Minoxidil inhibits proliferation and migration of cultured vascular smooth muscle cells and neointimal formation after balloon catheter injury

The goal of the study was to investigate the in vitro and in vivo inhibition of minoxidil on smooth muscle cell (SMC) proliferation and migration as well as neointimal formation. The in vitro effect of minoxidil was investigated by Boyden chamber assay and cell-cycle analysis. To evaluate the in vivo effect, we treated the animals with minoxidil in their drinking water before and after balloon catheter injury to carotid artery. Results showed that minoxidil inhibited SMC migration across type I collagen membrane in a dose-related manner (13.5% by 0.01 mg/ml; p < 0.05; 16.8% by 0.05 mg/ml: p < 0.01; 40.4% by 0.25 mg/ml; p < 0.001; and 65.8% by 1.25 mg/ml; p < 0.001). Minoxidil (0.8 mg/ml) increased the number of SMCs in G1 phase (p < 0.05) and decreased the number of SMCs in S phase (p < 0.001). In vivo minoxidil treatment reduced neointimal mass by 31.7% (120 mg/L) and 42.3% (200 mg/L), respectively. Data demonstrate that minoxidil inhibits vascular SMC proliferation and migration both in vitro and in vivo, and therefore may be useful to inhibit SMC hyperplasia that occurs in restenosis and other vascular diseases.     

Ajoene,a garlic compound,inhibits protein prenylation and arterial smooth muscle cell proliferation

(1) Ajoene is a garlic compound with anti-platelet properties and, in addition, was shown to inhibit cholesterol biosynthesis by affecting 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase and late enzymatic steps of the mevalonate (MVA) pathway. (2) MVA constitutes the precursor not only of cholesterol, but also of a number of non-sterol isoprenoids, such as farnesyl and geranylgeranyl groups. Covalent attachment of these MVA-derived isoprenoid groups (prenylation) is a required function of several proteins that regulate cell proliferation. We investigated the effect of ajoene on rat aortic smooth muscle cell proliferation as related to protein prenylation. (3) Cell counting, DNA synthesis, and cell cycle analysis showed that ajoene (1-50 micro M) interfered with the progression of the G1 phase of the cell cycle, and inhibited rat SMC proliferation. (4) Similar to the HMG-CoA reductase inhibitor simvastatin, ajoene inhibited cholesterol biosynthesis. However, in contrast to simvastatin, the antiproliferative effect of ajoene was not prevented by the addition of MVA, farnesol (FOH), and geranylgeraniol (GGOH). Labelling of smooth muscle cell cellular proteins with [3H]-FOH and [3H]-GGOH was significantly inhibited by ajoene. (5) In vitro assays for protein farnesyltransferase (PFTase) and protein geranylgeranyltransferase type I (PGGTase-I) confirmed that ajoene inhibits protein prenylation. High performance liquid chromatography (HPLC) and mass spectrometry analyses also demonstrated that ajoene causes a covalent modification of the cysteine SH group of a peptide substrate for protein PGGTase-I. (6) Altogether, our results provide evidence that ajoene interferes with the protein prenylation reaction, an effect that may contribute to its inhibition of SMC proliferation.     

Dissecting histone deacetylase role in pulmonary arterial smooth muscle cell proliferation and migration

Pulmonary Arterial Hypertension (PAH) is a rare and devasting condition characterized by elevated pulmonary vascular resistance and pulmonary artery pressure leading to right-heart failure and premature death.     

Zoledronate inhibits the proliferation, adhesion and migration of vascular smooth muscle cells

Bisphosphonates, which are extensively used in bone-related disorders, have been reported to inhibit atherosclerosis and neointimal hyperplasia. In the present study, we investigated the effects of a bisphosphonate, zoledronate, on the proliferation, adhesion, migration and microstructure of vascular smooth muscle cells (VSMCs) from Sprague-Dawley rats. It was shown that zoledronate suppressed VSMCs proliferation after 48 h cultivation in a dose depend manner, most obviously at concentrations above 10 µM. Cell cycle analysis indicated that zoledronate inhibited the proliferation of VSMCs via cell cycle arrest at S/G2/M phase. This inhibition was not associated with cell death. In a modified Boyden chamber model, it was shown that zoledronate dose-dependently inhibited VSMCs adhesion to collagen and migration stimulated by platelet-derived growth factor-BB. Western blot analysis suggested that zoledronate significantly inhibited the phosphorylation of focal adhesion kinase. Furthermore, we observed that more and more VSMCs changed from a bipolar appearance to a globular shape under inverted light microscope as zoledronate concentration increased from 0.1 to 100 µM. Images under transmission electron microscope confirmed this morphological change, and many electron density bodies were observed in zoledronate-treated VSMCs. These findings indicated that bisphosphonates' effects of suppressing atherosclerosis and neointimal hyperplasia might be due to inhibition of VSMCs, at least for zoledronate.     

