β-hydroxybutyrate-induced growth inhibition and collagen production in HK-2 cells are dependent on TGF-β and Smad3

被引:39
作者
Guh, JY
Chuang, TD
Chen, HC
Hung, WC
Lai, YH
Shin, SJ
Chuang, LY
机构
[1] Kaohsiung Med Univ, Dept Biochem, Kaohsiung 807, Taiwan
[2] Kaohsiung Med Univ, Dept Internal Med, Kaohsiung 807, Taiwan
[3] Kaohsiung Med Univ, Sch Technol Med Sci, Kaohsiung 807, Taiwan
关键词
diabetic nephropathy; beta-hydroxybutyrate; HK-2; cells; transforming growth factor-beta; Smad;
D O I
10.1046/j.1523-1755.2003.00330.x
中图分类号
R5 [内科学]; R69 [泌尿科学(泌尿生殖系疾病)];
学科分类号
1002 ; 100201 ;
摘要
Background. Ketonuria is common in diabetes. The major form of ketone body is beta-hydroxybutyrate (beta-HB), which is metabolized by the proximal tubule. Transforming growth factor beta (TGF-beta) and tubulopathy are important in diabetic nephropathy. Thus, the role of TGF-beta and the downstream Smad3 in beta-HB-induced effects in the human proximal tubule (HK-2 cell) was studied. Methods. Effects of beta-HB (0.1 to 10 mmol/L) on HK-2 cells were determined for: proliferation, cell cycle distribution, collagen production, tubular transdifferentiation [expression of alpha-smooth muscle actin (alpha-SMA) protein], TGF-beta, Smad2/3, p21(WAF1), and p27(kip1). Results. beta-HB (0.1 to 10 mmol/L) dose dependently decreased proliferation, arrested the cells in G(0)/G(1) phase of the cell cycle, and increased p21(WAF1)/p27(kip1) protein expression at 48 hours (without affecting p21(WAF1)/p27(kip1) mRNA and transcription). beta-HB (1 mmol/L) increased p21(WAF1)/p27(kip1) protein half-lives. beta-HB (1 mmol/L) increased TGF-beta transcription at 24 hours and TGF-beta1 mRNA/bioactivity at 48 hours. beta-HB (1 mmol/L) increased nuclear Smad2/3 protein expression and increased collagen production (without affecting tubular transdifferentiation), which were reversed by Smad7, dominant-negative Smad3, and N-acetylcysteine. Dominant-negative Smad3 reversed beta-HB-induced TGF-beta transcription at 24 hours, and reversed TGF-beta1 bioactivity at 48 hours. Dominant-negative Smad3 reversed beta-HB-induced p21(WAF1)/p27(kip1) protein expression at 48 hours. Finally, N-acetylcysteine, TGF-beta antibody, Smad7, and dominant-negative Smad3 reversed beta-HB (1 mmol/L)-induced growth inhibition at 48 hours. Conclusion. beta-HB activated Smad 2/3 by oxidative stress. TGF-beta and Smad3 mediate beta-HB-induced cell cycle-dependent growth inhibition while Smad3 mediate beta-HB-induced collagen production and p21(WAF1)/p27(kip1) protein expression in HK-2 cells. Moreover, beta-HB increased p21(WAF1)/p27(kip1) protein expression by increasing p21(WAF1)/p27(kip1) protein stability.
引用
收藏
页码:2041 / 2051
页数:11
相关论文
共 51 条
[1]  
*AM DIAB ASS, 2002, DIABETES CARE S1, V25, pS100
[2]   Mice lacking Smad3 show accelerated wound healing and an impaired local inflammatory response [J].
Ashcroft, GS ;
Yang, X ;
Glick, AB ;
Weinstein, M ;
Letterio, JJ ;
Mizel, DE ;
Anzano, M ;
Greenwell-Wild, T ;
Wahl, SM ;
Deng, CX ;
Roberts, AB .
NATURE CELL BIOLOGY, 1999, 1 (05) :260-266
[3]   Signal transduction by the TGF-β superfamily [J].
Attisano, L ;
Wrana, JL .
SCIENCE, 2002, 296 (5573) :1646-1647
[4]   High blood ketone body concentration in Type 2 non-insulin dependent diabetic patients [J].
Avogaro, A ;
Crepaldi, C ;
Miola, M ;
Maran, A ;
Pengo, V ;
Tiengo, A ;
DelPrato, S .
JOURNAL OF ENDOCRINOLOGICAL INVESTIGATION, 1996, 19 (02) :99-105
[5]   Transforming growth factor beta signal transduction in the kidney [J].
Böttinger, EP ;
Miao, JH ;
Schiffer, M ;
Bitzer, M ;
Roberts, ISD .
KIDNEY & BLOOD PRESSURE RESEARCH, 1998, 21 (2-4) :259-261
[6]   EFFECT OF 3-HYDROXYBUTYRATE INFUSION ON URINARY PROTEIN EXCRETION IN HEALTHY MAN [J].
CHRISTENSEN, CK ;
SCHMITZ, O ;
PEDERSEN, EB ;
ALBERTI, KGMM ;
MOGENSEN, CE .
SCANDINAVIAN JOURNAL OF CLINICAL & LABORATORY INVESTIGATION, 1986, 46 (03) :239-243
[7]   Extracellular signals and intracellular pathways in diabetic nephropathy [J].
Chuang, LY ;
Guh, JY .
NEPHROLOGY, 2001, 6 (04) :165-172
[8]   Infection of HepG2 cells with recombinant adenovirus encoding the HCV core protein induces p21WAF1 down-regulation -: effect of transforming growth factor β [J].
Dubourdeau, M ;
Miyamura, T ;
Matsuura, Y ;
Alric, L ;
Pipy, B ;
Rousseau, D .
JOURNAL OF HEPATOLOGY, 2002, 37 (04) :486-492
[9]   WAF1, A POTENTIAL MEDIATOR OF P53 TUMOR SUPPRESSION [J].
ELDEIRY, WS ;
TOKINO, T ;
VELCULESCU, VE ;
LEVY, DB ;
PARSONS, R ;
TRENT, JM ;
LIN, D ;
MERCER, WE ;
KINZLER, KW ;
VOGELSTEIN, B .
CELL, 1993, 75 (04) :817-825
[10]   Transforming growth factor-β regulates tubular epithelial-myofibroblast transdifferentiation in vitro [J].
Fan, JM ;
Ng, YY ;
Hill, PA ;
Nikolic-Paterson, DJ ;
Mu, W ;
Atkins, RC ;
Lan, HY .
KIDNEY INTERNATIONAL, 1999, 56 (04) :1455-1467