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Whitmania Pigra Whitman Extracts Inhibit Lipopolysaccharide Induced Rat Vascular Smooth Muscle Cells Migration and their Adhesion Ability to THP-1 and RAW 264.7 Cells

View Article: PubMed Central - PubMed

ABSTRACT

Aim:: Atherosclerosis is a kind of chronic inflammatory disease. A crucial pathology change of atherosclerosis is the migration of activated VSMCs to the intima where they interact with leukocytes by expressing adhesion molecules, including intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Moreover, monocyte chemoattractant protein-1 (MCP-1) expressed by VSMCs plays an important role in recruiting monocytes and macrophages. Leech (Whitmania pigra Whitman) is a traditional Chinese medicine to treat cardiovascular diseases including atherosclerosis, however previous research has rarely reported the molecular mechanism for its curative effect. Thus, our study focuses on the effects of leech extracts on the expression of inflammatory factors, adhesion molecules and MCP-1 in rat VSMCs.

Methods:: In our present study, wound-healing assay and Boyden chamber model were applied to evaluate the anti-migration effect of LEE (Leech Enzyme Extracts) on LPS induced VSMCs. The anti-adhesion effect was assessed using DiI-labeled THP-1 and RAW264.7.

Results:: LEE suppressed LPS-induced VSMCs migration and decreased the chemotaxis and adhesive capacity of THP-1 and RAW264.7 to LPS-stimulated VSMCs. LEE also attenuated the upregulation of a variety of pro-atherosclerotic factors by inhibiting the phosphorylation of p38 MAPK. LEE was also observed to prevent NF-κB p65 nuclear localization using immune-fluorescent staining.

Conclusions:: In conclusion, LEE suppresses LPS-induced upregulation of inflammatory factors, adhesion molecules and MCP-1 in rat VSMCs mainly via inhibiting the p38 MAPK/NF-κB pathways, thus partly uncovered LEE's molecular mechanisms for its therapeutic effect on atherosclerosis.

No MeSH data available.


The anti-chemotaxis effects of LEE(A) Inhibition of chemotaxis of THP-1 cells to LPS-stimulated VSMCs by LEE in a Boyden chamber assay. Bar = 100 µm. (B) Inhibition of chemotaxis of RAW264.7 cells to LPS-stimulated VSMCs by LEE in a Boyden chamber assay. Bar = 100 µm. (C) Number of THP-1 migration. (D) Number of RAW264.7 migration. #P <0.05 vs control, *P <0.05 vs LPS, **P <0.01 vs LPS. Data are from three independent experiments.
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Figure 2: The anti-chemotaxis effects of LEE(A) Inhibition of chemotaxis of THP-1 cells to LPS-stimulated VSMCs by LEE in a Boyden chamber assay. Bar = 100 µm. (B) Inhibition of chemotaxis of RAW264.7 cells to LPS-stimulated VSMCs by LEE in a Boyden chamber assay. Bar = 100 µm. (C) Number of THP-1 migration. (D) Number of RAW264.7 migration. #P <0.05 vs control, *P <0.05 vs LPS, **P <0.01 vs LPS. Data are from three independent experiments.

Mentions: In order to study whether LEE has an influence to the chemotaxis of THP-1 or RAW264.7 to LPS-stimulated VSMCs, we proposed a parameter on the chemotaxis inhibition rate, which was a function of the cell migration rate. The calculation method for MIR (migration inhibition rate) = 1 − (the number of cell migration in X group − the number of cell migration in blank control)/(the number of cell migration in model group − the number of cell migration in blank control). LEE (200 µg/mL) could decrease the chemotaxis of THP-1 and RAW264.7 to LPS-stimulated VSMCs, and the inhibition rate of those two experiments were 60% and 80% respectively (Fig. 2).


Whitmania Pigra Whitman Extracts Inhibit Lipopolysaccharide Induced Rat Vascular Smooth Muscle Cells Migration and their Adhesion Ability to THP-1 and RAW 264.7 Cells
The anti-chemotaxis effects of LEE(A) Inhibition of chemotaxis of THP-1 cells to LPS-stimulated VSMCs by LEE in a Boyden chamber assay. Bar = 100 µm. (B) Inhibition of chemotaxis of RAW264.7 cells to LPS-stimulated VSMCs by LEE in a Boyden chamber assay. Bar = 100 µm. (C) Number of THP-1 migration. (D) Number of RAW264.7 migration. #P <0.05 vs control, *P <0.05 vs LPS, **P <0.01 vs LPS. Data are from three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383546&req=5

Figure 2: The anti-chemotaxis effects of LEE(A) Inhibition of chemotaxis of THP-1 cells to LPS-stimulated VSMCs by LEE in a Boyden chamber assay. Bar = 100 µm. (B) Inhibition of chemotaxis of RAW264.7 cells to LPS-stimulated VSMCs by LEE in a Boyden chamber assay. Bar = 100 µm. (C) Number of THP-1 migration. (D) Number of RAW264.7 migration. #P <0.05 vs control, *P <0.05 vs LPS, **P <0.01 vs LPS. Data are from three independent experiments.
Mentions: In order to study whether LEE has an influence to the chemotaxis of THP-1 or RAW264.7 to LPS-stimulated VSMCs, we proposed a parameter on the chemotaxis inhibition rate, which was a function of the cell migration rate. The calculation method for MIR (migration inhibition rate) = 1 − (the number of cell migration in X group − the number of cell migration in blank control)/(the number of cell migration in model group − the number of cell migration in blank control). LEE (200 µg/mL) could decrease the chemotaxis of THP-1 and RAW264.7 to LPS-stimulated VSMCs, and the inhibition rate of those two experiments were 60% and 80% respectively (Fig. 2).

View Article: PubMed Central - PubMed

ABSTRACT

Aim:: Atherosclerosis is a kind of chronic inflammatory disease. A crucial pathology change of atherosclerosis is the migration of activated VSMCs to the intima where they interact with leukocytes by expressing adhesion molecules, including intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Moreover, monocyte chemoattractant protein-1 (MCP-1) expressed by VSMCs plays an important role in recruiting monocytes and macrophages. Leech (Whitmania pigra Whitman) is a traditional Chinese medicine to treat cardiovascular diseases including atherosclerosis, however previous research has rarely reported the molecular mechanism for its curative effect. Thus, our study focuses on the effects of leech extracts on the expression of inflammatory factors, adhesion molecules and MCP-1 in rat VSMCs.

Methods:: In our present study, wound-healing assay and Boyden chamber model were applied to evaluate the anti-migration effect of LEE (Leech Enzyme Extracts) on LPS induced VSMCs. The anti-adhesion effect was assessed using DiI-labeled THP-1 and RAW264.7.

Results:: LEE suppressed LPS-induced VSMCs migration and decreased the chemotaxis and adhesive capacity of THP-1 and RAW264.7 to LPS-stimulated VSMCs. LEE also attenuated the upregulation of a variety of pro-atherosclerotic factors by inhibiting the phosphorylation of p38 MAPK. LEE was also observed to prevent NF-&kappa;B p65 nuclear localization using immune-fluorescent staining.

Conclusions:: In conclusion, LEE suppresses LPS-induced upregulation of inflammatory factors, adhesion molecules and MCP-1 in rat VSMCs mainly via inhibiting the p38 MAPK/NF-&kappa;B pathways, thus partly uncovered LEE's molecular mechanisms for its therapeutic effect on atherosclerosis.

No MeSH data available.