Modulation of smooth muscle cell proliferation and migration: role of smooth muscle cell heterogeneity

Proliferation and migration of smooth muscle cells (SMCs) from the media towards the intima are key events in atherosclerosis and restenosis. During these processes, SMC undergo phenotypic modulations leading to SMC dedifferentiation. The identification and characterization of factors controlling these phenotypic changes are crucial in order to prevent the formation of intimal thickening. One of the questions which presently remains open, is to know whether any SMCs of the media are capable of accumulating into the intima or whether only a predisposed medial SMC subpopulation is involved in this process. The latter hypothesis implies that arterial SMCs are phenotypically heterogenous. In this chapter, we will describe the distinct SMC phenotypes identified in arteries of various species, including humans. Their role in the formation of intimal thickening will be discussed.     

Effects of cerivastatin on human arterial smooth muscle cell proliferation and migration in transfilter cocultures

Statins competitively inhibit 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity reducing mevalonate synthesis. In this study, antiproliferative and antimigratory effects of the new compound cerivastatin were analyzed and compared with classic statins of the first and second generation using mono- and cocultures of human arterial smooth muscle (haSMC) and endothelial (haEC) cells. Effects on the mitotic index and mitochondrial activity of haEC and haSMC monocultures were tested using BrdU enzyme-linked immunosorbent assay (ELISA) and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) tests, respectively. In lactate dehydrogenase (LDH) assays, cytotoxicity of statins was studied. Transfilter cocultures were performed for 14 days to evaluate haSMC growth under the stimulatory effect of proliferating haEC, which release growth factors [e.g., platelet-derived growth factor (PDGF)]. The hydrophobic statins simvastatin, lovastatin, and atorvastatin significantly inhibited haSMC and haEC growth in monocultures at 0.5-50 microM. However, most potent effects were exerted by cerivastatin in 10- to 30-fold lower doses without any significant cytotoxicity. More important, cerivastatin showed also significant effects on haSMC proliferation and migration in transfilter cocultures at extremely low doses (IC50, 0.04-0.06 microM), even when applied exclusively to the endothelial side and in the presence of low-density lipoprotein (LDL). Addition of mevalonate abolished the effects of cerivastatin completely. Even in the presence of growth-stimulating haEC and LDL, cerivastatin was found to be the most potent inhibitor of haSMC proliferation and migration in doses that also can be reached in human serum after oral drug administration. The results support the concept that statins seems to influence additional cellular mechanisms beyond cholesterol reduction, which might also have a relevance for the prevention of restenosis.     

Sauchinone inhibits angiotensin II-induced proliferation and migration of vascular smooth muscle cells

Hypertension is a common type of cardiovascular disease that remains a major cause of death in the world. Vascular remodelling is an important complication of hypertension, and vascular smooth muscle cells (VSMCs) play a major role in vascular remodelling. Sauchinone is one of the active lignins which has been found to possess vascular protective effects. However, the functional role of sauchinone in hypertension has not been investigated. The aim of this study was to evaluate the role of sauchinone in the angiotensin II (Ang II)-induced vascular remodelling model in VSMCs. The results showed that treatment of sauchinone inhibited Ang II-induced VSMCs proliferation and migration in VSMCs. Sauchinone treatment suppressed the reactive oxygen species (ROS) production and NADPH oxidase (NOX) activity in Ang II-induced VSMCs. The inhibitory effects of Ang II on expressions of VSMCs phenotype markers including α-smooth muscle actin (α-SMA), calponin, osteopontin were mitigated by sauchinone treatment. Furthermore, sauchinone inhibited Ang II-induced over-activation of TGF-β1/Smad3 signalling pathway in VSMCs. Taken together, this study identified sauchinone as a potential agent for preventing vascular remodelling in hypertension.     

