Anti-Fibrotic Effects by Moringa Root Extract in Rat Kidney Fibroblast

국문 초록 (Abstract)

신장섬유화는 내 외부적인 요인들에 의해 발생하며, 그 요인들에 의해 염증이 생기고 지속적인 손상이 일어날 경우 신기능의 상실이 유발된다. 또한 신장섬유화는 세포 외 기질의 과다축적, TGF-β나, TNF-α, IL-1과 같은 사이토카인에 의해 발생하며, TGF-β는 신장 섬유화의 과정과 Type ? collagen과 fibronectin, PAI-1을 포함한 섬유화 관련 인자들의 발현 유도에 중요한 역할을 한다. 본 연구에서는 TGF-β를 처리한 신장섬유화 유도 모델에서 Moringa oleifera Lam 추출물에 대한 섬유화 관련 인자들의 영향을 확인하였다. 실험 결과 TGF-β로 유도된 신장 섬유화 세포에서 모링가 추출물이 fibronectin, Type ? collagen과 PAI-1의 단백질 및 mRNA 발현을 저해하였으며, 모링가 추출물 중 모링가 뿌리추출물이 가장 영향이 있는 것으로 확인 되었다. 모링가 뿌리추출물이 어떠한 기전을 통하여 섬유화 관련 인자들의 발현을 조절하는지 알아보기 위한 TGF-β로 유도된 TβR?? 및 그 하위 기전의 인산화 정도를 확인한 실험에서 모링가 뿌리추출물이 TGF-β로 유도된 TβR??과 그 하위기전의 Smad4, ERK의
인산화를 저해하였다. 그러나 TGF-β에 의해 유도된 JNK와 p38, PI3K/AKT의 인산화에는 영향이 없었다. 따라서 모링가 뿌리추출물이 TGF-β로 유도된 신장 섬유아세포에서 TβR??와 그 하위 기전인 Smad4, ERK를 통해서 Type ? collagen 과 fibronectin, PAI-1의 발현을 조절하여 섬유화를 저해 한다는 것을 예상할 수 있다. 결론적으로 모링가 뿌리추출물이 섬유화 치료 및 완화에 좋은 물질이 될 수 있을 것으로 생각된다.

다국어 초록 (Multilingual Abstract)

Fibrosis in kidney by internal and external factors causes progressive loss of renal function. Renal fibrosis is the inevitable consequence of an excessive accumulation of the extracellular matrix. TGF-β plays an important role in the process of renal fibrosis and stimulates the synthesis of profibrotic factors, including collagens, fibronectin, and plasminogen activator inhibitor (PAI-1). We examined the effect of Moringa oleifera Lam (moringa) extracts in a rat kidney fibrosis model. We found that moringa root extract suppresses protein expression/mRNA levels of Type ? collagen, fibronectin, and PAI-1 induced by TGF-β in renal fibroblasts. Moringa root extract selectively inhibited phosphorylation of TGF-β-induced TβR?? and the downstream signaling pathway (e.g., Smad4), and phospho-ERK, but not JNK, p38, or PI3K/AKT. These results suggest that moringa root extract can act against TGF-β -induced renal fibrosis in rat kidney fibroblast cells by a mechanism related to its antifibrotic activity, which regulates expression of fibronectin, Type ? collagen, and PAI-1 through TβR???Smad2/3?Smad4 and ERK. Therefore, moringa root extract is an effective substance for fibrosis therapy and provides a new therapeutic strategy for diseases associated with elevated profibrotic factor synthesis.

목차 (Table of Contents)

• ABSTRACT
• 서론
• 재료 및 방법
• 결과
• 고찰
• References
• 초록

참고문헌 (Reference)

1 Meulders, Q., "Tumor necrosis factor alpha increases antifibrinolytic activity of cultured human mesangial cells" 42 : 327-334, 1992

2 Zhu, B., "Triptolide inhibits extracellular matrix protein synthesis by suppressing the Smad2 but not the MAPK pathway in TGF-beta1-stimulated NRK-49F cells" 25 : 3180-3191,

3 Lund, L. R., "Transforming growth factor-beta is a strong and fast acting positive regulator of the level of type-1 plasminogen activator inhibitor mRNA in WI-38 human lung fibroblasts" 6 : 1281-1286, 1987

4 Border, W. A., "Transforming growth factor beta in tissue fibrosis" 331 : 1286-1292, 1994

5 Kornblihtt, A. R., "The fibronectin gene as a model for splicing and transcription studies" 10 : 248-257, 1996

6 Hayashi, H., "The MAD-related protein Smad7 associates with the TGFbeta receptor and functions as an antagonist of TGFbeta signaling" 89 : 1165-1173, 1997

7 Ma, F. Y., "TGF-{beta}1 activated kinase-1 (TAK1) regulates inflammation and fibrosis in the obstructed kidney" 6 : 1410-1421,

8 Samarakoon, R., "TGF-beta1-induced plasminogen activator inhibitor-1 expression in vascular smooth muscle cells requires pp60(c-src)/EGFR(Y845) and Rho/ROCK signaling" 44 : 527-538, 2008

9 Kutz, S. M., "TGF-beta1-induced PAI-1 gene expression requires MEK activity and cell-to-substrate adhesion" 114 : 3905-3914, 2001

