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Inhibitory effects of lithospermic acid on proliferation and migration of rat vascular smooth muscle cells.

Chen L, Wang WY, Wang YP - Acta Pharmacol. Sin. (2009)

Bottom Line: To understand the effects of lithospermic acid (LA), a potent antioxidant from the water-soluble extract of Salvia miltiorrhiza, on the migration and proliferation of rat thoracic aorta vascular smooth muscle cells (VSMCs).Furthermore, LA attenuated LPS-induced VSMC migration by inhibiting MMP-9 expression and its enzymatic activity.LA is able to inhibit FBS-induced VSMC proliferation and LPS-induced VSMC migration, which suggests that LA may have therapeutic effects in the prevention of atherosclerosis, restenosis and neointimal hyperplasia.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of New Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.

ABSTRACT

Aim: To understand the effects of lithospermic acid (LA), a potent antioxidant from the water-soluble extract of Salvia miltiorrhiza, on the migration and proliferation of rat thoracic aorta vascular smooth muscle cells (VSMCs).

Methods: VSMC migration, proliferation, DNA synthesis and cell cycle progression were investigated by transwell migration analysis, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, bromodeoxyuridine (BrdU) incorporation assay, and flow cytometric detection, respectively. Intracellular reactive oxygen species (ROS) generation was detected using 2',7'-dichlorofluorescin diacetate (DCFH-DA). The expression of cyclin D1 protein and matrix metalloproteinase-9 (MMP-9) protein, as well as the phosphorylation state of ERK1/2, were determined using Western blots. The activity of MMP-9 and the expression of MMP-9 mRNA were assessed by gelatin zymography analysis and RT-PCR, respectively.

Results: LA (25-100 micromol/L) inhibited both lipopolysaccharide (LPS)- and fetal bovine serum (FBS)-induced ROS generation and ERK1/2 phosphorylation. By down-regulating the expression of cyclin D(1) and arresting cell cycle progression at the G(1) phase, LA inhibited both VSMC proliferation and DNA synthesis as induced by 5% FBS. Furthermore, LA attenuated LPS-induced VSMC migration by inhibiting MMP-9 expression and its enzymatic activity.

Conclusion: LA is able to inhibit FBS-induced VSMC proliferation and LPS-induced VSMC migration, which suggests that LA may have therapeutic effects in the prevention of atherosclerosis, restenosis and neointimal hyperplasia.

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Inhibitory effect of LA on FBS-stimulated cyclin D1 protein expression. Growth-arrested VSMCs were pretreated with LA (50 or 100 μmol/L) for 2 h and then stimulated with 5% FBS for 24 h. Expression of cyclin D1 protein was examined by Western blotting. β-actin was used as a loading control. Values are presented as mean±SEM. n=3. bP<0.05 vs control. eP<0.05, fP<0.01 vs FBS.
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fig5: Inhibitory effect of LA on FBS-stimulated cyclin D1 protein expression. Growth-arrested VSMCs were pretreated with LA (50 or 100 μmol/L) for 2 h and then stimulated with 5% FBS for 24 h. Expression of cyclin D1 protein was examined by Western blotting. β-actin was used as a loading control. Values are presented as mean±SEM. n=3. bP<0.05 vs control. eP<0.05, fP<0.01 vs FBS.

Mentions: It is known that cyclins, CDKs, and their inhibitors regulate cell cycle progression. Changes in the cyclin D1/CDK complex, for example, can either stop or promote cell proliferation25. We investigated the potential effect of LA on cyclin D1 protein expression. Our results showed that FBS significantly increased the expression of cyclin D1, while LA completely abolished this serum action (Figure 5).


Inhibitory effects of lithospermic acid on proliferation and migration of rat vascular smooth muscle cells.

Chen L, Wang WY, Wang YP - Acta Pharmacol. Sin. (2009)

Inhibitory effect of LA on FBS-stimulated cyclin D1 protein expression. Growth-arrested VSMCs were pretreated with LA (50 or 100 μmol/L) for 2 h and then stimulated with 5% FBS for 24 h. Expression of cyclin D1 protein was examined by Western blotting. β-actin was used as a loading control. Values are presented as mean±SEM. n=3. bP<0.05 vs control. eP<0.05, fP<0.01 vs FBS.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4007173&req=5

fig5: Inhibitory effect of LA on FBS-stimulated cyclin D1 protein expression. Growth-arrested VSMCs were pretreated with LA (50 or 100 μmol/L) for 2 h and then stimulated with 5% FBS for 24 h. Expression of cyclin D1 protein was examined by Western blotting. β-actin was used as a loading control. Values are presented as mean±SEM. n=3. bP<0.05 vs control. eP<0.05, fP<0.01 vs FBS.
Mentions: It is known that cyclins, CDKs, and their inhibitors regulate cell cycle progression. Changes in the cyclin D1/CDK complex, for example, can either stop or promote cell proliferation25. We investigated the potential effect of LA on cyclin D1 protein expression. Our results showed that FBS significantly increased the expression of cyclin D1, while LA completely abolished this serum action (Figure 5).

Bottom Line: To understand the effects of lithospermic acid (LA), a potent antioxidant from the water-soluble extract of Salvia miltiorrhiza, on the migration and proliferation of rat thoracic aorta vascular smooth muscle cells (VSMCs).Furthermore, LA attenuated LPS-induced VSMC migration by inhibiting MMP-9 expression and its enzymatic activity.LA is able to inhibit FBS-induced VSMC proliferation and LPS-induced VSMC migration, which suggests that LA may have therapeutic effects in the prevention of atherosclerosis, restenosis and neointimal hyperplasia.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of New Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.

ABSTRACT

Aim: To understand the effects of lithospermic acid (LA), a potent antioxidant from the water-soluble extract of Salvia miltiorrhiza, on the migration and proliferation of rat thoracic aorta vascular smooth muscle cells (VSMCs).

Methods: VSMC migration, proliferation, DNA synthesis and cell cycle progression were investigated by transwell migration analysis, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, bromodeoxyuridine (BrdU) incorporation assay, and flow cytometric detection, respectively. Intracellular reactive oxygen species (ROS) generation was detected using 2',7'-dichlorofluorescin diacetate (DCFH-DA). The expression of cyclin D1 protein and matrix metalloproteinase-9 (MMP-9) protein, as well as the phosphorylation state of ERK1/2, were determined using Western blots. The activity of MMP-9 and the expression of MMP-9 mRNA were assessed by gelatin zymography analysis and RT-PCR, respectively.

Results: LA (25-100 micromol/L) inhibited both lipopolysaccharide (LPS)- and fetal bovine serum (FBS)-induced ROS generation and ERK1/2 phosphorylation. By down-regulating the expression of cyclin D(1) and arresting cell cycle progression at the G(1) phase, LA inhibited both VSMC proliferation and DNA synthesis as induced by 5% FBS. Furthermore, LA attenuated LPS-induced VSMC migration by inhibiting MMP-9 expression and its enzymatic activity.

Conclusion: LA is able to inhibit FBS-induced VSMC proliferation and LPS-induced VSMC migration, which suggests that LA may have therapeutic effects in the prevention of atherosclerosis, restenosis and neointimal hyperplasia.

Show MeSH
Related in: MedlinePlus