The effects of pharmacological PAI-1 inhibition on thrombus formation and neointima formation after arterial injury

Objective: Plasminogen activator inhibitor (PAI)-1 plays a role in neointimal formation after percutaneous coronary intervention (PCI), the effect of overexpression or lack of PAI-1 is controversial. Murine arterial injury models develop neointimal hyperplasia similar to that observed in clinical coronary arterial restenosis after PCI. Methods and results: To clarify the role of PAI-1 in thrombus formation and neointimal formation after arterial injury, we used a specific PAI-1 inhibitor (IMD-1622) in a rat aorta–vein shunt model and a mouse arterial injury model. While the non-treated shunt model showed massive thrombus formation, IMD-1622 administration suppressed this. Injured arteries with vehicles showed significant neointimal formation with enhancement of adhesion molecules, fibrinogen accumulation and cell proliferation on day 28 after injury. However, intimal thickening and expression of these factors were suppressed in PAI-1 recipients. Conclusion: A specific PAI-1 inhibitor prevents thrombus formation and arterial neointimal formation after arterial injury. Thus, PAI-1 plays a critical role in arterial remodeling after mechanical injury. PAI-1 regulation may be useful to prevent thrombus and neointimal formation after PCI.     

川芎嗪对家兔主动脉损伤平滑肌细胞增殖的抑制作用(英文)

目的:探讨川芎嗪对主动脉球囊损伤平滑肌细胞增殖的抑制作用。方法:20只新西兰大白兔行主动脉球囊术。以川芎嗪(40mg·kg~(-1)·d~(-1),iv)共21天(术前7天,术后14天)处理的兔血清,或直接加入培养基,检测对原代培养的主动脉平滑肌细胞[~3H]胸腺嘧啶核苷酸掺入的影响。并对血管壁组织切片显微测量内膜厚度和管径。结果:球囊扩张处内膜厚度(77±23)μm和管腔狭窄(877±118)μm均较邻近正常处显著增加[分别为,(41±13)μm,P<0.01;(1033±175)μm,P<0.05],而川芎嗪显著降低损伤后内膜的增厚(56±16)μm,(P<0.05)和管腔的变窄(1023±157)μm,(P<0.05)。用川芎嗪治疗后血清培养主动脉平滑肌细胞,显示对[~3H]胸腺嘧啶核苷酸掺入明显抑制,直接加川芎嗪入培养基对掺入抑制呈剂量依赖[(40-4000)μg/well]。结论:川芎嗪对家兔主动脉球囊损伤平滑肌细胞增殖有显著抑制。     

Cardamonin inhibits angiotensin II-induced vascular smooth muscle cell proliferation and migration by downregulating p38 MAPK,Akt, and ERK phosphorylation

Cardamonin is a chalconoid isolated from various herbs, such as Alpinia katsumadai and Carya cathayensis Sarg. This study examined the effect of cardamonin on angiotensin II (Ang II)-induced proliferation and migration in rat vascular smooth muscle cells (VSMCs) as well as its underlying mechanisms. The results showed that cardamonin significantly inhibited Ang II-induced proliferation and migration in rat VSMCs in a concentration-dependent manner. Moreover, cardamonin suppressed Ang II-induced phosphorylation of p38 MAPK, Akt, and extracellular regulated protein kinase (ERK). These findings indicate that the downregulation of p38 MAPK, Akt, and ERK phosphorylation might be, at least in part, involved in cardamonin-suppressed proliferation and migration induced by Ang II in rat VSMCs. As proliferation and migration of VSMCs play critical roles in the pathogenesis of atherosclerosis, cardamonin might be a potential candidate for atherosclerosis treatment.     

