Effects of rosiglitazone on the proliferation of vascular smooth muscle cell induced by high glucose.
OBJECTIVE
To investigate the effects of the sensitizer rosiglitazone on the proliferation of vascular smooth muscle cell (VSMC) induced by high glucose administration.


METHODS
VSMCs were isolated from rat thoracic aortas and cultured in 10% fetal bovine serum (FBS). VSMC proliferation was evaluated by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and cell counting. The cell cycle was examined by flow cytometry. The protein expressions of proliferating cell nuclear antigen (PCNA) and matrix metalloproteinases-2 (MMP-2) were evaluated by Western blotting. MMP-2 mRNA expression was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and gelatinolytic activity was determined by zymography.


RESULTS
Promoted VSMC proliferation significantly increased the number of VSMCs in the S phase, the expressions of PCNA and MMP-2, and MMP-2 activity, as well as decreased the proportion of VSMCs in the G(0)/G(1) phase. Rosiglitazone at a concentration of 10 mumol/L markedly inhibited glucose-induced VSMC proliferation (1.869 +/- 0.22 vs 0.820 +/- 0.15, P < 0.01). Concomitantly, rosiglitazone inhibited PCNA expression (0.96 +/- 0.07 vs 0.75 +/- 0.06, P < 0.05) and cell cycle progression from G(0)/G(1) to S phase (the proportion of VSMCs in the G(0)/G(1) and S phase were 69.6 +/- 3.96% vs 84.3 +/- 1.73% and 25.2 +/- 1.73% vs 10.1 +/- 1.42% (P < 0.01), respectively). Furthermore, rosiglitazone significantly decreased MMP-2 mRNA expression (0.98 +/- 0.08 vs 0.71 +/- 0.05, P < 0.05), protein expression (0.80 +/- 0.04 vs 0.64 +/- 0.03, P < 0.05) and MMP-2 activity (320 +/- 25% vs 248 +/- 21%, P < 0.05).


CONCLUSIONS
Rosiglitazone significantly inhibited VSMC proliferation, at least in part by inhibiting high glucose-induced G(1)-->S phase transition, PCNA expression and MMP-2 synthesis.
Ling
Hong-Yan
HY
Department of Pharmacology, School of Pharmaceutical Sciences, Central South University, Changsha 410078, China.
Hu
Bi
B
Wang
Bing-Xiang
BX
Zu
Xu-Yu
XY
Feng
Shui-Dong
SD
Ou
He-Sheng
HS
Zhou
Shou-Hong
SH
Liao
Duan-Fang
DF
D000818:Animals; D001013:Aorta, Thoracic / Q000166:cytology; D002417:Cattle; D002452:Cell Count; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry / Q000379:methods; D005947:Glucose / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D007004:Hypoglycemic Agents / Q000494:pharmacology; D008297:Male; D008358:Mannose / Q000494:pharmacology; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects*; D018809:Proliferating Cell Nuclear Antigen / Q000187:drug effects / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077154:Rosiglitazone; D016196:S Phase / Q000187:drug effects; D012867:Skin / Q000187:drug effects / Q000293:injuries; D013778:Tetrazolium Salts; D013844:Thiazoles; D045162:Thiazolidinediones / Q000494:pharmacology*; D014343:Trypan Blue; D014945:Wound Healing / Q000187:drug effects / Q000502:physiology
D007004:Hypoglycemic Agents; D061965:Matrix Metalloproteinase Inhibitors; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D013778:Tetrazolium Salts; D013844:Thiazoles; D045162:Thiazolidinediones; D000077154:Rosiglitazone; D020778:Matrix Metalloproteinase 2; C022616:thiazolyl blue; D014343:Trypan Blue; D005947:Glucose; D008358:Mannose; D004121:Dimethyl Sulfoxide