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Calycosin-7-O-β-D-glucoside promotes oxidative stress-induced cytoskeleton reorganization through integrin-linked kinase signaling pathway in vascular endothelial cells.

Jiang YH, Sun W, Li W, Hu HZ, Zhou L, Jiang HH, Xu JX - BMC Complement Altern Med (2015)

Bottom Line: The effects of calycosin on LPS-induced changes in cell viability, apoptosis rate, cell migration, nitric oxide synthase (NOS), generationof intracellular reactive oxygen species (ROS) and cytoskeleton organization were determined.Calycosin improved cell viability, suppressed apoptosis and protected the cells from LPS-induced reduction in cell migration and generation of ROS, protein level of NOS at a comparable magnitude to that of Y27632 and valsartan.Calycosin are considered to be able to activate MLCP through promoting the generation of NO, decreasing PMLC, suppressing the cytoskeleton remodeling caused by activation of Rho/ROCK pathway and inhibiting AKT pathway by decreasing VEGF, VEGF R2 and PI3K level.

View Article: PubMed Central - PubMed

Affiliation: Affiliated Hospital of Shandong University of Traditional Chinese Medicine, West Wenhua road #42, Jinan, Shandong, 250011, PR China. jiang_yuehua@hotmail.com.

ABSTRACT

Background: Dysfunction of vascular endothelium is implicated in many pathological situations. Cytoskeleton plays an importance role in vascular endothelial permeability barrier and inflammatory response. Many Chinese herbs have the endothelial protective effect, of which, "Astragalus membranaceus" is a highly valued herb for treatment of cardiovascular and renal diseases in traditional Chinese medicine, In this study, we tested whether calycosin-7-O-β-D-glucoside (Calycosin), a main effective monomer component of "Astragalus membranaceus", could protect endothelial cells from bacterial endotoxin (LPS)-induced cell injury.

Methods: Endothelial cell injury was induced by exposing human umbilical vein endothelial cells (HUVECs) to LPS. The effects of calycosin on LPS-induced changes in cell viability, apoptosis rate, cell migration, nitric oxide synthase (NOS), generationof intracellular reactive oxygen species (ROS) and cytoskeleton organization were determined. Microarray assay was employed to screen the possible gene expression change. Based on the results of microarray assay, the expression profile of genes involved in Rho/ROCK pathway and AKT pathway were further evaluated with quantitative real-time RT-PCR or western blot methods.

Results: Calycosin improved cell viability, suppressed apoptosis and protected the cells from LPS-induced reduction in cell migration and generation of ROS, protein level of NOS at a comparable magnitude to that of Y27632 and valsartan. Similar to Y27632 and valsartan, Calycosin, also neutralized LPS-induced actomyosin contraction and vinculin protein aggregation. Microarray assay, real-time PCR and western blot results revealed that LPS induced expression of FN, ITG A5, RhoA, PI3K (or PIP2 in western blotting), FAK, VEGF and VEGF R2, and inhibited expression of MLCP. We believed multiple pathways involved in the regulation of calycosin on HUVECs. Calycosin are considered to be able to activate MLCP through promoting the generation of NO, decreasing PMLC, suppressing the cytoskeleton remodeling caused by activation of Rho/ROCK pathway and inhibiting AKT pathway by decreasing VEGF, VEGF R2 and PI3K level.

Conclusion: Calycosin protected HUVEC from LPS-induced endothelial injury, possibly through suppression of Rho/ROCK pathway and regulation of AKT pathway.

No MeSH data available.


Related in: MedlinePlus

Effect of Calycosin on LPS-induced the change of gene expresses induced by LPS. Results of up- or down-regulated molecules in microarray (a) and quantitive real-time RT-PCR (b). △P < 0.05, compared with normal HUVECs; ★P < 0.05, compared with LPS-induced inflammatory and injured HUVECs
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Fig6: Effect of Calycosin on LPS-induced the change of gene expresses induced by LPS. Results of up- or down-regulated molecules in microarray (a) and quantitive real-time RT-PCR (b). △P < 0.05, compared with normal HUVECs; ★P < 0.05, compared with LPS-induced inflammatory and injured HUVECs

Mentions: According to the findings from microarray, we performed quantitative real-time RT-PCR (Fig. 6) and western blotting (Fig. 7) analysis. LPS indeed increased the mRNA expression of FN, ITG A5, RhoA, PI3K FAK, VEGF and VEGFR2, and decreased the mRNA expression of MLCP, indicating an activation of Rho/ROCK pathway. This activation was blocked by Y27632 and valsartan. After treatment with calycosin, FN, ITG A5, VEGF, VEGF R2, PI3K and FAK were significantly decreased, while MLCP was significantly increased (P < 0.05); as for RhoA, calycosin only slightly elevated its levels (P > 0.05) (Fig. 6). Consistent with the elevated mRNA expression, the protein expression of ITG A5, PIP2, RhoA and PMLC was also elevated in LPS-induced HUVECs, which was suppressed by Y27632 or valsartan. Whereas, RhoA (Fig. 7a) were decreased slightly, and, ITG A5 (Fig. 7b), PIP2 (Fig. 7c) and PMLC (Fig. 7d) was decreased after intervention with calycosin.Fig. 6


Calycosin-7-O-β-D-glucoside promotes oxidative stress-induced cytoskeleton reorganization through integrin-linked kinase signaling pathway in vascular endothelial cells.