川芎嗪对家兔主动脉损伤平滑肌细胞增殖的抑制作用

AIM: To study the inhibitory effect of ligustrazine (Lig) on growth of cultured rabbit aortic vascular smooth muscle cells (VSMC) after balloon injury. METHODS: Twenty New Zealand white rabbits were subjected to arterial injury with a balloon catheter (810 kPa for three consecutive inflations, 1 min each time). The uptake of [3H]thymidine in primary cultural VSMC incubated with rabbit serum, which obtained from the animals treated without or with Lig (40 mg.kg-1.d-1, i.v.) for 21 d (7 d before and 14 d after the injury procedure) was determined. The determination was performed in direct addition of TMP to culture as well. And histological cross-sections of the blood wall were also analyzed. RESULTS: After balloon injury the intimal thickening (77 +/- 23) microns and lumen diameter narrowing (877 +/- 118) microns in dilated sites were increased significantly than the normal adjacent wall [(41 +/- 13) microns, P < 0.01; (1033 +/- 175) microns, P < 0.05, respectively]. Treatment with Lig decreased both intimal thickening (56 +/- 16) microns (P < 0.05) and lumen diameter narrowing (1023 +/- 157) microns (P < 0.05). Lig inhibited [3H]thymidine uptake in VSMC incubated with the serum obtained from these rabbits. Direct addition of Lig inhibited [3H]thymidine uptake in cultured VSMC in a dose-dependent manner (40-4000) micrograms/well. CONCLUSION: Lig shows a pronounced inhibitory effect on VSMC proliferation after balloon injury.     

TSPO ligands prevent the proliferation of vascular smooth muscle cells and attenuate neointima formation through AMPK activation

Abnormal growth of the intimal layer of blood vessels(neointima formation)contributes to the progression of atherosclerosis and in-stent restenosis.Recent evidence shows that the 18-kDa translocator protein(TSPO),a mitochondrial membrane protein,is involved in diverse cardiovascular diseases.In this study we investigated the role of endogenous TSPO in neointima formation after angioplasty in vitro and in vivo.We established a vascular injury model in vitro by using platelet-derived growth factor-BB(PDGF-BB)to stimulate rat thoracic aortic smooth muscle cells(A10 cells).We found that treatment with PDGF-BB(1-20 ng/mL)dose-dependently increased TSPO expression in A10 cells,which was blocked in the presence of PKC inhibitor or MAPK inhibitor.Overexpression of TSPO significantly promoted the proliferation and migration in A10 cells,whereas downregulation of TSPO expression by siRNA or treatment with TSPO ligands PK11195 or Ro5-4864(104 nM)produced the opposite effects.Furthermore,we found that PK11195(10−104 nM)dose-dependently activated AMPK in A10 cells.PK11195-induced inhibition on the proliferation and migration of PDGF-BB-treated A10 cells were abolished by compound C(an AMPK-specific inhibitor,103 nM).In rats with balloon-injured carotid arteries,TSPO expression was markedly upregulated in the carotid arteries.Administration of PK11195(3 mg/kg every 3 days,ip),starting from the initial balloon injury and lasting for 2 weeks,greatly attenuated carotid neointima formation by suppressing balloon injury-induced phenotype switching of VSMCs(increasedα-SMA expression).These results suggest that TSPO is a vascular injury-response molecule that promotes VSMC proliferation and migration and is responsible for the neointima formation after vascular injury,which provides a novel therapeutic target for various cardiovascular diseases including atherosclerosis and restenosis.     

Aminoguanidine induces constrictive vascular remodeling and inhibits smooth muscle cell death after balloon injury.

We examined the effects of aminoguanidine, an inhibitor of inducible nitric oxide synthase, in the rat model of balloon injury. Arteries were assessed by histomorphometry, and vascular smooth muscle cell death and proliferation were examined 24 h and 14 days after balloon injury by in situ terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) of fragmented DNA and expression of proliferating cell nuclear antigen, respectively. Aminoguanidine decreased the luminal area 14 days after balloon injury (0.19+/-0.04 mm2 vs. 0.35+/-0.02 mmr2; P < 0.005), and this effect was attributable to reduction of the total vessel area, i.e., constrictive vascular remodeling (0.42+/-0.03 mm2 vs. 0.55+/-0.03 mm2; P < 0.005). At 24 h after injury, the percentage of TUNEL-positive cells in the medial layer was reduced by aminoguanidine (2.0+/-1.0% vs. 17.3+/-5.4%; P < 0.05), and the percentage of proliferating cells was increased (18.4+/-5.5% vs. 4.9+/-2.2%; P < 0.05). Aminoguanidine did not influence the density of VSMC nuclei in the injured artery wall, systemic blood pressure or endothelium-dependent vasorelaxation. We conclude, that in the rat model of balloon injury, aminoguanidine induces luminal loss by constrictive vascular remodeling in association with reduced early VSMC death and increased proliferation.     

