Yonsei Med J.  2015 Nov;56(6):1597-1603. 10.3349/ymj.2015.56.6.1597.

All-Trans Retinoic Acid Has a Potential Therapeutic Role for Diabetic Nephropathy

• Kim CS
• Park JS
• Ahn CW
• Kim KR

• KMID: 2345888
• DOI: http://doi.org/10.3349/ymj.2015.56.6.1597

Abstract

PURPOSE
The aim of this study was to examine the effects of all-trans retinoic acid (ATRA) on diabetic nephropathy.
MATERIALS AND METHODS
We measured amounts of urinary albumin excretion (UAE) after administrating ATRA to Otsuka Long-Evans Tokushima Fatty (OLETF) rats. In order to understand the mechanism of action for ATRA, we administrated ATRA to examine its inhibitory action on the production of transforming growth factor-beta1 (TGF-beta1), protein kinase C (PKC), and reactive oxidative stress (ROS) in cultured rat mesangial cells (RMCs).
RESULTS
After 16 weeks of treatment, UAE was lower in the ATRA-treated OLETF rats than in the non-treated OLETF rats (0.07+/-0.03 mg/mgCr vs. 0.17+/-0.15 mg/mgCr, p<0.01). After incubation of RMCs in media containing 30 or 5 mM of glucose, treatment with ATRA showed time- and dose-dependent decreases in TGF-beta1 levels and ROS. Moreover, ATRA treatment showed a dose-dependent decrease in PKC expression.
CONCLUSION
ATRA treatment suppressed UAE and TGF-beta1 synthesis, which was mediated by significant reductions in PKC activity and ROS production. Our results suggest that ATRA has a potential therapeutic role for diabetic nephropathy.

Keyword

All-trans retinoic acid; diabetic nephropathy; transforming growth factor-beta1; protein kinase-C; reactive oxygen species

MeSH Terms

Animals
Diabetes Mellitus, Type 2/*complications
Diabetic Nephropathies/*complications/*drug therapy/pathology
Mesangial Cells/*metabolism
Oxidative Stress/drug effects
Rats
Rats, Inbred OLETF
Reactive Oxygen Species/metabolism
Transforming Growth Factor beta1/analysis/pharmacology
Tretinoin/*pharmacology/therapeutic use
Reactive Oxygen Species
Transforming Growth Factor beta1
Tretinoin

Figure

• 

  Fig. 1 Body weight of rats from 10 to 44 weeks of age. There was no significant difference in weights among the three groups prior to 28 weeks. After 28 weeks of age, the OLETF rats weighed more than the LETO rats of the same age, while there was no statistical difference in weights between the non-treated OLETF rat group and the ATRA-treated OLETF rat group. A statistical difference was determined between groups with the same duration of treatment period. White circle: non-treated OLETF rat, black circle: ATRA-treated OLETF rat, white square: LETO rat. *p<0.01 versus the LETO rats. OLETF, Otsuka Long-Evans Tokushima Fatty; LETO, Long-Evans-Tokushima-Otsuka; ATRA, all-trans retinoic acid.

• 

  Fig. 2 Effect of ATRA on TGF-β1 protein. (A) After the incubation of quiescent mesangial cells in media containing 30 (high) or 5 mM (control) glucose and different concentrations of ATRA. Treatment with ATRA under both 30 and 5 mM glucose conditions was associated with dose-dependent decreases in TGF-β1 levels (p<0.05 by a test for a trend). (B) After the incubation of quiescent rat mesangial cells with 10-5 M ATRA for the given periods (6, 24, and 48 hrs) in media containing 30 or 5 mM of glucose. Treatment with ATRA under both 30 and 5 mM glucose conditions showed time-dependent decreases in TGF-β1 levels (p<0.05 by a test for a trend). Values are expressed as mean±SEM of three separate experiments. *p<0.05 compared to the vehicle, i.e., without ATRA, †p<0.05 compared with 0 hour. TGF-β1, transforming growth factor-β1; ATRA, all-trans retinoic acid; SEM, standard error of the mean.

• 

  Fig. 3 Effect of all-trans retinoic acid (ATRA) on protein kinase C (PKC)-α (A), β (B), δ (C) expression at 24 hrs in rat mesangial cells (RMCs) in media containing 30 (high) or 5 (control) mM of glucose. Equal amounts of total cell lysate were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted with an anti-PKC isoforms. The RMCs were treated with a vehicle (without ATRA, C, Lane 1), 10-8 M (Lane 2), 10-7 M (Lane 3), 10-6 M (Lane 4), or 10-5 M (Lane 5) of ATRA. *p<0.05 versus the control.

• 

  Fig. 4 Effect of ATRA on dichlorofluorescein (DCF)-sensitive cellular reactive oxygen species (ROS). (A) Synchronized quiescent rat mesangial cells (RMCs) grown on cover glass were incubated in media containing 30 (high) or 5 (control) mM glucose for 6 hrs. After the incubation of the quiescent RMCs under different experimental conditions, DCF-sensitive cellular ROS were measured as described in the text. DCF-sensitive cellular ROS in the RMCs showed a dose-dependent decrease after ATRA administration (p<0.05 by a test for a trend). (B) After the incubation of the quiescent RMCs with 10-5 M of ATRA for the given periods (6, 24, and 48 hrs) in media containing 30 or 5 mM of glucose, a relative decrease of ROS was seen. DCF-sensitive cellular ROS in the RMCs showed a time-dependent decrease after ATRA administration (p<0.05 by a test for a trend). Values are expressed as mean±SEM of three separate experiments. *p<0.05 compared to the vehicle, †p<0.05 compared with 0 hour. ATRA, all-trans retinoic acid; SEM, standard error of the mean.

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