10 Heldin, C. H., "TGF-beta signalling from cell membrane to nucleus through SMAD proteins" 390 : 465-471, 1997

11 Leask, A., "TGF-beta signaling and the fibrotic response" 18 : 816-827, 2004

12 Hocevar, B. A., "TGF-beta induces fibronectin synthesis through a c-Jun N-terminal kinase-dependent, Smad4-independent pathway" 18 : 1345-1356, 1999

13 Border, W. A., "TGF-beta in kidney fibrosis: a target for gene therapy" 51 : 1388-1396, 1997

14 Verrecchia, F., "TGF-beta and TNF-alpha: antagonistic cytokines controlling type I collagen gene expression" 16 : 873-880, 2004

15 Cho, H. J., "Suppression of PAI-1 expression through inhibition of the EGFR-mediated signaling cascade in rat kidney fibroblast by ascofuranone" 107 : 335-344, 2009

16 Imamura, T., "Smad6 inhibits signalling by the TGF-beta superfamily" 389 : 622-626, 1997

17 Attisano, L, "Signal transduction by the TGF-beta superfamily" 296 : 1646-1647, 2002

18 Eddy, A. A., "Serine proteases, inhibitors and receptors in renal fibrosis" 101 : 656-664, 2009

19 Lebrun, J. J., "Roles of pathway-specific and inhibitory Smads in activin receptor signaling" 13 : 15-23, 1999

20 강덕희, "Role of ERK1/2 and p38 mitogen-activated protein kinases in the regulation of thrombospondin-1 by TGF-beta 1 in rat proximal tubular cells and mouse fibroblasts" AMER SOC NEPHROLOGY 16 : 899-904, 200504

21 Liu, Y., "Renal fibrosis: new insights into the pathogenesis and therapeutics" 69 : 213-217, 2006

22 Bhogal, R. K., "Regulatory effect of extracellular signal-regulated kinases (ERK) on type I collagen synthesis in human dermal fibroblasts stimulated by IL-4 and IL-13" 27 : 472-496, 2008

23 Rerolle, J. P., "Plasminogen activator inhibitor type 1 is a potential target in renal fibrogenesis" 58 : 1841-1850, 2000

24 Ponticos, M., "Pivotal role of connective tissue growth factor in lung fibrosis: MAPK-dependent transcriptional activation of type I collagen" 60 : 2142-2155, 2009

25 Souchelnytskyi, S., "Physical and functional interaction of murine and Xenopus Smad7 with bone morphogenetic protein receptors and transforming growth factor-beta receptors" 273 : 25364-25370, 1998

26 Lu, Y, "Phosphatidylinositol-3-kinase/akt regulates bleomycin-induced fibroblast proliferation and collagen production" 42 : 432-441,

27 Eitzman, D. T., "Of mice and men. The function of plasminogen activator inhibitors (PAIs) in vivo" 425 : 131-141, 1997

28 Anwar, F., "Moringa oleifera: a food plant with multiple medicinal uses" 21 : 17-25, 2007

29 Hynes, R., "Molecular biology of fibronectin" 1 : 67-90, 1985

30 Guo, B., "MAPK/AP-1-dependent regulation of PAI-1 gene expression by TGF-beta in rat mesangial cells" 68 : 972-984, 2005

31 Nakao, A., "Identification of Smad7, a TGFbeta-inducible antagonist of TGF-beta signalling" 389 : 631-635, 1997

32 Wilson, H. M., "Effect of transforming growth factor-beta 1 on plasminogen activators and plasminogen activator inhibitor-1 in renal glomerular cells" 1 : 343-350, 1993

33 Mahajan, S. G., "Effect of Moringa oleifera Lam. seed extract on ovalbumin-induced airway inflammation in guinea pigs" 20 : 897-909, 2008

34 Kohan, M., "EDA-containing cellular fibronectin induces fibroblast differentiation through binding to alpha4beta7 integrin receptor and MAPK/Erk 1/2-dependent signaling" 24 : 4503-4512,

35 Gaedeke, J., "Curcumin blocks multiple sites of the TGF-beta signaling cascade in renal cells" 66 : 112-120, 2004

36 Bharali, R., "Chemomodulatory effect of Moringa oleifera, Lam, on hepatic carcinogen metabolising enzymes, antioxidant parameters and skin papillomagenesis in mice" 4 : 131-139, 2003

37 Sreelatha, S., "Antiproliferation and Induction of Apoptosis by Moringa oleifera leaf extract on Human Cancer cells" 6 : 1270-1275,

38 Siddhuraju, P., "Antioxidant properties of various solvent extracts of total phenolic constituents from three different agroclimatic origins of drumstick tree (Moringa oleifera Lam.) leaves" 51 : 2144-2155, 2003

39 Guevara, A. P., "An antitumor promoter from Moringa oleifera Lam" 440 : 181-188, 1999

40 Hamza, A. A., "Ameliorative effects of Moringa oleifera Lam seed extract on liver fibrosis in rats" 48 : 345-355,

41 Yue, J., "Activation of the mitogen-activated protein kinase pathway by transforming growth factor-beta" 142 : 125-131, 2000

42 Shi, Y., "A structural basis for mutational inactivation of the tumour suppressor Smad4" 388 : 87-93, 1997