Apamin inhibits PDGF-BB-induced vascular smooth muscle cell proliferation and migration through suppressions of activated Akt and Erk signaling pathway

The increased proliferation and migration of vascular smooth muscle cells (VSMC) are key process in the development of atherosclerosis lesions. Platelet-derived growth factor (PDGF) initiates a multitude of biological effects that contribute to VSMC proliferation and migration. Apamin, a component of bee venom, has been known to block the Ca^2 +-activated K⁺ channels. However, the effects of apamin in the regulation PDGF-BB-induced VSMC proliferation and migration has not been identified. In this study, we investigate the inhibitory effect of apamin on PDGF-BB-induced VSMC proliferation and migration. Apamin suppressed the PDGF-BB-induced VSMC proliferation and migration with no apparent cytotoxic effect. In accordance with these findings, apamin induced the arrest of cell cycle progression at G0/G1 phase. Apamin also decreased the expressions of G0/G1 specific regulatory proteins including proliferating cell nuclear antigen (PCNA), cyclin D1, cyclin-dependent kinases (CDK) 4, cyclin E and CDK2, as well as increased the expression of p21^Cip1 in PDGF-BB-induced VSMC. Moreover, apamin inhibited PDGF-BB-induced phosphorylation of Akt and Erk1/2. These results suggest that apamin plays an important role in prevention of vascular proliferation and migration through the G0/G1 cell cycle arrest by PDGF signaling pathway. Thus, apamin may be a promising candidate for the therapy of atherosclerosis.     

Oestradiol inhibits smooth muscle cell proliferation of pig coronary artery.

1. The effect of oestradiol 17 beta on vascular smooth muscle proliferation was examined in segments of the pig left anterior descending coronary artery (LAD). It was established by cytochemical techniques that out-growth from the segments was composed of vascular smooth muscle cells. 2. [3H]-thymidine uptake by pig LAD segments was used as an index of vascular smooth muscle cell proliferation. Nitroprusside and forskolin significantly inhibited [3H]-thymidine uptake and were used as positive controls. 3. Oestradiol 17 beta (180-360 nM) inhibited thymidine uptake by pig LAD segments (P < 0.05). The inhibition was observed only in the absence of phenol red, which is a weak oestrogen receptor agonist. The anti-oestrogens tamoxifen and its more potent metabolite 4-hydroxytamoxifen, both of which are partial oestrogen receptor agonists, also significantly inhibited thymidine uptake. However, pretreatment with either tamoxifen or 4-hydroxytamoxifen did not significantly block oestradiol 17 beta-induced inhibition of thymidine uptake. 4. The LAD segments bound [3H]-oestradiol 17 beta in a time-dependent manner and about 20 to 30% was displaced by an excess of unlabelled oestradiol 17 beta. Autoradiography showed [3H]-oestradiol 17 beta was evenly distributed in the cytosol and nuclei of cells in the three layers of the vessel wall. 5. The data suggest that oestradiol 17 beta inhibits smooth muscle cell proliferation in porcine LAD segments, possibly through an oestrogen receptor mechanism. This in vitro effect suggests an in vivo role for oestradiol 17 beta in directly protecting coronary arteries against myointimal proliferation in premenopausal women.     

Ligustilide inhibits vascular smooth muscle cells proliferation

Proliferation and migration of vascular smooth muscle cells (VSMCs) are believed to develop atherosclerosis and venous bypass graft disease. Ligustilide is widely used to treat some pathological settings such as atherosclerosis and hypertension. The aim of this study was to examine the effect of ligustilide on VSMCs proliferation. The results show that ligustilide significantly inhibited VSMCs proliferation and cell cycle progression. Further analysis shows that ligustilide suppressed reactive oxygen species production and extracellular signal-related kinases (ERK), c-Jun N-terminal protein kinase (JNK), and p38 MAP kinase. Cells were treated with antioxidant, superoxide dismutase, catalase, and DPI, respectively, leading to repress ERK, JNK, and p38 activation. The inhibitors of mitogen activated protein kinase (MAPK), PD98059, SB203580, and Sp600125, inhibited cell proliferation. These findings suggest the antiproliferative effect of ligustilide was associated with the decrement of reactive oxygen species resulting in the suppression of MAPK pathway. Thus, ligustilide contribute to be the effective agent in preventing cardiovascular diseases.     

