식이에 의한 미토콘드리아 H2O2의 배출을 감소시킬 수 있는지 확인하는 것이다. 생쥐는 8 주간 고지방 식이, 지구성운동 또는 지구성 운동과 고지방 식이를 병행 실시하였으며, 포화지방산에 의한 근세포(myotube) 미토콘드리아의 H2O2 배출과 항산화 효소에 대한 PPARβ의 효과를 살펴보기 위해 PPARβ, sodium palmitate 또는 PPARβ와 sodium palmitate를 병행 처치하였다. 8주간 고지방 식이는 골격근 미토콘드리아로부터 H2O2 의 배출을 증가시키며 glucose transporter 4 (GLUT4)와 superoxide dismutase (SOD)2의 발현을 유의하게 감소시켰다. 그러나 8주간 지구성 운동은 SOD2, catalase 및 GLUT4의 발현을 증가시켰으며, 고지방 식이에 의한 골격근 미토콘드리아의 H2O2 배출을 감소시켰다. 또한 근세포에 PPARβ의 과발현은 SOD2, catalase 및 GLUT4를 증가시켰으며, 포화지방산에 의한 미토콘드리아 H2O2의 배출을 감소시켰다. 따라서 지구성 운동에 의하여 증가된 PPARβ는 SOD2, catalase 를 증가시키며, 이는 고지방 식이에 의한 골격근 미토콘드리아로부터 H2O2의 배출을 감소시켜 GLUT4의 발현 감소를차단한다. Chronic high fat diet (HFD) induced excessive hydrogen peroxide (H2O2) emission from mitochondria in skeletal muscle results in an increase insulin resistance. The aim of this study was to identify the effects of PPARβ induced by endurance exercise on excessive H2O2 emission from mitochondria in skeletal muscle by HFD. Studies of HFD, endurance treadmill exercise (Ex) and HFD+Ex in mice were conducted for 8 weeks. PPARβ transfection, sodium palmitate or PPARβ + sodium palmitate treatments in C2C12 myotubes were conducted to identify PPARβ effects on mitochondrial H2O2 emission induced by saturated fatty acids. HFD fed mice for 8 weeks result in an increase mitochondrial H2O2 emission and a decrease GLUT and SOD2, but endurance exercise for 8 weeks result in an increase SOD2, catalase and GLUT4 expression, and a decrease mitochondrial H2O2 emission in skeletal muscle. SOD2, catalase and GLUT4 expression were increased in myotubes by PPARβ overexpression result in a decrease mitochondrial H2O2 emission induced by palmitate. We concluded that an increased PPARβ by endurance exercise increases SOD2 and catalase that results in a decrease mitochondrial H2O2 emission, and blocks a decrease in GLUT4 expression.

Structural basis for differential activities of enantiomeric PPARγ agonists: Binding of S35 to the alternate site

Jang, J.Y.,Koh, M.,Bae, H.,An, D.R.,Im, H.N.,Kim, H.S.,Yoon, J.Y.,Yoon, H.J.,Han, B.W.,Park, S.B.,Suh, S.W. Elsevier Science 2017 Biochimica et biophysica acta. Proteins and proteo Vol.1865 No.6

<P>Peroxisome proliferator-activated receptor gamma (PPAR gamma) is a member of the nuclear receptor superfamily. It functions as a ligand-activated transcription factor and plays important roles in the regulation of adipocyte differentiation, type 2 diabetes mellitus, and inflammation. Many PPAR gamma agonists bind to the canonical ligand-binding pocket near the activation function-2 (AF-2) helix (i.e., helix H12) of the ligand-binding domain (LBD). More recently, an alternate ligand-binding site was identified in PPAR gamma LBD; it is located beside the 52 loop between the helices H2 ' and H3. We reported previously that the chirality of two optimized enantiomeric PPAR gamma ligands (S35 and R35) differentiates their PPAR gamma transcriptional activity, binding affinity, and inhibitory activity toward Cdk5 (cyclin-dependent kinase 5)-mediated phosphorylation of PPAR gamma at Ser245 (in PPAR gamma 1 numbering; Ser273 in PPAR gamma 2 numbering). S35 is a PPAR gamma phosphorylation inhibitor with promising glucose uptake potential, whereas R35 behaves as a potent conventional PPAR gamma agonist. To provide a structural basis for understanding the differential activities of these enantiomeric ligands, we have determined crystal structures of the PPAR gamma LBD in complex with either S35 or R35. S35 and R35 bind to the PPAR gamma LBD in significantly different manners. The partial agonist S35 occupies the alternate site near the Omega loop, whereas the full agonist R35 binds entirely to the canonical LBP. Alternate site binding of S35 affects the PPAR gamma transactivation and the inhibitory effect on PPAR gamma Ser245 phosphorylation. This study provides a useful platform for the development of a new generation of PPAR gamma ligands as anti-diabetic drug candidates.</P>

소풍순기원(疏風順氣元)이 mouse의 NMu2Li 간세포와 C2C12 골격근세포에서 PPARs 조절의 분자기전에 미치는 영향

오영진 ( Young Jin Oh ),신순식 ( Soon Shik Shin ),윤미정 ( Mi Chung Yoon ),김보경 ( Bo Kyung Kim ) 대한한방신경정신과학회 2009 동의신경정신과학회지 Vol.20 No.1

Objectives: We investigated the effects of Sopungsungi-won(Shufengshunqiyuan) (SSEx1, SSEx2) to treat the metabolic syndrome by the molecular mechanism of regulation of PPAR and modulation of mitochondrial MCAD, VLCAD mRNA expression. Methods: Mouse NMu2Li liver cells and C2C12 skeletal muscle myogenic progenital cells were transiently transfected with expression plasmids for PPAR(PPARα, PPARδ), a luciferase reporter gene construct containing 3 copies of the PPRE from the rat acyl-CoA oxidase gene and β-galactosidase gene. Cells were treated with several concentrated kinds of SSEx1, SSEx2 at the initial time of culture and analyzed PPARα, PPARδ reporter gene activity using spectrophotometer (405 nm). Total RNA was extracted from SSEx1, SSEx2 and measured mRNA levels of mitochondrial MCAD, VLCAD. Representative RT-PCR bands are shown. Results: 1. SSEx1 increased the expression of PPARα reporter gene activities at 0.1 μg/ml (p<0.01) and 10 μg/ml (p<0.05), SSEx2 at 0.1 μg/ml (p<0.01) and 1 μg/ml (p<0.05) significantly in NMu2Li liver cell lines. 2. SSEx1 increased the expression of PPARα reporter gene activities at 1 μg/ml (p<0.01), SSEx2 at 0.1 μg/ml (p<0.01) significantly in C2C12 skeletal muscle cells. 3. SSEx1, SSEx2 both showed no significant changes of the expression of PPARδ reporter gene activities in C2C12 skeletal muscle cells. 4. SSEx1 increased the modulation of mitochondrial MCAD mRNA expression (p<0.05) significantly in NMu2Li liver cell lines. 5. SSEx1, SSEx2 both increased the modulation of mitochondrial MCAD mRNA expression (p<0.05) significantly in C2C12 skeletal muscle cells. Conclusions: These results show the SSEx1, SSEx2 can be used as therapeutic agent for metabolic syndrome and it`s molecular mechanisms of PPAR more contribute to the activation of PPARα then PPARδ reporter gene activities and it`s total RNA more contribute to the modulation of mitochondrial MCAD then VLCAD mRNA expression.