Jiang YH, Sun W, Li W, Hu HZ, Zhou L, Jiang HH, Xu JX - BMC Complement Altern Med (2015)

Effect of Calycosin on LPS-induced the change of gene expresses induced by LPS. Results of up- or down-regulated molecules in microarray (a) and quantitive real-time RT-PCR (b). △P < 0.05, compared with normal HUVECs; ★P < 0.05, compared with LPS-induced inflammatory and injured HUVECs
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4562353&req=5

Fig6: Effect of Calycosin on LPS-induced the change of gene expresses induced by LPS. Results of up- or down-regulated molecules in microarray (a) and quantitive real-time RT-PCR (b). △P < 0.05, compared with normal HUVECs; ★P < 0.05, compared with LPS-induced inflammatory and injured HUVECs
Mentions: According to the findings from microarray, we performed quantitative real-time RT-PCR (Fig. 6) and western blotting (Fig. 7) analysis. LPS indeed increased the mRNA expression of FN, ITG A5, RhoA, PI3K FAK, VEGF and VEGFR2, and decreased the mRNA expression of MLCP, indicating an activation of Rho/ROCK pathway. This activation was blocked by Y27632 and valsartan. After treatment with calycosin, FN, ITG A5, VEGF, VEGF R2, PI3K and FAK were significantly decreased, while MLCP was significantly increased (P < 0.05); as for RhoA, calycosin only slightly elevated its levels (P > 0.05) (Fig. 6). Consistent with the elevated mRNA expression, the protein expression of ITG A5, PIP2, RhoA and PMLC was also elevated in LPS-induced HUVECs, which was suppressed by Y27632 or valsartan. Whereas, RhoA (Fig. 7a) were decreased slightly, and, ITG A5 (Fig. 7b), PIP2 (Fig. 7c) and PMLC (Fig. 7d) was decreased after intervention with calycosin.Fig. 6

Bottom Line: The effects of calycosin on LPS-induced changes in cell viability, apoptosis rate, cell migration, nitric oxide synthase (NOS), generationof intracellular reactive oxygen species (ROS) and cytoskeleton organization were determined.Calycosin improved cell viability, suppressed apoptosis and protected the cells from LPS-induced reduction in cell migration and generation of ROS, protein level of NOS at a comparable magnitude to that of Y27632 and valsartan.Calycosin are considered to be able to activate MLCP through promoting the generation of NO, decreasing PMLC, suppressing the cytoskeleton remodeling caused by activation of Rho/ROCK pathway and inhibiting AKT pathway by decreasing VEGF, VEGF R2 and PI3K level.

View Article: PubMed Central - PubMed

Affiliation: Affiliated Hospital of Shandong University of Traditional Chinese Medicine, West Wenhua road #42, Jinan, Shandong, 250011, PR China. jiang_yuehua@hotmail.com.

ABSTRACT

Background: Dysfunction of vascular endothelium is implicated in many pathological situations. Cytoskeleton plays an importance role in vascular endothelial permeability barrier and inflammatory response. Many Chinese herbs have the endothelial protective effect, of which, "Astragalus membranaceus" is a highly valued herb for treatment of cardiovascular and renal diseases in traditional Chinese medicine, In this study, we tested whether calycosin-7-O-β-D-glucoside (Calycosin), a main effective monomer component of "Astragalus membranaceus", could protect endothelial cells from bacterial endotoxin (LPS)-induced cell injury.

Methods: Endothelial cell injury was induced by exposing human umbilical vein endothelial cells (HUVECs) to LPS. The effects of calycosin on LPS-induced changes in cell viability, apoptosis rate, cell migration, nitric oxide synthase (NOS), generationof intracellular reactive oxygen species (ROS) and cytoskeleton organization were determined. Microarray assay was employed to screen the possible gene expression change. Based on the results of microarray assay, the expression profile of genes involved in Rho/ROCK pathway and AKT pathway were further evaluated with quantitative real-time RT-PCR or western blot methods.

Results: Calycosin improved cell viability, suppressed apoptosis and protected the cells from LPS-induced reduction in cell migration and generation of ROS, protein level of NOS at a comparable magnitude to that of Y27632 and valsartan. Similar to Y27632 and valsartan, Calycosin, also neutralized LPS-induced actomyosin contraction and vinculin protein aggregation. Microarray assay, real-time PCR and western blot results revealed that LPS induced expression of FN, ITG A5, RhoA, PI3K (or PIP2 in western blotting), FAK, VEGF and VEGF R2, and inhibited expression of MLCP. We believed multiple pathways involved in the regulation of calycosin on HUVECs. Calycosin are considered to be able to activate MLCP through promoting the generation of NO, decreasing PMLC, suppressing the cytoskeleton remodeling caused by activation of Rho/ROCK pathway and inhibiting AKT pathway by decreasing VEGF, VEGF R2 and PI3K level.

Conclusion: Calycosin protected HUVEC from LPS-induced endothelial injury, possibly through suppression of Rho/ROCK pathway and regulation of AKT pathway.

No MeSH data available.


Related in: MedlinePlus