大黄素通过自噬抑制血管平滑肌细胞的增殖和迁移

目的研究大黄素对大鼠胸主动脉血管平滑肌细胞(VSMC)增殖迁移的调控作用,并探讨其是否通过自噬激活实现。方法体外培养大鼠胸主动脉血管平滑肌细胞,采用CCK8法、免疫印迹法和划痕实验法分别检测大黄素对VSMC增殖和迁移的抑制作用;免疫印迹法检测自噬相关蛋白LC3和Beclin1表达水平。结果大黄素质量浓度依赖性地抑制VSMC的增殖迁移,同时上调细胞自噬。抑制细胞自噬后,大黄素对VSMC增殖迁移抑制作用减弱。结论大黄素通过细胞自噬抑制VSMC的增殖迁移。     

Onychin inhibits proliferation of vascular smooth muscle cells by regulating cell cycle

AIM: To investigate the effects of onychin on the proliferation of cultured rat artery vascular smooth muscle cells (VSMCs) in the presence of 10% new-born calf serum (NCS). METHODS: Rat VSMCs were incubated with onychin 150 micromol/L or genistein 10 micromol/L in the presence of 10% NCS for 24 h. The proliferation of VSMCs was measured by cell counting and MTS/PMS colorimetric assays. Cell cycle progression was evaluated by flow cytometry. Retinoblastoma (Rb) phosphorylation, and expression of cyclin D1 and cyclin E were measured by Western blot assays. The tyrosine phosphorylation of ERK1/2 was examined by immunoprecipitation techniques using anti-phospho-tyrosine antibodies. RESULTS: The proliferation of VSMCs was accelerated significantly in the presence of 10% NCS. Onychin reduced the metabolic rate of MTS and the cell number of VSMCs in the presence of 10% NCS in a dose-dependent manner. Flow cytometry analysis revealed that the G1-phase fraction ratio in the onychin group was higher than that in the 10% NCS group (85.2% vs 70.0%, P<0.01), while the S-phase fraction ratio in the onychin group was lower than that in 10% NCS group (4.3% vs 16.4%, P<0.01). Western blot analysis showed that onychin inhibited Rb phosphorylation and reduced the expression of cyclin D1 and cyclin E. The effects of onychin on proliferation, the cell cycle and the expression of cyclins in VSMCs were similar to those of genistein, an inhibitor of tyrosine kinase. Furthermore immunoprecipitation studies showed that both onychin and genistein markedly inhibited the tyrosine phosphorylation of ERK1/2 induced by 10% NCS. CONCLUSION: Onychin inhibits the proliferation of VSMCs through G1 phase cell cycle arrest by decreasing the tyrosine phosphorylation of ERK1/2, and the expression of cyclin D1 and cyclin E, and sequentially inhibiting Rb phosphorylation.     

Ferulic acid inhibits vascular smooth muscle cell proliferation induced by angiotensin II

The aim of this study was to determine the effects of ferulic acid on the proliferation and molecular mechanism in cultured vascular smooth muscle cell (VSMC) induced by angiotensin II. It was shown that ferulic acid significantly inhibited angiotensin II-induced VSMC proliferation in a dose-dependent manner. Western blotting analyses suggest that the antiproliferative effect of ferulic acid was involved in the mitogen-activated protein kinases (MAPKs) pathway. While no effect on p38, ferulic acid markedly inactivated the extracellular signal-regulated kinases (ERK1/2) and c-Jun N-terminal kinases (JNK), indicating that the inhibition of ferulic acid on VSMC proliferation was associated with ERK1/2 and JNK rather than p38 pathway. On the expression of cell cycle regulatory proteins, ferulic acid elevated the protein content of p21(waf1/cip1), decreased expression of cyclin D1 and inhibited phosphorylation of retinoblastoma protein, suggesting that ferulic acid inhibited VSMC proliferation by regulating the cell progression from G1 to S phase. The inactivation of MAPKs and modulation of cell cycle proteins of ferulic acid may be of importance in preventing cardiovascular disease.