Betulinic acid inhibits high glucose-induced vascular smooth muscle cells proliferation and migration.
The proliferation of vascular smooth muscle cells may perform a crucial role in the pathogenesis of diabetic vascular disease. The principal objective of this study was to determine the effects of betulinic acid (BA) on human aortic smooth muscle cell (HASMC) proliferation induced by high glucose (HG). In this study, [(3) H]-thymidine incorporation under 25 mM HG was accelerated significantly as compared with 5.5 mM glucose, and this increase was inhibited significantly by BA treatment. We utilized Western blotting analysis to evaluate the effects of BA on cell-cycle regulatory proteins. HG induced the expression of cyclins/CDKs and reduced the expression of p21(waf1/cip1) /p27(kip1). However, BA also attenuated the expression of HG-induced cell-cycle regulatory proteins. The results of gelatin zymography demonstrated that the HG-treated HASMC secreted gelatinases, probably including MMP-2/-9, which may be involved in the invasion and migration of HASMC. Additionally, BA suppressed the protein and mRNA expression levels of MMP-2/-9 in a dose-dependent manner. BA inhibited the production of HG-induced hydrogen peroxide (H(2)O(2)) and the formation of DCF-sensitive intracellular reactive oxygen species (ROS). Further, BA suppressed the nuclear translocation and phosphorylation of IκB-α of NF-κB under HG conditions. Our results showed that BA exerts multiple effects on HG-induced HASMC proliferation and migration, including the inhibition of both MMP-2 and MMP-9 transcription, protein activity, and the downregulation of ROS/NF-κB signaling, thereby suggesting that BA may be a possible therapeutic approach to the inhibition of diabetic vascular disease.
Yoon
Jung Joo
JJ
College of Oriental Medicine and Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan, Chonbuk 570-749, Republic of Korea.
Lee
Yun Jung
YJ
Kim
Jin Sook
JS
Kang
Dae Gill
DG
Lee
Ho Sub
HS
D001485:Basement Membrane / Q000187:drug effects / Q000378:metabolism; D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D053978:Pentacyclic Triterpenes; D011448:Prostaglandin Antagonists / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014315:Triterpenes / Q000494:pharmacology*; D000094062:Betulinic Acid
D053978:Pentacyclic Triterpenes; D011448:Prostaglandin Antagonists; D017382:Reactive Oxygen Species; D014315:Triterpenes; D006861:Hydrogen Peroxide; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D005947:Glucose; D000094062:Betulinic Acid